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B-factor Guided Proline Substitutions in Chromobacterium violaceum Amine Transaminase: Evaluation of the Proline Rule as a Method for Enzyme Stabilization.

Chembiochem. 2019-01; 
LandHenrik, Campillo-BrocalJonatan C, Svedendahl HumbleMaria, Berglun
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PCR Cloning and Subcloning … in Chemistry, Biotechnology and Health (CBH), Department of Industrial Biotechnology, AlbaNova University … 751 20, Uppsala, Sweden [c] Dr. M. Svedendahl Humble Pharem Biotech AB, Biovation … and cloned into pET28a(+) using restriction sites NcoI and BamHI by GenScript Get A Quote

摘要

Biocatalysis is attracting interest in the chemical industry as a sustainable alternative in large-scale chemical transformations. However, low operational stability of naturally evolved enzymes is a challenge and major efforts are required to engineer protein stability, usually by directed evolution. The development of methods for protein stabilization based on rational design is of great interest, as it would minimize the efforts needed to generate stable enzymes. Here we present a rational design strategy based on proline substitutions in flexible areas of the protein identified by analyzing B-factors. Several proline substitutions in the amine transaminase from Chromobacterium violaceum were shown to ... More

关键词

biocatalysis,enzyme stabilization,enzymes,molecular modeling,protein enginee