Abstract
A Gram-negative, rod-shaped, non-spore forming, non-motile and moderate halophilic bacteria designated as strain CMC-5 was isolated from decomposing seaweeds by enrichment culture. The growth of strain CMC-5 was assessed in synthetic seawater-based medium containing polysaccharide. The bacterium degraded and utilized agar, alginate, carrageenan, xylan, carboxymethyl cellulose and chitin. The strain was characterized using a polyphasic approach for taxonomic identification. Cellular fatty acid analysis showed the presence of iso-C15:0 as major fatty acid and significant amounts of iso-C17:1ω9c and C18:1ω7c . Phylogenetic analysis based on 16S rDNA sequence indicated that strain CMC-5 is phylogenetically related to Microbulbifer genus and 99% similar to type strain Microbulbifer elongatus DSM6810T. However in contrast to Microbulbifer elongatus DSM6810T, strain CMC-5 is non-motile, utilizes glucose, galactose, inositol and xylan, does not utilize fructose and succinate nor does it produce H2S. Further growth of bacterial strain CMC-5 was observed when inoculated in seawater-based medium containing sterile pieces of Gracilaria corticata thalli. The bacterial growth was associated with release of reducing sugar in the broth suggesting its role in carbon recycling of polysaccharides from seaweeds in marine ecosystem.
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References
Andrykovich G, Marx I (1988) Isolation of a new polysaccharide digesting bacteria from salt marsh. Appl Env Microbiol 54:1061–1062
Anzai Y, Kim H, Park JY (2000) Phylogenetic affiliation of the Pseudomonads based on 16S rDNA sequence. Int J Syst Evol Microbiol 50:1563–1589
Aoki Y, Kamei Y (2006) Preparation of recombinant polysaccharide degrading enzymes from the marine bacterium, Pseudomonas sp. ND137 for the production of protoplasts from Porphyra yezoensis. Eur J Phycol 41:321–328
Camacho PA, Salinias JM, Delgado M, Fuertes C (2007) Use of single cell detritus (SCD) produced from Laminaria saccharina in the feeding of the clam Ruditapes decussatus (Linnaeus, 1758). Aquaculture 1–4:211–218
Chen LCM, McCracken I (1993) An antibiotic protocol for preparing axenic culture of Porphyra linearis. Botanica Marina 36:29–33
Ekborg NA, Gonzalez JM, Howard MB, Taylor LE et al (2005) Saccharophagus degradans gen. nov., a versatile marine degrader of complex polysaccharides. Int J Syst Evol Microbiol 55:1545–1549
Ensor LA, Stosz SK, Weiner RM (1999) Expression of multiple complex polysaccharide degrading enzyme systems by marine bacterium strain 2–40. J Ind Microbiol Biotechnol 23:123–126
Felsenstein J (2006) PHYLIP (Phylogenetic Inference Package) version 3.66. Department of Genetics, University of Washington, Seattle, USA
Gacesa P, Wustman FS (1990) Plate assay for simultaneous detection of alginate lyases and determination of substrate specificities. Appl Environ Microbiol 56:2265–2267
Ghadi SC, Muraleedharan UD, Jawaid S (1997) Screening for agarolytic bacteria and development of a novel method for in situ detection of agarase enzyme. J Mar Biotechnol 5:194–200
Gonzalez JM, Mayer F, Moran MA, Hodson RE et al (1997) Microbulbifer hydrolyticus gen. nov., sp. nov., and Marinobacterium georgiense gen. nov., two marine bacteria from a lignin rich pulp mill waste enrichment community. Int J Syst Bacteriol 47:369–376
Hodgson DA, Chater KF (1981) A chromosomal locus controlling extracellular agarase production by Streptomyces coelicolor A3(2) and inactivation by chromosomal integration of plasmid SCP1. J Gen Microbiol 124:339–348
Hosoda A, Sakai M, Kanazawa S (2003) Isolation and characterization of agar-degrading Paenibacillus spp associated with the rhizosphere of spinach. Biosci Biotechnol Biochem 67:1048–1055
Ivanova EP, Bakunina IY, Sawabe T et al (2002) Two species of culturable bacteria associated with degradation of brown algae. Fucus evanescens. Microbiol Ecol 43:242–249
Kloareg B, Quatrano RS (1988) Structure of the cell walls of marine algae and ecophysiological functions of the matrix polysaccharides. Oceanogr Mar Biol Ann Rev 26:259–315
