Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gelsCitation formats

  • External authors:
  • Kristína Markošová
  • Igor Dolejš
  • Radek Stloukal
  • Leonardo Rios-Solis
  • Michal Rosenberg
  • Martina Micheletti
  • Gary J. Lye
  • Martin Rebroš

Standard

Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels. / Markošová, Kristína; Dolejš, Igor; Stloukal, Radek; Rios-Solis, Leonardo; Rosenberg, Michal; Micheletti, Martina; Lye, Gary J.; Turner, Nicholas; Rebroš, Martin.

In: Journal of Molecular Catalysis - B Enzymatic, Vol. 129, 01.07.2016, p. 69-74.

Research output: Contribution to journalArticle

Harvard

Markošová, K, Dolejš, I, Stloukal, R, Rios-Solis, L, Rosenberg, M, Micheletti, M, Lye, GJ, Turner, N & Rebroš, M 2016, 'Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels', Journal of Molecular Catalysis - B Enzymatic, vol. 129, pp. 69-74. https://doi.org/10.1016/j.molcatb.2016.04.009

APA

Markošová, K., Dolejš, I., Stloukal, R., Rios-Solis, L., Rosenberg, M., Micheletti, M., ... Rebroš, M. (2016). Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels. Journal of Molecular Catalysis - B Enzymatic, 129, 69-74. https://doi.org/10.1016/j.molcatb.2016.04.009

Vancouver

Markošová K, Dolejš I, Stloukal R, Rios-Solis L, Rosenberg M, Micheletti M et al. Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels. Journal of Molecular Catalysis - B Enzymatic. 2016 Jul 1;129:69-74. https://doi.org/10.1016/j.molcatb.2016.04.009

Author

Markošová, Kristína ; Dolejš, Igor ; Stloukal, Radek ; Rios-Solis, Leonardo ; Rosenberg, Michal ; Micheletti, Martina ; Lye, Gary J. ; Turner, Nicholas ; Rebroš, Martin. / Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels. In: Journal of Molecular Catalysis - B Enzymatic. 2016 ; Vol. 129. pp. 69-74.

Bibtex

@article{00e3af88b8c249a0a3afee9a4e503c5b,
title = "Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels",
abstract = "This study focused on the production and immobilisation of the crude enzyme extract of recombinant monoamine oxidase (EC 1.4.3.4), originating from Aspergillus Niger (MAO-N-D5) and expressed in Escherichia coli, in PVA gel using the LentiKats{\circledR} technique. MAO-Ns are important enzymes in the chemical industry due to their stereoselectivity and they are often used for the deracemisation of non-optically pure mixtures of amines. Biomass production, enzyme preparation, enzyme immobilisation, process parameters for the immobilised enzyme and characterisation of the enzyme are described in detail here. The biomass was prepared in laboratory bioreactors, and two different disruption techniques were compared. The activity of the enzyme was determined by biotransformation with secondary amine 3-azabicyclo [3,3,0] octane as a substrate. The crude enzyme extract showed 61.5{\%} of the whole cell activity and the immobilised enzyme showed a wider optimum pH and temperature ranges than the free enzyme. The initial specific activity of the immobilised monoamine oxidase crude enzyme extract remained at 80{\%} after 12 repeated biotransformations. For the first time, the full kinetic parameters of an immobilised MAO-N-D5 were obtained based on a ping-pong bi-bi reaction mechanism. The specific activity was 0.29 U g-1 (Lentikats) and the Km was 7.31 mM, which were similar in comparison to whole cell MAO-N-D5. Characterisation of immobilised MAO-N-D5 showed particular benefits in terms of activity and stability in comparison with free and whole cell MAO-N-D5; therefore, the immobilisation of this enzyme is very suitable for industrial applications.",
keywords = "Immobilisation, Kinetic parameters, Monoamine oxidase, Polyvinyl alcohol",
author = "Krist{\'i}na Markošov{\'a} and Igor Dolejš and Radek Stloukal and Leonardo Rios-Solis and Michal Rosenberg and Martina Micheletti and Lye, {Gary J.} and Nicholas Turner and Martin Rebroš",
year = "2016",
month = "7",
day = "1",
doi = "10.1016/j.molcatb.2016.04.009",
language = "English",
volume = "129",
pages = "69--74",
journal = "Journal of Molecular Catalysis B: Enzymatic",
issn = "1381-1177",
publisher = "Elsevier BV",

}

RIS

TY - JOUR

T1 - Immobilisation and kinetics of monoamine oxidase (MAO-N-D5) enzyme in polyvinyl alcohol gels

AU - Markošová, Kristína

AU - Dolejš, Igor

AU - Stloukal, Radek

AU - Rios-Solis, Leonardo

AU - Rosenberg, Michal

AU - Micheletti, Martina

AU - Lye, Gary J.

