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Rhizomucor miehei triglyceride lipase is synthesized as a precursor

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Lipids

Abstract

ARhizomucor miehie cDNA library constructed inEscherichia coli was screened with synthetic oligonucleotides designed from knowledge of a partial amino acid sequence of the secreted triglyceride lipase (triacyl-glycerol acylhydrolase EC 3.1.1.3) from this fungus. Lipase-specific recombinants were isolated and their insert sequenced. Unlike characterized bacterial and mammalian triglyceride lipases, the fungal enzyme is synthesized as a precursor, including a 70 amino acid residue propeptide between the 24 amino acid residues of the signal peptide and the 269 residues of the mature enzyme. The precursor processing mechanism, which involves cleavage between a methionine and a serine residue, is unknown. By sequence comparison with other lipases, a serine residue involved in substrate binding was identified in the fungal lipase. The sequence around this residue is well-conserved among characterized lipases. Conservation of an intron in an isolated cDNA recombinant and immunoprecipitation of in vitro synthesizedR. miehei translation products indicates that the expression of the lipase gene might involve inefficient mRNA splicing.

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Abbreviations

RML:

Rhizomucor miehei lipase

SDS:

sodium dodecylsulphate

bp:

base pair

HIC:

hydrophobic interaction chromatography

HPLC:

high performance liquid chromatography

kb:

kilobase

LU:

lipase units.

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Authors and Affiliations

  1. Novo Research Institute, Novo Allé, DK-2880, Bagsvaerd-Copenhagen, Denmark

    Esper Boel, Birgitte Huge-Jensen, Mogens Christensen, Lars Thim & Niels P. Fiil

Authors
  1. Esper Boel
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  2. Birgitte Huge-Jensen
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  3. Mogens Christensen
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  4. Lars Thim
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  5. Niels P. Fiil
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Boel, E., Huge-Jensen, B., Christensen, M. et al. Rhizomucor miehei triglyceride lipase is synthesized as a precursor. Lipids 23, 701–706 (1988). https://doi.org/10.1007/BF02535672

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  • DOI: https://doi.org/10.1007/BF02535672

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