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Studies on agar degrading Salegentibacter sp. and characterization of its agarase

R Vijayaraghavan, S Rajendran

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Int. J. Biosci.1( 5), 56-64, October 2011


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The phenotypic and agar degrading features of an unidentified marine bacteria was investigated. The strain was Gram-negative, obligatory aerobic and non motile. On the basis of several morphological features and a phylogenetic analysis of the genes coding for the 16S rDNA, this strain was identified as Salegentibacter sp. On solid agar medium, this isolate produced extracellular agarase which causes agarliquefaction around the colonies. An extracellular agarase was purified by ammonium sulfate precipitation, gel filtration and ion-exchange chromatography on DEAE-cellulose. The purified protein migrated as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it had a molecular weight of 28 kDa. The enzyme exhibited maximal activity at pH 7.5. The kinetic parameters of the enzyme were Km = 3 ± 0.19 mM, Kcat = 160 ± 10 S -1 and Kcat / Km = 53 ± 10 S -1.mM -1. The purified agarase may be used for application in protoplast formation of agarophytes which can subsequently be used in algal biotechnology to evolve a superior strain of Gracilaria sp. yielding superior quality agar.


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Studies on agar degrading Salegentibacter sp. and characterization of its agarase

Allouch J, Jam M, Helbert W, Barbeyron T, Kloareg B., Henrissat B, Czjzek M. 2003. The three-dimensional structures of two beta-agarases. J. Biol Chem 278, 47171-80.

Anderson NS, Rees DA. 1965. Porphyran: a polysaccharide with a marked repeating structure. J. Chem. Soc 10, 5880-5887.

Baumann P, Gauthier MJ, Baumann L. 1984. Genus Alteromonas Baumann, Baumann, Mandel and Allen 1972, p. 343–352. In N. R. Krieg and J. G. Holt (ed.), Bergey’s manual of systematic bacteriology. vol. The Williams & Wilkins Co., Baltimore.

Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem 72, 248-254.

Chiura HX, Tsukamoto K. 2000. Purification and characterization of novel agarase secreted by marine bacterium, Pseudoalteromonas sp. strain CKT1. Microb. Environ 15, 11–22.

Duckworth M, Turvey JR. 1969. The specificity of an agarase from a Cytophaga species. Biochem. J. 113, 693-697.

Ghadi SC, Muraleedharan UD, Jawaid S. 1997. Screening for agarolytic bacteria and development of a novel method for in situ detection of agarase. J. Mar. Biotechnol 5, 194-200.

Hirase S. 1969. Glactan from red algae. In Biochemistry of Polysaccharide, ed. Fujio, E., Kyouritu-shuppan, Tokyo.

Ivanova EP, Kiprianova EA, Mikhailov VV, Levanova GF, Garagulya AD, Gorshkova NM, Vysotskii MV, Nicolau DV, Yumoto N, Taguchi T, Yoshikawa S. 1998. Phenotypic diversity of Pseudoalteromonas citrea from different marine habitats and emendation of description. Int. J. Syst. Bacteriol 48, 247–256.

Izumi Yamaura, Toshihiko Matsumoto, Masaru Funatsu, Hisaji Shigeiri and Teruhiko Shibata. 1991. Purification and some properties of agarases from Pseudomonas sp. PT – 5. J. Biol. Chem 55 (10), 2531-2536.

Kendall K, Cullum J. 1984. Cloning and expression of an extracellular-agarase from Streptomyces coelicolor A3 (2) in streptomyces lividans 66. Gene 29, 315-321.

Kim BJ, Kim HJ, Ha SD, Hwang SH, Byun DS, Lee TH, Kong JY. 1999. Purification and characterization of β-agarase from marine bacterium Bacillus cereus ASK202. Biotechnol. Lett 21, 1011– 1015.

Kong JY, Hwang SH, Kim BJ, Bae SK, Kim JD. 1997. Cloning and expression of an agarase gene from a marine bacterium Pseudomonas sp. W7. Biotechnol. Lett 19, 23–26.

Lineweaver H, Bruk D. 1934. The determination of enzyme dissociation constants. J. Am. Chem. Soc 56, 658-666.

