XIAOZHOU (JOHN) ZHANG

Phone: (404)-293-6888 Website: www.xiaozhouzhang.com E-mail: [email protected] 226 Mountain Breeze Drive, Blacksburg, VA 24060

CURRICULUM VITAE

EDUCATION

 Ph.D. in Environmental Microbiology & Engineering, 2006, Nanjing Agricultural University, .  B.Eng. in Food Science & Engineering, 2001, Agricultural University, China.

WORKING EXPERIENCES

Principal Investigator and Chief Technology Officer (2011 - present) Gate Fuels, Inc., Blacksburg, VA.  Served as the Principal Investigator and managed several Small Business Innovation Research (SBIR) projects from NSF and DOE for one-step biochemical production from lignocellulosic biomass by engineered microorganisms;  Established Bacillus subtilis platforms through enzyme engineering and expression, mutagenesis approach, genome and metabolic engineering for commodity chemicals production from biomass;  Established new multi DNA fragments assembling system in yeast with new temperature sensitive yeast-B. subtilis-E. coli shuttle vectors for the reliable and fast genome and metabolic engineering in microorganisms; Research Scientist (2010 - 2013), Senior Research Associate (2009 - 2010), Postdoc (2008-2009) Biological Systems Engineering Department, Virginia Tech, Blacksburg, VA.  Isolated, identified, and characterized cellulolytic strains from different environmental soil samples;  Established a novel powerful and highly efficient Bacillus subtilis platform for protein engineering (especially for cellulase) by directed evolution and enzyme performance improvement;  Developed a new universal, simple, fast and enzyme-free cloning system in E. coli and Bacillus subtilis;  Created a new Bacillus subtilis strain for the super-competence development and transformation with an extremely easy protocol;  Cloned, characterized and engineered cellulases for cellulolytic microorganism Clostridium phytofermentans ISDg;  Constructed the first genetically engineered cellulolytic Bacillus subtilis strain and realized one-step low cost biofuels & biochemicals production from pretreated biomass; Postdoctoral Research Associate (2006 - 2008) Department of Biology, Georgia State University, Atlanta, GA.  Elucidated the mechanism of non-classical protein secretion pathways in Bacillus subtilis;  Constructed serial B. subtilis mutants and performed systematic analysis of their secretome. Assistant Engineer (2004 - 2006), Graduate Research Assistant (2001 -2006) Center of Research & Design in Environmental and Energy Source Engineering, Nanjing Agricultural University, Nanjing, China.  Developed a novel universal system for unmarked genetic engineering in Bacillus subtilis;  Achieved the high-level expression of the degradative gene and the efficient secretion of enzyme in Bacillus;  Discovered and engineered microorganisms from environment soil or waste water samples for in situ biodegradation of the pesticides and herbicides pollutions on farm produce or the biocontrol of plant pathogens;  Performed the field study of biodegradation by bacterial strains and applied molecular methods to monitor the changes of the microbial community. Senior Technical Consultant (2004 - 2006) Nanjing SciGene Science & Technology Co. Ltd., Nanjing, China.  Provided technical assistance, support, and advice to sales representatives and customers.

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TEACHING & MENTORING EXPERIENCES

 Mentor, Scieneering Program for interdisciplinary undergraduate studies and research, Virginia Tech, 2011.  Co-supervised two Master graduate students and two postdocs, Virginia Tech, 2008 - 2011.  Co-supervised one Ph.D. graduate student and three undergraduate students, Nanjing Agricultural University, 2005 - 2006.  Teaching Assistant, taught fundamental, principles and laboratory skills in microbiological research to undergraduate students, Nanjing Agricultural University, 2002

PROFESSIONAL SERVICES

AD HOC GRANT REVIEWERS: NSF SBIR Proposal Panelist • Maryland Technology Transfer and Commercialization Fund

