NREL is a national laboratory of the U.S. Department of Energy, Office of Energy 

Efficiency & Renewable Energy, operated by the Alliance for Sustainable Energy, LLC.

Contract No. DE-AC36-08GO28308

Technical Support to SBIR Phase 

II Project: Improved Conversion 

of Cellulose Waste to Ethanol 

Using a Dual Bioreactor System 

Cooperative Research and Development 

Final Report

CRADA Number: CRD-08-310

NREL Technical Contact: Min Zhang

CRADA Report

NREL/TP-7A10-53592

April 2013
NOTICE

This report was prepared as an account of work sponsored by an agency of the United States government. 

Neither the United States government nor any agency thereof, nor any of their employees, makes any warranty, 

express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of 

any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately 

owned rights. Reference herein to any specific commercial product, process, or service by trade name, 

trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, 

or favoring by the United States government or any agency thereof. The views and opinions of authors 

expressed herein do not necessarily state or reflect those of the United States government or any agency thereof.

Available electronically at http://www.osti.gov/bridge

Available for a processing fee to U.S. Department of Energy

and its contractors, in paper, from:

U.S. Department of Energy

Office of Scientific and Technical Information

P.O. Box 62

Oak Ridge, TN 37831-0062

phone: 865.576.8401

fax: 865.576.5728

email: mailto:reports@adonis.osti.gov

Available for sale to the public, in paper, from:

U.S. Department of Commerce

National Technical Information Service

5285 Port Royal Road

Springfield, VA 22161

phone: 800.553.6847

fax: 703.605.6900

email: orders@ntis.fedworld.gov

online ordering: http://www.ntis.gov/help/ordermethods.aspx

Cover Photos: (left to right) PIX 16416, PIX 17423, PIX 16560, PIX 17613, PIX 17436, PIX 17721

Printed on paper containing at least 50% wastepaper, including 10% post consumer waste. 
1 

Cooperative Research and Development Final Report

In accordance with Requirements set forth in Article XI.A(3) of the CRADA document, this document is 

the final CRADA report, including a list of Subject Inventions, to be forwarded to the Office of Science

and Technical Information as part of the commitment to the public to demonstrate results of federally 

funded research.

CRADA number: CRD-08-310

CRADA Title: Technical Support to SBIR Phase II project: Improved Conversion of Cellulose Waste to 

Ethanol Using a Dual Bioreactor System 

Parties to the Agreement: Technova Corporation

Joint Work Statement Funding Table showing DOE commitment:

Estimated Costs NREL Shared Resources

Year 1 $ 00.00

Year 2 $ 00.00

Year 3 $ 00.00

TOTALS $ 00.00

Abstract of CRADA work:

Over-dependence on fossil fuel has spurred research on alternative energy. Inedible plant materials such 

as grass and corn stover represent abundant renewable natural resources that can be transformed into 

biofuel. Problems in enzymatic conversion of biomass to sugars include the use of incomplete synergistic 

enzymes, end-product inhibition, and adsorption and loss of enzymes necessitating their use in large 

quantities.

Technova Corporation will develop a defined consortium of natural microorganisms that will efficiently 

break down biomass to energy-rich soluble sugars, and convert them to cleaner-burning ethanol fuel. The 

project will also develop a novel biocatalytic hybrid reactor system dedicated to this bioprocess, which 

embodies recent advances in nanotechnology. NREL will participate to develop a continuous 

fermentation process.

Summary of Research Results:

1) Fermentation of Technov sugar streams

Six samples of AFEX-treated corn stover hydrolyzed with fungi mixed culture were provided by 

Technova. NREL conducted HPLC analysis to determine the sugar concentrations of these samples. 

Fermentation of three selected Technov sugar streams to ethanol was performed using Zymomonas 

mobilis 8b. The fermentation profiles for sugar feeds streams was obtained. As a control, fermentation of 

Zymomonas mobilis 8b in RM with 1.1% glucose and 1% xylose is conducted. All sugars are converted 
2 

with 24 hrs as expected. The gluose and xylose in all three sugar streams are converted to ethanol within 

24 hours. These results showed that the sugar streams generated by AFEX-treated corn stover hydrolyzed 

with fungi mixed culture are easily fermentable and no toxicity was observed.

2) Prepared the xylose-fermenting strain Zymomonas mobilis 8b under MTA for providing Technova for 

immobilization.

3) Provided technical support and literature information to Technova for immobilization of Zymomonas 

mobilis 8b.

Subject Inventions listing: None

Report Date: Dec. 12, 2012 Responsible Technical Contact at Alliance/NREL: Min Zhang

This document contains NO confidential, protectable, or proprietary information.