Overexpression of thermoalkalophilic lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae

Jung Oh Ahn, Hyung Wook Jang, Hong Weon Lee, Eui Sung Choi, Seungjoo Haam, Tae Kwang Oh, Joon Ki Jung

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

An expression vector system was developed for the secretory production of recombinant Bacillus stearothermophilus L1 lipase in Saccharomyces cerevisiae. The mature L1 lipase gene was fused to α-amylase signal sequence from Aspergillus oryzae for the effective secretion into the culture broth and the expression was controlled under GAL10 (the gene coding UDP-galactose epimerase of S. cerevisiae) promoter. S. cerevisiae harboring the resulting plasmid successfully secreted L1 lipase into the culture broth. To examine an optimum condition for L1 lipase expression in the fed-batch culture, L1 lipase expression was induced at three different growth phases (early, mid, and late-exponential growth phases). Maximum production of L1 lipase (1,254,000 U/l, corresponding to 0.65/l) was found when the culture was induced at an early growth phase. Secreted recombinant L1 lipase was purified only through CM-Sepharose chromatography, and the purified enzyme showed 1,963 U/mg of specific activity and thermoalkalophilic properties similar to those reported for the enzyme expressed in Escherichia coli.

Original languageEnglish
Pages (from-to)451-456
Number of pages6
JournalJournal of Microbiology and Biotechnology
Volume13
Issue number3
Publication statusPublished - 2003 Jun 1

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Geobacillus stearothermophilus
Lipase
Saccharomyces cerevisiae
UDPglucose 4-Epimerase
Growth
Aspergillus oryzae
Agarose Chromatography
Batch Cell Culture Techniques
Enzymes
Amylases
Protein Sorting Signals
Genes
Plasmids
Escherichia coli

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Ahn, Jung Oh ; Jang, Hyung Wook ; Lee, Hong Weon ; Choi, Eui Sung ; Haam, Seungjoo ; Oh, Tae Kwang ; Jung, Joon Ki. / Overexpression of thermoalkalophilic lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae. In: Journal of Microbiology and Biotechnology. 2003 ; Vol. 13, No. 3. pp. 451-456.
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Overexpression of thermoalkalophilic lipase from Bacillus stearothermophilus L1 in Saccharomyces cerevisiae. / Ahn, Jung Oh; Jang, Hyung Wook; Lee, Hong Weon; Choi, Eui Sung; Haam, Seungjoo; Oh, Tae Kwang; Jung, Joon Ki.

In: Journal of Microbiology and Biotechnology, Vol. 13, No. 3, 01.06.2003, p. 451-456.

Research output: Contribution to journalArticle

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