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Department of Biotechnology Lund University, Sweden Annual Report 2009
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Department of Biotechnology Lund University, Sweden · 2011-02-08 · Lund University, Sweden Annual Report 2009 . Biotechnology at a glance The Department of Biotechnology at Lund

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Page 1: Department of Biotechnology Lund University, Sweden · 2011-02-08 · Lund University, Sweden Annual Report 2009 . Biotechnology at a glance The Department of Biotechnology at Lund

Department of BiotechnologyLund University, Sweden

Annual Report 2009

Page 2: Department of Biotechnology Lund University, Sweden · 2011-02-08 · Lund University, Sweden Annual Report 2009 . Biotechnology at a glance The Department of Biotechnology at Lund

Biotechnology at a glance

The Department of Biotechnology at Lund University was found-ed in 1985 by Professor Bo Mattiasson and is located at the Cen-ter for Chemistry and Chemical Engineering in Lund. During the past 25 years, both the number of research areas and number of people have grown. Today the department consists of about 60 researchers, PhD students and staff, and is visited by around 10 master students and 40 international guest researchers every year. The research activities at the department are focused on various aspects of environmental-, analytical-, food- medical- and indus-trial biotechnology and consist of both basic and applied research. The department is heavily involved in teaching at the Biotechnol-ogy programme and takes active part in the International Master’s programme in Biotechnology at Faculty of Engineering (LTH), Lund University. Department of Biotechnology is actively initiating collab-orative projects with industry and other academic institutions and spreading information about ongoing activities to the surrounding society.

Foreword

The need to inform about activities in academic research laboratories is es-calating. There is a need felt from the society in general, from the funding bodies and from present and future collaborators. This is the fifth year we are publishing an annual report compiling the activities within Department of Biotechnology.

As you will see in this report, the activities in the department cover a range of fields, from the very basic issues to some very applied. It is indeed a privi-lege to be active in a field with so close contact between basic science and applications, and at the same time in fields where the development is very fast.We hope that the information presented in this report will stimulate new thoughts and a wish to know more about some specific issues. Please use the contacts given to the individual scientists, or contact the editor of this document.

Cover photo: Energy crops cultivated at SLU Alnarp and used for biofuel production. Photo by Emma Kreuger.

Page 3: Department of Biotechnology Lund University, Sweden · 2011-02-08 · Lund University, Sweden Annual Report 2009 . Biotechnology at a glance The Department of Biotechnology at Lund

Dept. of Biotechnology Annual Report 2009 3

Head of Department´s report 4External communication 5Services for external customers 6 Research activities 8 Protein structure and function 9 Green chemicals and materials 10 Bioseparation 12 New materials 13 Bioanalysis 14 Biodiversity 15 Bioremediation 16 Functional foods 17 Renewable energy 18 Education 19 Courses 19 International Master’s programme in Biotechnology 20 Master thesis projects 20International collaborations 21Staff 22Post Docs 23 PhD students 24Visiting researchers and students 26Funding and grants 27Publications 28 Journal articles 28 Doctoral degrees and theses 30Conferences 30

CONTENTS

271815

965

Industry8%

Research councils19%

Education7%

Faculty15%

Public authorities17%

Foundations34%

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Dept. of Biotechnology Annual Report 20094

Head of Department´s reportThis annual report is meant to provide you with a glimpse of the activities being pursued at Department of Biotechnology.

The department is a vibrant, multi-cultural and multi-lingual place of work. The economy has been stable due to good external funding obtained by several of our researchers. We did however had to make com-promises during 2009 due to the need for increased co-financing from our faculty funds that did not leave much of those funds for other expenses.

Rajni Hatti-Kaul at the biorefinery. Photo by Bo Mattiasson.

Research in industrial biotechnology for the produc-tion of energy, chemicals and materials from renewable, surplus biomass or wastes, and waste-water treatment have evolved as the strong themes at the department and will continue to be so for some years to come. This is in line with the global concern for greenhouse gas emissions and climate change; the climax being the Climate change conference in neighbouring Copen-hagen held at the end of the year. The department is among the participants in the LU Biofuels network. Other important activities are in the areas of materials for separation of cells, tissue engineering, etc., func-tional foods and bioanalysis. Many of the projects are interdisciplinary and involve collaborations with other departments at Lund University, other universities and industry in the region. We continue to be active in in-ternational collaborations; in 2009 we became partners in a new European Marie Curie network dealing with biocatalysis. Nonetheless, search for new funds to sup-port our activities and to renew our research is always ongoing.

As during earlier years, the Department has been heav-ily involved in undergraduate courses both for Biotech-nology program and International Master Program in Biotechnology and Food Technology at the Engineer-ing Faculty. Seven of our graduate students defended their theses during 2009 and six new PhD students joined the department.

We have observed some fluctuations in our staff dur-ing the year. Dr Mihaela Nistor´s group dealing with bioanalysis moved from Analytical Chemistry to our department. A new Senior Lecturer position with spe-cialization in bioenergy was added to the Department staff, while Dr Igor Galaev, a senior lecturer of many years left us at the year end to join the industry.

We have tried to implement means for improving com-munication and dissemination of information among our personnel as well as to the outside world. The re-search station in Svalöv municipality initially equipped with biogas pilot plant is being developed into a bi-orefinery and we look forward to increasing activities there. Even a few start-up companies emerging from the department are slowly getting established. We are also proud of Martin Hedström who won the Venture Cup award given for innovative ideas.

I would like to thank the Biotechnology staff for their support and co-operation during the year and would like to encourage all to strive for scientific excellence and also in making the department a fun place to work. I hope that you will enjoy reading this annual report of our department and provide us with a feedback.

Rajni Hatti-Kaul, Lund, 18 February 2010

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Dept. of Biotechnology Annual Report 2009 5

External communicationThe Department of Biotechnology is a scientific institution providing an inspiring environment where scientific work and education go hand in hand with interactions with society. We consider engage-ment with business and industry as well as interactions with the wider community as very important in being able to meet the needs and expectations from the surroundings.

Besides the academic staff, the department has two technology coordinators with the task of be-ing ambassadors for entrepreneurial activities at the department and encouraging the interactions between academia and industry. We actively make contacts with industries and other potential col-laborators outside the university. Among the activ-ities we engage ourselves in are collective research programmes such as Mistra financed Greenchem and Vinnova supported Industrial biotechnology projects, bilateral agreements including contract research, training of company personnel by e.g. industrial PhD students or specialized courses and scientific collaborations of different kinds. Depart-ment of Biotechnology recognises the importance of allowing valuable research ideas being tested for their commercial importance and supports the start-up of businesses such as “Bioprocess control” and “Devenz”.

We inform about our ongoing research in the un-dergraduate courses given by the department as well as in other courses and workshops held at the university. During summer 2009, the department hosted a much appreciated BEST-course about the role of biotechnology in the sustainable society for 30 students coming from other countries in Eu-rope. BEST is an acronym for Board of European Students of Technology. As during earlier years, external communication in our research pro-gramme Greenchem, has been quite extensive. The programme was presented in the issue about the Year of Sustainability 2008/2009 in the Newslet-ter from “Universitas 21” which is a Network for International Higher Education. Several lectures at conferences, institutions (both in Sweden and abroad) and companies, interviews with various science magazines, Greenchem newsletter and the Greenchem homepage (www.greenchem.lu.se), have helped to spread the knowledge about indus-trial biotechnology to scientists, politicians, busi-ness people and the general public. Some examples for 2009 were inspiration lecture in innovation project for undergraduate students at the faculty of Engineering, and invited lectures in a workshop

organised by LU Biofuels network also at the faculty, a workshop on bioenergy in China and at two universi-ties in Malaysia.

The Department is involved in three different EU projects based on applications on superporous gels. Within one of these programs, we organized a sum-mer school “Porous Hydrogels for Biomedical Applica-tions: from Cytapheresis to Tissue Engineering” Sep. 29 – Oct. 2 in Antalya, Turkey.

The Biogas area continued to attract attention dur-ing 2009. The Department is a member of Biogas Syd – an organization where all stake-holders in the biogas area in southern Sweden meet. This organisation was formed as a result of activities initiated by the Depart-ment. During 2009 Biogas Syd carried out a lot of ac-tivities for promoting interactions with the surround-ing society and for that scientists from the Department were often involved. The research station at Anneberg attracts many visits of study groups and during such events biogas as well as biorefinery concepts are dis-cussed, often in relation to agricultural activities.

Several of our researchers have been invited to give lec-tures at different seminars, conferences and meetings as well as industries, both in Sweden and abroad, which can be read in later pages of this annual report.

Visit the home page of the Department of Biotechnology at www.biotek.lu.se!

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Dept. of Biotechnology Annual Report 20096

Services for external customersDepartment of Biotechnology offers services to other academic institutions and industry in different areas where the department has expertise and possesses special equipment or research facilities. We have the possibility to run pilot scale fermentations upto 400 litres scale, design and perform biogas production and biorefinery processes, assist in running mass spectrometry experiments or perform other type of evaluations and optimizations of biotech-nological processes or technologies.

Contact persons are indicated under the description of each service below. For general inquiries please contact technology coordinator Josefin Ahlqvist, [email protected], phone: 046-222 48 38.

