Monthly Report for May 2011

1. Summary of MAJOR research and service activities of the month with the following appendices:

1) Oral presentation at the 10th Annual Conference on Carbon Capture & Sequestration May 2-5, 2011, Pittsburgh, Pennsylvania. 2) Re-submitted the opinion paper titled with Bicarbonate Produced from Carbon Capture for Algae Culture to Trends in Biotechnology 3) Take part in developing proposal to Jinjing Group in China 4) Take part in developing LOI for proposal to DOE BRDI program 5) Prepared form for quarterly report to Algal Biofuel Research project.

2. Updated progress against the Expectation list:

Expected date Status Paper Oleaginous Yeast Cryptococcus curvatus Culture for 3/31/10 In press Biodiesel Feedstock with Wastewater from Anaerobic Bio-hydrogen Production Integrated process of Food waste and wastewater 5/30/10 In press treatment for biodiesel feedstock production Paper: High efficient CO2 delivery system for algae pond 5/29 Re-submitted (Trends in Biotechnology) High density culture of Aurantiochytrium limacinum 4/30/10 Add some SR21 for DHA production with crude glycerol from data biodiesel industry as feedstock Experiment paper on the above concept, paper 1 8/31 Under development Experiment paper on the above concept, paper 2 10/1/11 Under development

Proposal CO2 delivery to algae culture process with carbonate or SBIR To be bicarbonate, integration of algae culture with AD process developed CO2 absorption with alkali lake, and algae culture, NSF 9/1/10 To be developed

Invention disclosure and Patent

The formal patent on CO2 delivery 11/30/2011 To be developed

Experiment DHA production with glucose 3/1—3/31 To be conducted

Experiment work on CO2 delivery 4/1- 9/30 To be conducted 3. Updated BBEL member accomplishment summary (template attached)

Bioprocessing and Bioproduct Engineering Laboratory (BBEL) Group Member Accomplishments Name: Zhanyou Chi Date started with BBEL: 6/14/2005 Date of Update: 5/30/2011

Papers: Published: 1. Zhanyou Chi, Yan Liu, Craig Frear, Shulin Chen. Study of a two-stage growth of DHA producing marine algae Schizochytrium limacinum SR21 with shifting dissolved oxygen level. Applied Microbiology and Biotechnology, 2009, 81: 1141-1148 2. Zhanyou Chi, Denver Pyle, Zhiyou Wen, Craig Frear, Shulin Chen. A laboratory study of producing docosahexaenoic acid from biodiesel-waste glycerol by microalgal fermentation. Process Biochemistry, 2007, 42:1537-1545 3. Zhanyou Chi, Bo Hu, Yan Liu, Craig Frear, Zhiyou Wen, Shulin Chen. Production of Omega-3 Polyunsaturated Fatty Acids from Cull Potato Using an Algae Culture Process. Applied Biochemistry and Biotechnology, 2007, 137: 805-815 4. Shulin Chen, Zhanyou Chi, James V. O’Fallon, Yubin Zheng, Moumita Chakraborty, Dhrubojyoti D. Laskar. System Integration for Producing Microalgae as Biofuel Feedstock. Biofuels, 2010, 1: 889-910 5. Zhimin Li, Yan Liu, Zhanyou Chi, Shulin Chen, Effects of Pluronic F68 on Manganese Peroxidase Production by Pelletized Phanerochaete chrysosporium, Applied Biochemistry and Biotechnology, 2011, 164:487–496 6. Bo Hu, Yan Liu, Zhanyou Chi, Shulin Chen, biological hydrogen production via bacteria immobilized in calcium alginate gel beads, Biological Engineering, 2008, 1: 25-37

In press: 7. Zhanyou Chi, Yubin Zheng, Jingwei Ma, Shulin Chen. Yeast Cryptococcus curvatus Culture with Dark Fermentation Hydrogen Production Effluent as Feedstock for Microbial Lipid Production. International Journal of Hydrogen Energy 8. Zhanyou Chi, Yubin Zheng, Anping Jiang, Shulin Chen. Lipid Production by Culturing Oleaginous Yeast and Algae with Food Waste and Municipal Wastewater in an Integrated Process. Applied Biochemistry and Biotechnology

Submitted: 9. Zhanyou Chi, James V O’Fallon, Shulin Chen. Bicarbonate Produced from Carbon Capture for Algae Culture. Trends in Biotechnology. (submitted on May 28th 2011)

