Sustainable production of photosynthetic isobutanol and 3-methyl-1-butanol in the cyanobacterium Synechocystis sp. PCC 6803.

2-Keto acid pathway 3-Methyl-1-butanol Cyanobacteria Isobutanol Metabolic engineering Solar fuel Synechocystis sp. PCC 6803

Journal

Biotechnology for biofuels and bioproducts

ISSN: 2731-3654

Titre abrégé: Biotechnol Biofuels Bioprod

Pays: England

ID NLM: 9918300888906676

Informations de publication

Date de publication:
09 Sep 2023

Historique:

received: 21 04 2023

accepted: 24 08 2023

medline: 9 9 2023

pubmed: 9 9 2023

entrez: 8 9 2023

Statut: epublish

Résumé

Cyanobacteria are emerging as green cell factories for sustainable biofuel and chemical production, due to their photosynthetic ability to use solar energy, carbon dioxide and water in a direct process. The model cyanobacterial strain Synechocystis sp. PCC 6803 has been engineered for the isobutanol and 3-methyl-1-butanol production by introducing a synthetic 2-keto acid pathway. However, the achieved productions still remained low. In the present study, diverse metabolic engineering strategies were implemented in Synechocystis sp. PCC 6803 for further enhanced photosynthetic isobutanol and 3-methyl-1-butanol production. Long-term cultivation was performed on two selected strains resulting in maximum cumulative isobutanol and 3-methyl-1-butanol titers of 1247 mg L Enhanced isobutanol and 3-methyl-1-butanol production was successfully achieved in Synechocystis sp. PCC 6803 strains through diverse metabolic engineering strategies. The maximum cumulative isobutanol and 3-methyl-1-butanol titers, 1247 mg L

Sections du résumé

BACKGROUND BACKGROUND

RESULTS RESULTS

Long-term cultivation was performed on two selected strains resulting in maximum cumulative isobutanol and 3-methyl-1-butanol titers of 1247 mg L

CONCLUSIONS CONCLUSIONS

Enhanced isobutanol and 3-methyl-1-butanol production was successfully achieved in Synechocystis sp. PCC 6803 strains through diverse metabolic engineering strategies. The maximum cumulative isobutanol and 3-methyl-1-butanol titers, 1247 mg L

Identifiants

DOI: 10.1186/s13068-023-02385-1 PMID: 37684613 PMC: PMC10492371

pubmed: 37684613

doi: 10.1186/s13068-023-02385-1

pii: 10.1186/s13068-023-02385-1

pmc: PMC10492371

doi:

Types de publication

Journal Article

Langues

eng

Pagination

134

Subventions

Organisme : Energimyndigheten

ID : P46607-1

Organisme : NordForsk

ID : 82845

Informations de copyright

Références

US Energy Information Administration (EIA). International energy outlook 2021: with projections to 2050. 2021.

Keasling J, Garcia Martin H, Lee TS, Mukhopadhyay A, Singer SW, Sundstrom E. Microbial production of advanced biofuels. Nat Rev Microbiol. 2021;19(11):701–15.

pubmed: 34172951

Zhou YJ, Kerkhoven EJ, Nielsen J. Barriers and opportunities in bio-based production of hydrocarbons. Nat Energy. 2018;3(11):925–35.

Biofuels GJ. In-flight insights. Nat Energy. 2017;2(4):17065.

Lu J, Brigham CJ, Gai CS, Sinskey AJ. Studies on the production of branched-chain alcohols in engineered Ralstonia eutropha. Appl Microbiol Biotechnol. 2012;96(1):283–97.

pubmed: 22864971

Stockhardt JS, Hull CM. Vapor-liquid equilibria and boiling-point composition relations for systems n-butanol-water and isobutanol-water. Ind Eng Chem. 1931;23(12):1438–40.

Atsumi S, Hanai T, Liao JC. Non-fermentative pathways for synthesis of branched-chain higher alcohols as biofuels. Nature. 2008;451(7174):86–9.

pubmed: 18172501

Lee WH, Seo SO, Bae YH, Nan H, Jin YS, Seo JH. Isobutanol production in engineered Saccharomyces cerevisiae by overexpression of 2-ketoisovalerate decarboxylase and valine biosynthetic enzymes. Bioprocess Biosyst Eng. 2012;35(9):1467–75.

pubmed: 22543927

Li S, Wen J, Jia X. Engineering Bacillus subtilis for isobutanol production by heterologous Ehrlich pathway construction and the biosynthetic 2-ketoisovalerate precursor pathway overexpression. Appl Microbiol Biotechnol. 2011;91(3):577–89.

