PTT 104 Biotechnology and Industry Week 7: 24 October 2013 Department of Chemical Engineering Technology, UniMAP [email protected].

PTT 104Biotechnology and

IndustryWeek 7: 24 October 2013

Department of Chemical Engineering Technology, [email protected]

Course Outcome

CO3:Ability to differentiate scopes and importance of various biotechnological

streams.

Topics covered:Illustrate scopes of industrial

biotechnology and examine commercial production of microorganisms and product

from microorganisms.

BIOTECHNOLOGY POLICYTHRUSTS THRUST 1 : Agricultural Biotechnology

Development THRUST 2 : Healthcare Biotechnology

Development THRUST 3 : Industrial Biotechnology Development THRUST 4 : R&D and Technology Acquisition THRUST 5 : Human Capital Development THRUST 6 : Financial Infrastructure THRUST 7 : Legislative and Regulatory Framework THRUST 8 : Strategic Development THRUST 9 : Government Support and

Commitment

Industrial Biotechnology: applications

Create new products, such as plant-based biodegradable plastics;

Replace petroleum-based feedstocks by processing biomass using biorefineries to generate electricity, transport fuels or chemicals; 

Modify and develop new industrial processes, such as by using enzymes to reduce the amount of harsh chemicals used in textiles and the pulp and paper industry;

Reduce the environmental impact of manufacturing; for example by treating industrial wastewater onsite using biological mediums such as microbes;

Provide energy savings by adding enzymes in detergents, allowing clothes to be washed in lower temperatures; and

Provide water savings through more efficient processes such as using enzymes to break down chemicals and reduce subsequent washing steps in the textile industry

Economic

EnvironmentSocial

INDUSTRIAL BIOTECHNOLOGY: IMPACT ON SUSTAINIBILITY• Higher profits – lower costs (raw materials, process costs, investments, …)• Developing new products• Finding new uses for ag crops

• Creating new jobs and opportunities• Rural economic diversification and

growth• Lower risk for workers (lower temp.)• Less negative perception

Processes are carbon neutral – no contribution to global warming Products and byproducts are in most cases biodegradable Reduction of greenhouse gas emissions, and emissions to water and air Using renewable resources as feedstock help conserve fossil fuels

Industrial Biotechnology

Practices that use living cells: bacteria,

yeast or algae) Or component cells like

enzyme

Feedstocks/ biomass based materials

Generate industrial products and

processes

- products: Fuels/chemicals

- process: wastewater treatment

Microorganisms as Tools

Microbes have been used in food sectors: beer and wine, bread (yeasts).

Making decontamination processes for industrial wastes product removal more efficient

Leaching of oil and minerals from the soil to increase mining efficiency

Also used to clone and produce batch amounts of important proteins used in human medicine including insulin and growth hormone.

Fermentation

Animal, plant cells and microbes obtain energy from carbohydrates (glucose) using electrons from these sugar to create ATP.

ATP occurs in series. 1st step is glycolysis: convert sugar molecule into 2

mol of pyruvates. During the reaction, electrons are transferred from

glucose to NAD+ producing NADH. Results in ATP production.

O2 is important in this electron transport process. For anaerobes, they derived their energy from

sugars in the absence of O2 FERMENTATIONEg: alcohol fermentation and lactic acid fermentation

Microbial enzymes

Enzymes: increasing the speed of reaction by lowering the activation energy of a reaction.

Cheese making: using enzyme rennin

Eg: cellulase has been used widely in the industry.

In food industry: make animal food more digestible

Stone-washed jeans: the denim is treated with cellulases from T. reesei and A. niger. These microbes produce cellulase that can digest the cotton fibre softer fabric.

• Enzyme subtilisin: isolated from B. subtilis used in laundry detergent- degrade the protein stains.

• enzyme amylase: used to degrade starches for making corn syrup.

Industrial Biotechnology: In Malaysia

The industrial Biotechnology sector is broad.

3 main areas in Malaysia:- Biofuels- Biocatalyst- Fine and Specialty Chemicals

Biofuel represent an alternative fuel source to non-renewable petroleum-based fuels.

‘First-generation' or conventional biofuels are biofuels made from sugar, starch, and vegetable oil.

Biofuel

2nd generation biofuels: using lignocellulosic materials from wheat straw, sugarcane baggase, corn husks, discarded rice hulls and trees.

Breaking the lignocellulose into cellulose and hemicellulose using enzyme from fungi fermentable sugars/glucose

Glucose will be used by yeasts to produce ethanol.

Name and Description Source Application

Bioethanol

Ethanol produces by breakdown of biomass

Corn, sugarcane, molasses, wheat, barley

Motor vehicle transport

Biodiesel

Produced from vegetable oils from trans esterification process

Soybean oilPalm Oil

Blend with petroleum diesel

Biogas

Principally Methane, generated by biodegradation of feedstock

Landfill biomassWaste waterOther biomass and feedstock

Turbine based electricity generation

Principle Biotechnology Generated Biofuels

Bioethanol are not currently produced in Malaysia.

