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Cloning and characterization of β-agarase AgaYT from Flammeovirga yaeyamensis strain YT

Paper ID Volume ID Publish Year Pages File Format Full-Text
21573 43229 2011 8 PDF Available
Title
Cloning and characterization of β-agarase AgaYT from Flammeovirga yaeyamensis strain YT
Abstract

A bacterium with potent agar-degrading capability was isolated from the surface of a red algae, Gracilaria tenuistipitata. Based on phenotypic characteristics, 16S rDNA gene sequence and a phylogenetic analysis, this bacterium was identified and named as Flammeovirga yaeyamensis strain YT. PCR using homology-based degenerate primers was employed to clone any agarase gene belonging to GH16 family encoded in F. yaeyamensis strain YT. The resolved 1512 nucleotides revealed that the cloned gene, namely AgaYT, encodes a protein of 503 amino acids comprising a signal peptide, a glycosyl hydrolase catalytic module and a C-terminal domain with an unknown function. The recombinant protein r-AgaYT is an endo-type β-agarase hydrolyzing agarose to yield neoagarobiose and neoagarotetraose as the main hydrolytic products. The specific activity of r-AgaYT was determined about 178.6 U mg− 1 at 40°C and pH 8.0.

► The agaYT belonging to GH16 family has been cloned from F. yaeyamensis strain YT. ► The AgaYT has a catalytic module and a C-terminal domain with an unknown function. ► TLC analysis of hydrolytic products of agarose confirmed that AgaYT is a β- agarase.

Keywords
Flammeovirga yaeyamensis; Agarase; Thin-layer chromatography; Glycosyl hydrolase catalytic module; Degenerate primer
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Cloning and characterization of β-agarase AgaYT from Flammeovirga yaeyamensis strain YT
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 112, Issue 3, September 2011, Pages 225–232
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us