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Chromohalobacter salixigens uronate dehydrogenase: Directed evolution for improved thermal stability and mutant CsUDH-inc X-ray crystal structure

Process Biochemistry. 2020; 
Kurt Wagschala, *, Victor J. Chana, Jose H. Pereirab, Peter H. Zwartc, Banumathi Sankarand, *
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Molecular Biology Reagents The resulting DNA sequences were synthesized (Genscript USA Inc., Piscataway, NJ) and subcloned into pET29b(+) expression vector (EMD Millipore, Burlington, MA).  Get A Quote

摘要

Chromohalobacter salixigens contains a uronate dehydrogenase termed CsUDH that can convert uronic acids to their corresponding C1,C6-dicarboxy aldaric acids, an important enzyme reaction applicable for biotechnological use of sugar acids. To increase the thermal stability of this enzyme for biotechnological processes, directed evolution using gene family shuffling was applied, and the hits selected from 2-tier screening of a shuffled gene family library contained in total 16 mutations, only some of which when examined individually appreciably increased thermal stability. Most mutations, while having minimal or no effect on thermal stability when tested in isolation, were found to exhibit synergy when combined;... More

关键词

Uronate dehydrogenase Directed evolution Pectin Alginate Gene shuffling Protein thermal stability