Open AccessCharacterization of an Extracellular Lipase Encoded by LIP2 in Yarrowia lipolytica
Author(s) -
Georges Pignède,
H. Wang,
Franck Fudalej,
Claude Gaillardin,
Michel Séman,
JeanMarc Nicaud
Publication year - 2000
Publication title -
journal of bacteriology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.652
H-Index - 246
eISSN - 1067-8832
pISSN - 0021-9193
DOI - 10.1128/jb.182.10.2802-2810.2000
Subject(s) - yarrowia , lipase , biochemistry , biology , farnesyltransferase , yeast , extracellular , enzyme , prenylation
We isolated theLIP2 gene from the lipolytic yeastYarrowia lipolytica . It was found to encode a 334-amino-acid precursor protein. The secreted lipase is a 301-amino-acid glycosylated polypeptide which is a member of the triacylglycerol hydrolase family (EC3.1.1.3 ). The Lip2p precursor protein is processed by theKEX2 -like endoprotease encoded byXPR6 . Deletion of theXPR6 gene resulted in the secretion of an active but less stable proenzyme. Thus, the pro region does not inhibit lipase secretion and activity. However, it does play an essential role in the production of a stable enzyme. Processing was found to be correct inLIP2 A (multipleLIP2 copy integrant)-overexpressing strains, which secreted 100 times more activity than the wild type, demonstrating thatXPR6 maturation was not limiting. No extracellular lipase activity was detected with thelip2 knockout (KO) strain, strongly suggesting that extracellular lipase activity results from expression of theLIP2 gene. Nevertheless, thelip2 KO strain is still able to grow on triglycerides, suggesting an alternative pathway for triglyceride utilization inY. lipolytica .