AD HOC JOURNAL REVIEWERS: Biodegradation • Nucleic Acids Research • Bioresource Technology • Plant Soil & Environment • Current Microbiology • Microbiology China • Letters in Applied Microbiology • Microbial Cell Factories • Journal of Applied Microbiology • Chinese Journal of Biotechnology • Journal of Agricultural and Food Chemistry • European Food Research and Technology • Journal of Food Science & Technology • Protein & Peptide Letters• Frontiers in Cellulose Biotechnology (Review Editor)

GRANT AWARDED

 Principal Investigator, Small Business Innovation Research Phase I: High-yield production of fumarate from cellulosic biomass by recombinant cellulolytic Bacillus subtilis. US Department of Energy, 2012.  Principal Investigator, Small Business Innovation Research Phase I, IB & II: One-step production of lactic acid from lignocellulosic biomass by recombinant cellulolytic Bacillus subtilis. Division of Industrial Innovation and Partnerships, US National Science Foundation, 2011~2015.  Principal Investigator, The directed evolution of cellulase based on Bacillus subtilis expression and secretion system and the screening towards nature cellulose substrate. Platform Development Project, Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education, Jilin University, 2011.  Co-Principal Investigator, One-step production of microdiesel from lignocellulosic biomass by recombinant cellulolytic thermophilic Geobacillus thermoglucosidasius. Integrated Internal Competitive Grants Program, College of Agriculture and Life Sciences, Virginia Tech, 2010-2011.  Co-Principal Investigator, One-step production of ethanol from pretreated biomass by recombinant cellulose-utilizing Bacillus subtilis strain. The Biodesign and Bioprocessing Research Center, Virginia Tech, 2010-2012. SELECTED AWARDS

 Outstanding Graduate Student Award & Outstanding Ph.D. Dissertation Award, 2006.  Outstanding Academia Model of Nanjing Agricultural University, 2005.  Excellent Paper Awards, Annual Meeting of Jiangsu Society for Microbiology, 2004 & 2005.  Excellent Paper Award, Doctoral Academic Forum of Jiangsu Province, 2004.  Excellent Paper Award, Symposium on Microbiology Research for the Yong Scientists in Jiangsu, 2003.  Outstanding Undergraduate Student Award of Province, 2000.

PATENTS

 Zhang, X.-Z., et al. U.S. Patent Application 61/815,115, Method for creating recombinant microorganism for production of biofuels and chemicals directly from methane, 2013.

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 Zhang, X.-Z., et al. U.S. Patent Application 61/729,937, Method of creating recombinant cellulolytic Bacillus subtilis strain for low cost one-step fumarate production from lignocellulosic biomass, 2012.  Zhang, X.-Z., et al. U.S. Patent Application 61/479,640, One-Step production of lactate from cellulose as sole carbon source without other organic nutrients by recombinant cellulolytic Bacillus subtilis, 2012.  Zhang, X.-Z., et al. Virginia Tech Intellectual Properties (VTIP) Case Number 09-095, Recombinant Gram- positive microorganism growing on cellulose as the sole carbon source in defined media, 2009.

