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Substrate specificity of glycoside hydrolase family 1 β-glucosidase AtBGlu42 from Arabidopsis thaliana and its molecular mechanism
| Content Provider | Scilit |
|---|---|
| Author | Horikoshi, Shu Saburi, Wataru Yu, Jian Matsuura, Hideyuki Cairns, James R. Ketudat Yao, Min Mori, Haruhide |
| Copyright Year | 2021 |
| Description | Plants possess many glycoside hydrolase family 1 (GH1) β-glucosidases, which physiologically function in cell wall metabolism and activation of bioactive substances, but most remain uncharacterized. One GH1 isoenzyme AtBGlu42 in Arabidopsis thaliana has been identified to hydrolyze scopolin using the gene deficient plants, but no enzymatic properties were obtained. Its sequence similarity to another functionally characterized enzyme Os1BGlu4 in rice suggests that AtBGlu42 also acts on oligosaccharides. Here, we show that the recombinant AtBGlu42 possesses high $k_{cat}/K_{m}$ not only on scopolin, but also on various β-glucosides, cellooligosaccharides, and laminarioligosaccharides. Of the cellooligosaccharides, cellotriose was the most preferred. The crystal structure, determined at 1.7 Å resolution, suggests that Arg342 gives unfavorable binding to cellooligosaccharides at subsite +3. The mutants R342Y and R342A showed the highest preference on cellotetraose or cellopentaose with increased affinities at subsite +3, indicating that the residues at this position have an important role for chain length specificity. |
| Related Links | https://academic.oup.com/bbb/advance-article-pdf/doi/10.1093/bbb/zbab200/41974766/zbab200.pdf |
| Ending Page | 245 |
| Page Count | 15 |
| Starting Page | 231 |
| ISSN | 09168451 |
| e-ISSN | 13476947 |
| DOI | 10.1093/bbb/zbab200 |
| Journal | Bioscience, Biotechnology, and Biochemistry |
| Issue Number | 2 |
| Volume Number | 86 |
| Language | English |
| Publisher | Informa UK Limited |
| Publisher Date | 2021-11-22 |
| Access Restriction | Open |
| Subject Keyword | Journal: Bioscience, Biotechnology, and Biochemistry Biochemistry and Molecular Biology Β-glucosidase Glycoside Hydrolase Family 1 Substrate Specificity X-ray Crystallography Arabidopsis Thaliana |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Medicine Analytical Chemistry Molecular Biology Biochemistry Applied Microbiology and Biotechnology Biotechnology |