LC-MS/MS Mass spectrometerThe mass spectrometer at the Department of Biotech-nology is of the type Quadrupole-Time Of Flight, Q-TOF (Q-Star, Applied Biosystems), which is connect-ed to a LC-system. The mass spectrometer is equipped with an electrospray (Turbospray) ionization source and a nanospray system. The nanospray system can be connected to syringe pumps in order to get a continu-ous nano-system equipped with capillary columns for separation on a column packed with C

18 or other pack-

ing material of interest. It has also a Maldi ionization interface. Responsible: Martin Hedström ([email protected], phone: 046-222 75 78).

The station for Biogas research at Anneberg is situated close to an agricultural farm in the municipality of Svalöv. It is an important site for demonstration and for getting increased acceptance of farm based biogas technology.

Research station for biogas research

The research station is equipped with biogas reactors of different design in 1-80 m3 scale, with good facilities for on-line monitoring and data collection. A fully equipped control laboratory is also available on-site.Responsible: Lovisa Björnsson ([email protected], phone: 046-222 83 24).

Photos from the Department of Biotechnology research station at Anneberg. The photographed equipment is mainly used for research on biogas production. Photos by Josefin Ahlqvist.

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Dept. of Biotechnology Annual Report 2009 7

Pilot plant fermentorThe fermentation pilot plant was renovated in 2004 and located in close contact with the de-partment’s course lab. The pilot plant is equipped with fermentors (from 1 litre up to 400 litres scale), equipment for downstream processing of microorganisms (filters and cell disintegrator), as well as for downstream processing of biologi-cal molecules. The whole facility is approved for work with class 2 organisms. Responsible: Christina Wennerberg ([email protected], phone: 046-222 46 81).

To utilize biomass for production of a plethora of chemicals while the parts of the biomass that cannot efficiently be converted into specific chemicals can be transformed into bioenergy is a trend in industrial biotechnology today.

The department is involved in building a pilot plant biorefinery in conjunction with the research station for biogas production at Anneberg Farm, Svalövs municipality. The intentions are to operate in good pilot plant scale (several 100 litres up to a few m3). Besides suitable vessels for the conversions, there is equipment for isolation and purifi-cation of the compounds produced. A well equipped service lab is available. The unit is now in operation with the smaller reactors. The larger ones will successively be connected.

A few processes will be taken from the research labs in Lund and scaled up. At the same time a more holistic view on yields and energy efficiency will be applied. A few of the unit operations that will be essential in the future will initially be established in this new unit. Responsible: Bo Mattiasson ([email protected], phone: 046-222 82 64) and Martin Hedström ([email protected], phone: 046-222 75 78).

DSC - Differential Scanning CalorimeterThe differential scanning calorimeter (VP-DSC) at the department is a sensitive, easy to use instrument, with an active cell volume of approximately 0.5 ml used for studying samples in solution. The operating temperature of the instrument is in the range of –10ºC to +130ºC, and the instrument has a selectable scan rate in the range of 0ºC to 90ºC per hour (upscan), allowing studies of fast or slow transition processes.

It can be used to measure the intramolecular stability of a broad spectrum of biomolecules, including proteins, nucleic acids, lipids and detergent micellar systems. The VP-DSC provides fast, accurate transition midpoint (Tm) determination, as well as a thermodynamic profile that can provide insight into the factors that affect conformation and stability. Responsible: Eva Nordberg Karlsson ([email protected], phone: 046-222 46 26).

BiorefineryMartin Hedström and Marie Andersson at the scaling up process for production of platform chemicals. Photo by Bo Mattiasson.

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Dept. of Biotechnology Annual Report 20098

Research activities

Biotechnology for sustainable development is coming into focus at the same time as the awareness of climate change and shortage of water for a growing population are becoming focal points among politicians around the world. Over the years the research at Department of Biotechnology has evolved towards a sustainability thinking and has become increasingly valuable and important for the surrounding society. In the light of the global devel-opments, it appears that earlier decisions to direct the research towards the development of environment friendly processes have proven right.

The wide knowledge base at the department within enzyme technology, bioseparation, cell cultivation techniques and gene technology is used to direct research activities in diverse areas. ENVIRONMENTAL biotechnology involving wastewater treatment, bioremediation and energy production has been an important area and has at-tracted a lot of financing during 2009. INDUSTRIAL biotechnology or “white” biotechnology is seen as another important field related to environmental sustainability to position our research in, especially since we believe that biotechnology will play an important role in the future when it comes to replacing chemicals and materials based on fossil carbon with renewable feedstocks. The other areas of interest at the department are MEDICAL biotech-nology involving projects on development of materials for tissue engineering, production of pharmaceutical pro-teins and process analytical technology, and FOOD biotechnology involving antioxidants and production of food components with improved properties.

Overview of the research activities at the Department of Biotechnology. Illustration by Maria Andersson.

Renewable energy

BioremediationAntioxidantsBacteriocins

Feed additivesPolysaccharides

Green chemicalsBiomaterialsChiral products

Tissue engineeringPharmaceutical proteins

Plasmids/DNA

Enzyme technology

Protein engineering

Bioorganic synthesis

Enzyme stability and specificity

Microbial technology

Cell cultivation techniques

Extremophiles and extremozymes

Bioseparation

Smart polymersAffinity interaction

Aqueous two-phase systems

Cryogels

Simulated moving beds

Process analytical technology (PAT)

Biosensors

Bioanalysis

Flow injection analysis

Industrial biotechnology

Food biotechnology

Environmental biotechnology

Medical biotechnology

Protein structure and function

Biodiversity

Figure (top): SEM of Candida antarctica lipase B immobilized on macroporous polyacrylate beads (Novozym435). Image by Ulrika Törnvall.

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Dept. of Biotechnology Annual Report 2009 9

Protein structure and function

Creating functionality and stabilityStable and selective catalysts are prerequisites for efficient biocatalytic processes, and development of novel enzymes paired with a good understanding of the target proteins is thus an area of importance for creation of future biocatalysts.

ExtremozymesWe have specialized in research on enzymes from extremophiles, i.e. microorgan-isms adapted to extreme ecological conditions (heat or cold, high or low pH, or high salt concentration). Our research is based on use of recombinant enzymes, allowing genetic development of the biocatalysts selected based on properties like stability, reaction specificity or production yield. Production is a necessity for use and characterization of created variants, is achieved by usage of genetically modi-fied microorganisms. As a natural complement to enzyme development, batch and fed-batch strategies for their heterologous production are utilized. Our aim is de-velopment of biocatalysts for environmental friendly processing, e.g. for the pro-duction of green chemicals.

Carbohydrate modifying enzymesCarbohydrates are essential components in life, and biomass contains an array of structural and storage polysaccharides (e.g. starch, cellulose and hemicellulose) also important in industrial applications. Enzymes acting on these materials are gly-coside hydrolases (GHs), which hydrolyse glycosidic bonds between two or more carbohydrates or between a carbohydrate and a non-carbohydrate moiety. These enzymes are subject of growing interest due to a wide range of possible applications contributing to environmental-friendly processes. In a project supported by VR, in the research program Sustain-X-Enz, and in the research program Greenchem, rational and random engineering strategies are utilized to transform a number of GHs into more perfect biocatalysts also utilizing structural biology resources to un-derstand the molecular reasons for the changed properties. Carbohydrate binding modules are other interesting targets for specificity development. Use of wild type and developed xylanase is explored in a project with a South African University.

Molecular basis of oxidative inactivation of lipase Lack of stability during process conditions is the main bottleneck in the industrial application of biocatalysts. Candida antarctica lipase B is the most frequently used biocatalyst for synthesis of chemicals. The enzyme undergoes oxidative inactiva-tion in the presence of hydrogen peroxide used during chemo-enzymatic epoxida-tion of alkenes. In a project supported by the Swedish Research Council, we have attempted to understand the structural changes occurring in the enzyme when exposed to oxidizing conditions, with the help of various techniques such as mass spectrometry, circular dichroism and dynamic light scattering. The aim is to use the knowledge for designing stable biocatalyst preparations.

Thermostable alkaline phytase for animal feedPhytase is an enzyme used to degrade the antinutritional compound, phytate in animal feed with concominant increase in availability of phosphorus. Our studies involve a highly thermostable alkaline phytase from a Bacillus sp. isolated by us. The high thermal stability allows the enzyme to be included in the feed during pelleting. A number of enzyme variants have been generated by site-directed mu-tagenesis to understand the structure-function relationship of the phytase and to improve the activity and stability under acidic conditions.

ProjectsSpecificity design of glycosidasesSustain-X-Enz (see also Green chemicals and Materials)GreenChem (see also Green chemicals and Materials)A thermostable carbohydrate binding module as a general scaffold for diversity and specificityTo explore and understand how xylanases gain high potential as industrial biocatalystsHigh pH catalytic adaptation of extremozymesEnhancing the thermal stability of alkaline active enzymesImproving oxidative stability of lipaseStructure-function of Bacillus sp. phytase

PhD studentsSamiullah Khan, Tania Pozzo, Thuy Tran Thi, Ulrika Törnvall

Post DocsSuhaila Hashim, Catherine Paul

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Figure (top): Native structure of Candida ant-arctica lipase B with the catalytic triad resi-dues shown in blue. Residues found to be oxidized by hydrogen peroxide are two tryp-tophans (red), all four methionines (green), and all three disulfide bridges (yellow). Im-age by Ulrika Törnvall.

