Cofactor engineering of ketol-acid reductoisomerase (IlvC) and alcohol dehydrogenase (YqhD) improves the fusel alcohol yield in algal protein anaerobic fermentation

Wu, Weihua; Tran-Gyamfi, Mary Bao; Jaryenneh, James Dekontee; Davis, Ryan W.

doi:10.1016/j.algal.2016.08.013

Title: Cofactor engineering of ketol-acid reductoisomerase (IlvC) and alcohol dehydrogenase (YqhD) improves the fusel alcohol yield in algal protein anaerobic fermentation

Journal Article · 24 August 2016 · Algal Research

DOI: https://doi.org/10.1016/j.algal.2016.08.013 · OSTI ID:1338336

Wu, Weihua ^[1]; Tran-Gyamfi, Mary Bao ^[1]; Jaryenneh, James Dekontee ^[1]; Davis, Ryan W. ^[1]

Sandia National Lab. (SNL-CA), Livermore, CA (United States)

Recently the feasibility of conversion of algal protein to mixed alcohols has been demonstrated with an engineered E.coli strain, enabling comprehensive utilization of the biomass for biofuel applications. However, the yield and titers of mixed alcohol production must be improved for market adoption. A major limiting factor for achieving the necessary yield and titer improvements is cofactor imbalance during the fermentation of algal protein. To resolve this problem, a directed evolution approach was applied to modify the cofactor specificity of two key enzymes (IlvC and YqhD) from NADPH to NADH in the mixed alcohol metabolic pathway. Using high throughput screening, more than 20 YqhD mutants were identified to show activity on NADH as a cofactor. Of these 20 mutants, the top five of YqhD mutants were selected for combination with two IlvC mutants with NADH as a cofactor for the modification of the protein conversion strain. The combination of the IlvC and YqhD mutants yielded a refined E.coli strain, subtype AY3, with increased fusel alcohol yield of ~60% compared to wild type under anaerobic fermentation on amino acid mixtures. When applied to real algal protein hydrolysates, the strain AY3 produced 100% and 38% more total mixed alcohols than the wild type strain on two different algal hydrolysates, respectively. The results indicate that cofactor engineering is a promising approach to improve the feasibility of bioconversion of algal protein into mixed alcohols as advanced biofuels.

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

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: AC04-94AL85000; ACO4-94AL85000; 26336

OSTI ID:: 1338336

Alternate ID(s):: OSTI ID: 1402509

Report Number(s):: SAND2016-2002J; PII: S2211926416302715

Journal Information:: Algal Research, Vol. 19, Issue C; ISSN 2211-9264

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

References (21)

