Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/68639
標題: Replacement of methionine 208 in a truncated Bacillus sp TS-23 at alpha-amylase with oxidation-resistant leucine enhances its resistance to hydrogen peroxide
作者: Lin, L.L.
Lo, H.F.
Chiang, W.Y.
Hu, H.Y.
Hsu, W.H.
Chang, C.T.
關鍵字: alkalopsychrotrophic micrococcus sp;enzymatic-properties;protein;oxidation;escherichia-coli;alkaliphilic bacillus;amino-acids;residues;sequence;strain;gene
Project: Current Microbiology
期刊/報告no:: Current Microbiology, Volume 46, Issue 3, Page(s) 211-216.
摘要: 
The methionine residues at positions 17, 104, 208, 214, 292, 315, 324, and 446 in the primary amino acid sequence of a truncated Bacillus sp. TS-23 alpha-amylase (HiS(6)-tagged BLADeltaNC) was changed to oxidative-resistant leucine by site-directed mutagenesis. The mutant enzymes with an apparent molecular mass of approximately 54 kDa were overexpressed in recombinant Escherichia coli. The specific activity for Met315Leu and Met446Leu was decreased by more than 76%, while Met17Leu, Met104Leu, Met208Leu, Met214Leu, Met292Leu, and Met324Leu showed 247, 128, 37, 260, 232, and 241%, respectively, higher activity than the wild-type enzyme. In comparison with wild-type enzyme, a lower K-m value was observed for all mutant enzymes. The 3.2- and 4.5-fold increases in the catalytic efficiency (k(cat)/K-m) for Met208Leu and Met324Leu, respectively, were partly contributed by a 68% and 38% decrease in K-m values. Wild-type enzyme was sensitive to chemical oxidation, but Met208Leu was stable even in the presence of 500 mM H2O2. Except for Met214Leu, which was quite sensitive to H2O2, the other mutants showed a profile of oxidative inactivation similar to that of the wild-type enzyme. These observations indicate that the oxidative stability of HiS(6)-tagged BLADeltaNC can be improved by replacement of the critical methionine residue with leucine.
URI: http://hdl.handle.net/11455/68639
ISSN: 0343-8651
DOI: 10.1007/s00284-002-3846-y
Appears in Collections:期刊論文

Show full item record
 

Google ScholarTM

Check

Altmetric

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.