Unravelling the hidden power of esterases for biomanufacturing of short-chain esters

Sarnaik, Aditya P.; Shinde, Somnath; Mhatre, Apurv; Jansen, Abigail; Jha, Amit Kumar; McKeown, Haley; Davis, Ryan; Varman, Arul M.

doi:10.1038/s41598-023-37542-x

Title: Unravelling the hidden power of esterases for biomanufacturing of short-chain esters

Journal Article · 03 July 2023 · Scientific Reports

DOI: https://doi.org/10.1038/s41598-023-37542-x · OSTI ID:1989071

Sarnaik, Aditya P.; Shinde, Somnath; Mhatre, Apurv; Jansen, Abigail; Jha, Amit Kumar; McKeown, Haley; Davis, Ryan; Varman, Arul M.

Abstract Microbial production of esters has recently garnered wide attention, but the current production metrics are low. Evidently, the ester precursors (organic acids and alcohols) can be accumulated at higher titers by microbes like Escherichia coli . Hence, we hypothesized that their ‘direct esterification’ using esterases will be efficient. We engineered esterases from various microorganisms into E. coli , along with overexpression of ethanol and lactate pathway genes. High cell density fermentation exhibited the strains possessing esterase-A (SSL76) and carbohydrate esterase (SSL74) as the potent candidates. Fed-batch fermentation at pH 7 resulted in 80 mg/L of ethyl acetate and 10 mg/L of ethyl lactate accumulation by SSL76. At pH 6, the total ester titer improved by 2.5-fold, with SSL76 producing 225 mg/L of ethyl acetate, and 18.2 mg/L of ethyl lactate, the highest reported titer in E. coli . To our knowledge, this is the first successful demonstration of short-chain ester production by engineering ‘esterases’ in E. coli .

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Research Organization:: Sandia National Laboratories (SNL-CA), Livermore, CA (United States)

Sponsoring Organization:: USDOE; USDOE National Nuclear Security Administration (NNSA); USDOE Office of Energy Efficiency and Renewable Energy (EERE), Office of Sustainable Transportation. Bioenergy Technologies Office (BETO)

Grant/Contract Number:: NA0003525

OSTI ID:: 1989071

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 13; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United Kingdom

Language:: English

References (61)

