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The hydrolysis of new lipid-like substrates and trilaurin in monolayers catalyzed with the lipase fromPseudomonas fluorescens

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Abstract

A simple method for determining the enzymic hydrolysis parameters of lipid-like substrates and trilaurin assembled in monolayers at the water-air interface was suggested. At a surface pressure of 10 mN/m, the initial rates of lipolysis were found to be proportional to the decrease in area of the substrate monolayer caused by the enzymic hydrolysis in a single-compartment Langmuir balance. The kinetic parameters for the hydrolysis of trilaurin and three 1,3-dilaurylpseudoglycerides acetylated in position 2 with an amino acid (phenylalanine, leucine, or valine) catalyzed with lipase fromPseudomonas fluorescens were determined. Unlike models of enzymic hydrolysis that neglect the thickness of the substrate monolayer, our method allows the determination of kinetic parameters in standard dimensions. The values ofk cat for the synthetic pseudoglycerides were found to be significantly higher than that for trilaurin, while the values ofK m(app) were close. This may be due to the presence of positively charged primary amino groups in the molecules of pseudoglycerides.

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Abbreviations

A:

specific area (i.e., the area occupied by a molecule in monolayer)

DMAP:

N,N-dimethyl-4-aminopyridine

PG:

pseudoglyceride

Leu-PG:

1,3-O-dilauryl-2-O-leucylglycerol

Phe-PG:

1,3-O-dilauryl-2-O-phenylalanylglycerol

Val-PG:

1,3-O-dilauryl-2-O-valylglycerol

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, 117871, GSP-7 Moscow, Russia

    S. Yu. Zaitsev, T. A. Volchenkova, S. V. Belov & A. E. Ivanov

  2. Bergische Universität Gesamthochschule, Gauss-Strasse 20, 42097, Wuppertal, Germany

    B. Aha & M. P. Schneider

Authors
  1. S. Yu. Zaitsev
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  2. B. Aha
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  3. T. A. Volchenkova
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  4. S. V. Belov
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  5. M. P. Schneider
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  6. A. E. Ivanov
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Correspondence to A. E. Ivanov.

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Zaitsev, S.Y., Aha, B., Volchenkova, T.A. et al. The hydrolysis of new lipid-like substrates and trilaurin in monolayers catalyzed with the lipase fromPseudomonas fluorescens . Russ J Bioorg Chem 26, 201–206 (2000). https://doi.org/10.1007/BF02786346

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  • DOI: https://doi.org/10.1007/BF02786346

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