Bioscience, Biotechnology, and Biochemistry
Online ISSN : 1347-6947
Print ISSN : 0916-8451
Microbiology & Fermentation Technology Notes
Construction of Protease-deficient Candida boidinii Strains Useful for Recombinant Protein Production: Cloning and Disruption of Proteinase A Gene (PEP4) and Proteinase B Gene…
Toshihiro KOMEDAYasuyoshi SAKAINobuo KATOKeiji KONDO
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JOURNAL FREE ACCESS

2002 Volume 66 Issue 3 Pages 628-631

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Abstract

  The yeast Candida boidinii PEP4 and PRB1 genes, encoding proteinase A (PrA) and proteinase B (PrB), respectively, have been cloned and their primary structures were analyzed. The open reading frames of the PEP4 gene (1263 bp encoding a protein of 420 amino acids) and the PRB1 gene (1683 bp encoding a protein of 560 amino acids) were found. The deduced amino acid sequences of PrA and PrB are very similar to Saccharomyces cerevisiae PrA and PrB (64% and 61% identities, respectively). Both PEP4 and PRB1 genes were disrupted in the C. boidinii genome by one-step gene disruption. The resultant pep4Δ and the pep4Δ prb1Δ strains lost protease activity when compared with the wild-type original strain. The constructed C. boidinii strains are expected to be useful hosts for heterologous protein production.

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© 2002 by Japan Society for Bioscience, Biotechnology, and Agrochemistry
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