Flavors and Fragrances, 1. General Aspects Panten, Johannes; Surburg, Horst Ullmann's Encyclopedia of Industrial Chemistry https://doi.org/10.1002/14356007.a11_141.pub2	reference-book	September 2015
A membrane‐bound esterase PA2949 fromPseudomonas aeruginosais expressed and purified fromEscherichia coli Kovacic, Filip; Bleffert, Florian; Caliskan, Muttalip FEBS Open Bio, Vol. 6, Issue 5 https://doi.org/10.1002/2211-5463.12061	journal	April 2016
Metabolic analysis of adaptive evolution for in silico-designed lactate-producing strains Hua, Qiang; Joyce, Andrew R.; Fong, Stephen S. Biotechnology and Bioengineering, Vol. 95, Issue 5 https://doi.org/10.1002/bit.21073	journal	January 2006
Evaluating Factors That Influence Microbial Synthesis Yields by Linear Regression with Numerical and Ordinal Variables Colletti, Peter F.; Goyal, Yogesh; Varman, Arul M. Biotechnology and Bioengineering, Vol. 108, Issue 4 https://doi.org/10.1002/bit.22996	journal	November 2010
The temperature dependent functionality ofBrettanomyces bruxellensisstrains in wort fermentations Tyrawa, Caroline; Preiss, Richard; Armstrong, Meagan Journal of the Institute of Brewing, Vol. 125, Issue 3 https://doi.org/10.1002/jib.565	journal	January 2019
Molecular and biochemical aspects ofBrettanomycesin brewing Menoncin, Marcelo; Bonatto, Diego Journal of the Institute of Brewing, Vol. 125, Issue 4 https://doi.org/10.1002/jib.580	journal	January 2019
Low-Copy Plasmids can Perform as Well as or Better Than High-Copy Plasmids for Metabolic Engineering of Bacteria Jones, Kristala L.; Kim, Seon-Won; Keasling, J. D. Metabolic Engineering, Vol. 2, Issue 4 https://doi.org/10.1006/mben.2000.0161	journal	October 2000
Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of thelac promoter Donovan, R. S.; Robinson, C. W.; Glick, B. R. Journal of Industrial Microbiology, Vol. 16, Issue 3 https://doi.org/10.1007/BF01569997	journal	March 1996
Microbial formation of esters Park, Yong Cheol; Shaffer, Catherine Emily Horton; Bennett, George N. Applied Microbiology and Biotechnology, Vol. 85, Issue 1 https://doi.org/10.1007/s00253-009-2170-x	journal	August 2009
Production of isopropyl and butyl esters by Clostridium mono-culture and co-culture Cui, Yonghao; He, Jianzhong; Yang, Kun-Lin Journal of Industrial Microbiology & Biotechnology, Vol. 47, Issue 6-7 https://doi.org/10.1007/s10295-020-02279-3	journal	May 2020
Effect of temperature and pH on the formation of higher alcohols, fatty acids and esters in the malt whisky fermentation Ramsay, C. M.; Berry, D. R. Food Microbiology, Vol. 1, Issue 2 https://doi.org/10.1016/0740-0020(84)90021-2	journal	April 1984
Metabolic engineering in the biotechnological production of organic acids in the tricarboxylic acid cycle of microorganisms: Advances and prospects Yin, Xian; Li, Jianghua; Shin, Hyun-dong Biotechnology Advances, Vol. 33, Issue 6 https://doi.org/10.1016/j.biotechadv.2015.04.006	journal	November 2015
Microbial production of short and medium chain esters: Enzymes, pathways, and applications Kruis, Aleksander J.; Bohnenkamp, Anna C.; Patinios, Constantinos Biotechnology Advances, Vol. 37, Issue 7 https://doi.org/10.1016/j.biotechadv.2019.06.006	journal	November 2019
The raise of Brettanomyces yeast species for beer production Serra Colomer, Marc; Funch, Birgitte; Forster, Jochen Current Opinion in Biotechnology, Vol. 56 https://doi.org/10.1016/j.copbio.2018.07.009	journal	April 2019
Towards renewable flavors, fragrances, and beyond Lee, Jong-Won; Trinh, Cong T. Current Opinion in Biotechnology, Vol. 61 https://doi.org/10.1016/j.copbio.2019.12.017	journal	February 2020
High-throughput screening for efficient microbial biotechnology Sarnaik, Aditya; Liu, Arren; Nielsen, David Current Opinion in Biotechnology, Vol. 64 https://doi.org/10.1016/j.copbio.2020.02.019	journal	August 2020
