DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1:
DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM

D 2010

DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM

FILIPI, T., P. MAZURA, Radka DOPITOVÁ, Lubomír JANDA, Jiří DAMBORSKÝ et. al.

Základní údaje

Originální název

DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM

Autoři

FILIPI, T., P. MAZURA, Radka DOPITOVÁ, Lubomír JANDA, Jiří DAMBORSKÝ, N. S. KIRAN a B. BRZOBOHATY

Vydání

BRNO, MENDELNET 2010, od s. 821-823, 3 s. 2010

Nakladatel

MENDEL UNIV BRNO, FAC AGRONOMY

Další údaje

Jazyk

čeština

Typ výsledku

Stať ve sborníku

Obor

40101 Agriculture

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Forma vydání

tištěná verze "print"

Odkazy

URL

Organizační jednotka

Přírodovědecká fakulta

ISBN

978-80-7375-453-2

UT WoS

000341786800106

Klíčová slova anglicky

Zm-p60.1; beta-glucosidase; maize; tZOG; cZOG; cytokinin; protein evolution; protein mutagenesis

Změněno: 10. 9. 2020 11:08, Mgr. Marie Šípková, DiS.

Anotace

V originále

Maize beta-glucosidase Zm-p60.1 is one of many enzymes which are important for plant development. It liberates free zeatin from its transport and/or storage form zeatin-O-glucoside. Using an adapted site specific non-saturated random mutagenesis approach, it were prepared five multi-site mutants surrounding the active site (W373K/M376L, W373K/P372S/M376L, W373K/P372T/M376L, W373K/P372S and W373K/P372T) derived from the single mutant W373K to study the effect(s) of amino-acid changes on substrate specificity towards natural (trans-zeatin-O-beta-D-glucopyranoside and cis-zeatin-O-beta-D-glucopyranoside) and artificial (4-nitorophenyl-O-beta-D-glucopyranoside and 4-methylumbellyferyl O-beta-D-glucopyranoside) substrates. Kinetic and substrate specificity studies confirmed large differences among set of mutated enzymes. All enzymes surprisingly preferred cis-zeatin-O-beta-D-glucopyranoside over trans-zeatin-O-beta-D-glucopyranoside, whereas differences in hydrolytic efficiencies are considerable. Quantitative TLC confirmed the best cZOG/tZOG hydrolysis ratio toward cis-zeatin-O-beta-D-glucopyranoside in the triple mutant W373K/P372T/M376L. Moreover, it was also confirmed that only wild-type hydrolyzed trans-zeatin-N9-beta-D-glucopyranoside. No known plant beta-glucosidase hydrolyzes this substrate. Hydrolysis of trans-zeatin-N7-beta-D-glucopyranoside was not observed at all.

Návaznosti

LC06034, projekt VaV

Název: Regulace morfogeneze rostlinných buněk a orgánů

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Regulace morfogeneze rostlinných buněk a orgánů

Citovat

FILIPI, T., P. MAZURA, Radka DOPITOVÁ, Lubomír JANDA, Jiří DAMBORSKÝ, N. S. KIRAN a B. BRZOBOHATY. DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM. In Skarpa, P. MENDELNET 2010. BRNO: MENDEL UNIV BRNO, FAC AGRONOMY, 2010, s. 821-823. ISBN 978-80-7375-453-2.

@inproceedings{1678121,
   author = {Filipi, T. and Mazura, P. and Dopitová, Radka and Janda, Lubomír and Damborský, Jiří and Kiran, N. S. and Brzobohaty, B.},
   address = {BRNO},
   booktitle = {MENDELNET 2010},
   editor = {Skarpa, P.},
   keywords = {Zm-p60.1; beta-glucosidase; maize; tZOG; cZOG; cytokinin; protein evolution; protein mutagenesis},
   howpublished = {tištěná verze "print"},
   language = {cze},
   location = {BRNO},
   isbn = {978-80-7375-453-2},
   pages = {821-823},
   publisher = {MENDEL UNIV BRNO, FAC AGRONOMY},
   title = {DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM},
   url = {https://mnet.mendelu.cz/mendelnet2010/articles/19_filipi_284.pdf},
   year = {2010}
}

TY  - JOUR
ID  - 1678121
AU  - Filipi, T. - Mazura, P. - Dopitová, Radka - Janda, Lubomír - Damborský, Jiří - Kiran, N. S. - Brzobohaty, B.
PY  - 2010
TI  - DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM
PB  - MENDEL UNIV BRNO, FAC AGRONOMY
CY  - BRNO
SN  - 9788073754532
KW  - Zm-p60.1
KW  - beta-glucosidase
KW  - maize
KW  - tZOG
KW  - cZOG
KW  - cytokinin
KW  - protein evolution
KW  - protein mutagenesis
UR  - https://mnet.mendelu.cz/mendelnet2010/articles/19_filipi_284.pdf
N2  - Maize beta-glucosidase Zm-p60.1 is one of many enzymes which are important for plant development. It liberates free zeatin from its transport and/or storage form zeatin-O-glucoside. Using an adapted site specific non-saturated random mutagenesis approach, it were prepared five multi-site mutants surrounding the active site (W373K/M376L, W373K/P372S/M376L, W373K/P372T/M376L, W373K/P372S and W373K/P372T) derived from the single mutant W373K to study the effect(s) of amino-acid changes on substrate specificity towards natural (trans-zeatin-O-beta-D-glucopyranoside and cis-zeatin-O-beta-D-glucopyranoside) and artificial (4-nitorophenyl-O-beta-D-glucopyranoside and 4-methylumbellyferyl O-beta-D-glucopyranoside) substrates. Kinetic and substrate specificity studies confirmed large differences among set of mutated enzymes. All enzymes surprisingly preferred cis-zeatin-O-beta-D-glucopyranoside over trans-zeatin-O-beta-D-glucopyranoside, whereas differences in hydrolytic efficiencies are considerable. Quantitative TLC confirmed the best cZOG/tZOG hydrolysis ratio toward cis-zeatin-O-beta-D-glucopyranoside in the triple mutant W373K/P372T/M376L. Moreover, it was also confirmed that only wild-type hydrolyzed trans-zeatin-N9-beta-D-glucopyranoside. No known plant beta-glucosidase hydrolyzes this substrate. Hydrolysis of trans-zeatin-N7-beta-D-glucopyranoside was not observed at all.
ER  -

FILIPI, T., P. MAZURA, Radka DOPITOVÁ, Lubomír JANDA, Jiří DAMBORSKÝ, N. S. KIRAN a B. BRZOBOHATY. DESIGNING RECOMBINANT MAIZE beta-GLUCOSIDASE Zm-p60.1: DEVELOPMENT OF NOVEL ENZYMES MODULATING CYTOKININ METABOLISM. In Skarpa, P. \textit{MENDELNET 2010}. BRNO: MENDEL UNIV BRNO, FAC AGRONOMY, 2010, s.~821-823. ISBN~978-80-7375-453-2.

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