Role of L-alanine for redox self-sufficient amination of alcohols

Klatte S, Wendisch VF (2015)
Microbial Cell Factories 14(1): 9.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
Background In white biotechnology biocatalysis represents a key technology for chemical functionalization of non-natural compounds. The plasmid-born overproduction of an alcohol dehydrogenase, an L-alanine-dependent transaminase and an alanine dehydrogenase allows for redox self-sufficient amination of alcohols in whole cell biotransformation. Here, conditions to optimize the whole cell biocatalyst presented in (Bioorg Med Chem 22:5578–5585, 2014), and the role of L-alanine for efficient amine functionalization of 1,10-decanediol to 1,10-diaminodecane were analyzed. Results The enzymes of the cascade for amine functionalization of alcohols were characterized in vitro to find optimal conditions for an efficient process. Transaminase from Chromobacterium violaceum, TaCv, showed three-fold higher catalytic efficiency than transaminase from Vibrio fluvialis, TaVf, and improved production at 37°C. At 42°C, TaCv was more active, which matched thermostable alcohol dehydrogenase and alanine dehydrogenase and improved the 1,10-diaminodecane production rate four-fold. To study the role of L-alanine in the whole cell biotransformation, the L-alanine concentration was varied and 1,10.diaminodecane formation tested with constant 10 mM 1,10- decanediol and 100 mM NH4Cl. Only 5.6% diamine product were observed without added L-alanine. L-alanine concentrations equimolar to that of the alcohol enabled for 94% product formation but higher L-alanine concentrations allowed for 100% product formation. L-alanine was consumed by the E. coli biocatalyst, presumably due to pyruvate catabolism since up to 16 mM acetate accumulated. Biotransformation employing E. coli strain YYC202/pTrc99a-ald-adh-taCv, which is unable to catabolize pyruvate, resulted in conversion with a selectivity of 42 mol-%. Biotransformation with E. coli strains only lacking pyruvate oxidase PoxB showed similar reduced amination of 1,10-decanediol indicating that oxidative decarboxylation of pyruvate to acetate by PoxB is primarily responsible for pyruvate catabolism during redox self-sufficient amination of alcohols using this whole cell biocatalyst. Conclusion The replacement of the transaminase TaVf by TaCv, which showed higher activity at 42°C, in the artificial operon ald-adh-ta improved amination of alcohols in whole cell biotransformation. The addition of L-alanine, which was consumed by E. coli via pyruvate catabolism, was required for 100% product formation possibly by providing maintenance energy. Metabolic engineering revealed that pyruvate catabolism occurred primarily via oxidative decarboxylation to acetate by PoxB under the chosen biotranformation conditions.
Phosphate acetyltransferase; Pyruvate oxidase; Acetate kinase; Acetate formation; Energy maintenance; Chromobacterium violaceum; Transaminase; Escherichia coli; Whole cell biotransformation; Redox self-sufficient amination
Microbial Cell Factories
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Klatte S, Wendisch VF. Role of L-alanine for redox self-sufficient amination of alcohols. Microbial Cell Factories. 2015;14(1): 9.
Klatte, S., & Wendisch, V. F. (2015). Role of L-alanine for redox self-sufficient amination of alcohols. Microbial Cell Factories, 14(1), 9. doi:10.1186/s12934-014-0189-x
Klatte, Stephanie, and Wendisch, Volker F. 2015. “Role of L-alanine for redox self-sufficient amination of alcohols”. Microbial Cell Factories 14 (1): 9.
Klatte, S., and Wendisch, V. F. (2015). Role of L-alanine for redox self-sufficient amination of alcohols. Microbial Cell Factories 14:9.
Klatte, S., & Wendisch, V.F., 2015. Role of L-alanine for redox self-sufficient amination of alcohols. Microbial Cell Factories, 14(1): 9.
S. Klatte and V.F. Wendisch, “Role of L-alanine for redox self-sufficient amination of alcohols”, Microbial Cell Factories, vol. 14, 2015, : 9.
Klatte, S., Wendisch, V.F.: Role of L-alanine for redox self-sufficient amination of alcohols. Microbial Cell Factories. 14, : 9 (2015).
Klatte, Stephanie, and Wendisch, Volker F. “Role of L-alanine for redox self-sufficient amination of alcohols”. Microbial Cell Factories 14.1 (2015): 9.
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7 Zitationen in Europe PMC

