Suppression of water as a nucleophile in Candida antarctica lipase B catalysis

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Suppression of water as a nucleophile in Candida antarctica lipase B catalysis. / Larsen, Marianne Wittrup; Zielinska, Dorota F; Martinelle, Mats; Hidalgo, Aurelio; Jensen, Lars Juhl; Bornscheuer, Uwe T; Hult, Karl.

In: ChemBioChem, Vol. 11, No. 6, 12.04.2010, p. 796-801.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Larsen, MW, Zielinska, DF, Martinelle, M, Hidalgo, A, Jensen, LJ, Bornscheuer, UT & Hult, K 2010, 'Suppression of water as a nucleophile in Candida antarctica lipase B catalysis', ChemBioChem, vol. 11, no. 6, pp. 796-801. https://doi.org/10.1002/cbic.200900743

APA

Larsen, M. W., Zielinska, D. F., Martinelle, M., Hidalgo, A., Jensen, L. J., Bornscheuer, U. T., & Hult, K. (2010). Suppression of water as a nucleophile in Candida antarctica lipase B catalysis. ChemBioChem, 11(6), 796-801. https://doi.org/10.1002/cbic.200900743

Vancouver

Larsen MW, Zielinska DF, Martinelle M, Hidalgo A, Jensen LJ, Bornscheuer UT et al. Suppression of water as a nucleophile in Candida antarctica lipase B catalysis. ChemBioChem. 2010 Apr 12;11(6):796-801. https://doi.org/10.1002/cbic.200900743

Author

Larsen, Marianne Wittrup ; Zielinska, Dorota F ; Martinelle, Mats ; Hidalgo, Aurelio ; Jensen, Lars Juhl ; Bornscheuer, Uwe T ; Hult, Karl. / Suppression of water as a nucleophile in Candida antarctica lipase B catalysis. In: ChemBioChem. 2010 ; Vol. 11, No. 6. pp. 796-801.

Bibtex

@article{2cb9974faf4f41ce85930bc374f0b01b,
title = "Suppression of water as a nucleophile in Candida antarctica lipase B catalysis",
abstract = "A water tunnel in Candida antarctica lipase B that provides the active site with substrate water is hypothesized. A small, focused library created in order to prevent water from entering the active site through the tunnel was screened for increased transacylation over hydrolysis activity. A single mutant, S47L, in which the inner part of the tunnel was blocked, catalysed the transacylation of vinyl butyrate to 20 mM butanol 14 times faster than hydrolysis. The single mutant Q46A, which has a more open outer end of the tunnel, showed an increased hydrolysis rate and a decreased hydrolysis to transacylation ratio compared to the wild-type lipase. Mutants with a blocked tunnel could be very useful in applications in which hydrolysis is unwanted, such as the acylation of highly hydrophilic compounds in the presence of water.",
keywords = "Amino Acid Substitution, Binding Sites, Biocatalysis, Candida, Catalytic Domain, Computer Simulation, Hydrolysis, Lipase, Mutagenesis, Site-Directed, Recombinant Proteins, Water",
author = "Larsen, {Marianne Wittrup} and Zielinska, {Dorota F} and Mats Martinelle and Aurelio Hidalgo and Jensen, {Lars Juhl} and Bornscheuer, {Uwe T} and Karl Hult",
year = "2010",
month = apr,
day = "12",
doi = "10.1002/cbic.200900743",
language = "English",
volume = "11",
pages = "796--801",
journal = "ChemBioChem",
issn = "1439-4227",
publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",
number = "6",

}

RIS

TY - JOUR

T1 - Suppression of water as a nucleophile in Candida antarctica lipase B catalysis

AU - Larsen, Marianne Wittrup

AU - Zielinska, Dorota F

AU - Martinelle, Mats

AU - Hidalgo, Aurelio

AU - Jensen, Lars Juhl

AU - Bornscheuer, Uwe T

AU - Hult, Karl

PY - 2010/4/12

Y1 - 2010/4/12

N2 - A water tunnel in Candida antarctica lipase B that provides the active site with substrate water is hypothesized. A small, focused library created in order to prevent water from entering the active site through the tunnel was screened for increased transacylation over hydrolysis activity. A single mutant, S47L, in which the inner part of the tunnel was blocked, catalysed the transacylation of vinyl butyrate to 20 mM butanol 14 times faster than hydrolysis. The single mutant Q46A, which has a more open outer end of the tunnel, showed an increased hydrolysis rate and a decreased hydrolysis to transacylation ratio compared to the wild-type lipase. Mutants with a blocked tunnel could be very useful in applications in which hydrolysis is unwanted, such as the acylation of highly hydrophilic compounds in the presence of water.

AB - A water tunnel in Candida antarctica lipase B that provides the active site with substrate water is hypothesized. A small, focused library created in order to prevent water from entering the active site through the tunnel was screened for increased transacylation over hydrolysis activity. A single mutant, S47L, in which the inner part of the tunnel was blocked, catalysed the transacylation of vinyl butyrate to 20 mM butanol 14 times faster than hydrolysis. The single mutant Q46A, which has a more open outer end of the tunnel, showed an increased hydrolysis rate and a decreased hydrolysis to transacylation ratio compared to the wild-type lipase. Mutants with a blocked tunnel could be very useful in applications in which hydrolysis is unwanted, such as the acylation of highly hydrophilic compounds in the presence of water.

KW - Amino Acid Substitution

KW - Binding Sites

KW - Biocatalysis

KW - Candida

KW - Catalytic Domain

KW - Computer Simulation

KW - Hydrolysis

KW - Lipase

KW - Mutagenesis, Site-Directed

KW - Recombinant Proteins

KW - Water

U2 - 10.1002/cbic.200900743

DO - 10.1002/cbic.200900743

M3 - Journal article

C2 - 20235107

VL - 11

SP - 796

EP - 801

JO - ChemBioChem

JF - ChemBioChem

SN - 1439-4227

IS - 6

ER -

ID: 33748856