Microscopic Monitoring Provides Information on Structure and
Properties During Biocatalyst Immobilisation.

J 2014

Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation.

BIDMANOVÁ, Šárka, Eva HRDLIČKOVÁ, Josef JAROŠ, Ladislav ILKOVICS, Aleš HAMPL et. al.

Basic information

Original name

Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation.

Authors

BIDMANOVÁ, Šárka (203 Czech Republic, belonging to the institution), Eva HRDLIČKOVÁ (203 Czech Republic), Josef JAROŠ (203 Czech Republic, belonging to the institution), Ladislav ILKOVICS (203 Czech Republic, belonging to the institution), Aleš HAMPL (203 Czech Republic, belonging to the institution), Jiří DAMBORSKÝ (203 Czech Republic, belonging to the institution) and Zbyněk PROKOP (203 Czech Republic, guarantor, belonging to the institution)

Edition

Biotechnology Journal, WEINHEIM, GERMANY, WILEY-V C H VERLAG GMBH, 2014, 1860-6768

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 3.490

RIV identification code

RIV/00216224:14310/14:00074208

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1002/biot.201300049

UT WoS

000337700000015

Keywords in English

Cross-linked enzyme aggregates; Immobilization; Microscopy; Polyvinyl alcohol; Structure

Abstract

V originále

Enzymes have a wide range of applications in different industries owing to their high specificity and efficiency. Immobilisation is often used to improve biocatalyst properties, operational stability and reusability. However, changes in the structure of biocatalysts during immobilisation and under process conditions are still largely uncertain. Here, three microscopy techniques – bright-field, confocal and electron microscopy – were applied to determine the distribution and structure of an immobilised biocatalyst. Free enzyme (haloalkane dehalogenase), cross-linked enzyme aggregates (CLEAs) and CLEAs entrapped in polyvinyl alcohol lenses (lentikats) were used as model systems. Electron microscopy revealed that sonicated CLEAs underwent morphological changes that strongly correlated with increased catalytic activity compared to less structured, non-treated CLEAs. Confocal microscopy confirmed that loading of the biocatalyst was not the only factor affecting the catalytic activity of the lentikats. Confocal microscopy also showed a significant reduction in the pore size of lentikats exposed to 25% tetrahydrofuran and 50% dioxane. Narrow pores appeared to provide protection to CLEAs from the detrimental action of cosolvents, which significantly correlated with higher activity of CLEAs compared to free enzyme. The results showed that microscopy can provide valuable information about the structure and properties of a biocatalyst during immobilisation and under process conditions.

Links

EE2.3.20.0183, research and development project

Name: Centrum experimentální biomedicíny

GAP207/12/0775, research and development project

Name: Strukturně-funkční vztahy haloalkan dehalogenas

Investor: Czech Science Foundation

LO1214, research and development project

Name: Centrum pro výzkum toxických látek v prostředí (Acronym: RECETOX)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

BIDMANOVÁ, Šárka, Eva HRDLIČKOVÁ, Josef JAROŠ, Ladislav ILKOVICS, Aleš HAMPL, Jiří DAMBORSKÝ and Zbyněk PROKOP. Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation. Biotechnology Journal. WEINHEIM, GERMANY: WILEY-V C H VERLAG GMBH, 2014, vol. 9, No 6, p. 852-860. ISSN 1860-6768. Available from: https://dx.doi.org/10.1002/biot.201300049.

@article{1213465,
   author = {Bidmanová, Šárka and Hrdličková, Eva and Jaroš, Josef and Ilkovics, Ladislav and Hampl, Aleš and Damborský, Jiří and Prokop, Zbyněk},
   article_location = {WEINHEIM, GERMANY},
   article_number = {6},
   doi = {http://dx.doi.org/10.1002/biot.201300049},
   keywords = {Cross-linked enzyme aggregates; Immobilization; Microscopy; Polyvinyl alcohol; Structure},
   language = {eng},
   issn = {1860-6768},
   journal = {Biotechnology Journal},
   title = {Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation.},
   volume = {9},
   year = {2014}
}

TY  - JOUR
ID  - 1213465
AU  - Bidmanová, Šárka - Hrdličková, Eva - Jaroš, Josef - Ilkovics, Ladislav - Hampl, Aleš - Damborský, Jiří - Prokop, Zbyněk
PY  - 2014
TI  - Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation.
JF  - Biotechnology Journal
VL  - 9
IS  - 6
SP  - 852-860
EP  - 852-860
PB  - WILEY-V C H VERLAG GMBH
SN  - 18606768
KW  - Cross-linked enzyme aggregates
KW  - Immobilization
KW  - Microscopy
KW  - Polyvinyl alcohol
KW  - Structure
N2  - Enzymes have a wide range of applications in different industries owing to their high specificity and efficiency. Immobilisation is often used to improve biocatalyst properties, operational stability and reusability. However, changes in the structure of biocatalysts during immobilisation and under process conditions are still largely uncertain. Here, three microscopy techniques – bright-field, confocal and electron microscopy – were applied to determine the distribution and structure of an immobilised biocatalyst. Free enzyme (haloalkane dehalogenase), cross-linked enzyme aggregates (CLEAs) and CLEAs entrapped in polyvinyl alcohol lenses (lentikats) were used as model systems. Electron microscopy revealed that sonicated CLEAs underwent morphological changes that strongly correlated with increased catalytic activity compared to less structured, non-treated CLEAs. Confocal microscopy confirmed that loading of the biocatalyst was not the only factor affecting the catalytic activity of the lentikats. Confocal microscopy also showed a significant reduction in the pore size of lentikats exposed to 25% tetrahydrofuran and 50% dioxane. Narrow pores appeared to provide protection to CLEAs from the detrimental action of cosolvents, which significantly correlated with higher activity of CLEAs compared to free enzyme. The results showed that microscopy can provide valuable information about the structure and properties of a biocatalyst during immobilisation and under process conditions.
ER  -

BIDMANOVÁ, Šárka, Eva HRDLIČKOVÁ, Josef JAROŠ, Ladislav ILKOVICS, Aleš HAMPL, Jiří DAMBORSKÝ and Zbyněk PROKOP. Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation. \textit{Biotechnology Journal}. WEINHEIM, GERMANY: WILEY-V C H VERLAG GMBH, 2014, vol.~9, No~6, p.~852-860. ISSN~1860-6768. Available from: https://dx.doi.org/10.1002/biot.201300049.

Detailed Information on Publication Record