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Applied Microbiology and Biotechnology

Applied Microbiology and Biotechnology

IF: 3.89
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Mutations at subsite ?3 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhancing α-cyclodextrin specificity

Published:1 June 2009 DOI: 10.1007/s00253-009-1865-3
Zhaofeng Li, Jiayu Zhang, Miao Wang, Zhengbiao Gu, Guocheng Du, Jianke Li, Jing Wu, Jian Chen

Abstract

A major disadvantage of cyclodextrin production is the limited cyclodextrin product specificity of cyclodextrin glycosyltransferase (CGTase). Here, we described mutations of Asp372 and Tyr89 at subsite ?3 in the CGTase from Paenibacillus macerans strain JFB05-01. The results showed that Asp372 and Tyr89 played important roles in cyclodextrin product specificity of CGTase. The replacement of Asp372 by lysine and Tyr89 by aspartic acid, asparagine, lysine, and arginine resulted in a shift in specificity towards the production of α-cyclodextrin, which was most apparent for the mutants D372K and Y89R. Furthermore, the changes in cyclodextrin product specificity for the single mutants D372K and Y89R could be combined in the double mutant D372K/Y89R, which displayed a 1.5-fold increase in the production of α-cyclodextrin, with a concomitant 43% decrease in the production of β-cyclodextrin when compared to the wild-type CGTase. Thus, the D372K and Y89R single and double mutants were much more suitable for the industrial production of α-cyclodextrin than the wild-type enzyme. The enhanced α-cyclodextrin specificity of these mutants might be a result of stabilizing the bent conformation of the intermediate in the cyclization reaction.

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Related products
Procduct Name CAS Molecular Formula Supplier Price
L-Lysine 56-87-1 C6H14N2O2 855 suppliers $5.00-$4310.00
β-Cyclodextrin 7585-39-9 C42H70O35 645 suppliers $13.50-$1810.00
α-Cyclodextrin 10016-20-3 C36H60O30 438 suppliers $5.00-$3120.00
D-(-)-Asparagine monohydrate 2058-58-4 C4H8N2O3 273 suppliers $15.00-$7500.00
L-ASPARTIC ACID 6899-03-2 C4H7NO4 222 suppliers Inquiry
Arginine 7004-12-8 C40H39F5N4O7S 21 suppliers Inquiry
Phospho-c-Abl (Tyr89) (61A6) Rabbit mAb - Inquiry

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