dc.contributor.author | Zhang, J | |
dc.contributor.author | Lin, S | |
dc.contributor.author | Zeng, R | |
dc.date.accessioned | 2019-01-30T15:00:49Z | |
dc.date.issued | 2007-04-01 | |
dc.description.abstract | A psychrotrophic strain 7195 showing extracellular lipolytic activity towards tributyrin was isolated from deep-sea sediment of Prydz Bay and identified as a Psychrobacter species. By screening a genomic DNA library of Psychrobacter sp. 7195, an open reading frame of 954 by coding for a lipase gene, lipA1, was identified, cloned, and sequenced. The deduced LipA1 consisted of 317 amino acids with a molecular mass of 35,210 kDa. It had one consensus motif, G-N-S-M-G (GXSXG), containing the putative active-site serine, which was conserved in other cold-adapted lipolytic enzymes. The recombinant LipA1 was purified by column chromatography with DEAE Sepharose CL-4B, and Sephadex G-75, and preparative polyacrylamide gel electrophoresis, in sequence. The purified enzyme showed highest activity at 30°C, and was unstable at temperatures higher than 30°C, indicating that it was a typical cold-adapted enzyme. The optimal pH for activity was 9.0, and the enzyme was stable between pH 7.0-10.0 after 24 h incubation at 4°C. The addition of Ca2+and Mg2+enhanced the enzyme activity of LipA1, whereas the Cd2+, Zn2+, Co2+, Fe3+, Hg2+, Fe2+, Rb2+, and EDTA strongly inhibited the activity. The LipA1 was activated by various detergents, such as Triton X-100, Tween 80, Tween 40, Span 60, Span 40, CHAPS, and SDS, and showed better resistance towards them. Substrate specificity analysis showed that there was a preference for trimyristin and p-nitrophenyl myristate (C14acyl groups). © The Korean Society for Microbiology and Biotechnology. | en_GB |
dc.description.sponsorship | National Natural Science Funds of China | en_GB |
dc.identifier.citation | Vol. 17 (4), pp. 604 - 610 | en_GB |
dc.identifier.grantnumber | 40406029 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/35664 | |
dc.language.iso | en | en_GB |
dc.publisher | Korean Society for Microbiology and Biotechnology | en_GB |
dc.relation.url | http://www.jmb.or.kr/journal/main.html?mod=vol&tops=&year=2007 | en_GB |
dc.rights | © 2007 Korean Society for Microbiology and Biotechnology | en_GB |
dc.subject | Deep-sea sediment | en_GB |
dc.subject | cold-adapted lipase | en_GB |
dc.subject | Psychrobacter | en_GB |
dc.subject | recombinant protein | en_GB |
dc.title | Cloning, expression, and characterization of a cold-adapted lipase gene from an antarctic deep-sea psychorotrophic bacterium, Psychobacter sp. 7195 | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2019-01-30T15:00:49Z | |
dc.identifier.issn | 1017-7825 | |
dc.description | This is the final version. Available from Korean Society for Microbiology and Biotechnology via the link in this record | en_GB |
dc.identifier.journal | Journal of Microbiology and Biotechnology | en_GB |
dc.rights.uri | http://www.rioxx.net/licenses/all-rights-reserved | en_GB |
dcterms.dateAccepted | 2007-04-01 | |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2007-04-01 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2019-01-30T14:57:28Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2019-01-30T15:00:51Z | |
refterms.panel | A | en_GB |