Journal
PROCESS BIOCHEMISTRY
Volume 48, Issue 9, Pages 1317-1323Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2013.06.021
Keywords
Acinetobacter radioresistens CMC-1; ARL; Lipase; Secretion expression; Pichia pastoris
Categories
Funding
- National Natural Science Foundation of China [2010A010500003]
- Guangdong Key Technology RD Program [2012B010300008]
Ask authors/readers for more resources
In this study, a series of strategies was developed to enhance the expression of an alkaline lipase from Acinetobacter radioresistens (ARL) in Pichia pastoris. Activity of the lipase from recombinant strain carrying a single copy of codon-optimized ARL gene was 65 U/mL in shake flask culture with p-nitrophenyl caprylate as the substrate. The lipase yield was increased to 104 U/mL by introducing a short N-extension spacer peptide coding for the 10 amino acids (EEAEAEAEPK) between a-factor signal peptide and ARL. The N-terminal extension spacer did not affect the pH or temperature properties of the recombinant ARL. After the multi-copy constructs were identified by Q-PCR assay, a higher lipase activity of 180 U/mL was obtained. Further introduction of the spliced HAC1 gene into multi-copy integrants (>6 copies) extensively enhanced the ARL yield by 30-40%. As a result, the ARL yield reached 1.06 x 10(4) U/mL in a 10-L scaled-up fed-batch fermenter as well as the lipase showed some better properties compared to that wild one from A. radioresistens. (C) 2013 Elsevier Ltd. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available