title: Characterization of the 5' flanking region of lipase gene from Penicillium expansum and its application in molecular breeding.
abstract: A major challenge for further promotion of lipase productivity in Penicillium 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 (Ppel) 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 Ppel promoter and TtrpC 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.
authors: Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.
authors: Tian
authors: T
authors: Zhang
authors: Ying
authors: Y
authors: Peng
authors: Qingsheng
authors: Q
authors: Yu
authors: Jieliang
authors: J
authors: Wang
authors: Kexuan
authors: K
authors: Tang
mesh_terms: D024506:5' Flanking Region / Q000235:genetics*; D001483:Base Sequence; D032621:DNA Shuffling; D005800:Genes, Fungal / Q000235:genetics*; D008049:Lipase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D000069463:Olive Oil; D010407:Penicillium / Q000201:enzymology / Q000235:genetics*; D010938:Plant Oils / Q000378:metabolism
chemical_list: D000069463:Olive Oil; D010938:Plant Oils; D008049:Lipase
keywords: 5′ flanking region of lipase gene; Penicillium expansum; glucose repression; olive oil induction; promoter activity; strain improvement