2017 SIMB Annual Meeting and Exhibition Sunday, July 30 7:00 AM - 8:00 AM Workshops 1 and 2 Continental Breakfast Plaza Court 1 - Concourse Level 7:00 AM - 8:00 AM Workshops 1 and 2 Registration Plaza Registration - Concourse Level 8:00 AM - 3:00 PM SIMB Board of Directors Meeting Governors Square 17, Plaza Concourse Level 8:00 AM - 3:00 PM Workshop 1 - Fermentation Basics Plaza Court 2 - Concourse Level 8:00 AM - 3:30 PM Workshop 2 - Mining Microbial Genomes and Metagenomes for Biological Applications Plaza Court 3 - Concourse Level 9:00 AM - 6:00 PM SIMB Annual Meeting Registration Plaza Registration - Concourse Level 4:00 PM - 5:00 PM Opening Remarks and Keynote: George Garrity, Michigan State Univ, SIMB President; Hal Alper, Univ of Texas-Austin, 2017 Program Chair Keynote Speaker: Jack Gilbert, Univ of Chicago Plaza Ballroom A & B - Concourse Level 5:00 PM - 6:00 PM Science Slam Plaza Ballroom A & B - Concourse Level 6:00 PM - 8:00 PM Opening Reception/Exhibits Open Plaza Exhibit - Concourse Level 6:00 PM - 8:00 PM Session: PS1: Poster Session 1: Biocatalysis and Metabolic Engineering Plaza Exhibit - Concourse Level P1 Spatial constraints of a chaperone and its substrates for the efficient solubilisation of recombinant proteins S.C. Jung*, L.M. Bui, A. Geraldi, J.H. Lee, S. Lim, M.S. Kim, B.K. Han and S.C. Kim, Korea Advanced Institute of Science and Technology, Daejeon, Korea, Republic of (South) The production of functional recombinant proteins in soluble forms represents a challenging frontier in engineering bacterial hosts. Natural molecular chaperone systems have been widely used to solubilise various recombinant proteins, albeit with limited successes. Here, to facilitate the folding activity of a chaperone via enhancing the interaction with its substrate protein, we either anchored the bacterial chaperone DnaJ to the 3’ untranslated region of a target mRNA by fusion with an RNA binding domain in chaperone-recruiting mRNA scaffold (CRAS) system, or coupled the expression of DnaJ and a target recombinant protein using the overlapping stop-start codons 5’-TAATG-3’ between the two genes in a chaperone-substrate co-localised expression (CLEX) system. By spatially constraining bacterial molecular chaperones to the location of protein translation, we are able to surpass the in vivo solubilisation efficiency of the native chaperone system and to overexpress aggregation-prone recombinant proteins, producing up to 95% in functionally active soluble forms. P3 Engineering Yarrowia lipolytica for triacetic acid lactone (TAL) production C. Palmer*, K. Markham and H. Alper, The University of Texas at Austin, Austin, TX, USA Yarrowia lipolytica, an industrially attractive, non-conventional yeast, boasts a high innate capacity to produce acyl-CoA derived molecules such as triacylglycerides. Here we demonstrate the potential of rewiring Y. lipolytica to divert this precursor pool away from lipids and into alternative molecules of interest. Specifically, we explored the production of the simple polyketide, triacetic acid lactone (TAL). TAL has been proposed as a biorenewable platform chemical that can be converted into many downstream products including sorbic acid. Previous efforts to produce TAL in hosts such as E. coli and S. cerevisiae have been limited by the availability of acyl-CoA precursors. We enabled TAL production in the precursor rich host, Y. lipolytica, through heterologous expression of 2-pyrone synthase, an enzyme from Gerbera hybrida that catalyzes the formation of TAL by condensation of acetyl-CoA and malonyl- CoA. We next performed a series of strain engineering approaches to boost TAL production by metabolically rewiring Y. lipolytica for enhanced precursor accumulation. Final strain characterization was conducted in bioreactors to further optimize production titer, rate, and yield. Ultimately, we established a strain that produced the highest titer of TAL reported to date in any host. Here we present the details of these genetic engineering efforts as well as the production characterization of the resulting strain. P5 Enzymatic syntheses of (S)-ethyl 1-((S)-2-(4-cyanophenyl)-2- hydroxyethyl)-piperidine-3-carboxylate and (S)-4-(oxiran-2-yl)benzonitrile A. Singh*, Z. Guo and A. Goswami, Bristol-Myers Squibb, New Brunswick, NJ, USA BMS-960, (S)-1-((S)-2-Hydroxy-2-(4-(5-(3-phenyl-4-(trifluoromethyl)isoxazol-5-yl)-1,2,4-oxadiazol-3- yl)phenyl)ethyl)piperidine-3-carboxyl acid) is a S1P1 receptor agonist. At the beginning of the program, KRED-NADPH-114 enzyme catalyzed reduction was identified for the synthesis of the advanced intermediate (S)-ethyl 1-((S)-2-(4-cyanophenyl)-2-hydroxyethyl)-piperidine-3-carboxylate (1). The absolute stereochemistry was assigned based on X-ray crystallography. In order to