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Identification, Cloning, and Expression of L-Amino Acid Oxidase from Marine Pseudoalteromonas sp. B3
Joint Authors
Yu, Zhiliang
Zhou, Ning
Qiao, Hua
Qiu, Juanping
Source
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-8, 8 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-01-09
Country of Publication
Egypt
No. of Pages
8
Main Subjects
Medicine
Information Technology and Computer Science
Abstract EN
L-amino acid oxidase (LAAO) is attracting more attentions due to its broad and important biological functions.
Recently, an LAAO-producing marine microorganism (strain B3) was isolated from the intertidal zone of Dinghai sea area, China.
Physiological, biochemical, and molecular identifications together with phylogenetic analysis congruously suggested that it belonged to the genus Pseudoalteromonas.
Therefore, it was designated as Pseudoalteromonas sp.
B3.
Its capability of LAAO production was crossly confirmed by measuring the products of H2O2, a-keto acids, and N H 4 + in oxidization reaction.
Two rounds of PCR were performed to gain the entire B3-LAAO gene sequence of 1608 bps in length encoding for 535 amino acid residues.
This deduced amino acid sequence showed 60 kDa of the calculated molecular mass, supporting the SDS-PAGE result.
Like most of flavoproteins, B3-LAAO also contained two conserved typical motifs, GG-motif and β α β-dinucleotide-binding domain motif.
On the other hand, its unique substrate spectra and sequence information suggested that B3-LAAO was a novel LAAO.
Our results revealed that it could be functionally expressed in E.
coli BL21(DE3) using vectors, pET28b(+) and pET20b(+).
However, compared with the native LAAO, the expression level of the recombinant one was relatively low, most probably due to the formation of inclusion bodies.
Several solutions are currently being conducted in our lab to increase its expression level.
American Psychological Association (APA)
Yu, Zhiliang& Zhou, Ning& Qiao, Hua& Qiu, Juanping. 2014. Identification, Cloning, and Expression of L-Amino Acid Oxidase from Marine Pseudoalteromonas sp. B3. The Scientific World Journal،Vol. 2014, no. 2014, pp.1-8.
https://search.emarefa.net/detail/BIM-1051850
Modern Language Association (MLA)
Yu, Zhiliang…[et al.]. Identification, Cloning, and Expression of L-Amino Acid Oxidase from Marine Pseudoalteromonas sp. B3. The Scientific World Journal No. 2014 (2014), pp.1-8.
https://search.emarefa.net/detail/BIM-1051850
American Medical Association (AMA)
Yu, Zhiliang& Zhou, Ning& Qiao, Hua& Qiu, Juanping. Identification, Cloning, and Expression of L-Amino Acid Oxidase from Marine Pseudoalteromonas sp. B3. The Scientific World Journal. 2014. Vol. 2014, no. 2014, pp.1-8.
https://search.emarefa.net/detail/BIM-1051850
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1051850