Cofactor specificity engineering of a long-chain secondary alcohol dehydrogenase from Micrococcus luteus for redox-neutral biotransformation of fatty acids

2019 ◽  
Vol 55 (96) ◽  
pp. 14462-14465 ◽  
Author(s):  
Eun-Ji Seo ◽  
Hye-Ji Kim ◽  
Myeong-Ju Kim ◽  
Jeong-Sun Kim ◽  
Jin-Byung Park
Keyword(s):  
Fatty Acids ◽  
Alcohol Dehydrogenase ◽  
Micrococcus Luteus ◽  
Secondary Alcohol ◽  
Cofactor Specificity ◽  
Secondary Alcohol Dehydrogenase ◽  
Long Chain

Structure-based cofactor specificity engineering of an alcohol dehydrogenase (mLSADH) enables a redox-neutral biotransformation of C18 fatty acids into C9 fatty acids.

scholarly journals A novel secondary alcohol dehydrogenase from Micrococcus luteus WIUJH20: purification, cloning, and properties

2010 ◽  
Vol 24 (S1) ◽  
Author(s):  
Jenq‐Kuen Huang ◽  
Jung Ki Park ◽  
Babu Ram Dhungana ◽  
Nicholas D Youngblut ◽  
Chi‐Tsai Lin ◽  
...  
Keyword(s):  
Alcohol Dehydrogenase ◽  
Micrococcus Luteus ◽  
Secondary Alcohol ◽  
Secondary Alcohol Dehydrogenase

scholarly journals Physiological Function of Alcohol Dehydrogenases and Long-Chain (C30) Fatty Acids in Alcohol Tolerance of Thermoanaerobacter ethanolicus

2002 ◽  
Vol 68 (4) ◽  
pp. 1914-1918 ◽  
Author(s):  
D. S. Burdette ◽  
S.-H. Jung ◽  
G.-J. Shen ◽  
R. I. Hollingsworth ◽  
J. G. Zeikus
Keyword(s):  
Fatty Acids ◽  
Alcohol Dehydrogenase ◽  
Mutant Strain ◽  
Secondary Alcohol ◽  
Primary Alcohol ◽  
Alcohol Tolerance ◽  
Membrane Composition ◽  
Wild Type ◽  
Thermoanaerobacter Ethanolicus ◽  
Long Chain

ABSTRACT A mutant strain (39E H8) of Thermoanaerobacter ethanolicus that displayed high (8% [vol/vol]) ethanol tolerance for growth was developed and characterized in comparison to the wild-type strain (39E), which lacks alcohol tolerance (<1.5% [vol/vol]). The mutant strain, unlike the wild type, lacked primary alcohol dehydrogenase and was able to increase the percentage of transmembrane fatty acids (i.e., long-chain C30 fatty acids) in response to increasing levels of ethanol. The data support the hypothesis that primary alcohol dehydrogenase functions primarily in ethanol consumption, whereas secondary alcohol dehydrogenase functions in ethanol production. These results suggest that improved thermophilic ethanol fermentations at high alcohol levels can be developed by altering both cell membrane composition (e.g., increasing transmembrane fatty acids) and the metabolic machinery (e.g., altering primary alcohol dehydrogenase and lactate dehydrogenase activities).

scholarly journals Reconstruction of an Acetogenic 2,3-Butanediol Pathway Involving a Novel NADPH-Dependent Primary-Secondary Alcohol Dehydrogenase

2014 ◽  
Vol 80 (11) ◽  
pp. 3394-3403 ◽  
Author(s):  
Michael Köpke ◽  
Monica L. Gerth ◽  
Danielle J. Maddock ◽  
Alexander P. Mueller ◽  
FungMin Liew ◽  
...  
Keyword(s):  
Alcohol Dehydrogenase ◽  
Secondary Alcohol ◽  
Acetolactate Synthase ◽  
Chemical Production ◽  
Content Type ◽  
E Coli ◽  
Gas Fermentation ◽  
Secondary Alcohol Dehydrogenase ◽  
Clostridium Autoethanogenum ◽  
Butanediol Dehydrogenase

ABSTRACTAcetogenic bacteria use CO and/or CO2plus H2as their sole carbon and energy sources. Fermentation processes with these organisms hold promise for producing chemicals and biofuels from abundant waste gas feedstocks while simultaneously reducing industrial greenhouse gas emissions. The acetogenClostridium autoethanogenumis known to synthesize the pyruvate-derived metabolites lactate and 2,3-butanediol during gas fermentation. Industrially, 2,3-butanediol is valuable for chemical production. Here we identify and characterize theC. autoethanogenumenzymes for lactate and 2,3-butanediol biosynthesis. The putativeC. autoethanogenumlactate dehydrogenase was active when expressed inEscherichia coli. The 2,3-butanediol pathway was reconstituted inE. coliby cloning and expressing the candidate genes for acetolactate synthase, acetolactate decarboxylase, and 2,3-butanediol dehydrogenase. Under anaerobic conditions, the resultingE. colistrain produced 1.1 ± 0.2 mM 2R,3R-butanediol (23 μM h−1optical density unit−1), which is comparable to the level produced byC. autoethanogenumduring growth on CO-containing waste gases. In addition to the 2,3-butanediol dehydrogenase, we identified a strictly NADPH-dependent primary-secondary alcohol dehydrogenase (CaADH) that could reduce acetoin to 2,3-butanediol. Detailed kinetic analysis revealed that CaADH accepts a range of 2-, 3-, and 4-carbon substrates, including the nonphysiological ketones acetone and butanone. The high activity of CaADH toward acetone led us to predict, and confirm experimentally, thatC. autoethanogenumcan act as a whole-cell biocatalyst for converting exogenous acetone to isopropanol. Together, our results functionally validate the 2,3-butanediol pathway fromC. autoethanogenum, identify CaADH as a target for further engineering, and demonstrate the potential ofC. autoethanogenumas a platform for sustainable chemical production.

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