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)
STARS Citation
Singh, Harminder, "Identification Of Genes Encoding Acyl-coa Reductases And Aldehyde Reductases In Mycobacterial Genome By Characterization Of The Reductases Expressed In E. Coli" (2007). Electronic Theses and Dissertations. 3352.
https://stars.library.ucf.edu/etd/3352