Toward a Bio-based Economy: Biochemical Platform for Fuels ... · Dr. Dirk E. Maier Professor and Head . KSU Grain Science & Industry Faculty ... (Venugopal 2009) astaxanthin 40-70mg/kg

Toward a Bio-based Economy: Biochemical Platform for Fuels, Chemicals and Feed from Renewable Resources

Praveen V. Vadlani

Grain Science and Industry Department

Kansas State University

Manhattan, KS 66502

USA

Oil

Crops

Solvents

Molded parts

Refining Chemistry & Chemical Engineering

Biomass Biological Engineering

Current Feedstocks

New Feedstocks

Fuels

Fibers

Serving the Marketplace: A Bio-Based Economy

Source: www.dupont.com

Motivations for Bioeconomy

• Environmental quality – Local and regional (air quality,

solid waste disposal)

– Global climate change

• National security – Reduced reliance on foreign fuel

sources

• Rural development – Rural economies are not thriving

in many parts of the world

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KANSAS

• Located in the middle of the United States

KANSAS

• Economy:

–Agriculture

–Aviation

–Oil and Gas

– Service Sector

Kansas Agriculture

#1 Wheat Producing State

#1 Wheat Flour Producing State

#1 Grain Sorghum Producing State

#2 Cattle in Feedlots and Slaughter

#3 Land in Farms

#5 Cash Receipts from Farm Sales

#6 Exports of Farm Products

Manhattan, Kansas

• 2 ½ hours west of Kansas City

Manhattan, Kansas • Located in the Flint Hills of Kansas which consists of

limestone rock and prairie grass.

• Population of approximately 60,000, including students.

Kansas State University

• Beautiful campus with limestone buildings

• 23,300 students

College of Agriculture Our College is one of Nine at K-State:

• Agriculture

• Architecture, Planning and Design

• Arts and Sciences

• Business

• Education

• Engineering

• Human Ecology

• Technology and Aviation

• Veterinary Medicine

College of Agriculture

• 2,536 Agricultural students in total

• 2,087 Undergraduate students

• 449 Graduate students

• Partnering with businesses small and large (ADM, Cargill, John Deere, Rabobank) to provide real projects, internships, and jobs to our students

Special Targeted Programs

• Bioprocessing and Industrial Value-Added Program

• Sorghum Improvement Center

• Wheat Genetic and Genomic Resource Center

• National Plant Diagnostic Network

• National Agriculture Bio-security Center

• International Grains Program

• International Meat and Livestock Program

Department of

Grain Science & Industry - A Brief Overview

Dr. Dirk E. Maier Professor and Head

KSU Grain Science & Industry Faculty

August 2009

GSI North Complex on Kimball Ave

(View from top of East Stadium.)

Unique Departmental Programs

• Bioprocessing & Industrial Value Added Program

• “To develop bioprocessing technology and utilize agricultural-based materials to produce higher-value food, feed, and industrial uses products for economic development.”

Bioprocessing Lab

Value-Added Products: Biomaterials, Biopolymers, Enzymes, Biofuels,

Bioprocessing

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Fermentation from Latin verb ‘ fervere’ - action of yeast on fruit extracts Louis Pasteur – Father of Fermentation – ‘life without air’ Different types of fermentation by specific microorganisms Pure culture – Pure Product Bioprocessing – Integration of Biochemistry, Microbiology, Engineering principles Application of microbial means in an industrial process to produce specific products

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The Fuel Ethanol Process

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Break down starch to dissolved sugars

(Enzymes convert starch to simple sugar)

Fermentation of sugar to produce ethanol

(Yeast converts glucose into ethanol)

Purify ethanol to a concentrated form

(Distillation)

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FUEL ETHANOL INDUSTRY

Whole Corn Hammer Mill Slurry Tank

Jet Cooker

Steam

Fresh Water &

4 Recycled

Water Sources

Liquefaction

Enzymes

Mash Cooling

CO2

Beer

190 Proof

Ethanol

Syrup

Condensate

Evaporator

Thin

Stillage

DDGS Final

Product

5% Gasoline

200 Proof

Ethanol

Wet Grain

Whole

Stillage

Alcohol

Dehydration 200 Proof

Denatured

Ethanol Final

Product

Rotary Dryer

Distillation

System

Centrifuge

Continuous

Fermentation

Source: MGPI, Atchison, Kansas

A Bushel of Corn is 25kg (56lbs)

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Sugars 17 kg

Ethanol 8.1 kg(93% efficiency)

Wet Corn 25 kg

Starch 15.3 kg

DDG 7.6 kg

CO2 7.7 kg(93% efficiency)

ETHANOL – COST OF PRODUCTION

El e c t r i c i t y

4 . 1%

Fue l s

16 . 8 %

Wa st e ma na ge me nt

0 . 4 %

Wa t e r

0 . 1%

Enz y me s

2 . 4 %

Ye a st

0 . 4 %

Che mi c a l s

1. 9 %

De na t ur a nt

4 . 6 %

M a i nt e na nc e

4 . 0 %

La bor

4 . 1%

Admi ni st r a t i v e

c ost s

2 . 8 %

Ot he r c ost s

0 . 4 %

De p' n

12 . 0 %

Fe e dst oc k

4 6 . 1%

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Major Components In Lignocellulosic Biomass

