Overexpression of a thermostable lipase gene from Pseudomonas fluorescens in Escherichia coli |
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Authors: | Guk Hoon Chung Young Phil Lee Ook Joon Yoo Joon Shick Rhee |
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Institution: | (1) Department of Biological Science and Engineering, Korea Advanced of Institute of Science and Technology, Chongyang, P. O. Box 150, 130-650 Seoul, Korea |
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Abstract: | Summary A thermostable lipase gene from Pseudomonas fluorescens SIK W1 was overexpressed in Escherichia coli BL21 using expression vector pTTY2. The amount of lipase produced by E. coli BL21 with pTTY2 was more than 40% of the total cell proteins when induced with isopropyl--d-thiogalactopyranoside. The lipase was produced as inclusion bodies in the cytoplasm of E. coli. They were solubilized by 8 m urea and refolded into biologically active form. The refolded lipase showed high thermostability; the time required for 90% inactivation of the enzyme (D-value) was 4 h at 95°C and the increment of temperature to reduce heating times by 90% (z
d
value) was 76°C.Offprint requests to: J. S. Rhee |
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