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Characterization of recombinant FAD-independent catabolic acetolactate synthase from Enterococcus faecalis V583

Paper ID Volume ID Publish Year Pages File Format Full-Text
17156 42647 2013 6 PDF Available
Title
Characterization of recombinant FAD-independent catabolic acetolactate synthase from Enterococcus faecalis V583
Abstract

The catabolic acetolactate synthase (cALS) of Enterococcus faecalis V583 was cloned, expressed in Escherichia coli, and purified to homogeneity. The purified protein had a molecular weight of 60 kDa. The cALS of E. faecalis is highly homologous with other cALSs, while sharing low homology with its anabolic counterparts. The cALS of E. faecalis exhibits optimum activity at a temperature of 37 °C and pH 6.8. Based on the enzyme characterization, the apparent Km for pyruvate was calculated to be 1.37 mM, while the Kc for thiamin diphosphate (ThDP) and Mg2+ were found to be 0.031 μM and 1.27 mM, respectively. Negligible absorbance at 450 nm and lack of activity enhancement upon addition of flavin adenine dinucleotide (FAD) to the assay buffer suggest that the cALS of E. faecalis is not FAD-dependent. The enzyme showed extreme stability against the organic solvent dimethyl sulfoxide (DMSO), whereas the activity decreased to less than 50% in the presence of acetone and ethanol.

► Catabolic acetolactate synthase of Enterococcus faecalis was cloned and purified. ► The catabolic enzyme was characterized and found to be FAD-independent. ► The FAD-independent enzyme was extremely stabile against organic solvents.

Keywords
Enterococcus faecalis; Catabolic; Acetolactate synthase; ThDP; Flavin adenine dinucleotideAL, acetolactate; ALS, acetolactate synthase; cALS, catabolic acetolactate synthase; AHAS, acetohydroxyacid synthase; BCAA, branched-chain amino acid; FAD, flavin ad
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Characterization of recombinant FAD-independent catabolic acetolactate synthase from Enterococcus faecalis V583
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 52, Issue 1, 10 January 2013, Pages 54–59
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us