Rajni Hatti-KaulProfessor

Gashaw MamoResearch Engineer

Martin HedströmResearch Engineer

Eva Nordberg KarlssonAssociate Professor

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Dept. of Biotechnology Annual Report 200910

Green chemicals and materials

Using industrial biotechnology

Industrial biotechnology has a promising role as an enabling technology for production of chemicals, materials and energy from bio-based renew-able raw materials. The production processes are based on fermentation or biocatalysis using whole cells or enzymes, and are characterized by reduced energy consumption and waste generation, and biodegradable products. At Department of Biotechnology, activities in industrial biotechnology have proliferated during the past years. Biocatalytic processes are developed for synthesis of speciality- and fine chemicals and for modification of raw ma-terials; enzymes are screened primarily from extremophilic microorganisms and developed for use in the reactions. Fermentation is also used for pro-duction of chemicals and polymers.

Speciality chemicals from renewable resources - Greenchem The research program Greenchem financed by Mistra (Swedish Founda-tion for Strategic Environmental Research) focuses on the production of “green” chemicals targeted for applications in environment-friendly surface coatings, lubricants, consumer care and cleaning. The programme is inter-disciplinary involving cooperation between biotechnology, environment systems analysis and innovation systems analysis, and also with industries representing an integrated value chain from raw materials to the users of in-dustrial chemicals. Biocatalysis is used for synthesis of the chemicals, pref-erably under solvent-free conditions. Immobilized enzyme preparations are developed in-house whenever necessary.

Fatty epoxides, glycidyl ethers and polyester acrylates are synthesized for use in surface coatings. New enzymes were evaluated during the year for syn-thesis of glycidyl ethers. Development and evaluation of a promising new product for coating applications has been ongoing. Biolubricants based on complex esters of fatty acids with potentially superior properties than vegetable oils have been synthesized and evaluated. Optimization of the process parameters was done during 2009 for modelling and scaling up together with the industrial partner. Production of different biosurfactant compounds – alkyl glycosides and alkanolamides - has been investigated. Cyclodextrin glycosyltransferases (CGTases) and other enzymes have been used to elongate the carbohydrate part of alkyl glycosides, thereby giving products which were previously impossible to synthesize. During 2009, a patent application was submitted and the methodology is currently further improved to increase efficiency and to decrease process costs. Thermostable enzymes from hyperthermophiles have been evaluated for the synthesis of alkyl glycosides.

ProjectsGreenchemBiodegradable plasticsIndustrial biotechnology for pro-duction of platform chemicals Development of process technolo-gies for immobilized biocatalystsLignin based formaldehyde-free wood adhesivesExtraction and enzyme modifica-tion of compounds from agricul-tural waste (Sustain-X-Enz)

PhD studentsHugo Cavero, Georgina Chavéz, Thuoc Doan Van, Daniel Guzmán, Hector Guzmán, Anna Hagström, Victor Ibra-him, Samiullah Khan, Marlene Munoz, Tania Pozzo, Gustav Rehn, Ramin Sabet, David Svensson, Ulrika Törnvall

Project workersElin Brun, Tarek Dishisha, Pontus Lun-demo, Regine Wuttke

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Post DocsTeresa Alvarez Aliaga, Cecilia Orellana-Coca, Carl Grey, Ravi Kiran Purama, Javier Linares Pastén, Catherine Paul, Konstanze Stiba, Sang-Hyun Pyo, Suhaila Hashim, Daniela Zehentgruber

Patrick AdlercreutzProfessor

Rajni Hatti-KaulProfessor

Bo MattiassonProfessor

Olle HolstProfessor

Eva NordbergKarlssonAssociate Professor

Figure (top): Esterification reaction for the biolubricants synthesis catalysed by lipase. Photo by Cecilia Orellana-Coca.

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Dept. of Biotechnology Annual Report 2009 11

Development of process technologies for immobilized biocatalystsIn this VINNOVA funded project methodology for the immobilization of enzymes and whole cell catalysts is developed. The project is coordinated by Cambrex Karlskoga and involves three other companies. Dem-onstration processes for production of chiral amines, dihydroxyacetone and 3-hydroxypropionaldehyde are developed.

Halophilic bacteria as a source of biodegradable plastics and compatible solutesPolyhydroxyalkanoates (PHAs) are biodegradable polymers produced by several bacteria as energy reservoirs. PHAs can be used as disposable plastics in several areas ranging from packing commodities to medical devices. We have isolated a number of novel moderately halophilic PHA-producing bacteria from saline lakes in Bo-livia. Production of polyhydroxybutyrate (PHB) by Halomonas boliviensis has been investigated in detail. The productivity of the polymer has been improved and also combined with production of compatible solutes. In 2009, modification of PHB and preparation of polymeric gels with increased hydrophilicity has been done for application in tissue engineering applications. Also, a highly productive process for production of the com-patible solutes – ectoine and hydroxyectoine – has been developed. The products were evaluation for enzyme stabilization under extreme pH conditions.

Extraction and enzyme modification of compounds from agricultural wasteThis collaboration project (between Biotechnology, Environmental systems analysis (P Börjesson) at Lund University, and Analytical chemistry (C Turner, P Sjöberg, at Uppsala University) funded by Formas, aims to make an impact in recovery of high-value compounds from agricultural byproducts/waste using environ-mentally sustainable techniques. Fundamental studies are performed around supercritical carbon dioxide and pressurized hot water extraction, targeting industrially and nutritionally valuable compounds (a first target antioxidative molecules) in for example onion skin, and birch bark. Thermostable enzymes are used in high-temperature/pressure processes to convert/derivatize target compounds to facilitate analytical determination as well as increase the yield. Life cycle assessment (LCA) is applied to evaluate proposed processes and utilization of waste materials.

Lignin based formaldehyde-free wood adhesives This project aims to look at the possibility to modify the lignin enzymatically to make it more reactive for cross-linking with other molecules and to evaluate the modified structures for gluing properties. Cross-linking of the modified lignin with other molecules will be based on the knowledge on the structure of proteinaceous “super-glues” produced by marine organisms such as mussels and barnacles to anchor themselves to rocks. De-velopment of strong and environmentally benign wood adhesives from lignin would thus potentially result in a reduction in the use of formaldehyde based adhesives.

Speciality Chemicals from Renewable Resources

Greenchem

Figure (top): Rapeseed. Photo by Greenchem.

Platform chemicals from renewable resourcesIn a VINNOVA (The Swedish Governmental Agency for Innova-tion Systems) funded project, by-products of agriculture and biofuel industry are used as raw materials for production of organic acids using industrial biotechnology. The organic acids serve as building blocks for other important secondary chemicals. The project is co-ordinated by Perstorp AB and involves 5 other companies. Lab-scale processes based on free- and immobilized cells have been developed for production of propionic acid from glycerol.

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Isolation of biomolecules and environmen-tal pollutants - sorting molecules and cells

Our research in the area of bioseparation deals with development of proce-dures for isolation of specific biomolecules, and, for development of new separation media with emphasis on: The work on supermacroporous gels, so called cryogels is continuing. New modes of preparing gels and a range of applications are studied both with regard to isolation of biomolecules and to enrich environmental pollutants before destruction/final storage.

Development and applications of new materials for bioseparation- Cryogels encased into Kaldnes plastic carriers are developed for operation in stirred tank reactors combining the robustness of plastic carriers with high porosity and facilitated mass-transport within cryogels. Applications are both in protein separation and in environmental separation when target molecules shall be enriched from crude feed streams. - Tailor-made cryogels are developed for enzyme and cell immobilization allowing processing turbid feeds due to large interconnected pores within cryogels.- A new technology for affinity separation of cells is developed utilising elasticity of cryogels to facilitate detachment of bound cells.- Composite cryogels containing activated carbon are under development for environmental application and for hemoperfusion.- Endocrine disruptors, non-metabolised pharmaceuticals and pesticides which are present in very low concentration and hence escape traditional waste water treatment are selectively captured by molecular imprinted cryo-gels, concentrated after elution and later on degraded by microorganisms.

Efficient processes for purification of biomolecules- The process for isolation and purifi-cation of poly-unsaturated fatty acids from fish waste has been developed.- Isolation of surface active com-pounds after enzymatic synthesis presents a lot of separation challeng-es. A new chromatographic process has been developed and is now evalu-ated in simulated moving bed (SMB) mode.- A new platform, Chromatopan-ning, for integrated selection of target phages from libraries (panning) with concomitant infection of E. coli cells has been developed allowing to reduce 2-3 fold the total process time.

Bioseparation

Igor GalaevSenior Lecturer

Bo MattiassonProfessor

ProjectsSmart surfacesCarbohydrate-specific synthetic polymers as components of mu-coadhesive gels and stimuli-re-sponsive supports for cell separa-tion and analysis Smart polymers for selective recog-nition of biomoleculesSimulated moving bed technology for efficient protein purificationApplication of cryogels housed in plastic covers in wastewater treat-ment and biocatalysisIsolation of PUFA from industrial waste streamsEnvironmental separation

PhD studentsSolmaz Hajizadeh, Harald Kirsebom, Betty Mbatia, Wim Noppe, Zulma Perez, Deepti Sahoo, Linda Önnby

Post DocAarti Ozarkar

Visiting studentsAlly Mahadhi, Anna Scaglia

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on column infecting E. coli cells with bound phages phage amplification phage elution

bind to column and wash non-bound phages

The chromato-panning protocol with on-column infection of E.coli cells by specifically bound bacteriophages. Illustration by Igor Galaev.

