The effect of disulfide bond introduction and related Cys/Ser mutations on the stability of a cyclohexanone monooxygenase
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- 作者：Sandy Schmidt ; Maika Genz ; Kathleen Balke ; Uwe T. Bornscheuer ; uwe.bornscheuer@uni-greifswald.de" class="auth_mail" title="E-mail the corresponding author
- 关键词：BVMO ; Baeyer&ndash ; Villiger monooxygenase ; CHMO ; cyclohexanone monooxygenase ; PAMO ; phenyl acetone monooxygenase ; ABD-F ; [4-aminosulfonyl)-7-fluoro-2 ; 1 ; 3-benzoxadiazole] ; TCEP ; [Tris (2-carboxyethyl) phosphine hydrochloride] ; DS ; disulfide
- 刊名：Journal of Biotechnology
- 出版年：2015
- 出版时间：20 November 2015
- 年：2015
- 卷：214
- 期：Complete
- 页码：199-211
- 全文大小：1745 K
文摘
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Application of an efficient and easy method for the introduction of additional disulfide bonds without the need of a crystal structure.
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CHMO variants created with higher thermal as well as oxidative stability without any loss of catalytic activity.
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Additional creation and analysis of single Cys/Ser mutants.
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20">Major improvement of the enzyme half-life by only one exceptional cysteine mutation.
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25">ABD-F labeling as fast assay to verify the correct formation of disulfide bonds.