pubmed: 21533914

Baez A, Cho KM, Liao JC. High-flux isobutanol production using engineered Escherichia coli: a bioreactor study with in situ product removal. Appl Microbiol Biotechnol. 2011;90(5):1681–90.

pubmed: 21547458 pmcid: 3094657

Atsumi S, Higashide W, Liao JC. Direct photosynthetic recycling of carbon dioxide to isobutyraldehyde. Nat Biotechnol. 2009;27(12):1177–80.

pubmed: 19915552

Miao R, Liu X, Englund E, Lindberg P, Lindblad P. Isobutanol production in Synechocystis PCC 6803 using heterologous and endogenous alcohol dehydrogenases. Metab Eng Commun. 2017;5:45–53.

pubmed: 29188183 pmcid: 5699533

Varman AM, Xiao Y, Pakrasi HB, Tang YJ. Metabolic engineering of Synechocystis sp. strain PCC 6803 for isobutanol production. Appl Environ Microbiol. 2013;79(3):908–14.

pubmed: 23183979 pmcid: 3568544

Miao R, Xie H, Ho FM, Lindblad P. Protein engineering of alpha-ketoisovalerate decarboxylase for improved isobutanol production in Synechocystis PCC 6803. Metab Eng. 2018;47:42–8.

pubmed: 29501927

Xie H, Lindblad P. Expressing 2-keto acid pathway enzymes significantly increases photosynthetic isobutanol production. Microb Cell Fact. 2022;21(1):17.

pubmed: 35105340 pmcid: 8805274

Sherkhanov S, Korman TP, Chan S, Faham S, Liu H, Sawaya MR, Hsu WT, Vikram E, Cheng T, Bowie JU. Isobutanol production freed from biological limits using synthetic biochemistry. Nat Commun. 2020;11(1):4292.

pubmed: 32855421 pmcid: 7453195

Fu C, Li Z, Jia C, Zhang W, Zhang Y, Yi C, Xie S. Recent advances on bio-based isobutanol separation. Energy Convers Manag: X. 2021;10: 100059.

Englund E, Andersen-Ranberg J, Miao R, Hamberger B, Lindberg P. Metabolic Engineering of Synechocystis sp. PCC 6803 for Production of the Plant Diterpenoid Manoyl Oxide. ACS Synth Biol. 2015;4(12):1270–8.

pubmed: 26133196 pmcid: 4685428

Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol. 1979;111:1–61.

Miao R, Xie H, Lindblad P. Enhancement of photosynthetic isobutanol production in engineered cells of Synechocystis PCC 6803. Biotechnol Biofuels. 2018;11:267.

pubmed: 30275907 pmcid: 6158846

Lai MJ, Tsai JC, Lan EI. CRISPRi-enhanced direct photosynthetic conversion of carbon dioxide to succinic acid by metabolically engineered cyanobacteria. Bioresour Technol. 2022;366: 128131.

pubmed: 36252759

Kouhen OM, Joset F. Biosynthesis of the branched-chain amino acids in the cyanobacterium Synechocystis PCC 6803: existence of compensatory pathways. Curr Microbiol. 2002;45(2):94–8.

pubmed: 12070685

Maestri O, Joset F. Regulation by external pH and stationary growth phase of the acetolactate synthase from Synechocystis PCC 6803. Mol Microbiol. 2000;37(4):828–38.

pubmed: 10972805

Kaneko T, Sato S, Kotani H, Tanaka A, Asamizu E, Nakamura Y, Miyajima N, Hirosawa M, Sugiura M, Sasamoto S, Kimura T, Hosouchi T, Matsuno A, Muraki A, Nakazaki N, Naruo K, Okumura S, Shimpo S, Takeuchi C, Wada T, Watanabe A, Yamada M, Yasuda M, Tabata S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC 6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions. DNA Res. 1996;3(3):109–36.

pubmed: 8905231

Englund E, Liang F, Lindberg P. Evaluation of promoters and ribosome binding sites for biotechnological applications in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Sci Rep. 2016;6:36640.

pubmed: 27857166 pmcid: 5114575

Huang HH, Lindblad P. Wide-dynamic-range promoters engineered for cyanobacteria. J Biol Eng. 2013;7:10.

pubmed: 23607865 pmcid: 3724501

Mills LA, McCormick AJ, Lea-Smith DJ. Current knowledge and recent advances in understanding metabolism of the model cyanobacterium Synechocystis sp. PCC 6803. Biosci Rep. 2020;40:4.

