Evaluation of inducible promoters on the secretion of a ZZ-Proinsulin fusion protein

• Type: Article in International Scientific Journal
• Date: 2003

Authors

Filipe Mergulhão (Author) FEUP; Gabriel Monteiro (Author) Other; Gen Larsson (Author) Other; Maria Bostrom (Author) Other; Anne Farewell (Author) Other; Thomas Nystrom (Author) Other; Joaquim Cabral (Author) Other; Angela Taipa (Author) Other

Journal

Title: Biotechnology and Applied Biochemistry

Vol. 38 No. Part 1

Pages: 87-93

ISSN: 0885-4513

Publisher: Wiley-Blackwell

Indexing

ISI Web of Science

Scientific classification

CORDIS: Technological sciences > Technology > Biotecnology

Other information

DOI: 10.1042/BA20030043

Abstract (EN): Four inducible promoters, uspA, uspB, IacUV5 and malK, were evaluated in the expression of the fusion protein ZZ-proinsulin by Escherichia coli. The aim was to select for their effects on the most appropriate expression system (promoter and culture medium) for secretion of ZZ-proinsulin to the periplasmic space and culture medium. All the expression vectors contained the RNase III cleavage site to ensure that the mRNA translation rate remained independent of 5'-untranslated regions thus making promoter strength comparisons more accurate. The highest ZZ-proinsulin secretion yields were 6.2 mg/g of dry cell weight in the periplasmic space and 2.6 mg/g of dry cell weight in the culture medium using the malK promoter. It was also demonstrated that the use of M9 minimal medium favours secretion.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 7