An automated method to evaluate the enzyme kinetics ofglucosidases

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F17%3A00506682" target="_blank" >RIV/68081707:_____/17:00506682 - isvavai.cz</a>

Alternative codes found

RIV/62156489:43210/17:43912527

Result on the web

<a href="https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/pro.3078" target="_blank" >https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/pro.3078</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/pro.3078" target="_blank" >10.1002/pro.3078</a>

Result language

angličtina

Original language name

An automated method to evaluate the enzyme kinetics ofglucosidases

Original language description

Enzyme kinetic measurements are important for the characterization and engineering of biocatalysts, with applications in a wide range of research fields. The measurement of initial reaction velocity is usually slow and laborious, which motivated us to explore the possibilities for automating this process. Our model enzyme is the maizeglucosidase Zm-p60.1. Zm-p60.1 plays a significant role in plant growth and development by regulating levels of the active plant hormone cytokinin. Zm-p60.1 belongs to a wide group of hydrolytic enzymes. Members of this group hydrolyze several different types of glucosides, releasing glucose as a secondary product. Enzyme kinetic measurements using artificial substrates are well established, but burdensome and time-consuming. Thus, they are a suitable target for process automation. Simple optical methods for enzyme kinetic measurements using natural substrates are often impossible given the optical properties of the enzymatic reaction products. However, we have developed an automated method based on glucose detection, as glucose is released from all substrates of glucosidase reactions. The presented method can obtain 24 data points from up to 15 substrate concentrations to precisely describe the enzyme kinetics. The combination of an automated liquid handling process with assays that have been optimized for measuring the initial hydrolysis velocity ofglucosidases yields two distinct methods that are faster, cheaper, and more accurate than the established protocols.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

10608 - Biochemistry and molecular biology

Project

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Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Protein Science

ISSN

0961-8368

e-ISSN

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Volume of the periodical

26

Issue of the periodical within the volume

2

Country of publishing house

US - UNITED STATES

Number of pages

7

Pages from-to

382-388

UT code for WoS article

000393960300021

EID of the result in the Scopus database

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