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Dept. of Biotechnology Annual Report 2009 13

New materials

ProjectsFundamentals of cryopolymeriza-tionChromato-panning of phage li-braries using cryogelsCarbohydrate-specific synthetic polymers as sugar sensorsBiomaterials for wound healing and tissue regenerationMatiss – An EU-project for utilizing cryogels as scaffolds or as wound dressingsStarch based cryogelsCryogels from small particles e.g. bacterial cellsCarbosorb – An EU-project where cryogels and other adsorbents are to be used for enriching/treating pollutants present at low concen-trationMonaco – An EU-program for ad-sorption of compounds from the blood streamEnrichment of heavy metal ions and other impurities

PhD studentsDaniel Guzman, Solmaz Hajizadeh, Harald Kirsebom, Marina Kuzimenko-va, Marlene Munoz, Wim Noppe, Gry Ravn Jespersen

Post DocsLinda Elowsson, Aarti Ozarkar

Visiting researchers Dmitry Berillo, Junxian Yun

Visiting studentsSéverine Cozzi, Ilyas Inci, Oksana Zaushitsyna

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Bo MattiassonProfessor

Combining polymer technology and biotechnology yields a range of new materials with interesting properties for use in areas such as bioseparation, medical biotechnology and bioanalysis.

Cryogels Macroporous hydrogels, so called cryogels, are produced by polymerization in a partially frozen state when the ice crystals perform as porogens. After completing the polymerization and melting the ice a system of large and interconnected pores is formed. The high mechanical strength along with unique tissue-like elasticity and uniform porous structure make cryogels very promising materials for bioseparation of nanoparticles, as scaffolds for the cultivation of mammalian cells and implants in tissue engineering. The emphasis in the project in 2009 was on studying cryopolymerization using 1H-NMR. The technique allows following simultaneously the polymeriza-tion and the freezing processes and provides data on the mechanisms of cry-opolymerization. One project deals with investigating how the porosity and the properties of cryogels changes by adding inert solutes. A new project has been started on detailed modeling of mass-transport inside cryogels.

Design of biodegradable, biocompatible superporous structures has been a main target. A few leads are now under development.

Biomaterials Cryogels containing natural polymers like collagen or fibrinogen, have been intensively studied as scaffolds for tissue engineering within a few EU-col-laborations. Work has also been initiated on using microbially produced polyalkanoates as scaffolds for tissue engineering.

Ordered aggregation of particlesSuperporous structures can be formed from bacterial cells or small polymer particles by arranging them spatially via a cryostructuring process and then cross-linking in order to stabilize the structure formed. Such aggregation of particles offers certain advantages over conventional entrapment: the pores in the particles remain intact also in the aggregated form. This has been shown for activated carbon that is going to be used for wound dressing. Evaluation of biocatalytic and affinity-based applications are ongoing.

Stimuli-responsive polymersStimuli-responsive polymers change their macroscopic behaviour as a result of a small change in an environmental parameter. Superporous hydrogels made from such polymers do not show the expected dramatic changes in behaviour. If, however, stimuli-responsive polymers are first aggregated into small particles that later are ordered into a superporous structure, then dra-matic shifts in behaviour of the formed gels are seen when the temperature changes over the transition temperature.

Polymers in the service of Biotechnology and Biomedicine

Igor GalaevSenior Lecturer

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Dept. of Biotechnology Annual Report 200914

Bioanalysis

Martin HedströmResearch Engineer

Bioanalysis for process analytical technology

Quality control with regard to chemical composition involves analysis. The more sensitive analysis, the better control can be offered, provided that the analysis is simple and quick. There are several different applications that are studied within the department:

• continuous integrated sampling and monitoring of fermentations• quality control of food and feed• monitoring of trace amounts of microbial toxins• tracing viral infections via ultrasensitive assay of viral proteins• monitoring of trace impurities in biopharmaceutical production

A few basic technology platforms have been developed:• flow-injection binding assays• capacitive affinity biosensor for ultrasensitive monitoring• electrochemical affinity sensors

Trace amount of impurities are attracting a lot of interest today among pharmaceutical companies producing proteins/peptides for injection. Mon-itoring of endotoxin, host cell proteins and ligands that have leaked from affinity adsorbents used (e.g. protein A from purification of monoclonal an-tibodies) are examples of projects that are studied today. Furthermore, when producing recombinant proteins, there is always a risk that some of the target molecules become modified e.g. by proteolysis such that truncated proteins are mixed with the true target molecule. To monitor this is usually a tedious process, but a new technology developed in the department makes it possible to detect levels of truncated proteins quick enough so that it will be possible to harvest the valuable compound when the levels of truncated molecules start to rise.

Regardless of detection principle, it is within the concept of process analyti-cal technology (PAT) of great importance to be able to perform integrated sampling, transportation and processing of the sample taken from e.g. the fermentation vessel in a well-defined and controlled way. Hence, research activities within the bioanalytical area require sig-nificant automation which now is addressed clearly in the bioanalysis group.

ProjectsDevelopment of analytical systems for the trace-amount detection of bacterial toxinsAnalytical approach for the moni-toring of affinity column ligand leakageMonitoring of HIV capsid p24 pro-teinDevelopment of an affinity sen-sor based on application of intact membrane proteinsA process analytical technology (PAT) approach for the concomi-tant on-line monitoring of target proteins and bacterial contami-nants present during purificationMonitoring and control of host cell proteins (HCPs) during recom-binant production using a capaci-tive biosensorMonitoring and control of recom-binant human growth hormone during fermentation and down-stream processing using capillary-LC/MSDevelopments of cell-based flow devices for process controlSol-gel encapsulation of biomol-ecules for the use in diagnostic testsImproving profitability of dairy SMEs through developing, validat-ing and promoting the use of a biosensor for ovulation detection (Ovultest)Calpastatin biosensor for meat ten-derness prediction (Tendercheck)

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Working on a lab-version of a ca-pacitive biosensor. Photo by Ma-rika Murto.

Mihaela NistorResearch Scientist

PhD studentsMahmoud Labib, Mohammad Mazlomi, Kinga Zor

Visiting researchersShabana Basheer, Warakron Limbut

Visiting studentsMarie Andersson, Supaporn Dawan, Katarzyna Dymek, Audrey Faure, Saluma Samanman, Kosin Teeparuksa-pan, Alexandru Zabara

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Dept. of Biotechnology Annual Report 2009 15

Biodiversity

ProjectsBiodiversity of alkaline and saline environmentsIsolation and characterization of extremophiles from highly alkaline environmentsAnaerobic microorganisms for bio-technological applicationsScreening for antimicrobial sub-stances from thermophiles from hot spring in JordanScreening for polysaccharide pro-ducing LABs from Indian indig-enous foods

PhD studentsRawana Al Khalili, Rosa Aragão, Geor-gina Chavez, Carla Crespo, Daniel Guz-man, Victor Ibrahim, Laura Mendoza

Post DocsTeresa Alvarez Aliaga, Javier Linares Pasten

Visiting studentAmi Patel

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Laguna Verde, Bolivia, from which Halomonas andenense was isolated. Photo from Scribd.

Rajni Hatti-KaulProfessor

Gashaw MamoResearch Engineer

Bo MattiassonProfessor

Olle HolstProfessor

Nature - an infinite source of activities

Microbial diversity existing in nature is extremely vast. Only a small frac-tion of the natural microorganisms have so far been identified. Microorgan-isms inhabiting extreme environments, the so called extremophiles, have been of interest both for understanding the mechanisms used by them for survival as well as for exploiting their potential for applications requiring extreme conditions. Over the years, a diverse group of extremophiles have been isolated at Department of Biotechnology from samples of alkaline soda lakes in East African Rift Valley, hot springs in Iceland, and saline Al-tiplano region of Bolivia. A number of novel microorganisms belonging to genera Bacillus, Dietzia, Brevibacterium, Halomonas, Amphibacillus etc have been identified. The studies on biodiversity have involved collaborations with universities in the respective countries. Some interesting hydrolytic enzymes from extremophiles have been investigated for use in applications such as animal feed, production of chemicals, etc.

Some halophilic bacteria are being used for the production of biopolyester, polyhydroxybutyrate and compatible solutes. Some other novel microor-ganisms isolated from alkaline environments have been screened for the presence of monooxygenase activities, and an assay for rapid screening of cyclohexanone consuming bacteria has been developed.

Isolation and characterization of thermophiles from hot springs in Jordan is going on with special attention to production of antimicrobial substances. Several strains (Bacillus sp.) exhibiting activity have been found and their antimicrobial profiles are currently investigated.

Anaerobic microorganisms play an important role in many cycles of ele-ments in nature, however, they are poorly understood. During the year the work has continued to isolate new organisms producing enzymes with spe-cial properties. The isolation is based on the microdroplet technique which allows many more organisms to be isolated and identified than that achieved by classical techniques. Several new species e.g. some new Clostridium spe-cies, have been isolated. Genes from some anaerobes are being cloned and expressed for use in the production of platform chemicals.

White-rot fungi are often efficient at degradation of aromatic structures. Among several fungi isolated from the tropical regions of Bolivia good pro-ducers of oxidative enzymes including peroxidases and laccase have been found. One example is Galerina sp., that has been investigated for their capability of producing laccase. The enzyme has been characterized and evaluated for demethylation of lignin and decolorization of textile dyes. Indian local foods and plant material are being screened for polysaccharide producing lactic acid bacteria in collaboration with Agricultural University Anand, India. Currently ca 20 isolates (e.g. Weissella and Lactobacillus) out of several hundreds are selected for further characterization. The polysac-charides will be further investigated for applications in foods.

