Human pancreatic triglyceride lipase expressed in yeast cells: purification and characterization

Protein Expression and Purification
Y Yang, M E Lowe

Abstract

A cDNA clone encoding human pancreatic triglyceride lipase was cloned into a yeast expression vector so that the yeast PHO1 signal peptide replaced the native signal peptide. Pichia pastoris cells were transfected with the vector, and clones expressing human pancreatic triglyceride lipase were isolated. Recombinant human pancreatic lipase was expressed in broth cultures and was purified from the medium by DEAE blue Sepharose and hydroxyapatite chromatography. The highly purified lipase had specific activities for various triglyceride substrates identical to those of tissue-purified human pancreatic triglyceride lipase; it was inhibited by bile salts, required colipase for activity, and demonstrated interfacial activation. This expression system is suitable for the rapid, efficient production of human pancreatic triglyceride lipase in amounts adequate for biophysical studies.

References

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Citations

Dec 26, 2001·The Journal of Biological Chemistry·Dymphna D'AgostinoMark E Lowe
Jul 23, 2009·American Journal of Physiology. Endocrinology and Metabolism·Rita MillerMark E Lowe
Sep 3, 2003·Journal of Lipid Research·Takahiro TsujitaHoward L Brockman
Jun 1, 2010·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Brendan M McGuireBruce F Scharschmidt
Jan 6, 2007·Digestive Diseases and Sciences·David C Whitcomb, Mark E Lowe
Mar 12, 2003·International Journal of Obesity and Related Metabolic Disorders : Journal of the International Association for the Study of Obesity·C Sebban-KreuzerB Kerfelec
Oct 28, 2004·Biochemical Society Transactions·M T Andrews
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Sep 15, 2001·American Journal of Physiology. Regulatory, Integrative and Comparative Physiology·V W BauerM T Andrews
Jun 10, 2011·The Journal of Biological Chemistry·Xunjun XiaoMark E Lowe
Nov 5, 2019·Biochimica Et Biophysica Acta. Molecular and Cell Biology of Lipids·Ly VilloAivar Lookene
Apr 3, 2001·Protein Expression and Purification·J C Rotticci-MulderM Martinelle
Sep 29, 2005·Analytical Biochemistry·W E MomsenH L Brockman
Feb 17, 2006·Protein Expression and Purification·Ahmed AloulouFrédéric Carrière
Jul 25, 2006·Protein Expression and Purification·Corinne Sebban-KreuzerIsabelle Crenon

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