Patents and Invention disclosures filed 1. Zhanyou Chi, Baisuo Zhao, Shulin Chen, Halo-alkaliphilic algae or cyanobacteria culture with bicarbonate as feedstock and recycle the regenerated carbonate for carbon capture (Provisional) 2. Zhanyou Chi, Zhiyou Wen, Craig Frear, Shulin Chen, Heterotrophic algae high cell density production method and systems, US Patent, Application number 12/132,131 3. Zhanyou Chi, Shulin Chen, Craig Frear, Tianxi Zhang, Ben Lucker, Integrated Algae System for Production of Biofuel Feedstock, Application number: PCT/US2009/052234

Proposals Funded 1. Development of a Two-Stage Culture Process for Production of Omega 3 Rich Algae from Biodiesel Waste Glycerol for Use as an Aquaculture Fish Feed Supplement, NSF 2007 ($100,000) 2. Integrated algae production system for biofuel feedstock based on utilization of wastewater resources, Boeing company, 2008 ($150,000) 3. Biohydrogen and biodiesel co-production with treatment of high solid food waste, Washington State Department of Ecology, 2008 ($120,000) 4. Algae grown in aquaculture effluent for use as an Omega-3 fatty acid feed additive, The Northwest Center for Aquaculture Research and Education, 2009 ($35,000) 5. Production of Omega- 3 Polyunsaturated Fatty Acids from Cull Potato, Washington State Potato Commission, 2005~2008 ($30,000/year) 6. Production of Omega- 3 Polyunsaturated Fatty Acids from Cull Potato, IMPACT center in Washington State University, 2006~2007 ($30,000/year) 7. Production of DHA Rich Algae Biomass as Cattle Feed Supplement to Enhance Milk Quality, CSANR in Washington State University, 2006~2007 ($20,000/year)

Proposal development contribution Seagrant, Co-production of DHA and Carotenoids by high-density culture of 04/26/2006 marine algae Schizochytrium BIOAg program of CSANR, Production of DHA Rich Algae biomass as Cattle 07/21/2006 Feed Supplement to Enhance Milk Quality USDA-SBIR, Co-production of Carotenoids and DHA by high-density culture of 09/01/2006 marine algae Schizochytrium using crude glycerin from biodiesel waste UI/WSU Aquaculture Initiative, Growing PUFA-enriched algae (Omega-3 11/06/2006 fatty acids) from aquaculture nutrient discharge and using it as a fish feed ingredient (collaborate with Tony) NSF-SBIR, Development of a Two-Stage Culture Process for Production of 12/04/2006 Omega 3 Rich Algae from Biodiesel Waste Glycerol for Use as an Aquaculture Fish Feed Supplement WSPC, Production of Omega-3 Polyunsaturated Fatty Acids from Cull Potato 12/15/2006

NOAA-SBIR, Waste water using to produce DHA 1/17/07 US-Pakistan cooperation fund, Transfer EPA biosynthesis related genes to 6/30/07 algae Schizochytrium sp to improve EPA production BioAg , Production of DHA Rich Algae biomass as Cattle Feed Supplement 7/30/07 to Enhance Milk Quality Emerging research issues for Washington agriculture, Production of Omega- 9/30/07 3 Polyunsaturated Fatty Acids from biodiesel waste glycerol and Cull Potato WA department of ecology, Biohydrogen and biodiesel co-production with 10/5/07 treatment of high solid food waste dairy commission, Feeding DHA-Rich Algae Biomass to Dairy Cow to Enhance 10/15/07 Milk Quality NOAA-preproposal, production of PUFA-enriched marine algae 10/18/07 (Schizochytrium) grown in marine aquaculture effluent for use as an Omega-3 fatty acid feed additive

DARPA 2/19/08 Boeing, Integrated algae production system for biofuel feedstock based on 4/3/08 utilization of wastewater resources

Life Science Discovery Fund, Enhancing prevention of heart disease and cancer 4/16/08 using nutraceuticals derived from Washington Agriculture NSF-SBIR-phase II, a continuous three-stage culture process for heterotrophic 7/31/08 microalgae NSF-Interagency Opportunities in Metabolic Engineering, Molecular regulating 10/22/08 mechanism of dissolved oxygen effect on lipid synthesis in oleaginous microorganisms

Life Science Discovery Fund, Enhancing prevention of heart disease and cancer 4/15/09 using nutraceuticals derived from Washington Agriculture UI/WSU Aquaculture Initiative, Growing PUFA-enriched algae (Omega-3 2/16/09 fatty acids) from aquaculture nutrient discharge and using it as a fish feed ingredient NSF : Molecular regulating mechanism of dissolved oxygen effect on fatty acid 9/16/09 synthesis in oleaginous microorganisms USAID, Integrated System for Wastewater Treatment, H2 Production, and 11/16/09 Biodiesel Production

NSF-SBIR, Algae or Cyanobacteria Culture with Bicarbonate as Feedstock and 12/3/10 Recycle the Regenerated Carbonate for Carbon Capture

DOE, Utilizing All Sugars from Cereal Straws for Lipid-based “Drop-In” Fuel 2/17/2011 Production (partially involved).

