Amino acid substitutions of His296 alter the catalytic properties of Zymomonas mobilis 10232 levansucrase

文献类型: 外文期刊

第一作者: Wang, Jin

作者: Wang, Jin;Li, Shu Ying;Chen, Ming;Yan, Yong Liang;Yu, Hai Ying;Zhan, Yu Hua;Peng, Zi Xin;Lin, Min;Li, Gang

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关键词: Zymomonas mobilis;levansucrase;His296;hydrolysis activity;transfructosylation activity;levan;BACILLUS-SUBTILIS LEVANSUCRASE;SITE-DIRECTED MUTAGENESIS;LEVAN;RESIDUES

期刊名称:ACTA BIOCHIMICA POLONICA ( 影响因子:2.149; 五年影响因子:2.175 )

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收录情况: SCI

摘要: His296 of Zymomonas mobilis levansucrase (EC 2.4.1.10) is crucial for the catalysis of the trans-fructosylation reaction. The three-dimensional structures of levansucrases revealed the His296 is involved in the substrate recognition and binding. In this study, nine mutants were created by site-directed mutagenesis, in which His296 was substituted with amino acids of different polarity, charge and length. The substitutions of His296 with Arg or Trp retained partial hydrolysis and transfructosylation activities. The positively charged Lys substitution resulted in a 2.5-fold increase of sucrose hydrolysis. Substitutions with short (Cys or Ser), negatively charged (Glu) or polar (Tyr) amino acids virtually abolished both the activities. Analysis of transfructosylation products indicated that the mutants synthesized different oligosaccharides, suggesting that amino acid substitutions of His296 strongly affected both the enzyme activity and transfructosylation products.

分类号: Q5

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