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Title 

New thermostable D-methionine amidase from Brevibacillus borstelensis BCS-1 and its application for D-phenylalanine production

Authors 

Dae Heoun BaekJae Jun SongSeung Goo LeeSeok Joon KwonY AsanoMoon Hee Sung

Publisher 

Elsevier

Issue Date 

2003

Citation 

Enzyme and Microbial Technology, vol. 32, no. 1, pp. 131-139

Keywords 

brevibacillus borstelensis BCS-1d-phenylalanine productionthermophilic bacteriumthermostable d-methionine amidaseamino acidsdextro methionine amidasedextro phenylalaninedithiothreitoldodecyl sulfate sodiumedetic acid

Abstract 

A new thermostable D-methionine amidase was found in a cell-free extract of Brevibacillus borstelensis BCS-1. After five steps of purification, the specific activity increased approximately 207-fold and the purity was more than 98%. The molecular weight of the enzyme was estimated to be 199kDa by gel permeation chromatography and 30kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which indicates that the thermostable D-methionine amidase was a homo-hexamer consisting of a single subunit. The purified enzyme was stable up to 65°C within a broad pH range from 6.5 to 10.0, and its maximum activity was measured at pH 9.5 and 70°C. The enzyme activity increased about five-fold with the addition of Co2+, yet was strongly inhibited by Hg2+, 2-mercaptoethanol, dithiothreitol, and ethylenediaminetetracetic acid. The thermostable D-methionine amidase exhibited a high amidase activity and D-stereospecificity toward D-amino acid amides and esters, yet did not hydrolyze D-peptides. The catalytic efficiencies (kcat/Km, mM-1s-1) of the enzyme for D-methioninamide and D-alaninamide were 3086 and 21.5, respectively, and the enantiomeric excess (ee) and enantiomeric ratio of D-phenylalanine produced from DL-phenylalaninamide were 97.1 and 196%, respectively.

ISSN 

0141-0229

Link 

http://dx.doi.org/10.1016/S0141-0229(02)00268-5

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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