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Gene cloning, expression and characterisation of a new β-agarase, AgWH50C, producing neoagarobiose from Agarivorans gilvus WH0801.
- Source :
-
World Journal of Microbiology & Biotechnology . Jun2014, Vol. 30 Issue 6, p1691-1698. 8p. - Publication Year :
- 2014
-
Abstract
- agWH50C, a novel β-agarase gene, was cloned from Agarivorans gilvus WH0801 by degenerate PCR and nested PCR. The gene agWH50C comprized a 2,223-bp, encoding a protein of 740 amino acids. Sequencing results demonstrated that AgWH50C shared 45 % sequence identity with a well characterized β-agarase, Aga50D, from Saccharophagus degradans 2-40. The mature agarase was expressed in Escherichia coli and purified by affinity chromatography. The optimum pH and temperature for AgWH50C activity were 6.0 and 30 °C. The K and V values for agarose were 12.55 mg/ml and 1.17 U/mg. Analysis of the hydrolysis products using linear ion trap mass spectrometry, Fourier transform-nuclear magnetic resonance spectrometry and thin-layer chromatography confirmed that the reaction product of AgWH50c was α-neoagarobiose alone. Therefore, our novel agarase has the potential for industrial applications to produce neoagarobiose as well as provides a key β-agarase for fermentation of agar biomass. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 09593993
- Volume :
- 30
- Issue :
- 6
- Database :
- Academic Search Index
- Journal :
- World Journal of Microbiology & Biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 95975228
- Full Text :
- https://doi.org/10.1007/s11274-013-1591-y