B-esterases characterisation in the digestive tract of the common octopus and the European cuttlefish and their in vitro responses to contaminants of environmental concern Omedes, S.; Andrade, M.; Escolar, O. Environmental Research, Vol. 210 https://doi.org/10.1016/j.envres.2022.112961	journal	July 2022
Use of modular, synthetic scaffolds for improved production of glucaric acid in engineered E. coli Moon, Tae Seok; Dueber, John E.; Shiue, Eric Metabolic Engineering, Vol. 12, Issue 3 https://doi.org/10.1016/j.ymben.2010.01.003	journal	May 2010
Synergy between 13C-metabolic flux analysis and flux balance analysis for understanding metabolic adaption to anaerobiosis in E. coli Chen, Xuewen; Alonso, Ana P.; Allen, Doug K. Metabolic Engineering, Vol. 13, Issue 1 https://doi.org/10.1016/j.ymben.2010.11.004	journal	January 2011
Engineering modular ester fermentative pathways in Escherichia coli Layton, Donovan S.; Trinh, Cong T. Metabolic Engineering, Vol. 26 https://doi.org/10.1016/j.ymben.2014.09.006	journal	November 2014
Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels Atsumi, Shota; Hanai, Taizo; Liao, James C. Nature, Vol. 451, Issue 7174, p. 86-89 https://doi.org/10.1038/nature06450	journal	January 2008
Expanding ester biosynthesis in Escherichia coli Rodriguez, Gabriel M.; Tashiro, Yohei; Atsumi, Shota Nature Chemical Biology, Vol. 10, Issue 4 https://doi.org/10.1038/nchembio.1476	journal	March 2014
Intracellular ion concentrations and cation-dependent remodelling of bacterial MreB assemblies Szatmári, Dávid; Sárkány, Péter; Kocsis, Béla Scientific Reports, Vol. 10, Issue 1 https://doi.org/10.1038/s41598-020-68960-w	journal	July 2020
Acetate fluxes in Escherichia coli are determined by the thermodynamic control of the Pta-AckA pathway Enjalbert, Brice; Millard, Pierre; Dinclaux, Mickael Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/srep42135	journal	February 2017
The EstA esterase is responsible for the main capacity of Lactococcus lactis to synthesize short chain fatty acid esters in vitro Nardi, M.; Fiez-Vandal, C.; Tailliez, P. Journal of Applied Microbiology, Vol. 93, Issue 6 https://doi.org/10.1046/j.1365-2672.2002.01793.x	journal	December 2002
Optimization of culture conditions to improve the expression level of beta1–epsilon toxin ofClostridium perfringenstype B inEscherichia coli Lin, Chuwen; Cheng, Likun; Wang, Jing Biotechnology & Biotechnological Equipment, Vol. 30, Issue 2 https://doi.org/10.1080/13102818.2015.1126201	journal	January 2016
The molecular biology of fruity and floral aromas in beer and other alcoholic beverages Holt, Sylvester; Miks, Marta H.; de Carvalho, Bruna Trindade FEMS Microbiology Reviews, Vol. 43, Issue 3 https://doi.org/10.1093/femsre/fuy041	journal	November 2018
Use of signal sequences as an in situ removable sequence element to stimulate protein synthesis in cell-free extracts Ahn, J. -H.; Hwang, M. -Y.; Lee, K. -H. Nucleic Acids Research, Vol. 35, Issue 4 https://doi.org/10.1093/nar/gkl917	journal	January 2007
eQuilibrator--the biochemical thermodynamics calculator Flamholz, A.; Noor, E.; Bar-Even, A. Nucleic Acids Research, Vol. 40, Issue D1 https://doi.org/10.1093/nar/gkr874	journal	November 2011
Optimization of fermentation conditions for the production of curcumin by engineered Escherichia coli Couto, Márcia R.; Rodrigues, Joana L.; Rodrigues, Lígia R. Journal of The Royal Society Interface, Vol. 14, Issue 133 https://doi.org/10.1098/rsif.2017.0470	journal	August 2017
Production and biological function of volatile esters in Saccharomyces cerevisiae: Production and biological function of volatile esters in yeast Saerens, Sofie M. G.; Delvaux, Freddy R.; Verstrepen, Kevin J. Microbial Biotechnology, Vol. 3, Issue 2 https://doi.org/10.1111/j.1751-7915.2009.00106.x	journal	April 2009