Figure (top): Scanning electron microscopy of an alginate microbead (size approx 40 µm) used for entrapment of anaerobic single cells. Image by Rosa Peralta Aragão.

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Dept. of Biotechnology Annual Report 200916

Bioremediation

Bo MattiassonProfessor

ProjectsRemoval of heavy metal ions using sulphate reducing bacteriaEnrichment of heavy metals on chelating matrices before release and precipitationEnrichment and destruction of en-docrine disruptors in wastewaterDevelopment of separation tools and systems specially adapted to waste water treatmentDegradation of textile dyes in wastewater from textile industriesFungal bioreactors for degradation of POPsPhotochemical oxidation of or-ganic pollutants as a pre-step to biodegradationBiosensor for environmental analy-sis

PhD studentsMaria Jonstrup, Linda Önnby Visiting studentsMartha Jarquin, Marisa Punzi

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Figure (top): Textile dyes. Photo by Maria Jonstrup.

Pollutant removal

Persistent organic pollutants (POPs) include a large variety of substances such as pesticides, surfactants or wood preservatives (i.e. creosote mixture) as well as products released from industrial activities (i.e. dioxins, polyaro-matic hydrocarbons). Heavy metals belong to another group of compounds causing severe environmental problems. A new group of potentially hazard-ous chemicals have come into focus in recent years: pharmaceuticals and molecules with endocrine disrupting activity.

The department has focused much attention on developing biotechnologi-cal methods, either alone or in combination with physical treatment, to degrade or immobilize these hazardous compounds. Sometimes the con-centration at which the hazardous compound is present is very low, and then an enrichment step is used before treatment. This is used especially for endocrine disruptors and pharmaceuticals.

Dye remediation is another area where biotechnological treatment methods can be used for the treatment of textile dyes using biological and physico-chemical techniques. In textile industries, considerable amounts of water and chemicals are used. During the dyeing process about 20% of dye is lost to the wastewater and wastewaters from textile industries usually contain large amounts of dyes as well as organic matter, salts and other substances. The target of the project is to deliver a viable treatment method to the tex-tile industry for cleaner production.

Figure (above): Plastic carriers by Kaldnes (KPC) pre-pared with cryogel used for capture of pollutants and hormones. Figure (right): KPCs in column for copper capturing. Photos by Linda Önnby.

Marika MurtoAssistant Professor

Anaerobic decolourisation of azo dyes. Pho-to by Maria Jonstrup.

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Dept. of Biotechnology Annual Report 2009 17

Functional foods

Patrick AdlercreutzProfessor

Biotechnology for production of healthy food

Recent advances in biotechnology have increased the possibilities to make food components with improved properties. Enzymatic methods are used to tailor-make lipids containing health-promoting fatty acids, such as ome-ga-3 fatty acids. Likewise enzymes can be used for preparation of health-promoting carbohydrates from plant material.

The department is involved in one project within the FUNCFOOD program organised by the Functional Food Science Centre at Lund Univer-sity. The PhD student Julia Svensson is jointly supervised by Patrick Adler-creutz and by Åke Nilsson and Lena Ohlsson at the Medical Faculty. The project concerns lipids rich in the omega-3 fatty acid alfa-linolenic acid.

There is evidence that the preferential oxidation of alfa-linolenic acid may be beneficial in fighting obesity and diabetes. Starting from natural fats and oils such as rape seed oil, linseed oil and butter oil, enzymes are used as catalysts for making new products with the aim to optimise the health promoting properties of alfa-linolenic acid. The physiological effects of the new products in humans are studied as part of the project at the Medical Faculty.

The department is also running two projects within the Antidiabetic food center (AFC). In one project, we (Patrick Adlercreutz, Eva Nordberg Karls-son, and Henrik Stålbrand from the Biochemistry department) are using glycoside hydrolases for selective hydrolysis of plant material (focusing on agricultural byproducts). The carbohydrate products obtained are evalu-ated with respect to physiological effects in animals and ability to promote growth of probiotic bacteria.

Consumption of beta-glucans from e.g. oat has been shown to reduce cho-lesterols levels in blood thereby reducing the risk for coronary vascular dis-eases. Other soluble fibers from plant material have also been demonstrated to have similar effects. We are now exploring possible health beneficial ef-fects of microbial soluble fibers (polysaccharides) in a project together with Prof. Per Hellstrand and Dr Krisitina Andersson at Dept of Experimental Medical Science, Lund University.and Aventure AB and with support from Antidiabetic Food Center. Various commercially available products as well as our own isolates (see “Biodiversity”) are evaluated in animal models for their effects.

ProjectsSafe food products containing long-chain omega-3 fatty acidsHealth promoting lipids containing alfa-linolenic acidAntidiabetic carbohydrates by en-zymatic transformation of plant materialMicrobial dietary fibers for human health and well-being

PhD studentsCecilia Lindström, Julia Svensson

Post DocsMushtaq Ahmad, Carl Grey, Shiva Shanker Kaki

Project workerPeter Falck

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Olle HolstProfessor

“The development of more healthy food will be essen-tial for combating life style

related diseases, such as diabetes and heart disease”Eva Nordberg Karlsson

Associate Professor

Oat husks, is currently an agricultural waste, but is in the anti diabetic carbohydrate-project treated with enzymes to obtain oligosaccharides from the remaining hemicellulose. Photo by Eva Nordberg Karlsson.

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Dept. of Biotechnology Annual Report 200918

Renewable energy

Lovisa BjörnssonAssociate Professor

Jing LiuAssistant Professor

Biotechnical solutions will reduce our fos-sil fuel dependence

A sustainable bioenergy system should: 1. be resource efficient, 2. have a high energy efficiency, 3. have maximized environmental benefits and 4. be cost efficient. These four issues are addressed in the bioenergy research at Department of Biotechnology. Biomass based production of ethanol, hydrogen, methane or combinations hereof are studied, with focus on the microbial methane production process. Challenging research issues are how to improve microbial and operational efficiency of the systems, which in our research is addressed by:

Figure (top): Lab-scale experiment on biogas production from straw leach-ate. Photo by Valentine Nkemka.

Bo MattiassonProfessor

Marika MurtoAssistant Professor

ProjectsCo-production of ethanol and methane from hempChallenges in large scale production of bi-ogas from energy cropsWrams Gunnarstorp: optimising large scale co-digestionCrops 4 biogas: an interdisciplinary project on sustainable production of biogas from energy cropsEnzymatic upgrading of biogasCounteracting ammonia inhibition in anaer-obic digesters by struvite crystallizationThermophilic microorganism catalyses pro-duction of ethanol from glucose and/or xy-loseBiohythane: Biological hythane production from biomassDry anaerobic co-digestion of seaweed and cow manureAnaerobic digestion of biomass to promote local energy production in developing coun-triesProject for developing the concept of “Intel-ligent Biogas Plant”: 1) plant-wide compu-ter simulation for full-scale biogas plants; 2) development and evaluation of computer simulation models for anaerobic digestion processes; 3) evaluation of anaerobic CSTRs operated in series Measurement of biogas potential using a multichannel device for registration of po-tential and kinetics

PhD studentsMalik Badshah, Carla Crespo, Nges Ivo Achu, Emma Kreuger, Maryam Latifian

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Anaerobic digestion of agricultural residues and food industy waste in CSTRs. Photo by Lovisa Björnsson.

Improved energy efficiency, giving higher energy output. This is achieved by optimized process de-sign, improved process control, or pretreatment of substrates for higher conversion rates.Quantification and optimisation of the environ-mental benefits inherent in the process. Biorefinery concepts, where high-value products are produced, combined with energy production from the residuals to ensure sustainability.

Visiting researchersFederico Escobar, Xinmei Fu, Valentine Nkongndem Nkemka

Research engineersSofia Johannesson, Dalibor Jovanovic

Visiting studentsJorge Arenales Rivera, Sten Strömberg

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Dept. of Biotechnology Annual Report 2009 19

Education

Course Code Credits (hp)

No. of stu-dents

Responsible teacher

Programme related courses

Bioanalytical chemistry KBT050 7.5 38 Bo Mattiasson

Bioprocess technology KBT115 7.5 89 Olle Holst

Biotechnological separation processes KBT060 7.5 35 Igor Galaev

Biotechnology, process and plant design KBT042 15 18 Olle Holst

Biotechnology KKKA05 15 83 Olle Holst

Biotechnology, Advanced course KBT410 15 2 Olle Holst

Environmental biotechnology KBT080 7.5 27 Lovisa Björnsson

Enzyme technology KBK031 7.5 44 Patrick Adlercreutz

Green chemistry and biotechnology KBTF01 7.5 32 Eva Nordberg Karlsson

Independent courses

Degree project in biotechnology for a Bachelor of Science

KBTL01 15 1 Olle Holst

Entrepreneurship in biotechnology TNK021 7.5 13 Bo Mattiasson

Green chemistry and biotechnology TNK280 7.5 2 Eva Nordberg Karlsson

New solutions to old problems - biotech-nology for a sustainable society

TFRF35 3 28 Olle Holst

PhD courses

Mass spectrometry 3 3 Martin Hedström

Microbiology, Advanced course 7.5 4 Olle Holst

The department is responsible for giving nine courses at Masters level and four courses belonging to the LTH independent course programs. The number of students has increased over the past years which has resulted in a slightly higher work load for responsible teachers but also created a larger pool of students available for master thesis projects and as possible recruits for PhD positions. Courses offered during 2009 are listed below.

