NDLI: Characterization of alginate lyase gene using a metagenomic library constructed from the gut microflora of abalone

Content Provider	Springer Nature Link
Author	Sim, Su Jung Baik, Keun Sik Park, Seong Chan Choe, Han Na Seong, Chi Nam Shin, Tai Sun Woo, Hee Chul Cho, Jeong Yong Kim, Duwoon
Copyright Year	2011
Abstract	A metagenomic fosmid library was constructed using a genomic DNA mixture extracted from the gut microflora of abalone. The library gave an alginate lyase positive clone (AlyDW) harboring a 31.7-kbp insert. The AlyDW insert consisted of 22 open reading frames (ORFs). The deduced amino acid sequences of ORFs 11–13 were similar to those of known alginate lyase genes, which are found adjacent in the genome of Klebsiella pneumoniae subsp. aerogenes, Vibrio splendidus, and Vibrio sp. belonging to the phylum Gammaproteobacteria. Among the three recombinant proteins expressed from the three ORFs, alginate lyase activity was only observed in the recombinant protein (AlyDW11) coded by ORF 11. The expressed protein (AlyDW11) had the highest alginate lyase activity at pH 7.0 and 45°C in the presence of 1 mM AgNO3. The alginate lyase activity of ORF 11 was confirmed to be endolytic by thin-layer chromatography. AlyDW11 preferred poly(β-d-mannuronate) as a substrate over poly(α-l-guluronate). AlyDW11 contained three highly conserved regions, RSEL, QIH, and YFKAGVYNQ, which may act to stabilize the three-dimensional conformation and function of the alginate lyase.
Starting Page	585
Ending Page	593
Page Count	9
File Format	PDF
ISSN	13675435
Journal	Journal of Industrial Microbiology and Biotechnology
Volume Number	39
Issue Number	4
e-ISSN	14765535
Language	English
Publisher	Springer-Verlag
Publisher Date	2011-11-10
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Abalone Alginate lyase Metagenomic library β-d-Mannuronate α-l-Guluronate Inorganic Chemistry Microbiology Bioinformatics Genetic Engineering Biochemistry Biotechnology
Content Type	Text
Resource Type	Article
Subject	Medicine Bioengineering Applied Microbiology and Biotechnology Biotechnology

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