Kurita K (2006) Chitin and chitosan: functional biopolymers from marine crustaceans. Mar Biotechnol 8:203–226
Maloy SR (1989) Experimental techniques in bacterial genetics. Jones and Bartlett, Boston, USA
Mandel M, Marmur J (1968) Use of ultraviolet absorbance-temperature profile for determining the guanine plus cytosine content of DNA. Methods Enzymol 12B:195–206
Miller GL (1960) Measurement of carboxymethyl cellulase activity. Anal Biochem 1:127–132
Nishijima M, Takadera T, Imamura N et al (2009) Microbulbifer variabilis sp. nov. and Microbulbifer epialgicus sp. nov., isolated from Pacific marine algae, possess a rod–coccus cell cycle in association with the growth phase. Int J Syst Evol Microbiol 59:1696–1707
Palleroni NJ (1984) Genus Pseudomonas. Migula 1894. In: Krieg NR, Holt JG (eds) Bergey’s manual of systematic bacteriology, vol I. Williams and Wilkins, Baltimore, pp 141–199
Pidiyar V, Kaznowski A, Narayan NB et al (2002) Aeromonas culicicola sp. nov., from the midgut of Culex quinquefasciatus. Int J Syst Evol Microbiol 52:1723–1728
Quatrano RS, Cladwell BA (1978) Isolation of a unique marine bacterium capable of growth on wide of polysaccharides from macroalgae. Appl Environ Microbiol 36(6):979–981
Ruijssenaars HJ, Hartmans S (2001) Plate screening methods for the detection of polysaccharase producing microorganisms. Appl Microbiol Biotechnol 55:143–149
Ryu S, Cho S, Park S et al (2001) Cloning of cel9A gene and characterization of its gene product from marine bacterium Pseudomonas sp. SK 38. Appl Microbiol Biotechnol 57:138–145
Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt F (ed) Methods for general and molecular bacteriology. American Society for Microbiology, Washington D.C., pp 607–654
Tanaka T, Yan L, Burgess JG (2003) Microbulbifer arenaceous sp. nov., a novel endolithic bacterium isolated from the inside of red sand stone. Curr Microbiol 47:412–416
Tang SK, Wang Y, Cai M et al (2008) Microbulbifer halophilus sp. nov., a moderately halophilic bacterium from north-west China. Int J Syst Evol Microbiol 58:2036–2040
Tang JC, Taniguchi H, Chu H et al (2009) Isolation and characterization of alginate-degrading bacteria for disposal of seaweed wastes. Lett Appl Microbiol 48:38–43
Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680
Weiner R, Chakravorthy D, Whiteland L (1998) The architecture of degradative complex polysaccharide enzyme arrays in a marine bacterium has implications for bioremediation. In: Gal L, Halvorson (eds) New developments in marine biotechnology. Plenum Press, New York, pp 171–176
Yoon JH, Kim H, Kang KH et al (2003) Transfer of Pseudomonas elongata Humm 1946 to the genus Microbulbifer as Microbulbifer elongatus comb. nov. Int J Syst Evol Microbiol 53:1357–1361
Yoon JH, Kim IG, Shin DY et al (2003) Microbulbifer salipaludis sp. nov., a moderate halophile isolated from a Korean salt marsh. Int J Syst Evol Microbiol 53:53–57
Yoon JH, Kim IG, Oh TK et al (2004) Microbulbifer maritimus sp. nov., isolated from an intertidal sediment from the yellow sea, Korea. Int J Syst Evol Microbiol 54:1111–1116
Yoon JH, Jung YS, Kang SJ et al (2007) Microbulbifer celer sp. nov., isolated from a marine solar saltern of the yellow sea in Korea. Int. J Syst Evol Microbiol 57:2365–2369
Acknowledgement
The authors would like to thank Dr. Tapan Chakraborthy, Institute of Microbial Technology, Chandigarh, India, and Dr. Shanta Nair, National Institute of Oceanography, Goa, India, for DNA G + C and FAME analysis, respectively. This work was supported by Department of Science and Technology, Govt. of India, New Delhi (SERC Fast Track Scheme No SR/FTP/LS-264/2000).
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Jonnadula, R., Verma, P., Shouche, Y.S. et al. Characterization of Microbulbifer Strain CMC-5, a New Biochemical Variant of Microbulbifer elongatus Type Strain DSM6810T Isolated from Decomposing Seaweeds. Curr Microbiol 59, 600–607 (2009). https://doi.org/10.1007/s00284-009-9480-1
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DOI: https://doi.org/10.1007/s00284-009-9480-1