A Novel Culture Method for High Level Production of Heterologous Protein in Saccharomyces cerevisiae

Tadashi Nagashima, Yutaka Yamamoto, Katsuya Gomi, Katsuhiko Kitamoto, Chieko Kumagai

研究成果: Article査読

7 被引用数 (Scopus)

抄録

A high level production system for heterologous protein by cold culture of yeast transformants at 15°C was developed. The yeast transformants, carrying a plasmid containing cDNA for Aspergillus oryzae α-amylase (Taka-amylase A) or human lysozyme synthetic DNA, were cultivated in a selective medium for 1 or 2 days until full growth at 30°C. The yeast cells were harvested by centrifugation from the culture fluid and then were transferred to YPD medium. These inoculated broths were incubated for 2 days at 15°C and then for another 2 days at 30°C. By the cold culture method described above, higher amounts of Taka-amylase A (28.6 mg/liter) and human lysozyme (6.1 mg/liter) were produced by the yeast transformants compared to those by conventional methods. Heterologous protein productions using YEp, YCp, and Yip types of yeast expression vectors with AD HI or GAPDH promoter by the cold culture method showed effective productivity of about 2-fold compared to those by the conventional method of culture at 30°C. The high level production of heterologous protein by this method was not specific to the S. cerevisiae strains examined.

本文言語English
ページ(範囲)1292-1296
ページ数5
ジャーナルBioscience, Biotechnology, and Biochemistry
58
7
DOI
出版ステータスPublished - 1994 1 1
外部発表はい

ASJC Scopus subject areas

  • バイオテクノロジー
  • 分析化学
  • 生化学
  • 応用微生物学とバイオテクノロジー
  • 分子生物学
  • 有機化学

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