Leon O, Quintana L, Peruzzo G, Slebe JC. 1992. Purification and properties of an extracellular agarase from Alteromonas sp. strain C-1. Appl. Environ. Microbiol 58, 4060–4063.

Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem 193, 265-275.

Morrice LM, McLean MW, Williamson FB, Long WF. 1983a. Purification and some properties of β-agarase I and II from Pseudomonas atlantica. Eur. J. Biochem 135, 553-558.

Morrice LM, Mclean MW, Long WF, Williamson FB. 1983b. β-agarases I and I I from Pseudomonas atlantica. Eur. J. Biochem 137, 149–154.

Murielle JM, Didier Flament, Julie Allouch, Philippe Potin, Laurent Thion, Bernard Kloareg, Mirjam Czjzek, William Helbert, Gurvan Michel, Tristan Barbeyron. 2005. The endo-β-agarases AgaA and AgaB from the marine bacterium Zobellia galactanivorans: two paralogue enzymes with different molecular organizations and catalytic behaviours. J. Biochem 385, 703–713.

Nelson NA. 1994. Photometric adaptation of the Somogyi method for the determination of glucose. J. Biol. Chem 153, 375-380.

Potin P, Richard C, Rochas C, Kloareg B. 1993. Purification and characterization of the alpha-agarase from Alteromonas agarlyticus (Cataldi) comb. nov., strain GJ1B. Eur J Biochem 214, 599-607.

Ruimy R, Breittmayer V, Elbaze P, Lafay B, Boussemart O, Gauthier M, Christen R. 1994. Phylogenetic analysis and assessment of the genera Vibrio, Photobacterium, and Plesiomonas deduced from small-subunit rRNA sequences. Int. J. Syst. Bacteriol 44, 416–426.

Sambrook J, Russel DW. 2001. Molecular Cloning: A Laboratory Manual. 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.

Sneath PHA. 1994. In: W. M. Hensyl (ed.). Bergey’s Manual of Systematic Bacteriology. Ninth ed., Williams & Wilkins, Philadelphia, PA, USA.

Sugano Y, Matsumoto T, Kodama H, Noma M. 1993. Cloning and sequencing of agaA, a unique agarase 0107 gene from a marine bacterium, Vibriosp. strain JT0107. Appl Environ Microbiol 5, 3750-56.

Sugano Y, Matsumoto T, Noma M. 1994. Sequence analysis of the agaB gene encoding a new beta-agarase from Vibrio sp. strain JT0107. Biochem. Biophys Acta 1218, 105-108.

Van der Meulen HJ, Harder W. 1975. Production and characterization of the agarase of Cytoplaga flevensis. Antonie. Van. Leeuwenhoek 41, 431–447.

Vera J, Alvarez R, Murano E, Slebe JC, Leon O. 1998. Identification of a marine agarolytic pseudoalteromonas isolate and characterization of its extracellular agarase. Appl Environ Microbiol 64, 4378-83.

Weisberg WG, Barns SM, Pelletier DA, Lane DJ. 1991. 16S ribosomal DNA amplification for phylogenetic study. J. Bacteriol 173, 697-703.

Yaphe W. 1957. The use of agarase from Pseudomonas atlantica in the identification of agar in marine algae (Rhodophyceae) . Can. J. Microbiol 3, 987–993.

Yasushi Sugano, Ichiro Terada, Masatoshi Arita, Masana Noma and Takashi Matsumoto. 1993. Purification and Characterization of a new agarase from a marine bacterium, Vibrio sp. Strain JT0107. J. App. & Env. Microbiol p. 1549-1554.

Zhong Z, Toukdarian A, Helinski D, Knauf V, Sykes S, Wilkinson JE, O’Bryne C, Shea T, Loughery D, and Caspi R. 2001. Sequence analysis of a 101-kilobase plasmid required for agar degradation by a Microscilla isolate. Appl Environ Microbiol 67, 5771-5779.

Zhou J, Burns M, Tiedje J. 1996. DNA recovery from soils of diverse composition. J. Appl. Environ. Microbiol 62, 316-322.