1. Characterization of the 5′ flanking region of lipase gene fromPenicillium expansumand its application in molecular breeding
- Author
-
Tian Zhang, Qingsheng Yu, Ying Peng, Jieliang Wang, and Kexuan Tang
- Subjects
Molecular breeding ,biology ,Process Chemistry and Technology ,5' flanking region ,Biomedical Engineering ,food and beverages ,Bioengineering ,General Medicine ,biology.organism_classification ,Applied Microbiology and Biotechnology ,Genetically modified organism ,law.invention ,Terminator (genetics) ,Biochemistry ,law ,Drug Discovery ,biology.protein ,Recombinant DNA ,Molecular Medicine ,Lipase ,Penicillium expansum ,Gene ,Biotechnology - 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.
- Published
- 2014