accommodate a common intermediate approach, an earlier epoxide intermediate (S)-4-(oxiran-2-yl)benzonitrile (2) was synthesized by KRED-NADH-110 catalyzed reduction of the corresponding α-bromoketone to the chiral α- bromohydrin followed by cyclisation to the epoxide. P7 Upgrading biomass-derived sugars to HMF/furfural via ketose intermediates W. Wang*, A. Mittal and D.K. Johnson, National Renewable Energy Laboratory, Golden, CO, USA As lignocellulosic biomass is considered to be the only sustainable resource with the potential to deliver renewable fuels and biobased chemicals, exploring fermentative or chemical pathways that convert biomass-derived sugars to fuels/chemicals has attracted a lot of interests from many researchers. We are investigating a hydrocarbon pathway from mixed sugars via HMF/furfural. Current processes to produce HMF/furfural generally involve the use of acid catalysts in biphasic systems or solvents such as ionic liquids. However, the yield from transforming glucose to HMF is very low compared to xylose conversion to furfural. In this study, we present an efficient chemical transformation pathway to HMF/furfural via ketose sugars, i.e. fructose and xylulose, which were generated from glucose and xylose via enzymatic isomerization. In enzymatic isomerization, by adding sodium borate to complex with the ketoses, xylose conversion reached equilibrium after 2h with a conversion of 91% and glucose conversion reached 84% after 4h. At 120oC, pH 0.5, and 15 min reaction time, mixed sugars (predominantly ketoses) , were converted to HMF and furfural in yields of 77% and 96%, respectively (based on starting aldose concentrations). These results demonstrate that this combined biological and chemical process could be an effective pathway to simultaneously upgrade glucose and xylose to HMF and furfural intermediates in the production of hydrocarbons. P9 Characterization of the unique pectin deconstruction system of Paenibacillus amylolyticus 27C64 C. Keggi* and J.D. Peterson, University of Georgia, Athens, GA, USA P. amylolyticus 27C64, a Gram-positive bacterium originally isolated from the hindgut of a Tipula abdominalis (aquatic cranefly) larvae, is capable of degrading plant cell wall polysaccharides including pectin. Two previously characterized pectate lyases from this organism have demonstrated broad substrate specificity and one has been used successfully in place of a commercial pectinase cocktail to ferment pectin-rich biomass to ethanol. To identify additional pectinases in this organism, its genome was sequenced and mined using two automated carbohydrate active enzyme annotation tools. Fifteen new putative pectinases were identified including esterases, lyases, and hydrolases predicted to be active on both types of the pectic backbone. In contrast to other systems which typically demethylate pectin extracellularly, this system’s sole putative pectin methylesterase appears to be cytoplasmic. Also of note is an enzyme with both a putative RG lyase and RG acetylesterase domain. An enzyme with both of these activities has not yet been described. Two newly identified pectate lyases, pamy_1763 and pamy_4669, have been heterologously overexpressed in E. coli and are currently being characterized. Together these enzymes allow P. amylolyticus to use polygalacturonic acid (PGA) as a supplemental carbon source in tryptic soy broth. The system appears to be induced by PGA and susceptible to catabolite repression by a number of sugars. Quantitative real-time PCR is currently being used to confirm transcription of these 17 genes in inducing conditions. To our knowledge, this is the first attempt to characterize a complete pectinolytic system within the Paenibacillus genus. P11 The impact of stress-response related transcription factors on lignocellulosic hydrolysate inhibitor tolerance of Saccharomyces strains J. Mertens* and R.E. Hector, USDA-ARS, NCAUR, Peoria, IL, USA; C.D. Skory, United States Department of Agriculture, Agricultural Research Service, National Center for Agricultural Utilization Research, Peoria, IL, USA Plant biomass is a desirable feedstock for the production of renewable fuels and chemicals. Unfortunately, pretreatment processes to release sugars locked in plant biomass, or lignocellulosic feedstocks, lead to the production of fermentation inhibitors, such as furfural and hydroxymethyl furfural, resulting in the inefficient fermentation of the lignocellulosic derived sugars by Saccharomyces cerevisiae. Numerous transcription factor genes associated with stress response are upregulated in Saccharomyces cultures grown in the presence of these inhibitors and overexpression of these transcription factors offers a potential route to improved inhibitor tolerance. Overexpression of one of