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Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes
The β-glycoside hydrolases (LXYL-P1−1 and LXYL-P1−2) from Lentinula edodes (strain M95.33) can specifically hydrolyze 7-β-xylosyl-10-deacetyltaxol (XDT) to form 10-deacetyltaxol for the semi-synthesis of Taxol. Our previous study showed that both the I368T mutation in LXYL-P1−1 and the T368E mutatio...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6337304/ https://www.ncbi.nlm.nih.gov/pubmed/30586935 http://dx.doi.org/10.3390/molecules24010059 |
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author | Chen, Jing-Jing Liang, Xiao Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping |
author_facet | Chen, Jing-Jing Liang, Xiao Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping |
author_sort | Chen, Jing-Jing |
collection | PubMed |
description | The β-glycoside hydrolases (LXYL-P1−1 and LXYL-P1−2) from Lentinula edodes (strain M95.33) can specifically hydrolyze 7-β-xylosyl-10-deacetyltaxol (XDT) to form 10-deacetyltaxol for the semi-synthesis of Taxol. Our previous study showed that both the I368T mutation in LXYL-P1−1 and the T368E mutation in LXYL-P1−2 could increase the enzyme activity, which prompted us to investigate the effect of the I368E mutation on LXYL-P1−1 activity. In this study, the β-xylosidase and β-glucosidase activities of LXYL-P1−1(I368E) were 1.5 and 2.2 times higher than those of LXYL-P1−1. Most importantly, combination of I368E and V91S exerted the cumulative effects on the improvement of the enzyme activities and catalytic efficiency. The β-xylosidase and β-glucosidase activities of the double mutant LXYL-P1−1(V91S/I368E) were 3.2 and 1.7-fold higher than those of LXYL-P1−1(I368E). Similarly, the catalytic efficiency of LXYL-P1−1(V91S/I368E) on 7-β-xylosyl-10-deacetyltaxol was 1.8-fold higher than that of LXYL-P1−1(I368E) due to the dramatic increase in the substrate affinity. Molecular docking results suggest that the V91S and I368E mutation might positively promote the interaction between enzyme and substrate through altering the loop conformation near XDT and increasing the hydrogen bonds among Ser(91), Trp(301), and XDT. This study lays the foundation for exploring the relationship between the structure and function of the β-glycoside hydrolases. |
format | Online Article Text |
id | pubmed-6337304 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-63373042019-01-25 Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes Chen, Jing-Jing Liang, Xiao Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping Molecules Article The β-glycoside hydrolases (LXYL-P1−1 and LXYL-P1−2) from Lentinula edodes (strain M95.33) can specifically hydrolyze 7-β-xylosyl-10-deacetyltaxol (XDT) to form 10-deacetyltaxol for the semi-synthesis of Taxol. Our previous study showed that both the I368T mutation in LXYL-P1−1 and the T368E mutation in LXYL-P1−2 could increase the enzyme activity, which prompted us to investigate the effect of the I368E mutation on LXYL-P1−1 activity. In this study, the β-xylosidase and β-glucosidase activities of LXYL-P1−1(I368E) were 1.5 and 2.2 times higher than those of LXYL-P1−1. Most importantly, combination of I368E and V91S exerted the cumulative effects on the improvement of the enzyme activities and catalytic efficiency. The β-xylosidase and β-glucosidase activities of the double mutant LXYL-P1−1(V91S/I368E) were 3.2 and 1.7-fold higher than those of LXYL-P1−1(I368E). Similarly, the catalytic efficiency of LXYL-P1−1(V91S/I368E) on 7-β-xylosyl-10-deacetyltaxol was 1.8-fold higher than that of LXYL-P1−1(I368E) due to the dramatic increase in the substrate affinity. Molecular docking results suggest that the V91S and I368E mutation might positively promote the interaction between enzyme and substrate through altering the loop conformation near XDT and increasing the hydrogen bonds among Ser(91), Trp(301), and XDT. This study lays the foundation for exploring the relationship between the structure and function of the β-glycoside hydrolases. MDPI 2018-12-24 /pmc/articles/PMC6337304/ /pubmed/30586935 http://dx.doi.org/10.3390/molecules24010059 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Chen, Jing-Jing Liang, Xiao Chen, Tian-Jiao Yang, Jin-Ling Zhu, Ping Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title | Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title_full | Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title_fullStr | Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title_full_unstemmed | Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title_short | Site-directed Mutagenesis of a β-Glycoside Hydrolase from Lentinula edodes |
title_sort | site-directed mutagenesis of a β-glycoside hydrolase from lentinula edodes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6337304/ https://www.ncbi.nlm.nih.gov/pubmed/30586935 http://dx.doi.org/10.3390/molecules24010059 |
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