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Biotechnological production of mono- and diamines using bacteria: recent progress, applications, and perspectives.
Wendisch VF, Mindt M, Pérez-García F., Appl Microbiol Biotechnol 102(8), 2018
PMID: 29520601
Whole-cell biocatalysts by design.
Lin B, Tao Y., Microb Cell Fact 16(1), 2017
PMID: 28610636
In vivo plug-and-play: a modular multi-enzyme single-cell catalyst for the asymmetric amination of ketoacids and ketones.
Farnberger JE, Lorenz E, Richter N, Wendisch VF, Kroutil W., Microb Cell Fact 16(1), 2017
PMID: 28754115
Biocatalytic hydrogen-borrowing cascades.
Knaus T, Mutti FG., Chim Oggi 35(5), 2017
PMID: 29515288
Enzymatic network for production of ether amines from alcohols.
Palacio CM, Crismaru CG, Bartsch S, Navickas V, Ditrich K, Breuer M, Abu R, Woodley JM, Baldenius K, Wu B, Janssen DB., Biotechnol Bioeng 113(9), 2016
PMID: 26915048

23 References

Daten bereitgestellt von Europe PubMed Central.

Crystal structures of unbound and aminooxyacetate-bound Escherichia coli gamma-aminobutyrate aminotransferase.
Liu W, Peterson PE, Carter RJ, Zhou X, Langston JA, Fisher AJ, Toney MD., Biochemistry 43(34), 2004
PMID: 15323550
Crystal structure of an (R)-selective ω-transaminase from Aspergillus terreus.
Lyskowski A, Gruber C, Steinkellner G, Schurmann M, Schwab H, Gruber K, Steiner K., PLoS ONE 9(1), 2014
PMID: 24498081
Cofactor regeneration at the lab scale.
Wichmann R, Vasic-Racki D., Adv. Biochem. Eng. Biotechnol. 92(), 2005
PMID: 15791939
Redox self-sufficient biocatalyst network for the amination of primary alcohols.
Sattler JH, Fuchs M, Tauber K, Mutti FG, Faber K, Pfeffer J, Haas T, Kroutil W., Angew. Chem. Int. Ed. Engl. 51(36), 2012
PMID: 22887645
Redox self-sufficient whole cell biotransformation for amination of alcohols.
Klatte S, Wendisch VF., Bioorg. Med. Chem. 22(20), 2014
PMID: 24894767

Pyruvate oxidase contributes to the aerobic growth efficiency of Escherichia coli
Abdel-Hamid AM, Attwood MM, Guest JR., 2001
Subtoxic product levels limit the epoxidation capacity of recombinant E. coli by increasing microbial energy demands.
Kuhn D, Fritzsch FS, Zhang X, Wendisch VF, Blank LM, Buhler B, Schmid A., J. Biotechnol. 163(2), 2012
PMID: 22922011
Substrate spectrum of omega-transaminase from Chromobacterium violaceum DSM30191 and its potential for biocatalysis
Kaulmann U, Smithies K, Smith MEB, HaileS HC, Ward JM., 2007
Purification, characterization, and molecular cloning of a novel amine:pyruvate transaminase from Vibrio fluvialis JS17.
Shin JS, Yun H, Jang JW, Park I, Kim BG., Appl. Microbiol. Biotechnol. 61(5-6), 2003
PMID: 12687298
Reduction of aerobic acetate production by Escherichia coli
Farmer WR, Liao JC., 1997
Acetate accumulation through alternative metabolic pathways in ackA (-) pta (-) poxB (-) triple mutant in E. coli B (BL21).
Phue JN, Lee SJ, Kaufman JB, Negrete A, Shiloach J., Biotechnol. Lett. 32(12), 2010
PMID: 20703804

Sambrook J, Russell D., 2001
Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.
Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H., Mol. Syst. Biol. 2(), 2006
PMID: 16738554
Studies on transformation of Escherichia coli with plasmids.
Hanahan D., J. Mol. Biol. 166(4), 1983
PMID: 6345791
Enzymatic assembly of DNA molecules up to several hundred kilobases.
Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA 3rd, Smith HO., Nat. Methods 6(5), 2009
PMID: 19363495

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