SELECTED PUBLICATIONS

1. Yang, C.-K., Zhang, X.-Z., Lu, C.-D., and Tai, P.-C. (2014) An internal hydrophobic helical domain of Bacillus subtilis enolase is essential but not sufficient as a non-cleavablesignal for its secretion, Biochemical and Biophysical Research Communications, in press. 2. Zhang, X.-Z., You, C., and Zhang, Y.-H.P. (2014) Transformation of Bacillus subtilis, In Lianhong Sun and Weying Shou (eds.), Engineering and Analyzing Multicellular Systems: Methods and Protocols, Methods in Molecular Biology, vol. 1151, DOI 10.1007/978-1-4939-0554-6_7, © Springer Science+Business Media New York 2014. 3. Ahmad, S., Ma, H., Akhtar, M. W., Zhang, Y.-H. P., and Zhang, X.-Z.* (2014) Directed evolution of Clostridium phytofermentans glycoside hydrolase family 9 endoglucanase for enhanced specific activity on solid cellulosic substrate, BioEnergy Research, DOI: 10.1007/s12155-013-9382-8. 4. You, C., Chen, H., Myung, S., Sathitsuksanoh, N., Ma, H., Zhang, X.-Z., Li, J., and Zhang, Y.-H. P. (2013) Enzymatic transformation of non-food biomass to starch, Proceedings of the National Academy of Sciences of the United States of America, in press. 5. You, C., Zhang, X.-Z.*, and Zhang, Y.-H. P. (2012) Simple Cloning: direct transformation of PCR product (DNA multimer) to Escherichia coli and Bacillus subtilis, Applied and Environmental Microbiology, 78, 1593- 1595. 6. You, C., Zhang, X.-Z., Sathitsuksanoh, N., Lynd, L., and Zhang, Y.-H.P. (2012) Enhanced microbial cellulose utilization of recalcitrant cellulose by an ex vivo cellulosome-microbe complex, Applied and Environmental Microbiology, 78, 1437-1444. 7. You, C., Zhang, X.-Z., and Zhang, Y.-H. P. (2012) Mini-scaffoldin enhanced mini-cellulosomehydrolysis performance on low-accessibility cellulose (Avicel) more than on high-accessibility amorphous cellulose, Biochemical Engineering Journal, 63, 57-65. 8. Wang, Q., Qian, C., Zhang, X.-Z., Liu, N., Yan, X., and Zhou, Z. (2012) Characterization of a novel thermostable β-glucosidase from a metagenomic library of termite gut, Enzyme and Microbial Technology, 51,319-324. 9. Yang, C.-K., Ewis, H. E., Zhang, X.-Z., Lu, C.-D., Hu, H.-J., Pan, Y., Abdelal, A. T., and Tai, P. C. (2011) Non-classical protein secretion of Bacillus subtilis in the stationary phase is not due to cell lysis. Journal of Bacteriology, 193, 5607-5615. 10. Zhang, X.-Z., Sathitsuksanoh, N., Zhu, Z. and Zhang, Y.-H. P. (2011) One-step production of lactate from cellulose as sole carbon source without any other organic nutrient by recombinant cellulolytic Bacillus subtilis, Metabolic Engineering, 13, 364-372. 11. Myung, S., Zhang, X.-Z., and Zhang, Y.-H. P. (2011) Ultra-stable phosphoglucose isomerase through immobilization of cellulosebinding module-tagged thermophilic enzyme on low-cost high-capacity cellulosic adsorbent, Biotechnology Progress, 27, 969-975. 12. Liao, H., Zhang, X.-Z., Rollin, J.A., and Zhang, Y.-H.P. (2011) Selection of a minimal set of bacterial cellulases for an expression in a host for consolidated bioprocessing of lignocellulose, Biotechnology Journal, DOI: 10.1002/biot.201100157. 13. Zhang, X.-Z., and Zhang, Y.-H. P. (2011) Simple, fast and high-efficiency transformation system for directed evolution of cellulase in Bacillus subtilis, Microbial Biotechnology 4, 98-105. 14. Zhang, X.-Z.*, and Zhang, Y.-H. P. (2011) Cellulases: characteristics, sources, production and applications, In Bioprocessing Technologies in Integrated Biorefinery for Production of Biofuels, Biochemicals,

