Keywords

M.tb Acyl-CoA reductase, Rv3391, Rv1543, Rv1544

Abstract

Mycobacterium tuberculosis has been long known to produce wax esters. However, the enzymes involved in their biosynthesis have not been identified. Here we report the identification of Rv3391 and Rv1543 as genes that encode fatty acyl-CoA reductases and Rv1544 as one that encodes an aldehyde reductase. When expressed in E.coli, the products of Rv3391 and Rv1543 catalyzed reduction of fatty acyl-CoA to fatty alcohol with the corresponding aldehyde as an intermediate with an optimal pH of 7.0. Both enzymes showed a strong preference for NADPH over NADH as a reductant. Apparent Km for NADPH was 38 [micro]M for Rv3391 product and 202 [micro]M for Rv1543 product. Both enzymes reduced saturated fatty acyl-CoA such as palmitoyl-CoA and stearyl-CoA but showed a preference for oleoyl-CoA. Apparent Km for oleoyl-CoA was 13 [micro]M for Rv3391 product and 7 [micro]M for Rv1543 product. The Rv1544 product catalyzed fatty aldehyde reduction to fatty alcohol but not acyl-CoA reduction. The optimal pH for aldehyde reduction was 8.0. This aldehyde reductase showed a strong preference for NADPH with an apparent Km of 83 [micro]M. All three reductases were inhibited by SH directed reagents.

Notes

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Graduation Date

2007

Semester

Fall

Advisor

Kolattukudy, Pappachan

Degree

Master of Science (M.S.)

College

Burnett College of Biomedical Sciences

Department

Molecular Biology and Microbiology

Degree Program

Molecular and Microbiology

Format

application/pdf

Identifier

CFE0001903

URL

http://purl.fcla.edu/fcla/etd/CFE0001903

Language

English

Release Date

November 2007

Length of Campus-only Access

None

Access Status

Masters Thesis (Open Access)

Restricted to the UCF community until November 2007; it will then be open access.

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