Ally Mahady och Mridul Jain are doing laboratory exercise in the course Environmental biotechnology, KBT080. Photo by Emma Kreuger.

Courses

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Dept. of Biotechnology Annual Report 200920

4

Jorge Arenales RiveraComparison of two-stage anaerobic digestion of seaweed and cow ma-nure

Nuno BimboOptimization of Poly(3-hydroxybu-tyrate) production by Halomonas bo-liviensis using fed-batch cultivation

Milica FidanoskaThe use of hydrogenase from Ralsto-nia eutropha for cofactor regenera-tion in biocatalytic redox reaction

Master thesis projects

As earlier years the department of Biotechnology took active part in teaching and supervising students on the In-ternational Master’s programme in Biotechnology during 2009. The objective of this programme is to offer courses focused on modern methods of Biotechnology. Prof. Olle Holst is the coordinator of the programme.

International Master’s programme in Biotechnology

Sofia JohannessonEvaluation of anaerobic CSTRs oper-ated in series

Ida JohanssonUtredning av en ny kolhydratbaserad agglutinationsanalys för detektion av xenoantikroppar med antikroppsspeci-fika Glycobeads

Watson Lima Afonso NetoImmobilization of spray dried E.coli containing recombinant aminotrans-ferase using hydrous titanium oxide

Camilla Melin FürstExploring the oxidative inactivation of lipase B from Candida antarctica

Catarina SousaExpression of a novel thermostable CGTase of Archaeal origin

Sophie WirsénEnzymatisk omestring - fria fettsyror och acylmigrering

Figure (right): Rajni Hatti-Kaul, Gunnar Lidén and Xue-Song Bai attending the workshop on “Biomass Energy” in Nanning, China, arranged within the IVA-CAE cooperation.

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Dept. of Biotechnology Annual Report 2009 21

International collaborations

North, Central and South AmericaNICARAGUA

Universidad Nacional Autonoma de Nica-

ragua, Managua

BOLIVIA

Universidad Mayor de San Andres, La Paz

Universidad Mayor de San Simón,

Cochabamba

ARGENTINA

University of La Pampa, Santa Rosa

EuropeICELAND

Prokaria Ltd. Reykjavik

GREAT BRITAIN

University of Newcastle, Newcastle upon

Tyne

University of Bath

University of Brighton

FRANCE

Foreign Affairs Ministry, Paris

ENSCR, Reinnes

Veolia Water, Paris

Anjou recherche

Veolia Environment, Maisons-Laffitte

DENMARK

Technical University of Denmark, Kgs.

Lyngby

University of Aarhus

GERMANY

Technical University, Braunschweig

Christian-Albrechts University, Kiel

University of Stuttgart

AUSTRIA

Technical University, Graz

PORTUGAL

University of Algarve, Faro

SPAIN

Valladolid University, Valladolid

ITALY

Politecnico di Milano

RUSSIA

Russian Academy of Sciences, Moscow

AfricaEGYPT

Suez Canal University, Ismailia

Cairo University, Cairo

Beni-Sueif University

Nuclear Research Centre, Cairo

KENYA

University of Nairobi, Nairobi

TANZANIA

University of Dar es Salaam

SOUTH AFRICA

Durban University of Technology

University of Limpopo

TUNISIA

National Institute of Applied Science and

Technology (INSAT)

AsiaTURKEY

Hacettepe University, Ankara

JORDAN

Philadelphia University, Amman

AZERBADIJAN

Institute of Polymer Materials, Baku

IRAN

Azad University, Tehran

Babol University of Technology, Babol

Isfahan University of technology, Isfahan

INDIA

Indian Institute of Technology, Delhi

Indian Institute of Technology, Kanpur

Agharkar Research Institute, Pune

Bhabha Atomic Research Centre, Mumbai

Central Food Technology Research Insti-

tute, Mysore

National Institute for Interdisciplinary Sci-

ence and Technology, Kerala

SRI LANKA

University of Jaffna, Jaffna

THAILAND

Asian Institute of Technology, Bangkok

Prince of Songkhla University, Hat Yai

VIETNAM

Hanoi University of Education, Hanoi

MALAYSIA

Universiti Putra Malaysia, Selangor

Universiti Teknologi Malaysia, Johor

CHINA

Institute of Process Engineering, Chinese

Academy of Sciences, Bejing

JAPAN

Miyazaki University, Miyazaki

During 2009, the department collaborated with universities, institutes and companies in about 30 countries world-wide. The collaboration included both minor initiatives such as student exchange as well as large research pro-grammes spanning over several years. The collaboration covered all major research areas at the department.

Figure (left): The department of Biotechnol-ogy has collaborations in about 30 coun-tries. Illustration by Dept of Biotechnology.

Figure (right): Rajni and Bo together with Dr Prashant Dhakepalkhar and his wife at the Indian Congress of Micro-biology in Pune in December 2009.

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Dept. of Biotechnology Annual Report 200922

Staff

Professor, Head of [email protected]+46-46-2224840

Rajni Hatti-Kaul

Research [email protected]+46-46-2227578

Martin Hedström

[email protected]+46-46-2229856

Siv Holmqvist

[email protected]+46-46-2229844

Olle Holst

Assistant [email protected]+46-46-2228347

Jing Liu

[email protected]+46-46-2224842

Patrick Adlercreutz

Programme SecretaryTechnology [email protected]+46-46-2224838

Josefin Ahlqvist

Technology [email protected]

Maria Andersson

Associate [email protected]+46-46-2228324

Lovisa Björnsson

Senior [email protected]+46-46-2220881

Igor Galaev

Research [email protected]+46-46-2224741

Gashaw Mamo

[email protected]+46-46-2228264

Bo Mattiasson

Assistant [email protected]+46-46-2228193

Marika Murto

Research [email protected]+46-46-2224948

Frans Peder Nilson

Associate [email protected] +46-46-2224626

Eva Nordberg Karlsson

Economy AdministratorGunilla Flodmark

Laboratory [email protected]+46-46-2228157

Tarek Dishisha

Post Doctoral [email protected]+46-46-2220858

Carl Grey

Laboratory [email protected]

Pontus Lundemo

Laboratory [email protected]+46-46-2227363

Elin Brun

Research [email protected]

Mihaela Nistor

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Dept. of Biotechnology Annual Report 2009 23

[email protected]

Suhaila Hashim

[email protected]

Javier Linares Pastén

[email protected]

Sindu Mathew

[email protected]

Aarti Ozarkar

[email protected]

Catherine Paul

Post Docs

[email protected]

Sang-Hyun [email protected]

Ravi Kiran Purama

[email protected]

Shabana Basheer

[email protected]

Mushtaq Ahmed

[email protected]

Teresa Alvarez Aliaga

Post Doctoral Scientist+46-46-2224741 [email protected]

Cecilia Orellana

Research Engineer [email protected]+46-46-2224681

Christina Wenner-berg

[email protected]

Konstanze Stiba

[email protected]

Shiva Shanker

[email protected]

Linda Elowsson

[email protected]+46-46-2229659

Kersti Skarin-Norén

Economy [email protected]+46-46-2229659

Jeanette Nylin

[email protected]

Daniela Zehentgruber

Adjunct [email protected]+46-06-2228157

Shukun Yu

Laboratory [email protected]+46-46-2228157

Regine Wuttke

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Dept. of Biotechnology Annual Report 200924

[email protected]

Rosa Peralta Aragão

[email protected]

Simon Bengtsson

[email protected]

Malik Badshah

[email protected]

Hugo Cavero

[email protected]

Georgina Chavez Lizarraga

[email protected]

Carla Melgar Crespo

[email protected]

Thuoc Doan Van

[email protected]

Daniel Guzmán

[email protected]

Hector Guzmán

[email protected]

Anna Hagström

[email protected]

Victor Ibrahim

[email protected]

Nges Ivo Achu

[email protected]

Maria Jonstrup

[email protected]

Samiullah Khan

[email protected]

Harald Kirsebom

[email protected]

Emma Kreuger

PhD students

[email protected]

Abolgashem Danesh

[email protected]

Solmaz Hajizadeh

[email protected]

Rawana Al-Khalili

[email protected]

Gry Ravn Jespersen

Marina Kuzimenkova

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Dept. of Biotechnology Annual Report 2009 25

Mahmoud Labib

[email protected]

Maryam Latifian

[email protected]

Cecilia Lindström

[email protected]

Betty Mbatia

[email protected]

Laura Mendoza Fernandez

[email protected]

Marlene Munoz Gaitan

[email protected]

Zulma Perez

[email protected]

Tania Pozzo

[email protected]

Gustav Rehn

[email protected]

Fabian Rundbäck

[email protected]

Deepti Sahoo

[email protected]

David Svensson

[email protected]

Julia Svensson

[email protected]

Thuy Tran Thi

[email protected]

Ulrika Törnvall

[email protected]

Linda Önnby

[email protected]

Kinga [email protected]

Ramin Sabet

[email protected]

Roya Rezaei

[email protected]

Mohammad Mazlomi

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Dept. of Biotechnology Annual Report 200926