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and Biopolymers from Biomass (Yang, S.-T., Enshasy, H. A. E., Thongchul, N., and Lo, Y. M., Eds.), John Wiley & Sons, Inc. ISBN-13: 978-0470541951. 15. Zhang, X.-Z., Zhang, Z., Zhu, Z., Sathitsuksanoh, N., Yang, Y., and Zhang, Y.-H. P. (2010) The non- cellulosomal family 48 cellobiohydrolase from Clostridium phytofermentans ISDg: heterologous expression, characterization, and processivity, Applied Microbiology and Biotechnology 86, 525-533. 16. Zhang, X.-Z.*, and Zhang, Y.-H. P. (2010) One-step production of biocommodities from lignocellulosic biomass by recombinant cellulolytic Bacillus subtilis: Opportunities and challenges, Engineering in Life Sciences 10, 398-406. 17. Zhang, X.-Z., Sathitsuksanoh, N., and Zhang, Y.-H. P. (2010) Glycoside hydrolase family 9 processive endoglucanase from Clostridium phytofermentans: heterologous expression, characterization, and synergy with family 48 cellobiohydrolase, Bioresource Technology 101, 5534-5538. 18. Liu, W., Zhang, X.-Z., Zhang, Z., and Zhang, Y.-H. P. (2010) Engineering of Clostridium phytofermentans endoglucanase Cel5A for improved thermostability, Appl. Environ. Microbiol. 76, 4914-4917. 19. Zhang, Z.-H., Hong, Q., Xu, J.-H., Zhang, X.-Z., and Li, S.-P. (2006) Isolation of fenitrothion-degrading strain Burkholderia sp. FDS-1 and cloning of mpd gene, Biodegradation 17, 275-283. 20. Zhang, X.-Z., Yan, X., Cui, Z.-L., and Li, S.-P. (2006) Recombinant expression and secretion of mpd gene using the promoters and signal peptide-encoding sequences of ytkA and ywoF gene from Bacillus subtilis, Chinese Journal of Biotechnology 22, 249-256. 21. Zhang, X.-Z., Yan, X., Cui, Z.-L., and Li, S.-P. (2006) Cloning of the strong promoters and signal peptide- encoding sequence from Brevibacillus brevis and construction of the shuttle expression-secretion vector, Acta Microbiologica Sinica 46, 526-530. 22. Zhang, X.-Z., Yan, X., Cui, Z. L., Hong, Q., and Li, S. P. (2006) mazF, a novel counter-selectable marker for unmarked chromosomal manipulation in Bacillus subtilis, Nucleic Acids Res 34, e71. 23. Zhang, R.-F., Cui, Z.-L., Zhang, X.-Z., Jiang, J.-D., Gu, J.-D., and Li, S.-P. (2006) Cloning of the organophosphorus pesticide hydrolase gene clusters of seven degradative bacteria isolated from a methyl parathion contaminated site and evidence of their horizontal gene transfer, Biodegradation 17, 467-472. 24. Zhang, X.-Z., Xu, J.-H., and Li, S.-P. (2005) Isolation, screening and preliminary identification of plant pathogenic fungi biocontrol strains of Bacillus spp., Soils 37, 85-88. 25. Zhang, X.-Z., Cui, Z.-L., Hong, Q., and Li, S.-P. (2005) High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis WB800, Appl. Environ. Microbiol. 71, 4101-4103. 26. Zhang, G.-S., Hong, Q., Ma, A.-Z., Zhang, X.-Z., and Li, S.-P. (2005) Cloning, expression of the gene of dehydrochlorination of BHC from enrichment and its degradation experiment, Acta Microbiologica Sinica 45, 44-47. 27. Jiang, J.-D., Zhang, R.-F., He, J., Zhang, X.-Z., Cui, Z.-L., and Li, S.-P. (2005) Bacterial chemotaxis to environmental pollutants and its significance in bioremediation, Acta Ecologica Sinica 25, 1764-1771. 28. Hong, Q., Zhang, Z.-H., Zhang, X.-Z., Xu, J.-H., and Li, S.-P. (2005) Cloning and sequencing of promoters of a moderate halophilic bacterium Halomonas sp. BY-1, Chinese Journal of Applied and Environmental Biology 11, 729-732. 29. Hong, Q., Xu, J.-H., Wu, J., Zhang, X.-Z., and Li, S.-P. (2005) Cloning of a salt-tolerance related DNA fragment of a moderate halophilic bacterium Halomonas sp. BYS-1, Chinese Journal of Applied and Environmental Biology 11, 729-732. 30. Zhang, G.-S., Hong, Q., Liu, Z., Zhang, X.-Z., and Li, S.-P. (2004) Isolation and preliminary mechanism of acid tolerance of an acidphilic yeast strain R1, Acta Microbiologica Sinica 44, 631-635. 31. Liu, Z., Zhang, X.-Z., He, J., Lin, S.-W., Pan, J.-Y., and Li, S.-P. (2004) Effects of nutrient substances and metal ions on degradation of p-nitrophenol by DLL-E4 (Pseudomonas putida), Acta Pedologica Sinica 41, 170-175. 32. Cui, Z.-L., Zhang, X.-Z., Zhang, Z.-H., and Li, S.-P. (2004) Construction and application of a promoter trapping vector with methyl parathion hydrolase gene mpd as the reporter, Biotechnology Letters 26, 1115- 1118. 33. Liu, Z., Zhang, X.-Z., and Li, S.-P. (2003) Use of methyl parathion (MP)-degrading strain DLL-E4 (Pseudomonas sp.) to remove MP residue on the surface of agriculture products, Chinese Journal of Applied Ecology 14, 1770-1774. 34. Liu, Z., Ma, A.-Z., Zhang, X.-Z., Zhang, X.-H., and Li, S.-P. (2003) Study on reaction system and characteristics of methyl parathion hydrolase, Journal of Nanjing Agricultural University 26, 1770-1774.

