Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 77, Issue 1, Pages 20-25Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2011.01.002
Keywords
Apostichopus japonicus; Pichia pastoris; Lysozyme; Expression; Antimicrobial activity
Categories
Funding
- National High-Tech Research and Development Program of China [2006AA10Z412]
- Medium or Small Science and Technology Enterprises in China [06C26222120113]
Ask authors/readers for more resources
Apostichopus japonicus (sea cucumber) is one of the economically important farmed echinoderm species in Northern China. As a crucial enzyme in innate immunity, lysozyme plays a key role in the overall defense against pathogens in A. japonicus. In the present study, a lysozyme gene from A. japonicus was cloned by PCR and expressed in Pichia past oris using the expression vector pPIC9K. The expressed lysozyme had a molecular mass of similar to 14 kD, as shown by SDS-PAGE and Western-blotting. The expression condition was optimized, and the highest expression level was achieved by induction with 1% methanol at pH 5.0 for 120 h. The recombinant lysozyme was purified by affinity chromatography using a Ni-NTA column. The specific activity of the purified lysozyme was 34,000 U/mg using Micrococcus lysodeikticus as substrates. It exhibited antimicrobial activity toward M. lysodeikticus, as detected by growth inhibition on agar plate and turbidity assay, suggesting a potential application of A. japonicus lysozyme as an antimicrobial agent in A. japonicus aquaculture. (c) 2011 Elsevier Inc. 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