Number of the records: 1

Engineering the cytokinin-glucoside specificity of the maize beta-D-glucosidase Zm-p60.1 using site-directed random mutagenesis

SYSNO ASEP	0440809
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Engineering the cytokinin-glucoside specificity of the maize beta-D-glucosidase Zm-p60.1 using site-directed random mutagenesis
Author(s)	Filipi, T. (CZ) Mazura, P. (CZ) Janda, L. (CZ) Kiran, N.S. (CZ) Brzobohatý, Břetislav (BFU-R)_{RID, ORCID}
Number of authors	5
Source Title	Phytochemistry. - : Elsevier - ISSN 0031-9422 Roč. 74, FEB2012 (2012), s. 40-48
Number of pages	9 s.
Publication form	Print - P
Language	eng - English
Country	GB - United Kingdom
Keywords	beta-Glucosidase ; cis-Zeatin-O-beta-D-glucopyranoside ; Cytokinin metabolism
Subject RIV	BO - Biophysics
Institutional support	BFU-R - RVO:68081707
UT WOS	000300815700004
DOI	10.1016/j.phytochem.2011.10.008
Annotation	The maize beta-D-glucosidase Zm-p60.1 releases active cytokinins from their storage/transport forms, and its over-expression in tobacco disrupts zeatin metabolism. The role of the active-site microenvironment in fine-tuning Zm-p60.1 substrate specificity has been explored, particularly in the W373K mutant, using site-directed random mutagenesis to investigate the influence of amino acid changes around the 373 position. Two triple (P372T/W373K/M376L and P372S/W373K/M376L) and three double mutants (P372T/W373K, P372S/W373K and W373K/M376L) were prepared. Their catalytic parameters with two artificial substrates show tight interdependence between substrate catalysis and protein structure. P372T/W373K/M376L exhibited the most significant effect on natural substrate specificity: the ratio of hydrolysis of cis-zeatin-O-beta-D-glucopyranoside versus the trans-zeatin-O-beta-D-glucopyranoside shifted from 1.3 in wild-type to 9.4 in favor of the cis- isomer.
Workplace	Institute of Biophysics
Contact	Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244
Year of Publishing	2015

Number of the records: 1