AU - Turner, Nicholas

AU - Rebroš, Martin

PY - 2016/7/1

Y1 - 2016/7/1

N2 - This study focused on the production and immobilisation of the crude enzyme extract of recombinant monoamine oxidase (EC 1.4.3.4), originating from Aspergillus Niger (MAO-N-D5) and expressed in Escherichia coli, in PVA gel using the LentiKats® technique. MAO-Ns are important enzymes in the chemical industry due to their stereoselectivity and they are often used for the deracemisation of non-optically pure mixtures of amines. Biomass production, enzyme preparation, enzyme immobilisation, process parameters for the immobilised enzyme and characterisation of the enzyme are described in detail here. The biomass was prepared in laboratory bioreactors, and two different disruption techniques were compared. The activity of the enzyme was determined by biotransformation with secondary amine 3-azabicyclo [3,3,0] octane as a substrate. The crude enzyme extract showed 61.5% of the whole cell activity and the immobilised enzyme showed a wider optimum pH and temperature ranges than the free enzyme. The initial specific activity of the immobilised monoamine oxidase crude enzyme extract remained at 80% after 12 repeated biotransformations. For the first time, the full kinetic parameters of an immobilised MAO-N-D5 were obtained based on a ping-pong bi-bi reaction mechanism. The specific activity was 0.29 U g-1 (Lentikats) and the Km was 7.31 mM, which were similar in comparison to whole cell MAO-N-D5. Characterisation of immobilised MAO-N-D5 showed particular benefits in terms of activity and stability in comparison with free and whole cell MAO-N-D5; therefore, the immobilisation of this enzyme is very suitable for industrial applications.

AB - This study focused on the production and immobilisation of the crude enzyme extract of recombinant monoamine oxidase (EC 1.4.3.4), originating from Aspergillus Niger (MAO-N-D5) and expressed in Escherichia coli, in PVA gel using the LentiKats® technique. MAO-Ns are important enzymes in the chemical industry due to their stereoselectivity and they are often used for the deracemisation of non-optically pure mixtures of amines. Biomass production, enzyme preparation, enzyme immobilisation, process parameters for the immobilised enzyme and characterisation of the enzyme are described in detail here. The biomass was prepared in laboratory bioreactors, and two different disruption techniques were compared. The activity of the enzyme was determined by biotransformation with secondary amine 3-azabicyclo [3,3,0] octane as a substrate. The crude enzyme extract showed 61.5% of the whole cell activity and the immobilised enzyme showed a wider optimum pH and temperature ranges than the free enzyme. The initial specific activity of the immobilised monoamine oxidase crude enzyme extract remained at 80% after 12 repeated biotransformations. For the first time, the full kinetic parameters of an immobilised MAO-N-D5 were obtained based on a ping-pong bi-bi reaction mechanism. The specific activity was 0.29 U g-1 (Lentikats) and the Km was 7.31 mM, which were similar in comparison to whole cell MAO-N-D5. Characterisation of immobilised MAO-N-D5 showed particular benefits in terms of activity and stability in comparison with free and whole cell MAO-N-D5; therefore, the immobilisation of this enzyme is very suitable for industrial applications.

KW - Immobilisation

KW - Kinetic parameters

KW - Monoamine oxidase

KW - Polyvinyl alcohol

UR - http://www.scopus.com/inward/record.url?scp=84964598902&partnerID=8YFLogxK

U2 - 10.1016/j.molcatb.2016.04.009

DO - 10.1016/j.molcatb.2016.04.009

M3 - Article

VL - 129

SP - 69

EP - 74

JO - Journal of Molecular Catalysis B: Enzymatic

JF - Journal of Molecular Catalysis B: Enzymatic

SN - 1381-1177

ER -