Seagrant, CO2 Capture and Conversion into Bicarbonate for Alkaliphilic 3/3/2011 Cyanobacteria Culture

Presentations 1. Zhanyou Chi, Shulin Chen, Carbon Capture and Conversion into Bicarbonate for Algae Culture, 10th Annual Conference on Carbon Capture & Sequestration May 2-5, 2011, Pittsburgh, Pennsylvania (Oral presentation) 2. Zhanyou Chi, Craig Frear, Shulin Chen, High density culture of Aurantiochytrium limacinum SR21 for DHA production with crude glycerol from biodiesel industry as feedstock, 2010 World Congress on Industrial Biotechnology and Bioprocessing, June 27-30, 2010, National Harbor, Washington, D.C. (Oral presentation) 3. Zhanyou Chi, Craig Frear, Shulin Chen, Fed batch two-stage culture of Aurantiochytrium limacinum SR21 for DHA production, 2010 ASABE Annual International Meeting from June 20 to 23, at Pittsburg, Pennsylvania (Oral presentation) 4. Zhanyou Chi, Yubin Zheng, Anping Jiang, Shulin Chen, Integrated Process of Food Waste and Wastewater Treatment for Biodiesel Feedstock Production, ASABE Bioenergy Engineering, October 11-14, 2009, Bellevue, WA (Poster) 5. Zhanyou Chi, Yan Liu, Craig Frear, Shulin Chen, Study of a two- stage culture of DHA producing marine algae Schizochytrium limacinum SR21 with shifting dissolved oxygen level, AIChE Annual Meeting, 2008, Philadelphia, PA (Oral presentation) 6. Zhimin Li, Yan Liu, Zhanyou Chi, and Shulin Chen, Effectiveness of Shear Protective Additives in Enzyme Production by Pelleted Phanerochaete Chrysosporium, AIChE Annual Meeting, 2008, Philadelphia, PA (Oral presentation) 7. Zhanyou Chi, Yubin Zheng, Chenlin Li, Zhimin Li, Bo Hu, Shulin Chen, Oleaginous yeast Cryptococcus curvatus culture for biodiesel feedstock with wastewater from dark fermentative hydrogen production, 30th Symposium on Biotechnology for Fuels and Chemicals, 2008, New Orleans, LA (Poster) 8. Zhanyou Chi, Yan Liu, Bo Hu, Craig Frear, Shulin Chen, DHA production from high biomass concentration culture of Schizochytrium sp. with a shift strategy, The World Congress on Industrial Biotechnology & Bioprocessing, 2007, Orlando, FL (Poster) 9. Zhanyou Chi, Yan Liu, Bo Hu, Craig Frear, Shulin Chen, High cell density culture of Schizochytrium to produce DHA with a “Shift” strategy, 29th Symposium on Biotechnology for Fuels and Chemicals, 2007, Denver, CO (Poster) 10. Zhanyou Chi, Bo Hu, Yan Liu, Shulin Chen, Production of Omega-3 Polyunsaturated Fatty Acids from Cull Potato Using an Algae Culture Process. 28th Symposium on Biotechnology for Fuels and Chemicals, 2006, Nashville, TN (Poster)

Project report Title Agency Date submitted WTC phase I report, Development of Omega 3 Rich Algae from WTC 11/01/2006 Biodiesel Waste for Use as an Animal Feed Supplement Potato commission annual report Potato 01/10/2007 commission Annual report of NSF SBIR NSF SBIR 1/15/08 Annual report of BioAg BioAg 6/30/07 Potato commission annual report Potato 1/15/08 commission WSDE quarterly report WSDE 3/31/2008 BioAg annual report BioAg 8/25/2008 WSDE quarterly report WSDE 6/30/2008 WSDE quarterly report WSDE 9/30/2008 Boeing project quarterly report Boeing 7/30/2008 Boeing project quarterly report Boeing 10/31/08 Department of Ecology Final report WSDE 6/30/2009 Boeing final report Boeing 12/29/09 4. Project status report (also applicable to thesis project)

5. Write up of the experiment work that can be used later in draft publication or report

6. Draft manuscript or patents (can be specified as a separate file)