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Characterization of the 5′ flanking region of lipase gene from P enicillium expansum and its application in molecular breeding
Author(s) -
Zhang Tian,
Peng Ying,
Yu Qingsheng,
Wang Jieliang,
Tang Kexuan
Publication year - 2014
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1002/bab.1214
Subject(s) - penicillium expansum , lipase , biology , promoter , gene , expression cassette , recombinant dna , microbiology and biotechnology , genetics , biochemistry , enzyme , botany , gene expression , postharvest , vector (molecular biology)
A major challenge for further promotion of lipase productivity in P enicillium expansum PE ‐12 is to find a suitable promoter that can function efficiently in this industrial strain. In this study, the 5′ flanking region of P . expansum lipase ( P pel) containing a putative novel promoter sequence was characterized by fusing to β ‐glucuronidase ( GUS ) and subsequently introducing into P . expansum . As a result, all the transformants showed blue color quickly after incubation in GUS detection buffer, suggesting a strong promoter activity of this fragment. Glucose repression was identified for the promoter, whereas olive oil acted as a positive regulator. Facilitated by this novel promoter, P . expansum PE ‐12 was genetically modified, with an improved lipase yield, via a recombinant plasmid with P . expansum lipase gene ( PEL ) under the control of P pel promoter and T trpC terminator. The highest lipase yield among the modified strains could attain 2,100 U/mL, which is more than twofold of the previous industrial strain (900 U/mL). The engineered strain through molecular breeding method as well as this new promoter has great value in lipase industry.