High level secretion of calf chymosin using a glucoamylase-prochymosin fusion gene in aspergillus oryzae

Kozo Tsuchiya, Tadashi Nagashima, Yutaka Yamamoto, Katsuya Gomi, Katsuhiko Kitamoto, Chieko Kumagai, Gakuzo Tamura

Research output: Contribution to journalArticle

Abstract

A recombinant chymosin was secreted at high levels using fusion genes with A. oryzae glucoamylase gene (glaA) and a wheat bran solid-state culture system. Two portions of the A. oryzae glucoamylase, one with almost the entire glucoamylase (GA1-603) lacking 9 amino acids at the carboxyl terminal, and the other (GA1-511) lacking the starch binding-domain, were fused in frame with prochymosin cDNA. Western blot analysis indicated that the mature chymosin was released from the secreted fusion protein by autocatalytic processing. The transformant harboring the GA1-511-prochymosin construct showed about 5-fold chymosin production of the transformant in which the chymosin gene was directly expressed under the control of the glaA promoter in submerged culture. Moreover, wheat bran solid-state culture gave about 500-fold higher yield of the chymosin (approximately 150 mg/kg wheat bran) compared with the submerged culture.

Original languageEnglish
Pages (from-to)895-899
Number of pages5
JournalBioscience, Biotechnology and Biochemistry
Volume58
Issue number5
DOIs
Publication statusPublished - 1994 Jan 1
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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