k 2006

High-throughput characterization of enzymes from genomic and proteomic projects-multivariate statistical approach

MONINCOVÁ, Marta, Andrea FOŘTOVÁ, Radka CHALOUPKOVÁ, Yukari SATO, Zbyněk PROKOP et. al.

Basic information

Original name

High-throughput characterization of enzymes from genomic and proteomic projects-multivariate statistical approach

Name in Czech

Rychlá charakteristika enzymů z genomických a proteomických projektů-využití vícerozměrné statistiky

Authors

MONINCOVÁ, Marta (203 Czech Republic, belonging to the institution), Andrea FOŘTOVÁ (203 Czech Republic, belonging to the institution), Radka CHALOUPKOVÁ (203 Czech Republic, belonging to the institution), Yukari SATO (392 Japan), Zbyněk PROKOP (203 Czech Republic, belonging to the institution) and Jiří DAMBORSKÝ (203 Czech Republic, guarantor, belonging to the institution)

Edition

Materials Structure in Chemistry, Biology, Physics and Technology, 2006

Other information

Language

English

Type of outcome

Prezentace na konferencích

Field of Study

10600 1.6 Biological sciences

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

RIV identification code

RIV/00216224:14310/06:00017056

Organization unit

Faculty of Science

Keywords in English

haloalkane dehalogenase; substrate specificity; screening; multivariate statistics
Změněno: 29/4/2011 10:54, prof. Mgr. Jiří Damborský, Dr.

Abstract

V originále

Hight throughput genomic and proteomic methods release a lot of novel enzymes every year which require systematic characterization and cataloguing of their properties. Here, we illustrate a novel approach, using multivariate statistics to characterize and describe a protein family with broad substrate specificity. Principle of the approach consists in multivariate statistic method, principle component analysis [1], which enabled firstly to choose sufficient set of 30 substrates from 194 halogenated compounds respecting maximum variability in physical-chemical properties [2]. Quick and reliable enzymatic assay follows the selection and produces an activity data of particular proteins with selected substrates. Third step is application of principal component analysis on enzyme activity data matrix to describe the difference in substrate specifities. In paralell, kinetic constants Km and kcat were measured. The obtained dataset, completed with phycical-chemical properties of halogenated hydrocarbons tested, was submitted to quantitave structure-activity relationship. The concept have been verified on screening of haloalkane dehalogenases. Members of this enzyme family cleave carbon-halogen bond in halogenated hydrocarbons. Haloalkane dehalogenases can be used in bioremediation, as industrial biocatalysts or as biosensors. To increase their applicability, the outcome from principle component and phylogenetic analysis can be combine to look for novel enzymes with promising biotechnological properties. Quantitave structure-activity relationship brought detail view into explanation of activity or inactivity of particular substrates tested. Such information is helpful in process of rational design and improvement of haloalkane dehalogenases planned to be applied in biotechnology. The methodology described here and applied on haloalkane dehalogenase family has potential to extend systematic knowledge of particular enzymes and protein families saving time, money and experimental capacity without loosing information. References: 1. S. Wold, K. Esbensen & P. Geladi, Chemometrics and Intelligent Laboratory Systems, 2 (1987) 37-52. 2. S. Marvanova, Y. Nagata, M. Wimmerova, J. Sykorova, K. Hynkova & J. Damborsky, Journal of Microbiological Methods, 44 (2001) 149-157.

In Czech

Prezentace Mgr. Monincové na konferenci Materials Structure in Chemistry, Biology, Physics and Technology s názvem "Rychlá charakteristika enzymů z genomických a proteomických projektů-využití vícerozměrné statistiky".

Links

MSM0021622412, plan (intention)
Name: Interakce mezi chemickými látkami, prostředím a biologickými systémy a jejich důsledky na globální, regionální a lokální úrovni (INCHEMBIOL) (Acronym: INCHEMBIOL)
Investor: Ministry of Education, Youth and Sports of the CR, Interactions among the chemicals, environment and biological systems and their consequences on the global, regional and local scales (INCHEMBIOL)