Improvement of alkaliphily of Bacillus alkaline xylanase by introducing amino acid substitutions both on catalytic cleft and protein surface

Hirohito Umemoto; Ihsanawati; Mayuko Inami; Rie Yatsunami; Toshiaki Fukui; Takashi Kumasaka; Nobuo Tanaka; Satoshi Nakamura

doi:10.1271/bbb.80869

Improvement of alkaliphily of Bacillus alkaline xylanase by introducing amino acid substitutions both on catalytic cleft and protein surface

Biosci Biotechnol Biochem. 2009 Apr 23;73(4):965-7. doi: 10.1271/bbb.80869. Epub 2009 Apr 7.

Authors

Hirohito Umemoto¹, Ihsanawati, Mayuko Inami, Rie Yatsunami, Toshiaki Fukui, Takashi Kumasaka, Nobuo Tanaka, Satoshi Nakamura

Affiliation

¹ Department of Bioengineering, Tokyo Institute of Technology, 4529 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.

PMID: 19352020
DOI: 10.1271/bbb.80869

Abstract

Xylanase J (XynJ) from alkaliphilic Bacillus sp. 41M-1 is an alkaline xylanase. The crystal structure has been solved with XynJ. Improvement of the alkaliphily of XynJ was attempted by amino acid substitutions. Reinforcing the characteristic salt bridge in the catalytic cleft and introducing excess Arg residues on the protein surface shifted the optimum pH of the wild-type enzyme from 8.5 to 9.5.

MeSH terms

Amino Acid Substitution*
Bacillus / enzymology*
Catalytic Domain*
Endo-1,4-beta Xylanases / chemistry*
Endo-1,4-beta Xylanases / genetics*
Endo-1,4-beta Xylanases / metabolism*
Hydrogen-Ion Concentration
Models, Molecular
Mutation
Protein Engineering / methods*
Temperature

Substances

Endo-1,4-beta Xylanases