Effect of substrate feed rate on recombinant protein secretion, degradation and invlusion body formation in Escherichia coli
2005 (English)In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 68, no 1, 82-90 p.Article in journal (Refereed) Published
The effect of changes in substrate feed rate during fedbatch cultivation was investigated with respect to soluble protein formation and transport of product to the periplasm in Escherichia coli. Production was transcribed from the P-malK promoter; and the cytoplasmic part of the production was compared with production from the P-lacUV5 promoter. The fusion protein product, Zb-MalE, was at all times accumulated in the soluble protein fraction except during high-feed-rate production in the cytoplasm. This was due to a substantial degree of proteolysis in all production systems, as shown by the degradation pattern of the product. The product was also further subjected to inclusion body fori-nation. Production in the periplasm resulted in accumulation of the full-length protein; and this production system led to a cellular physiology where the stringent response could be avoided. Furthermore, the secretion could be used to abort the diauxic growth phase resulting from use of the P-malK promoter. At high feed rate, the accumulation of acetic acid, due to overflow metabolism, could furthermore be completely avoided.
Place, publisher, year, edition, pages
2005. Vol. 68, no 1, 82-90 p.
fed-batch production, cultivations, expression, promoters, fusion, system
Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:kth:diva-6752DOI: 10.1007/s00253-004-1855-4ISI: 000230859900013ScopusID: 2-s2.0-23944489637OAI: oai:DiVA.org:kth-6752DiVA: diva2:11551
QC 201010082007-02-022007-02-022012-09-27Bibliographically approved