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Mutations at Subsite -3 in Cyclodextrin Glycosyltransferase From Paenibacillus Macerans Enhancing Alpha-Cyclodextrin Specificity

Mutations at Subsite -3 in Cyclodextrin Glycosyltransferase From Paenibacillus Macerans Enhancing Alpha-Cyclodextrin Specificity

Zhaofeng Li, Jiayu Zhang, Miao Wang, Zhengbiao Gu, Guocheng Du, Jianke Li, Jing Wu, Jian Chen

Appl Microbiol Biotechnol. 2009 Jun;83(3):483-90.

PMID: 19190904

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 alpha-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 alpha-cyclodextrin, with a concomitant 43% decrease in the production of beta-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 alpha-cyclodextrin than the wild-type enzyme. The enhanced alpha-cyclodextrin specificity of these mutants might be a result of stabilizing the bent conformation of the intermediate in the cyclization reaction.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP10016203	Alpha Cyclodextrin		10016-20-3	Price

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