Functional expression of the lipase gene Lip2 of Pleurotus sapidus in Escherichia coli.
The lipase Lip2 of the edible basidiomycete, Pleurotus sapidus, is an extracellular enzyme capable of hydrolysing xanthophyll esters with high efficiency. The gene encoding Lip2 was expressed in Escherichia coli TOP10 using the gene III signal sequence to accumulate proteins in the periplasmatic space. The heterologous expression under control of the araBAD promoter led to the high level production of recombinant protein, mainly as inclusion bodies, but partially in a soluble and active form. A fusion with a C-terminal His tag was used for purification and immunochemical detection of the target protein. This is the first example of a heterologous expression and periplasmatic accumulation of a catalytically active lipase from a basidiomycete fungus.
Zelena
Kateryna
K
Institut für Lebensmittelchemie, Zentrum Angewandte Chemie der Leibniz Universität Hannover, Callinstr. 5, 30167, Hannover, Germany.
Krügener
Sven
S
Lunkenbein
Stefan
S
Zorn
Holger
H
Berger
Ralf G
RG
D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D002479:Inclusion Bodies / Q000737:chemistry; D008049:Lipase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D019897:Periplasm / Q000737:chemistry; D020076:Pleurotus / Q000201:enzymology*; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D008049:Lipase