Visiting researchers and students

Adrien Dartiguelongue ENSIACET France

Ally Mahadhy University of Dar es Salaam Tanzania

Ami Patel Anand Agricultural University India

Anna Scaglia Padova Italy

Audrey Faure University of Perpignan France

Berillo Dmitriy Kazakh State University Kazakhstan

Caroline Pierre ENSCR France

Celesty Anapalagan Medical faculty University of Jaffna Sri Lanka

Cristina Teixeira University of Algarve Portugal

Daniel Zehentgruber Research Centre Julich Germany

Ilyas Inci Hacettepe University Turkey

Jargalan Gerelsaikhan Mongolia

Jorge Arenales Rivera University of Valladolid Spain

Junxian Yun Zhejiang University of Technology China

Katazyna Dymek A. Mickiewicz University in Pozna Poland

Kosin Teeparuksapun Prince of Songkla University Thailand

Laura Buxó Barnés Universitat de Barcelona Spain

Luca Rossoni University Bicocca di Milano Italy

Marisa Punzi Universita Di Pavia Italy

Milica Fidanoska Macedonia

Natalia Avaliani Ilia State University Georgia

Nokuthula Mchunu South Africa

Oksana Zaushitshyna Moscow State Academy M.V Lomonosov Russia

Osvaldo Delgado PROIMI Tucuman Argentina

Raguparan Pararasasingam Universtity of Jaffna Sri Lanka

Saluma Samanman Prince of Songkla University Thailand

Séverine Cozzi France

Stanislav Obruoc Brno University of Technology Czech Rep.

Subajini Jesuthasan University of Jaffna Sri Lanka

Supaporn Dawan Prince of Songkla University Thailand

Warakorn Limbut Prince of Songkla University Thailand

Watson Neto University of Algarve Portugal

Wilfred Tila Johnson University of Dar es Salaam Tanzania

Xinmei Fu South West University of Science and Technology China

Yasser Gaber Hasan Beni-Sueif University Egypt

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Dept. of Biotechnology Annual Report 2009 27

Funding and grantsAs many other departments and institutions at the university, the Department of Biotechnology is dependent on external sources for carrying out the research. The distribution of sources is shown below. Besides the funding given above, several visiting scientists and PhD students are supported directly from external funding agencies.

Source Funds (kSEK)

Faculty 6 001

Education 2 995

Research councils 7 530

Public authorities 7 005

Foundations 13 604

Industry 3 020

Total 40 155

Industry8%

Research councils19%

Education7%

Faculty15%

Public authorities17%

Foundations34%

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Dept. of Biotechnology Annual Report 200928

During 2009, 35 research papers were published in peer reviewed journals and seven PhD theses were published and defended. Several scientific results were also presented in several conferences. For reprints of papers contact Siv Holmqvist, [email protected].

Publications

Peer-reviewed articles

N. I. Achu and J. Liu Enhancement of anaerobic digestion of dewatered-sewage sludge through anaerobic pre-treatmentRenew. Energy 2009, 34: 1795-1800

G. Baydemir, N. Bereli, M. Andac¸ R. Say, I. Yu. Galaev and A. DenizliBilirubin recognition via molecularly imprinted supermacro-porous cryogelsColloid Surf. B: Biointerfaces 2009, 68: 33-38

G. Baydemir, N. Bereli, M. Andac¸ R. Say, I. Yu. Galaev and A. DenizliSupermacroporous poly(hydroxyethyl methacrylate) based cryogel with embedded bilirubin imprinted particlesReact. Funct. Polym. 2009, 69: 36-42

N. Bölgen, I. Vargel, P. Korkusuz, E. Güzel, F. M. Plieva, I. Yu. Galaev, B. Mattiasson and E. PiskinTissue responses to novel tissue engineering biodegradable cryogel-scaffolds: an animal modelJ. Biomed. Mater. Res. Part A 2009, 91: 60-68

E. N. Efremenko, I.V. Lyagin, F. M. Plieva, I. Yu. Galaev and B. MattiassonDried–reswollen immobilized biocatalysts for detoxification of organophosphorous compounds in the flow systemsAppl. Biochem. Biotechnol. 2009, 159: 251-260

T. Essam, M. A. Amin, O. El Tayeb, B. Mattiasson and B. GuieysseKinetics and metabolic versatility of highly tolerant phenol degrading Alcaligenes strain TW1J. Hazard. Mater. Online 6 September 2009

D. Guzman, J. Quillaguaman, M. Muñoz and R. Hatti-KaulHalomonas andenense sp. nov., a moderate halophile iso-lated from the saline lake Laguna Colorada in Bolivia Int. J. Syst. Evol. Microbiol. 2009, doi:10.1099/ijs.0.014522-0

H. Guzman, D. V. Thuoc, J. Martin, R. Hatti-Kaul and J. QuillaguamánA process for the production of ectoine and poly(3-hydroxy-butyrate) by Halomonas boliviensis Appl. Microbiol. Biotechnol. 2009, 84: 1069-1077

A. E. V. Hagström, M. Nordblad and P. AdlercreutzBiocatalytic polyester acrylation - process optimization and enzyme stabilityBiotechnol. Bioeng. 2009, 102: 693-699

A. Ivanov, J. Eccles, H. A. Panahi, A. Kumar, M. Kuzi-menkova, L. Nilsson, B. Bergenståhl, N. Long, G. J Phillips, S. V. Mikhalovsky, I. Y. Galaev and B. Mattias-sonBoronate-containing polymer brushes: Characterization, interaction with saccharides and mammalian cancer cellsJ. Biomed. Mater. Res. Part A 2009, 88: 213–225

H. Kirsebom, B. Mattiasson and I. Yu. GalaevBuilding macroporous materials from microgels and mi-crobes via a one-step cryogelationLangmuir 2009, 25: 8462–8465

H. Kirsebom, G. Rata, D. Topgaard, B. Mattiasson and I. Y. GalaevMechanism of cryopolymerization: diffusion-controlled polymerization in a nonfrozen microphase. An NMR studyMacromolecules 2009, 42: 5208–5214

P. Kueseng, M. Le Noir, B. Mattiasson, P. Thavarung-kul, P. KanatharanaMolecularly imprinted polymer for analysis of trace atrazine herbicide in waterJ. Environ. Sci. Health Part B-Pestic. Contam. Agric. Wastes 2009, 44: 772–780

M. Labib, M. Hedström, M. Amin and B. MattiassonA capacitive immunosensor for detection of cholera toxinAnal. Chim. Acta 2009, 634: 255–261

M. Labib, M. Hedström, M. Amin and B. MattiassonA multipurpose capacitive biosensor for assay and quality control of human immunoglobulin GBiotechnol. Bioeng. 2009, Online 20 May 2009

D. J. Leak, R. A. Sheldon, J. M. Woodley and P. Adler-creutzBiocatalysts for selective introduction of oxygen Biocatal. Biotransform. 2009 27: 1-26

M. Le Noir, F. M. Plieva and B. MattiassonRemoval of endocrine-disrupting compounds from water using macroporous molecularly imprinted cryogels in a moving-bed reactorJ. Sep. Sci. 2009, 32: 1471–1479

G. Mamo, M. Thunnissen, R. Hatti-Kaul and B. Mat-tiassonAn alkaline active xylanase: Insights into mechanisms of high pH catalytic adaptationBiochimie 2009, 91: 1187–1196

Journal articles

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Dept. of Biotechnology Annual Report 2009 29

S. Mathew and P. AdlercreutzMediator facilitated, laccase catalysed oxidation of granular potato starch and the physico-chemical characterization of the oxidized productsBioresour. Technol. 2009, 100: 3576-3584

B. Mattiasson, K. Teeparuksapun and M. HedströmImmunochemical binding assays for detection and quantifi-cation of trace impurities in biotechnological productionTrends Biotechnol. Online 5 November 2009

L. Murean, M. Nistor, Sz. Gáspár, I. C. Popescu and E. CsöregiMonitoring of glucose and glutamate using enzyme microstructures and scanning electrochemical microscopy, Bioelectrochemistry 2009, 76: 81-86

W. Noppe, F. Plieva, I. Yu. Galaev, H. Pottel, H. Deck-myn and Bo MattiassonChromato-panning: An efficient new mode of identifying suitable ligands from phage display librariesBMC Biotechnol. 2009, 9:21 (17 March 2009 – Open Ac-cess)

M. Nordblad, A. E. V. Hagström and P. AdlercreutzEnzymatic synthesis of polymer acrylates and their evalua-tion as wood coatingsIndust. Biotechnol. 2009, 5: 53-61

A. Numnuam, P. Kanatharana, B.Mattiasson, P. Asa-watreratanakul, B. Wongkittisuksa, C. Limsakul and P. ThavarungkulCapacitive biosensor for quantification of trace amounts of DNABiosens. Bioelectron. 2009, 24: 2559–2565

F. Plieva, A. Kumar, I. Y. Galaev and B. Mattiasson Design of biomaterials via gelation at subzero tempera-tures–cryogelationIn Advanced biomaterials: fundamentals, processing and applications. Eds. B. Basu, D.S. Katti and A. Kumar. Wiley, New Jersey, USA. 2009, pp 499-531

F. Plieva, E. De Seta, I. Y. Galaev and B. MattiassonMacroporous elastic polyacrylamide monolith columns: processing under compression and scale-upSep. Purif. Technol. 2009, 65: 110–116