Transformation of alcohols to esters promoted by hydrogen bonds using oxygen as the oxidant under metal-free conditions Liu, Mingyang; Zhang, Zhanrong; Liu, Huizhen Science Advances, Vol. 4, Issue 10 https://doi.org/10.1126/sciadv.aas9319	journal	October 2018
Production of Glucaric Acid from a Synthetic Pathway in Recombinant Escherichia coli Moon, T. S.; Yoon, S. -H.; Lanza, A. M. Applied and Environmental Microbiology, Vol. 75, Issue 3 https://doi.org/10.1128/AEM.00973-08	journal	December 2008
Metabolic Engineering of Synechocystis sp. Strain PCC 6803 for Isobutanol Production Varman, Arul M.; Xiao, Yi; Pakrasi, Himadri B. Applied and Environmental Microbiology, Vol. 79, Issue 3 https://doi.org/10.1128/AEM.02827-12	journal	November 2012
The Autotransporter Esterase EstA of Pseudomonas aeruginosa Is Required for Rhamnolipid Production, Cell Motility, and Biofilm Formation Wilhelm, Susanne; Gdynia, Aneta; Tielen, Petra Journal of Bacteriology, Vol. 189, Issue 18 https://doi.org/10.1128/JB.00023-07	journal	September 2007
pH of the Cytoplasm and Periplasm of Escherichia coli: Rapid Measurement by Green Fluorescent Protein Fluorimetry Wilks, J. C.; Slonczewski, J. L. Journal of Bacteriology, Vol. 189, Issue 15 https://doi.org/10.1128/JB.00615-07	journal	June 2007
Cytoplasmic pH Response to Acid Stress in Individual Cells of Escherichia coli and Bacillus subtilis Observed by Fluorescence Ratio Imaging Microscopy Martinez, Keith A.; Kitko, Ryan D.; Mershon, J. Patrick Applied and Environmental Microbiology, Vol. 78, Issue 10 https://doi.org/10.1128/aem.00354-12	journal	March 2012
Metabolic Engineering of Escherichia coli for Efficient Conversion of Glycerol to Ethanol Trinh, C. T.; Srienc, F. Applied and Environmental Microbiology, Vol. 75, Issue 21 https://doi.org/10.1128/aem.00670-09	journal	September 2009
Minimal Escherichia coli Cell for the Most Efficient Production of Ethanol from Hexoses and Pentoses Trinh, C. T.; Unrean, P.; Srienc, F. Applied and Environmental Microbiology, Vol. 74, Issue 12, p. 3634-3643 https://doi.org/10.1128/aem.02708-07	journal	April 2008
Expression Levels of the Yeast Alcohol Acetyltransferase Genes ATF1, Lg-ATF1, and ATF2 Control the Formation of a Broad Range of Volatile Esters Verstrepen, K. J.; Van Laere, S. D. M.; Vanderhaegen, B. M. P. Applied and Environmental Microbiology, Vol. 69, Issue 9 https://doi.org/10.1128/aem.69.9.5228-5237.2003	journal	September 2003
Photoautotrophic production of D-lactic acid in an engineered cyanobacterium Varman, Arul M.; Yu, Yi; You, Le Microbial Cell Factories, Vol. 12, Issue 1 https://doi.org/10.1186/1475-2859-12-117	journal	January 2013
Engineering of Serine-Deamination pathway, Entner-Doudoroff pathway and pyruvate dehydrogenase complex to improve poly(3-hydroxybutyrate) production in Escherichia coli Zhang, Yan; Lin, Zhenquan; Liu, Qiaojie Microbial Cell Factories, Vol. 13, Issue 1 https://doi.org/10.1186/s12934-014-0172-6	journal	December 2014
Parallel substrate supply and pH stabilization for optimal screening of E. coli with the membrane-based fed-batch shake flask Philip, P.; Kern, D.; Goldmanns, J. Microbial Cell Factories, Vol. 17, Issue 1 https://doi.org/10.1186/s12934-018-0917-8	journal	May 2018
Recent advances in lignin valorization with bacterial cultures: microorganisms, metabolic pathways, and bio-products Xu, Zhaoxian; Lei, Peng; Zhai, Rui Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1376-0	journal	February 2019
A review on commercial-scale high-value products that can be produced alongside cellulosic ethanol Rosales-Calderon, Oscar; Arantes, Valdeir Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1529-1	journal	October 2019
Microbial biosynthesis of lactate esters Lee, Jong-Won; Trinh, Cong T. Biotechnology for Biofuels, Vol. 12, Issue 1 https://doi.org/10.1186/s13068-019-1563-z	journal	September 2019
