Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 103, Issue 13, Pages 5231-5241Publisher
SPRINGER
DOI: 10.1007/s00253-019-09803-1
Keywords
Pectate lyase; Enzymatic assay; Cold-active; Specific activity; Mutation analysis
Categories
Funding
- Technical innovation special fund of Hubei Province [2017ACA171]
- Hubei Provincial Natural Science Foundation [2018CFA042, 2018CFB319]
- Wuhan Yellow Crane Talents (Science) Program
Ask authors/readers for more resources
Cold-active enzymes have become attractive biocatalysts in biotechnological applications for their ability to retain high catalytic activity below 30 degrees C, which allows energy reduction and cost saving. Here, a 1041 bp gene pel1 encoding a 34.7 KDa pectate lyase was cloned from a facultatively psychrophilic Antarctic bacterium Massilia eurypsychrophila and heterologously expressed in Escherichia coli. PEL1 presented the highest 66% identity to the reported mesophilic pectate lyase PLXc. The purified PEL1 exhibits the optimum temperature and pH of 30 degrees C and 10 toward polygalacturonic acid, respectively. PEL1 is a cold-active enzyme that can retain 60% and 25% relative activity at 10 degrees C and 0 degrees C, respectively, while it loses most of activity at 40 degrees C for 10 min. PEL1 has the highest specific activity (78.75 U mg(-1)) than all other reported cold-active pectinase, making it a better choice for use in industry. Based on the detailed sequence and structure comparison between PEL1 and PLXc and mutation analysis, more flexible structure and some loop regions may contribute to the cold activity and thermal instability of PEL1. Our investigations of the cold-active mechanism of PEL1 might guide the rational design of PEL1 and other related enzymes.
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