D. Sahoo, J. Andersson and B. MattiassonImmobilized metal affinity chromatography in open-loop simulated moving bed technology: purification of a heat stable histidine tagged beta-glucosidaseJ. Chromatogr. B 2009, 877: 1651–1656

I. S. Savina, M. B. Dainiak, H. Jungvid, S. V. Mikha-lovsky and I. Yu. GalaevBiomimetic macroporous hydrogels: Protein ligand distri-bution and cell response to the ligand architecture in the scaffoldJ. Biomater. Sci. 2009, 20: 1781-1795

D. Svensson, S. Ulvenlund and P. AdlercreutzEfficient synthesis of a long carbohydrate chain alkyl glyco-side catalysed by cyclodextrin glycosyltransferase (CGTase)Biotechnol. Bioeng. 2009, 104: 854-861

D. Svensson, S. Ulvenlund and P. AdlercreutzEnzymatic route to alkyl glycosides having oligomeric head groupsGreen Chem. 2009, 11: 1222-1226

K. Teeparuksapun, P. Kanatharana, W. Limbut, C. Thammakhet, P. Asawatreratanakul, B. Mattiasson, B. Wongkittisuksa, C. Limsakul and P. ThavarungkulDisposable electrodes for capacitive immunosensorElectroanalysis 2009, 21: 1066–1074

D. V. Thuoc, H. Guzmán, M. Thi Hang and R. Hatti-KaulEctoine production by Halomonas boliviensis: optimization using response surface methodology. Marine Biotechnol., 2009, DOI 10.1007/s10126-009-9246-6

T. Tran, G. Mamo, B. Mattiasson and R. Hatti-KaulA thermostable phytase from Bacillus sp. MD2: cloning, expression and high level production in Escherichia coli J. Indust. Microbiol. Biotechnol. 2009, DOI: 10.1007/s10295-009-0671-3

U. Törnvall, C. M. Furst, R. Hatti-Kaul and M. Hed-strömMass spectrometric analysis of peptides from an immobi-lized lipase: focus on oxidative modifications Rapid Commun. Mass Spec. 2009, 23: 2959-2964

U. Törnvall, L. M. Tufvesson, P. Börjesson and R. Hatti-Kaul Biocatalytic production of fatty epoxides from rapeseed and tall oil derivatives: process and environment evaluation Indust. Biotechnol. 2009, 5: 184-192

J. Yun, H. Kirsebom, I. Yu. Galaev and B. MattiassonModeling of protein breakthrough performance in cryogel columns by taking into account the overall axial dispersion J. Sep. Sci. 2009, 32: 2601-2607

K. Zór, R. Ortiz, E. Saatci, R. G. Bardsley, T. Parr, E. Csöregi and M. NistorLabel free capacitive immunosensor for detecting calpasta-tin - a meat tenderness biomarkerBioelectrochemistry 2009, 79: 93-99

Mathias Nordblad, Anna E V Hagström, and Patrick Adlercreutz*

Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden

*Corresponding authorEmail: [email protected]: +46 (46) 222 4842Fax: +46 (46) 222 4713

[email protected]@biotek.lu.se

Submitted: 1 Oct 2008; Revised: 11 March 2009, 6 May 2009;Accepted: 18 May 2009

KEYWORDSacrylation, lipase, polymer modification, radiation cure, wood coating

ABBREVIATIONSBEPD, butyl ethyl propanediol; CalB, Candida antarctica lipase B; CN435, commercially available polyol acrylate; Darocur® 1173, 2-hydroxy-2-methyl-1-phenylpropane-1-one, photoinitiator; DPGDA, dipropylene glycol diacrylate; EO, ethane diol residue; GC, gas chromatography; GPC, gel permeation chromatography; HHPA, hexahydrophthalic acid; LR 9004, commercially available polyester acrylate Laromer® LR 9004; Mn, number average molecular weight; Mw, weight average molecular weight; MEHQ, methoxyhydroquinone; NPG, neopentyl glycol; PD, pentanediol; PDDA, pentanediol diacrylate; PDMA, pentanediol monoacrylate; PPTTA, alkoxylated pentaerythritol tetraacrylate; R3215, polyol, ethoxylated TMP (15:1); TMP, trimethylol propane

AbstractUV-curable acrylates are the base for a large and growing product

group within the coatings industry. Traditional chemical catalysis is poorly suited for acrylation reactions since these require low tem-peratures to avoid polymerization. In this study, immobilized Candida antarctica lipase B has been used to efficiently acrylate four polymers through transesterification of ethyl acrylate, via a solvent-free process

using continuous distillation. The four products were based on three polyesters and one polyether, which represent two major classes of acrylates used in coatings today. The performance of the products was estimated with regard to curing rate, hardness, and chemical-resis-tance. While each product by itself had some weakness, a combina-tion of acrylated polyether and polyester produced an adequate curing rate, good hardness, and excellent chemical-resistance properties.

Introductionlmost all wood used for flooring and furniture is treated with some kind of surface coating. The coating has several purposes, one of which is to protect the surface from stains by substances such as water, grease, and beverages such as

coffee. It can also be of interest to make the surface harder to improve the scratch-resistance.

Lacquers are coatings that can cross-link after application on the intended surface. This type of coating gives considerably better chemical- and mechanical-resistance than more traditional alterna-tives such as oils and waxes. The basis for a lacquer is a binder molecule that contains functional groups for polymerization. Several compounds can be combined in a coating to achieve the desired properties. Since the binders, which are often based on polymers themselves, can be highly viscous, solvents, or reactive diluents (sol-vents that can be incorporated into the hardened lacquer) are needed. In addition to the cross-linkable materials, wetting agents are often used to aid in the application of the coating to a wood surface. Other additives alter the appearance of the coating with respect to texture, gloss, and color.

UV-curable lacquers cross-link when they are irradiated with UV-light. These coatings can be applied and cured in a matter of minutes, which is a great benefit in a production process. Acrylates (esters of acrylic acid) represent a large segment of the market for UV-curable coatings. They are used both as binders and as reactive diluents. Cross-linking of the acrylic functionalities occurs through radical polymerization of double bonds.

A thorough introduction to the considerations involved in formu-lating UV-coatings has been published by Zwanenburg.1 The polymer resin used in acrylic binders can be based on several different types of

Enzymatic synthesis of polymer acrylates and their evaluation as wood coatings

A

110 INDUSTRIAL BIOTECHNOLOGY SUMMER 2009

PEER REV I EWOR IG INAL R ESEARCH

Publication in Industrial Biotechnology.

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Dept. of Biotechnology Annual Report 200930

Doctoral degrees and theses

ConferencesBelow are listed the national and international conferences attended by Biotechnology co-workers during 2009.

5th IWA specialist conference on oxidation technolo-gies for water and wastewater treatmentBerlin, Germany, 30 Mar-1 Apr 2009

Carbohydrate Bioengineering Ischia, Italy, 10-13 May 2009

XXth International Symposium on Bioelectrochemistry and BioenergeticsSibiu, Romania, 10-14 May 2009

Lipidforum Helsingör, Denmark, 14-17 June 2009

Science ForumWageningen, The Netherlands, 16-17 June 2009

Biotrans 2009 Bern, Switzerland, 5-9 July 2009

14th European Congress on Biotechnology - SYMBIO-SIS - Science, Industry and Society (ECB 14) Barcelona, Spain, 13-16 September 2009

World Bioenergy: Clean Vehicles & Fuels Stockholm, 16-18 September, 2009

1st IWA Development Congress, Water and sanitation services, What works in developing countriesMexico City, Mexico, 15-19 November 2009

BioMicroWorldLisbon, Portugal, 2-4 December 2009

Fourth International Conference on Fermented Foods, Health Status and Social Well-being, 11-12 December 2009, AAU, Anand, India

50th Association of Microbiologists of India Confer-encePune, India, 15-18 December 2009

Simon BengtssonProduction of polyhydroxyalkanoates in biological treatment of industrial wastewaters

Thouc Doan VanProduction of poly(3-hydroxybutyrate) and ectoines using a halophilic bacterium

Torbjörn HåkanssonMicrobial remediation of heavy metal ion polluted soil and water

Marina KuzimenkovaCarbohydrate-specific, boronate-containing copoly-mers: From sugar-sensing to cell adhesion

Valentine Nkemka and Marika Murto attended the 1st IWA Development Congress, Water and sanita-tion services, What works in developing countries, in Mexico City, Mexico.

Mahmoud Aziz LabibDevelopment of capacitive biosensors for monitoring of bacterial toxins and other biomolecules

David SvenssonEnzymatic synthesis of alkyl glycosides - new applica-tions for glycoside hydrolases

Ulrika TörnvallChemo-enzymatic epoxidation - activity and stability of Candida antarctica lipase B

Thouc Doan Van (2nd to the left) defended his thesis “Production of poly(3-hydroxybutyrate) and ectoines using a halophilic bacterium” in Dec. 2009.

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Dept. of Biotechnology Annual Report 2009 31

Department spring tripMay 2009Immeln

Photos by Tarek Dishisha and Emma Kreuger

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Dept. of Biotechnology Annual Report 200932

Department of BiotechnologyCenter for Chemistry and Chemical Engineering

Lund UniversityP.O. Box 124

SE-221 00 LundSWEDEN

www.biotek.lu.se

Email addresses Personal addresses are constructed by

name and surname as follows:[email protected]

Annual report 2009

Publisher: Rajni Hatti-Kaul Editor: Marika Murto