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35. Liu, Z., Hong, Q., Zhang, X.-Z., Xu, J.-H., and Li, S.-P. (2003) Character of p-nitrophenol degradation by methyl parathion degrading strain DLL-E4, China Environmental Science 23, 435-439. 36. Liu, Z., Hong, Q., Xu, J.-H., Wu, J., Zhang, X.-Z., Zhang, X.-H., Ma, A.-Z., Zhu, J., and Li, S.-P. (2003) Cloning, analysis and fusion expression of methyl parathion hydrolase, Acta Genetica Sinica 30, 1020-1026.

INVITED & CONTRIBUTED TALKS

1. Zhang, X.-Z. (2011) Bacillus subtilis platforms for cellulase engineering and biocommodities production from cellulose in a single step. Institute of Plant Physiology and Ecology, SIBS, Chinese Academy of Sciences; Nanjing Agricultural University; Qingdao Agricultural University; Polytechnic; Qingdao Continent Biotech Co., Ltd. In May ~ June. 2. Zhang, X.-Z., and Zhang, Y.-H. P. (2010) Industrially-safe recombinant cellulolytic Bacillus subtilis strains for one-step ethanol production from pretreated biomass. The 239th American Chemical Society National Meeting. San Francisco, CA. 3. Zhang, X.-Z., and Zhang, Y.-H. P. (2010) Performance improvement of family 5 endoglucanase by directed evolution and selection on natural cellulosic substrate. The 239th American Chemical Society National Meeting. San Francisco, CA. 4. Zhang, X.-Z., Zhu, Z., and Zhang, Y.-H. P. (2009) Recombinant cellulolytic Bacillus subtilis grows on cellulose as the sole carbon source in defined media. The 238th American Chemical Society National Meeting. Washington, DC. Aug 16-20. 5. Zhang, X.-Z. (2006) Establishment and application of the new expression and genetic manipulation system in Bacillus subtilis. Institute of Plant Physiology and Ecology, Chinese Academy of Sciences. Shanghai, China. Jun 10, 2006. 6. Zhang, X.-Z. (2005) High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis WB800. Jiangsu Society for Microbiology 2005 Annual Meeting. China. Nov 12. 7. Zhang, X.-Z. (2004) High-level expression and secretion of methyl parathion hydrolase in Bacillus subtilis. Doctoral Academic Forum of Jiangsu Province. China. Dec 26. 8. Zhang, X.-Z. (2004) Isolation, screening and preliminary identification of plant pathogenic fungi biocontrol strains of Bacillus spp. Jiangsu Society for Microbiology 2004 Annual Meeting. China. Nov 21. 9. Zhang, X.-Z. (2003) Recombinant expression and secretion of methyl parathion hydrolase gene mpd in Bacillus subtilis. The 6th Symposium on Microbiology Research for the Yong Scientists in Jiangsu Province. China. Dec.