Lignin

Hemicellulose

Cellulose

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Bioethanol production from different biomass sources

Forage sorghum (Sorghum bicolor),

Switchgrass (Panicum virgatum),

Miscanthus (Miscanthus giganteus)

wheat straw (Triticum aestivum)

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Using Corn in Modified Corn Processing – David Johnston

Carotenoids in Animal Feed

• Anti-oxidants, enhance immune response in ruminants (Yang & Tume 1993)

• Precursors of Vitamin-A (van de Berg et al 2000)

• Reduce mastitis in cows (Chew 1995)

• Improve keeping quality of milk and meat (Noizere et al 2006)

• Improve reproductive efficiency (Hurley & Doane 1989, Chew 1993)

• β-carotene enhances color of flesh (salmon & trout) and egg yolk

• Fish feed-total carotenoids 30-120mg/kg (Venugopal 2009)

astaxanthin 40-70mg/kg (Decker 2000)

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Carotenoids

• Astaxanthin – Natural $7,000-15,000/kg

– Synthetic $2,000/kg

• β-carotene (Caswell & Zilberman 2000)

– Natural $1,000-2,000/kg

– Synthetic $400-800/kg

• Carotenoid supplementation to animal feed is expensive

• Natural product preferred

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Red yeasts fermentation • Monoculture

Phaffia rhodozyma (PR) Astaxanthin & β-carotene producer Sporobolomyces roseus (SR) β-carotene producer only

• Samples centrifuged, pellet freeze dried

• Carotenoids extracted • HPLC • MALDI/TOF MS confirmation

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PR SR

Seed flask

SR

Lab scale fermentation

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Carotenoid-enriched DDGS

Ananda N, Vadlani PV (2010) Journal of Agricultural and Food Chemistry 58 (24): 12585–12591 Ananda N, Vadlani PV (2010) Journal of Industrial Microbiology and Biotechnology 37(11): 1183-1192

Soybean Fiber and Oligosaccharides Bioconversion

Soybean Oligosaccharides • Sucrose 2.5 - 8.2% • Raffinose 0.1 – 0.9% • Stachyose 1.4 – 4.1% Soybean Hull • 8 – 10% of soybean (dry basis) • Hull is made of 75% non-starch carbohydrates • 12% pectin

Project Advantages

• Bioconversion of under-valued soluble carbohydrates to high-value protein

• Removal of oligosaccharides – decreases flatulence and abdominal discomfort, and increases nutritive value of animal feed

• Bioconversion of soy hulls (fiber) to protein

• Microbial bioprocessing of soybean meal and hull - better sulfur amino acid profile and additional nutrients (vitamin B12)

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Production of succinic acid

• A. succiniproducens

• A. succinogenes

• E. coli

– succinic acid minor fermentation product (~7.8% of total)

• Deletion of ldh

• overexpression of PEPC/ malic enzyme or PYC

• Deletion of ldh and pfl

– NZN111

• Deletion of ldh/pfl/ptsG

– APF111

Non-recombinant microorganisms :

Metabolically Engineered E. coli:

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Engineered Ecoli for Succinic Acid

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SBS550MG (pHL413)

Succinic acid produced from 20g/L (~110 moles) of glucose

E. coli Wild Type

succinate

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Feedstocks Technologies Markets

Residues:

Ag, MSW

Commodity: Corn, oil seed

Herbaceous: Switchgrass

Woody: Poplar,willow

Bio-Chemical

Hydrolysis Fermentation Biocataysis

Thermo-Chemical Combustion Gasification/Pyrolysis Chemistry/Catalysis Separations Tech.

Final Products

Electricity

Fuels

Chemicals Plastics

Heat/Steam

Intermediary

Products

Methanol

Biosynthesis gas

Hydrogen

Organic Acids

Ethanol

Biorefinery

Source: bioenergy.ornl.gov/02workshop/paster.ppt

S

p

e

c

i a

l

t

y

C

h

e

m

i c

a

l s

Process for Converting Starch to Biochemicals

Cu

rre

nt

Pro

ce

ss

Fermentation

Corn

Tapioca

Wheat

Water

Liquefaction Saccharification

Jet Cooker

110°C, 5-8min.

95° C

, 90m

in.

Jet Cooker

120°C, 5-8min.

60° C

, 36-7

2 h

rs.

Sta

rch

Ta

nk

Glucose

Sta

rch S

epara

tion

Purification

Ion Exchange

Cabin

Fructose Fuel Alcohol

Bioresource Refining

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Conclusions

• Enzymes and microbial systems will play a vital role in the new industrial biotechnology

• Cost of enzymes and dosage requirements will dictate the economics of commodity chemicals

• Efficient microbial systems essential for high product yield and concentration

• Integrated Biocatalysis & Fermentation– for ethanol and beyond

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• Integrated Biorefineries– broader product mix, cost efficiencies and for sustainability

• Integration will involve – physical, biochemical and thermo-chemical platforms

• Strategic partnership with established chemical and/or oil companies is essential

• Raw materials cost should become marginal

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Acknowledgments

• USDA (CSREES Award# 2004-33610-14308) and FAS Program

• NSF (BES 0420840)

• Grain Science and Industry Department, KSU

• MeadWestvaco Corporation

• Graduate Students, Bioprocessing Lab, BIVAP

• Center for Sustainable Energy, KSU

• Kansas Soybean Commission