However initiatives are underway to develop ethanol and other biofuels from non food agricultural crop sources such as Jatropha curcas and oil palm based (trunks, fronds, empty fruit bunch, shell and fiber)

Potential Crops in Biofuel Production

Production of Biodiesel

Biogas Production

Biogas production using anaerobic digestion (oxygen free) is a biological treatment process to reduce odor, produce energy and improve the storage and handling characteristics of manure.

Anaerobic digestion is the natural breakdown of organic materials into methane and carbon dioxide gas and fertiliser.  This process takes place naturally, or in an anaerobic digester.

A typical anaerobic digester is a sealed vessel, or series of vessels, in which bacteria act without oxygen.  The organic material contents need to be fully mixed and warmed, usually to blood temperature.

Biogas is the name given to the mixture of gases formed during the anaerobic digestion of organic wastes.

Biocatalyst are proteins that act to accelerate chemical reactions by bringing chemical compound s involved in a reaction.

Biocatalysts must be produced by living organisms and are typically derived from plant, animal, or microbial sources

Bio-Catalyst

Malaysia’s biological diversity offers developers of novel biocatalyst a significant opportunity to isolate novel biocatalysts.

A variety of different biocatalyst have been isolated from Malaysian isolated microorganisms.

Type of Biocatalyst Microorganisms

Lipase, lipoprotein lipase Hunicola lanuginosa, Aspergillus niger, Aspergilus flavus, Mucor miehei, Bacillus sp., Pseudomonas sp.

Protease Bacillus megaterium, Trichoderma sp., Aspergillus niger

Cellulase Aspergillus niger, Tricgoderma resei

Lignin degrading enzymes Phanerochate chyososporium, Humicola grisea

Tannase Aspergillus niger,

Mannase Aspergillus niger

Phytase Aspergillus niger

Chitinase Fusarium sp.

Biocatalyst Isolated from Malaysian Microorganisms

Production Process1. Fermentation2. Formulation 3. Recovery

Novozymes Company Technologies in Enzyme Production

Making microorganisms produce more enzymes. Novozymes is the world leader in developing new methods to optimize the amount of enzymes that microorganisms can produce. The result is cheaper products and faster delivery to their customers.

Enzyme production: Fermentation

Fermentation to produce industrial enzymes starts with a vial of dried or frozen microorganisms called a production strain. This production strain is selected to produce large amounts of the enzyme(s) of interest.

Sterilization – A key facilitator in the production of enzymes

A key element of fermentation science is sterilization. In order to cultivate a particular production strain it is necessary to start by eliminating all the native microorganisms present in the raw materials and equipment. If this is not done satisfactorily, the wild organisms will quickly outnumber the production strain, and no production will occur. Sterilization can be achieved by heat and/or special filters.

The cultivation process

The production strain is first cultivated in a small flask containing nutrients and agar. The flask is placed in an incubator that provides the optimal temperature for the previously frozen/dried cells to germinate.

Once the flask is ready, the cells are transferred to a seed fermentor, which is a large tank containing previously sterilized raw materials and water known as the medium. Seed fermentation allows the cells to reproduce and adapt to the environment and nutrients that they will encounter later on.

Following seed fermentation, the cells are transferred to a larger tank, the main fermentor, where temperature, pH, and dissolved O2 are carefully controlled to optimize enzyme production. Additional nutrients may be added to enhance productivity. When the main fermentation is complete, the mixture of cells, nutrients, and enzymes, referred to as the broth, is ready for filtration and purification.

2. Recovery

The purpose of the recovery process is to separate the enzyme from the biomass and to produce a solution that contains the enzyme at a purity that can be used for formulation of the final product.

The main factors that influence the design of an enzyme recovery process are:

1.The properties of the production organism2. The characteristics of the enzyme 3. Product quality demands4. The type of product to be produced5. The environmental impact of the process

Formulation of the enzymes is the third important process step after fermentation and recovery.

The nature of the enzyme protein is the starting point of all formulation work, and knowledge about parameters such as solubility and compatibility is indispensable.

A new enzyme molecule with excellent performance can fail in the market if the enzyme is not stable during transportation and storage.

Formulation

The industrial biotechnology sector is a key contributor to the production of vitamins, amino acids, and other biochemical such as lactic acid and glycerol

Industrial Biotechnology: Fine & Specialty Chemicals

Production of Amino Acid

Currently, the amino acids used in amino acid products are mainly manufactured by the fermentation method using natural materials, similar to yogurt, beer, vinegar, miso (bean paste), soy sauce, etc.

Coryneform bacteria

Kojic Acid Production from Mushroom

In 1989, kojic acid was first discovered as a natural by product from the Japanese mushroom. Since then it has been widely use as an effective skin lightening agent. Kojic acid works by blocking the production of skin melanin.

Production of PolyLactic Acid

Industrial Biotechnology: Microbial Biomass production

Production of single cell proteinProduction of mushrooms

Single cell protein

Single-cell protein (SCP) typically refers to sources of mixed protein extracted from pure or mixed cultures of algae, yeasts, fungi or bacteria (grown on agricultural wastes) used as a substitute for protein-rich foods, in human and animal feeds.

Mushrooms

In mushroom production: solid-state fermentation is employed.