CONFERENCE POSTERS

1. Zhang, X.-Z., et al. (2011) Bacillus subtilis platforms for cellulase engineering and biocommodities production from cellulose in a single step. The 2011 BioEnergy Science Center Retreat, Chattanooga, Tennessee, July 18-21; The 2011 Gordon Conference on Cellulosomes, Cellulases & Other Carbohydrate Modifying Enzymes, Stonehill College, Easton, MA, July 24-29, 2011; The 5th China Summit Forum on Industrial Biotechnology Development, Qingdao, May 25-27. 2. Zhang, X.-Z., and Zhang, Y.-H. P. (2010) A novel powerful and efficient Bacillus subtilis platform for cellulase engineering and performance improvement towards natural cellulose substrate. The 2010 Gordon Conference on Biocatalysis. Bryant University, Smithfield, RI. Jul 11-16. 3. Zhang, X.-Z., and Zhang, Y.-H. P. (2010) A novel powerful and efficient Bacillus subtilis platform for cellulase engineering and performance improvement towards natural cellulose substrate. The 2010 BioEnergy Science Center Retreat. Asheville, NC. Jun 21-23. 4. Zhang, Z., Zhang, X.-Z., Liu, W., and Zhang, Y.-H. P. (2009) Heterologous expression, purification and characterization of the endoglucanase Cel5B from Clostridium phytofermentans. The 2009 BioEnergy Science Center Retreat. Asheville, NC. Jun 24-26. 5. Zhang, X.-Z., Zhu, Z., and Y.-H. P. Zhang. . (2009) Over-expression of endoglucanase enables Bacillus subtilis to grow on cellulose as the sole carbon source in defined media. The 2009 AIChE Annual Meeting. Nashville, TN. Nov 8-12.

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6. Zhang, X.-Z., Zhu, Z., Sathitsuksanoh, N., and Zhang, Y.-H. P. (2009) Developing cellulolytic Bacillus subtilis by co-expression of cellulases for cellulosic ethanol production. The 2009 Gordon Conference on Cellulosomes, Cellulases & Other Carbohydrate Modifying Enzymes. Andover, NH. Jul 26-30. 7. Zhang, X.-Z., Zhu, Z., Sathitsuksanoh, N., and Zhang, Y.-H. P. (2009) Overexpression of endoglucanase enables Bacillus subtilis to grow on cellulose in defined media. The 2009 BioEnergy Science Center Retreat. Asheville, NC. Jun 24-26. 8. Zhang, X.-Z., Zhang, Z., Zhu, Z., Sathitsuksanoh, N., Yang, Y., and Zhang, Y.-H. P. (2009) A highly-active family 48 cellobiohydrolase from Clostridium phytofermentans ISDg: Heterologous expression in Escherichia coli and Bacillus subtilis, characterization, and processivity. The 2009 BioEnergy Science Center Retreat. Asheville, NC. Jun 24-26. 9. Liu, W., Zhang, X.-Z., Zhang, Z., and Zhang, Y.-H. P. (2009) Engineering cellulases on their natural substrates by directed evolution. The 31st Symposium on Biotechnology for Fuels and Chemicals. San Francisco, CA, May 3-6. 10. Liu, W., Zhang, X.-Z., Zhang, Z., and Zhang, Y.-H. P. (2009) Engineering cellulases on their natural substrates by directed evolution and rational design. The 238th American Chemical Society National Meeting. Washington, DC. Aug 16-20. 11. Zhang, X.-Z., Lu, C.-D., and Tai, P. C. (2008) A highly conserved hydrophobic α-helix and basic residues flanking the hydrophobic core region are important for the export of the Bacillus subtilis enolase in Escherichia coli. Aug. 2008. The 6th Annual Biotechnology Symposium at Georgia State University. Atlanta, GA. Aug 14-15.

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