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.

Základní údaje

Originální název

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

Autoři

BIDMANOVÁ, Šárka (203 Česká republika, domácí), Eva HRDLIČKOVÁ (203 Česká republika), Josef JAROŠ (203 Česká republika, domácí), Ladislav ILKOVICS (203 Česká republika, domácí), Aleš HAMPL (203 Česká republika, domácí), Jiří DAMBORSKÝ (203 Česká republika, domácí) a Zbyněk PROKOP (203 Česká republika, garant, domácí)

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Německo

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 3.490

Kód RIV

RIV/00216224:14310/14:00074208

Organizační jednotka

Přírodovědecká fakulta

DOI

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

UT WoS

000337700000015

Klíčová slova anglicky

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

Štítky

AKR, EL OK, podil, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 21. 3. 2017 08:05, prof. Mgr. Jiří Damborský, Dr.

Anotace

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.

Návaznosti

EE2.3.20.0183, projekt VaV

Název: Centrum experimentální biomedicíny

GAP207/12/0775, projekt VaV

Název: Strukturně-funkční vztahy haloalkan dehalogenas

Investor: Grantová agentura ČR, Structure-functional Relationships of Haloalkane Dehalogenases

LO1214, projekt VaV

Název: Centrum pro výzkum toxických látek v prostředí (Akronym: RECETOX)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Centrum pro výzkum toxických látek v prostředí

Citovat

BIDMANOVÁ, Šárka, Eva HRDLIČKOVÁ, Josef JAROŠ, Ladislav ILKOVICS, Aleš HAMPL, Jiří DAMBORSKÝ a Zbyněk PROKOP. Microscopic Monitoring Provides Information on Structure and Properties During Biocatalyst Immobilisation. Biotechnology Journal. WEINHEIM, GERMANY: WILEY-V C H VERLAG GMBH, 2014, roč. 9, č. 6, s. 852-860. ISSN 1860-6768. Dostupné z: 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 a 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, roč.~9, č.~6, s.~852-860. ISSN~1860-6768. Dostupné z: https://dx.doi.org/10.1002/biot.201300049.

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