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Relative Activities of the β-Ketoacyl-CoA and Acyl-CoA Reductases Influence the Product Profile and Flux in a Reversed β-Oxidation Pathway

Journal Article · · ACS Catalysis
 [1];  [2];  [2];  [2];  [2];  [2];  [2]
  1. University of Wisconsin−Madison, WI (United States); University of Wisconsin-Madison Department of Chemical and Biological Engineering
  2. University of Wisconsin−Madison, WI (United States)
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The β-oxidation pathway, normally involved in the catabolism of fatty acids, can be functionally made to act as a fermentative, iterative, elongation pathway when driven by the activity of a trans-enoyl-CoA reductase (TER). Here, the terminal acyl-CoA reduction to alcohol can occur on substrates with varied chain lengths, leading to a broad distribution of fermentation products in vivo. Tight control of the average chain length and product profile is desirable, as the chain length greatly influences the molecular properties and the commercial value. Lacking a termination enzyme with a narrow chain length preference, we sought alternative factors that could influence the product profile and pathway flux in the iterative pathway. In this study, we reconstituted the reversed β-oxidation (R-βox) pathway in vitro with a purified tri-functional complex (FadBA) responsible for the thiolase, enoyl-CoA hydratase, and hydroxyacyl-CoA dehydrogenase activities, a TER, and an acyl-CoA reductase. Using this system, we determined the rate-limiting step of the elongation cycle and demonstrated that by controlling the ratio of these three enzymes and the ratio of NADH and NADPH, we can influence the average chain length of the alcohol product profile.
Research Organization:
Great Lakes Bioenergy Research Center, Madison, WI (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
Grant/Contract Number:
SC0018409
OSTI ID:
2228434
Journal Information:
ACS Catalysis, Journal Name: ACS Catalysis Journal Issue: 9 Vol. 13; ISSN 2155-5435
Publisher:
American Chemical Society (ACS)Copyright Statement
Country of Publication:
United States
Language:
English

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Figures / Tables (5)

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Figure 3(p. 34)figure Figure 3
Figure 4(p. 36)figure Figure 4
Figure 5(p. 38)figure Figure 5

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Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
enzyme kinetics
fatty alcohols
metabolic engineering
thiolase
β-oxidation pathway