Abstract
Purpose of work
The purpose of this study is to report a ι-carrageenase which degrades ι-carrageenan yielding neo-ι-carratetraose as the main product in the absence of NaCl.
The gene for a new ι-carrageenase, CgiB_Ce, from Cellulophaga sp. QY3 was cloned and sequenced. It comprised an ORF of 1,386 bp encoding for a protein of 461 amino acid residues. From its sequence analysis, CgiB_Ce is a new member of GH family 82 and shared the highest identity of 32 % in amino acids with ι-carrageenase CgiA2 from Zobellia galactanovorans indicating that it is a hitherto uncharacterized protein. The recombinant CgiB_Ce had maximum specific activity (1,870 U/mg) at 45 °C and pH 6.5. It was stable between pH 6.0–9.6 and below 40 °C. Although its activity was enhanced by NaCl, the enzyme was active in the absence of NaCl. CgiB_Ce is an endo-type ι-carrageenase that hydrolyzes β-1,4-linkages of ι-carrageenan, yielding neo-ι-carratetraose as the main product (more than 80 % of the total product).
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References
Barbeyron T, Michel G, Potin P et al (2000) ι-Carrageenases constitute a novel family of glycoside hydrolases, unrelated to that of κ-carrageenases. J Biol Chem 275:35499–35505
Campo VL, Kawano DF, Silva DB Jr et al (2009) Carrageenans: biological properties, chemical modifications and structural analysis—a review. Carbohydr Polym 77:167–180
Cantarel BL, Coutinho PM, Rancurel C et al (2009) The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics. Nucleic Acids Res 37:D233–D238
Hatada Y, Mizuno M, Li Z et al (2011) Hyper-production and characterization of the iota-carrageenase useful for ι-carrageenan oligosaccharide production from a deep-sea bacterium, Microbulbifer thermotolerans JAMB-A94T, and insight into the unusual catalytic mechanism. Mar Biotechnol (NY) 13:411–422
Jouanneau D, Boulenguer P, Mazoyer J et al (2010) Complete assignment of 1H and 13C NMR spectra of standard neo-ι-carrabiose oligosaccharides. Carbohydr Res 345:547–551
Ren S, Li J, Wang W et al (2010) Protective effects of kappa-ca3000+CP against ultraviolet-induced damage in HaCaT and MEF cells. J Photochem Photobiol B 101:22–30
Tamura K, Dudley J, Nei M et al (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599
Tan G, Gao Y, Shi M et al (2005) SiteFinding-PCR: a simple and efficient PCR method for chromosome walking. Nucleic Acids Res 33:e122
Wang W, Zhang P, Hao C et al (2011) In vitro inhibitory effect of carrageenan oligosaccharide on influenza a H1N1 virus. Antivir Res 92:237–246
Yang B, Yu G, Zhao X et al (2009) Mechanism of mild acid hydrolysis of galactan polysaccharides with highly ordered disaccharide repeats leading to a complete series of exclusively odd-numbered oligosaccharides. FEBS J 276:2125–2137
Yuan H, Song J, Li X et al (2006) Immunomodulation and antitumor activity of κ-carrageenan oligosaccharides. Cancer Lett 243:228–234
Acknowledgments
We are very grateful to Prof. Guangli Yu (School of Medicine and Pharmacy, Ocean University of China, China) for interpretation of the NMR spectra. The research was supported by National Science Foundation of China (31070712 and 31000361) and Special Fund for Marine Scientific Research in the Public Interest (201105027-3 and 201005024).
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Ma, S., Tan, YL., Yu, WG. et al. Cloning, expression and characterization of a new ι-carrageenase from marine bacterium, Cellulophaga sp.. Biotechnol Lett 35, 1617–1622 (2013). https://doi.org/10.1007/s10529-013-1244-0
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DOI: https://doi.org/10.1007/s10529-013-1244-0