William J. Coleman

Designed a protein expression system for high-value carotenoids using a novel enzyme and engineered operon: Discovered and cloned a novel crtW beta-carotene ketolase gene, designed a six-gene synthetic DNA operon, and implemented a successful genomic integration and gene expression protocol for astaxanthin nutraceutical production in a novel host. Granted U.S. Patent No. 11,851,664. Co-developed and scaled-up a benchtop heterotrophic fermentation process optimization and CRO/CMO tech transfer to enable toll production of 1.4 metric tons of novel, high protein (>70%) product for testing: Produced high-quality material for commercial trials at 3x the milestone amount and 6 months ahead of schedule. Evaluated five potential host strains for their ability to produce high protein on sugar feedstocks: Performed over 90 10-liter scale fermentations on fructose to confirm 1-2 prime candidates. Optimized the media and process conditions to eliminate unwanted products.

Directed the engineering and development of novel bacterial strains that use H2 as an energy source to capture and convert industrial carbon dioxide emissions into valuable small molecule products: Co-founded the development of proprietary chemoautotrophic bacterial strains, engineered by synthetic biology and adaptive strain evolution, to produce proteins, carotenoids and other molecules using industrial flue gas and hydrogen as feedstocks. Granted U.S. Patent No. 11,439,172. Developed novel gas fermentation procedures and high-throughput analytical methods for quantitating the titer of the products.

• Co-founded the methane bioconversion project for biofuels (gas-to-liquids) and chemicals by introducing and improving methanotroph-based technology. Developed methods for strain evaluation, high-throughput transformation, random mutagenesis, assay development and high-throughput screening, shake flask metabolic studies, and fermentation improvement. Led the group responsible for intellectual property protection. Resulted in a $75 million R&D collaboration with Intrexon Energy Partners to produce isobutanol and farnesene. Granted U.S. Patent No. 11,821,019. • Developed the technical know-how and procedures to create a novel enzyme-based product for the Johnson & Johnson Consumer Products Company. Employed literature searching and genome mining to identify enzyme candidates. Designed combinatorial expression vectors tailored to appropriate microbial hosts, and developed new enzyme assays for the desired activity. Met all project milestones. • Provided technical guidance to the Genopaver, LLC project on using microbial synthetic biology to produce valuable active pharmaceutical ingredients (APIs). Used genome mining and sequence-structure-function analysis to identify uncharacterized sequences that could encode desirable enzyme activities. Developed customized enzyme assays and analytical methods for quantitating chiral alkaloid products. • Established, maintained and supervised a state-of-the-art 2,000 s.f. high-throughput screening and analytical lab providing services to all projects in the division. Instrumentation included robotic liquid handling (BioMek), QPix460, UHPLC, UPLC-MS, GC-MS, microplate readers and imaging workstations. • Provided scientific expertise to numerous business development projects that became multi-million dollar exclusive channel collaborations with large industrial, consumer products and pharmaceutical partners, including Sanofi Chimie. Participated in business presentations and technical discussions with potential clients.

• Supervised and participated in the company’s drug development and directed evolution programs for enhanced enzymes, antibody substitutes, and metabolic engineering of microorganisms: Designed and constructed novel chimeric ADC-type therapeutic proteins that incorporate engineered antibody-linker-payload constructs. Created enhanced xylanase for hemicellulose deconstruction and isoprenoid synthase for algal hydrocarbon biosynthesis in collaboration with Sapphire Energy using synthetic biology techniques. Characterized the enhanced activity and thermostability by improving existing detection techniques. Developed high-throughput fluorogenic/chemiluminescent enzyme screening assays, including pooled screening, for enzyme characterization and optimization.

Directed multidisciplinary research teams and obtained funding for directed-evolution based improvements in industrial and biofuel enzymes for corporate clients: • Identified appropriate enzyme genes, created heterologous expression constructs, designed mutagenized libraries and implemented high-throughput chromogenic and fluorogenic enzyme assays for glycosidases, cellulases and xylanases. • Awarded a $500,000 NSF Phase II SBIR grant for successfully enhancing the thermostability of an endoglucanase for biomass conversion and biofuels. • Collaborated with the Pfizer-La Jolla Biotransformations Group to develop an improved nitrilase for “green” pharmaceutical production. • Awarded NIH Phase I SBIR grants to enhance the fibrin-specificity of streptokinase and to reduce the inhibitor sensitivity of DNA polymerase. • Employed shotgun cloning and metagenomics sequencing for gene discovery and microbial ID of unknown microbial populations from human and environmental microbiomes. • Received NIH anti-bioterrorism funding for a fluorescence imaging-based system to identify Bacillus species. • Led the cross-functional molecular biology and assay development teams for commercializing the company’s $30 million “Kcat Technology” platform for high-throughput enzyme library and biocatalyst screening. • Researched and developed commercial applications for directed evolution and digital imaging spectroscopy: (1) industrial applications of randomized cassette mutagenesis for creating complex libraries of mutants, and (2) very high-throughput, solid-phase library screening for enzyme improvement, strain development, and metagenomics. • Engineered a novel galactose oxidase with altered substrate specificity for a commercial process at Hercules, Inc. • Developed PCR-shotgun cloning and sequencing methods for identifying unculturable environmental and human microbiome bacteria (using 16S/23S rRNA, etc.). • Wrote numerous SBIR grants and patents. • Assisted in patent litigation.

Ph.D. Thesis: Biochemical/biophysical studies of enzyme activation.

Photosynthesis; directed evolution; purple non-sulfur bacteria; photosynthetic reaction center; light reactions; renewable energy; solar energy conversion; protein engineering; membrane proteins; quinone binding; bacteriochlorophyll; mutagenesis; structure-function studies; protein chimeras; spectroscopy; electron transfer; kinetics; enzymology.