Recent advances in the valorization of plant biomass Ning, Peng; Yang, Guofeng; Hu, Lihong Biotechnology for Biofuels, Vol. 14, Issue 1 https://doi.org/10.1186/s13068-021-01949-3	journal	April 2021
Clean synthesis of biolubricant range esters using novel liquid lipase enzyme in solvent free medium Trivedi, Jayati; Aila, Mounika; Sharma, Chandra Dutt SpringerPlus, Vol. 4, Issue 1 https://doi.org/10.1186/s40064-015-0937-3	journal	April 2015
Production of medium-chain volatile flavour esters in Pichia pastoris whole-cell biocatalysts with extracellular expression of Saccharomyces cerevisiae acyl-CoA:ethanol O-acyltransferase Eht1 or Eeb1 Zhuang, Shiwen; Fu, Junshu; Powell, Chris SpringerPlus, Vol. 4, Issue 1 https://doi.org/10.1186/s40064-015-1195-0	journal	September 2015
Study on the Production of Bacterial Cellulose from Acetobacter Xylinum Using Agro - Waste Lestari, Puji; Elfrida, Nitariani; Suryani, Ani Jordan Journal of Biological Sciences, Vol. 7, Issue 1 https://doi.org/10.12816/0008218	journal	March 2014
Consistent Estimation of Gibbs Energy Using Component Contributions Noor, Elad; Haraldsdóttir, Hulda S.; Milo, Ron PLoS Computational Biology, Vol. 9, Issue 7 https://doi.org/10.1371/journal.pcbi.1003098	journal	July 2013
Biochemical Characterization of a First Fungal Esterase from Rhizomucor miehei Showing High Efficiency of Ester Synthesis Liu, Yu; Xu, Haibo; Yan, Qiaojuan PLoS ONE, Vol. 8, Issue 10 https://doi.org/10.1371/journal.pone.0077856	journal	October 2013
Purification and biochemical characterization of FrsA protein from Vibrio vulnificus as an esterase Wang, Xiaoqin; Li, Zhi-Min; Li, Qingyue PLOS ONE, Vol. 14, Issue 4 https://doi.org/10.1371/journal.pone.0215084	journal	April 2019
Novel perspective on a conventional technique: Impact of ultra-low temperature on bacterial viability and protein extraction Sarnaik, Aditya; Mhatre, Apurv; Faisal, Muhammad PLOS ONE, Vol. 16, Issue 5 https://doi.org/10.1371/journal.pone.0251640	journal	May 2021
Group Contribution Method for Thermodynamic Analysis of Complex Metabolic Networks Jankowski, Matthew D.; Henry, Christopher S.; Broadbelt, Linda J. Biophysical Journal, Vol. 95, Issue 3 https://doi.org/10.1529/biophysj.107.124784	journal	August 2008
DIFFERENTIAL EFFECTS OF TEMPERATURE AND pH ON THE ANTIBIOTIC RESISTANCE OF PATHOGENIC AND NON-PATHOGENIC STRAINS OF ESCHERICHIA COLI Chaudhry, Narender; Mukherjee, Tapan K.; Mukherjee, Tapan K. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 8, Issue 9 https://doi.org/10.22159/ijpps.2016v8i9.12664	journal	September 2016
Bacterial Valorization of Lignin: Strains, Enzymes, Conversion Pathways, Biosensors, and Perspectives Lee, Siseon; Kang, Minsik; Bae, Jung-Hoon Frontiers in Bioengineering and Biotechnology, Vol. 7 https://doi.org/10.3389/fbioe.2019.00209	journal	September 2019
Sustainable Production of Microbial Isoprenoid Derived Advanced Biojet Fuels Using Different Generation Feedstocks: A Review Walls, Laura Ellen; Rios-Solis, Leonardo Frontiers in Bioengineering and Biotechnology, Vol. 8 https://doi.org/10.3389/fbioe.2020.599560	journal	October 2020
Contribution of Eat1 and Other Alcohol Acyltransferases to Ester Production in Saccharomyces cerevisiae Kruis, Aleksander J.; Gallone, Brigida; Jonker, Timo Frontiers in Microbiology, Vol. 9 https://doi.org/10.3389/fmicb.2018.03202	journal	December 2018
Bacterial Signal Peptides- Navigating the Journey of Proteins Kaushik, Sharbani; He, Haoze; Dalbey, Ross E. Frontiers in Physiology, Vol. 13 https://doi.org/10.3389/fphys.2022.933153	journal	July 2022
Exploring Metagenomic Enzymes: A Novel Esterase Useful for Short-Chain Ester Synthesis Maester, Thaís Carvalho; Pereira, Mariana Rangel; Malaman, Aliandra M. Gibertoni Catalysts, Vol. 10, Issue 10 https://doi.org/10.3390/catal10101100	journal	September 2020

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