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

Title
Characterization of recombinant FAD-independent catabolic acetolactate synthase from Enterococcus faecalis V583
Author
김순길
Keywords
Enterococcus faecalis; Catabolic; Acetolactate synthase; ThDP; Flavin adenine dinucleotide; ACETOHYDROXYACID SYNTHASE; ESCHERICHIA-COLI; KLEBSIELLA-PNEUMONIAE; SERRATIA-MARCESCENS; FORMING ENZYME; AEROBACTER AEROGENES; POTENT INHIBITORS; BACILLUS-SUBTILIS; PURIFICATION; PH
Issue Date
2013-01
Publisher
Elsevier Science B.V., Amsterdam.
Citation
Enzyme and Microbial Technology, 10 January 2013, 52(1), P.54-59
Abstract
The catabolic acetolactate synthase (cALS) of Enterococcus faecalis V583 was cloned, expressed in Escherichia colt, 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 degrees C and pH 6.8. Based on the enzyme characterization, the apparent K-m for pyruvate was calculated to be 1.37 mM, while the K-c for thiamin diphosphate (ThDP) and Mg2+ were found to be 0.031 mu M 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. (C) 2012 Elsevier Inc. All rights reserved.
URI
https://www.sciencedirect.com/science/article/pii/S0141022912001743http://repository.hanyang.ac.kr/handle/20.500.11754/69595
ISSN
0141-0229
DOI
10.1016/j.enzmictec.2012.10.006
Appears in Collections:
COLLEGE OF MEDICINE[S](의과대학) > MEDICINE(의학과) > Articles
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