Microbial engineering for the production of advanced biofuels Peralta-Yahya, Pamela P.; Zhang, Fuzhong; del Cardayre, Stephen B. Nature, Vol. 488, Issue 7411 https://doi.org/10.1038/nature11478	journal	August 2012
Algal biofuels Razeghifard, Reza Photosynthesis Research, Vol. 117, Issue 1-3 https://doi.org/10.1007/s11120-013-9828-z	journal	April 2013
Algal biofuels: the eternal promise? Luque, Rafael Energy & Environmental Science, Vol. 3, Issue 3 https://doi.org/10.1039/b922597h	journal	January 2010
An overview of algae bioethanol production: An overview of algae bioethanol production Li, Kexun; Liu, Shun; Liu, Xianhua International Journal of Energy Research, Vol. 38, Issue 8 https://doi.org/10.1002/er.3164	journal	January 2014
Hydrogen Production. Green Algae as a Source of Energy Melis, Anastasios; Happe, Thomas Plant Physiology, Vol. 127, Issue 3, p. 740-748 https://doi.org/10.1104/pp.010498	journal	November 2001
Biofuels from microalgae: Lipid extraction and methane production from the residual biomass in a biorefinery approach Hernández, D.; Solana, M.; Riaño, B. Bioresource Technology, Vol. 170 https://doi.org/10.1016/j.biortech.2014.07.109	journal	October 2014
Assessing microalgae biorefinery routes for the production of biofuels via hydrothermal liquefaction López Barreiro, Diego; Samorì, Chiara; Terranella, Giuseppe Bioresource Technology, Vol. 174 https://doi.org/10.1016/j.biortech.2014.10.031	journal	December 2014
One-pot bioconversion of algae biomass into terpenes for advanced biofuels and bioproducts Wu, Weihua; Davis, Ryan W. Algal Research, Vol. 17 https://doi.org/10.1016/j.algal.2016.05.005	journal	July 2016
Biodiesel from algae: challenges and prospects Scott, Stuart A.; Davey, Matthew P.; Dennis, John S. Current Opinion in Biotechnology, Vol. 21, Issue 3 https://doi.org/10.1016/j.copbio.2010.03.005	journal	June 2010
Commercialization potential of microalgae for biofuels production Singh, Jasvinder; Gu, Sai Renewable and Sustainable Energy Reviews, Vol. 14, Issue 9 https://doi.org/10.1016/j.rser.2010.06.014	journal	December 2010
Conversion of proteins into biofuels by engineering nitrogen flux Huo, Yi-Xin; Cho, Kwang Myung; Rivera, Jimmy G Lafontaine Nature Biotechnology, Vol. 29, Issue 4, p. 346-351 https://doi.org/10.1038/nbt.1789	journal	March 2011
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
Activating transhydrogenase and NAD kinase in combination for improving isobutanol production Shi, Aiqin; Zhu, Xinna; Lu, Jiao Metabolic Engineering, Vol. 16 https://doi.org/10.1016/j.ymben.2012.11.008	journal	March 2013
Engineered ketol-acid reductoisomerase and alcohol dehydrogenase enable anaerobic 2-methylpropan-1-ol production at theoretical yield in Escherichia coli Bastian, Sabine; Liu, Xiang; Meyerowitz, Joseph T. Metabolic Engineering, Vol. 13, Issue 3, p. 345-352 https://doi.org/10.1016/j.ymben.2011.02.004	journal	May 2011
Site-saturation mutagenesis of formate dehydrogenase from Candida bodinii creating effective NADP+-dependent FDH enzymes Wu, Weihua; Zhu, Dunming; Hua, Ling Journal of Molecular Catalysis B: Enzymatic, Vol. 61, Issue 3-4 https://doi.org/10.1016/j.molcatb.2009.06.005	journal	December 2009
Flash hydrolysis of microalgae (Scenedesmus sp.) for protein extraction and production of biofuels intermediates Garcia-Moscoso, Jose Luis; Obeid, Wassim; Kumar, Sandeep The Journal of Supercritical Fluids, Vol. 82 https://doi.org/10.1016/j.supflu.2013.07.012	journal	October 2013
Kinetics of Peptides and Arginine Production from Microalgae ( Scenedesmus sp .) by Flash Hydrolysis Garcia-Moscoso, Jose L.; Teymouri, Ali; Kumar, Sandeep Industrial & Engineering Chemistry Research, Vol. 54, Issue 7 https://doi.org/10.1021/ie5047279	journal	February 2015
Crystal Structure of E.coli Alcohol Dehydrogenase YqhD: Evidence of a Covalently Modified NADP Coenzyme Sulzenbacher, Gerlind; Alvarez, Karine; van den Heuvel, Robert H. H. Journal of Molecular Biology, Vol. 342, Issue 2, p. 489-502 https://doi.org/10.1016/j.jmb.2004.07.034	journal	September 2004
Microbial synthesis of n-butanol, isobutanol, and other higher alcohols from diverse resources Lan, Ethan I.; Liao, James C. Bioresource Technology, Vol. 135 https://doi.org/10.1016/j.biortech.2012.09.104	journal	May 2013
An evolutionary strategy for isobutanol production strain development in Escherichia coli Smith, Kevin M.; Liao, James C. Metabolic Engineering, Vol. 13, Issue 6 https://doi.org/10.1016/j.ymben.2011.08.004	journal	November 2011
Genomic Encyclopedia of Bacteria and Archaea: Sequencing a Myriad of Type Strains Kyrpides, Nikos C.; Hugenholtz, Philip; Eisen, Jonathan A. PLoS Biology, Vol. 12, Issue 8 https://doi.org/10.1371/journal.pbio.1001920	journal	August 2014

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Bioconversion of distillers’ grains hydrolysates to advanced biofuels by an Escherichia coli co-culture Liu, Fang; Wu, Weihua; Tran-Gyamfi, Mary B. Microbial Cell Factories, Vol. 16, Issue 1 https://doi.org/10.1186/s12934-017-0804-8	journal	November 2017
Characterization of four endophytic fungi as potential consolidated bioprocessing hosts for conversion of lignocellulose into advanced biofuels Wu, Weihua; Davis, Ryan W.; Tran-Gyamfi, Mary Bao Applied Microbiology and Biotechnology, Vol. 101, Issue 6 https://doi.org/10.1007/s00253-017-8091-1	journal	January 2017
Development of algae biorefinery concepts for biofuels and bioproducts; a perspective on process-compatible products and their impact on cost-reduction Laurens, Lieve M. L.; Markham, Jennifer; Templeton, David W. Energy & Environmental Science, Vol. 10, Issue 8 https://doi.org/10.1039/c7ee01306j	journal	January 2017

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59 BASIC BIOLOGICAL SCIENCES
directed evolution
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Title: Cofactor engineering of ketol-acid reductoisomerase (IlvC) and alcohol dehydrogenase (YqhD) improves the fusel alcohol yield in algal protein anaerobic fermentation

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