Journal
JOURNAL OF BIOCHEMISTRY
Volume 164, Issue 6, Pages 407-414Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jb/mvy067
Keywords
Escherichia coli; human pancreatic lipase; lipolytic activity; refolding; Strep-tag II
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Funding
- Daiwa Securities Health Foundation
- Japan Foundation of Applied Enzymology
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An active recombinant human pancreatic lipase (recHPL) was successfully prepared for the first time from the Escherichia coli expression system using short Strep-tag II (ST II). The recHPL-ST II was solubilized using 8 M urea from E. coli lysate and purified on a Strep-Tactin-Sepharose column. After refolding by stepwise dialyses in the presence of glycerol and Ca2+ for 2 days followed by gel filtration, 1.8-6 mg of active recHPL-ST II was obtained from 1 L of culture. The recHPL was non-glycosylated, but showed almost equal specific activity, pH-dependency and time-dependent stability compared to those of native porcine pancreatic lipase (PPL) at 37 degrees C. However, the recHPL lost its lipolytic activity above 50 degrees C, showing a lower heat-stability than that of native PPL, which retained half its activity at this temperature.
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