Wang Y, Chen L, Zhang W. Proteomic and metabolomic analyses reveal metabolic responses to 3-hydroxypropionic acid synthesized internally in cyanobacterium Synechocystis sp. PCC 6803. Biotechnol Biofuels. 2016;9:209.

pubmed: 27757169 pmcid: 5053081

Yunus IS, Lee TS. Applications of targeted proteomics in metabolic engineering: advances and opportunities. Curr Opin Biotechnol. 2022;75: 102709.

pubmed: 35279538

Xu C, Wang B, Yang L, Zhongming HuL, Yi L, Wang Y, Chen S, Emili A, Wan C. Global landscape of native protein complexes in Synechocystis sp. PCC 6803. Genomics Proteomics Bioinform. 2022;20(4):715–27.

Cengic I, Canadas IC, Minton NP, Hudson EP. Inducible CRISPR/Cas9 allows for multiplexed and rapidly segregated single-target genome editing in Synechocystis sp. PCC 6803. ACS Synth Biol. 2022;11(9):3100–13.

pubmed: 35969224 pmcid: 9486961

Kojima K, Keta S, Uesaka K, Kato A, Takatani N, Ihara K, Omata T, Aichi M. A simple method for isolation and construction of markerless cyanobacterial mutants defective in acyl-acyl carrier protein synthetase. Appl Microbiol Biotechnol. 2016;100(23):10107–13.

pubmed: 27704180 pmcid: 5102962

Lea-Smith DJ, Vasudevan R, Howe CJ. Generation of marked and markerless mutants in model cyanobacterial species. J Vis Exp. 2016;111:54001.

Li H, Shen CR, Huang CH, Sung LY, Wu MY, Hu YC. CRISPR-Cas9 for the genome engineering of cyanobacteria and succinate production. Metab Eng. 2016;38:293–302.

pubmed: 27693320

Soh LMJ, Mak WS, Lin PP, Mi L, Chen FY, Damoiseaux R, Siegel JB, Liao JC. Engineering a thermostable keto acid decarboxylase using directed evolution and computationally directed protein design. ACS Synth Biol. 2017;6(4):610–8.

pubmed: 28052191

Liang F, Englund E, Lindberg P, Lindblad P. Engineered cyanobacteria with enhanced growth show increased ethanol production and higher biofuel to biomass ratio. Metab Eng. 2018;46:51–9.

pubmed: 29477858

Liang F, Lindblad P. Effects of overexpressing photosynthetic carbon flux control enzymes in the cyanobacterium Synechocystis PCC 6803. Metab Eng. 2016;38:56–64.

pubmed: 27328433

Nakahara K, Yamamoto H, Miyake C, Yokota A. Purification and characterization of class-I and class-II fructose-1,6-bisphosphate aldolases from the cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol. 2003;44(3):326–33.

pubmed: 12668779

Yang C, Hua Q, Shimizu K. Metabolic flux analysis in Synechocystis using isotope distribution from

pubmed: 12616690

Luinenburg I, Coleman JR. A requirement for phosphoenolpyruvate carboxylase in the cyanobacterium Synechococcus PCC 7942. Arch Microbiol. 1990;154(5):471–4.

Cheah YE, Xu Y, Sacco SA, Babele PK, Zheng AO, Johnson CH, Young JD. Systematic identification and elimination of flux bottlenecks in the aldehyde production pathway of Synechococcus elongatus PCC 7942. Metab Eng. 2020;60:56–65.

pubmed: 32222320 pmcid: 7217728

Liu W, Zhang R, Wei M, Cao Y, Xian M. Increasing the pyruvate pool by overexpressing phosphoenolpyruvate carboxykinase or triosephosphate isomerase enhances phloroglucinol production in Escherichia coli. Biotechnol Lett. 2020;42(4):633–40.

pubmed: 31965395

Satoh S, Ikeuchi M, Mimuro M, Tanaka A. Chlorophyll b expressed in Cyanobacteria functions as a light-harvesting antenna in photosystem I through flexibility of the proteins. J Biol Chem. 2001;276(6):4293–7.

pubmed: 11073958

Jazmin LJ, Xu Y, Cheah YE, Adebiyi AO, Johnson CH, Young JD. Isotopically nonstationary

pubmed: 28479191 pmcid: 5660605

Englund E, Shabestary K, Hudson EP, Lindberg P. Systematic overexpression study to find target enzymes enhancing production of terpenes in Synechocystis PCC 6803, using isoprene as a model compound. Metab Eng. 2018;49:164–77.

pubmed: 30025762