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

• Authors: Shu Ying Li , Ming Chen , Gang Li , Yong Liang Yan , Hai Ying Yu , Yu Hua Zhan , Zi Xin Peng , Jin Wang , Min Lin
• Languages of publication: EN

Abstracts

EN

His296 of Zymomonas mobilis levansucrase (EC 2.4.1.10) is crucial for the catalysis of the transfructosylation 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.

Keywords

EN

Zymomonas mobilis; His296; levansucrase; levan; transfructosylation activity; hydrolysis activity

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2008
• Volume: 55
• Issue: 1
• Pages: 201-206

Dates

published

2008

received

2007-10-23

revised

2008-01-29

accepted

2008-02-12

(unknown)

2008-03-07

Contributors

author

Shu Ying Li

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Ming Chen

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Gang Li

• Sichuan Habio Bioengineering Co. Ltd., Chengdu, Sichuan, P.R. China

author

Yong Liang Yan

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Hai Ying Yu

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Yu Hua Zhan

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Zi Xin Peng

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

author

Jin Wang

• Plant Genomics Research center, Mianyang Normal College, Mianyang, Sichuan, P.R. China

author

Min Lin

• Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, P.R. China

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