A novel psychrophilic alkaline phosphatase from the metagenome of tidal flat sediments = 갯벌 메타지놈 유래의 신규 저온성 알칼라인 포스파타아제

Cited 28 time in scopus
Metadata Downloads

Full metadata record

DC FieldValueLanguage
dc.contributor.authorDae-Hee Lee-
dc.contributor.authorSu-Lim Choi-
dc.contributor.authorEugene Rha-
dc.contributor.authorSoo Jin Kim-
dc.contributor.authorSoo Jin Yeom-
dc.contributor.authorJae Hee Moon-
dc.contributor.authorSeung Goo Lee-
dc.date.accessioned2017-04-19T10:01:58Z-
dc.date.available2017-04-19T10:01:58Z-
dc.date.issued2015-
dc.identifier.issn14726750-
dc.identifier.uri10.1186/s12896-015-0115-2ko
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/12457-
dc.description.abstractBackgroundAlkaline phosphatase (AP) catalyzes the hydrolytic cleavage of phosphate monoesters under alkaline conditions and plays important roles in microbial ecology and molecular biology applications. Here, we report on the first isolation and biochemical characterization of a thermolabile AP from a metagenome.ResultsThe gene encoding a novel AP was isolated from a metagenomic library constructed with ocean-tidal flat sediments from the west coast of Korea. The metagenome-derived AP (mAP) gene composed of 1,824 nucleotides encodes a polypeptide with a calculated molecular mass of 64 kDa. The deduced amino acid sequence of mAP showed a high degree of similarity to other members of the AP family. Phylogenetic analysis revealed that the mAP is shown to be a member of a recently identified family of PhoX that is distinct from the well-studied classical PhoA family. When the open reading frame encoding mAP was cloned and expressed in recombinant Escherichia coli, the mature mAP was secreted to the periplasm and lacks an 81-amino-acid N-terminal Tat signal peptide. Mature mAP was purified to homogeneity as a monomeric enzyme with a molecular mass of 56 kDa. The purified mAP displayed typical features of a psychrophilic enzyme: high catalytic activity at low temperature and a remarkable thermal instability. The optimal temperature for the enzymatic activity of mAP was 37°C and complete thermal inactivation of the enzyme was observed at 65°C within 15 min. mAP was activated by Ca2+ and exhibited maximal activity at pH 9.0. Except for phytic acid and glucose 1-phosphate, mAP showed phosphatase activity against various phosphorylated substrates indicating that it had low substrate specificity. In addition, the mAP was able to remove terminal phosphates from cohesive and blunt ends of linearized plasmid DNA, exhibiting comparable efficiency to commercially available APs that have been used in molecular biology.ConclusionsThe presented mAP enzyme is the first thermolabile AP found in cold-adapted marine metagenomes and may be useful for efficient dephosphorylation of linearized DNA.-
dc.publisherSpringer-BMC-
dc.titleA novel psychrophilic alkaline phosphatase from the metagenome of tidal flat sediments = 갯벌 메타지놈 유래의 신규 저온성 알칼라인 포스파타아제-
dc.title.alternativeA novel psychrophilic alkaline phosphatase from the metagenome of tidal flat sediments-
dc.typeArticle-
dc.citation.titleBMC Biotechnology-
dc.citation.number0-
dc.citation.endPage1-
dc.citation.startPage1-
dc.citation.volume15-
dc.contributor.affiliatedAuthorDae-Hee Lee-
dc.contributor.affiliatedAuthorEugene Rha-
dc.contributor.affiliatedAuthorSeung Goo Lee-
dc.contributor.alternativeName이대희-
dc.contributor.alternativeName최수림-
dc.contributor.alternativeName나유진-
dc.contributor.alternativeName김수진-
dc.contributor.alternativeName염수진-
dc.contributor.alternativeName문재희-
dc.contributor.alternativeName이승구-
dc.identifier.bibliographicCitationBMC Biotechnology, vol. 15, pp. 1-1-
dc.identifier.doi10.1186/s12896-015-0115-2-
dc.subject.keywordAlkaline phosphatase-
dc.subject.keywordEscherichia coli-
dc.subject.keywordMetagenome-
dc.subject.keywordPhoX-
dc.subject.localAlkaline phosphatase-
dc.subject.localEscherichia coli-
dc.subject.localMetagenome-
dc.subject.localPhoX-
dc.description.journalClassY-
Appears in Collections:
Division of Biomaterials Research > Synthetic Biology and Bioengineering Research Center > 1. Journal Articles
Files in This Item:

Items in OpenAccess@KRIBB are protected by copyright, with all rights reserved, unless otherwise indicated.