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Isolation and characterization of alginate-degrading bacteria Sinomicrobium oceani

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Abstract

This research work is focused on identifying alginate-degrading bacteria from various marine as well as alginate industrial substrates using basal salt medium with alginate as sole source of carbon. In the present study, alginate-degrading colonies formed soft pits on the surface of the medium with clear haloes around them gradually degrading the solid medium into liquid on day 7. The isolate positive for alginolytic activity formed a circular pale yellow zone around the colony. Out of the 23 isolates studied, 11 exclusively isolated from the alginate industrial wastes were chosen for the degradation study. On the basis of bacterial growth, total sugar, reducing sugar and total protein in the culture medium, isolates AAl 01, AAl 02, AAl 03 and AAl 04 were selected of which consortium AAl 02 + AAl 04 was found promising for maximum alginate lyase activity on day 4 of inoculation at 28 °C with pH 7.5. Batch culture studies showed that fresh biomass of Sargassum wightii produced 37.88 ± 1.37 % of reducing sugar by direct saccharification using bacterial consortium, whereas by acid pretreatment + bacterial consortium, the yield was 62.69 ± 2.90 %. In the case of pigment-phycocolloid spent, 82.41 ± 4.50 % of reducing sugar was released using bacterial consortium, whereas by mild acid pretreatment + bacterial consortium, 86.68 ± 7.11 % was achieved. Results of morphology, biochemical tests and 16S rRNA sequence confirmed the two promising isolates as Sinomicrobium oceani (AAl 02 and AAl 04) belonging to Flavobacteriaceae. Seaweed biomass degradation using biological methods is non-toxic and considered environmentally safe.

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References

  1. Percival E (1979) Polysaccharides of green, red and brown seaweeds: their basic structure, biosynthesis and function. Br Phycol J 14:103–117

    Article  Google Scholar 

  2. Arunkumar K, Selvapalam N, Rengasamy R (2005) The antibacterial compound sulpho glycerolipid 1-0 palmitoyl-3-0(6′-sulpho-α quinovopyranosyl)-glycerol from Sargassum wightii Greville (Phaeophyceae). Bot Mar 40:441–445

    Google Scholar 

  3. Torres MR, Sousa APA, Silva Filho EAT, Melo DF, Feitosa JPA, de Paula RCM, Lima MGS (2007) Extraction and physicochemical characterization of Sargassum vulgare alginate from Brazil. Carbohydr Res 342(14):2067–2074

    Article  Google Scholar 

  4. Moe ST, Draget KI, Skjåk-Braek G, Smidsrød O (1995) Alginates (chapter 9). In: A.M. S (ed) Food polysaccharide and their application. Marcel Dekker, New York, pp. 245–286

    Google Scholar 

  5. Rinaudo M (2007) Comprehensive glycoscience. In: Kamerling JP (ed) Seaweed polysaccharides (Vol. 2). Elsevier, Amsterdam, pp. 691–735

    Google Scholar 

  6. Chapman ARO, Chapman DJ (1980) Seaweeds and their uses. Can J Bot 61:1654–1659

    Google Scholar 

  7. Wilma JS (1990) Alginates. In: Harris P (ed) Food gel. Elsevier Applied Sciences Publisher Ltd, Essex, p. 54

    Google Scholar 

  8. McHugh DJ (1987) Production, properties and uses of alginates. In: McHugh, D.J. (Eds.), Production and utilization of products from commercial seaweed, FAO Fish. Teech. paper 288, pp 74

  9. Haug A, Larsen B, Smidsrod O (1974) Uronic acid sequence in alginate from different sources. Carbohydr Res 32:217–225

    Article  Google Scholar 

  10. Ci SX, Huynh TH, Louie LW, Yong A, Beals BJ, Ron N, Tsang WG, Soon-Shiang P, Desai NP (1999) Molecular mass distribution of sodium alginate by high-performance size-exclusion chromatography. J Chromatogr A 864:199–210

    Article  Google Scholar 

  11. Johnson FA, Craig DM, Mercer AD (1997) Characterization of the block structure and molecular weight of sodium alginates. J Pharm Pharmacol 49:639–643

    Article  Google Scholar 

  12. Kupper FC, Kloareg B, Guern J, Potin P (2001) Oligoguluronates elicit an oxidative burst in the brown algal kelp Laminaria digitata. Plant Physiol 125:278–291

    Article  Google Scholar 

  13. Kataoka K, Suzuki Y, Kitada M, Hashimoto T, Chou H, Bai H, Ohta M, Wu S, Suzuki K, Ide C (2004) Alginate enhances elongation of early regenerating axons in spinal cord of young rats. Tissue Eng 10(3–4):493–504

    Article  Google Scholar 

  14. Lee KY, Mooney DJ (2012) Alginate: properties and biomedical applications. Prog Polym Sci 37(1):106–126

    Article  Google Scholar 

  15. Zhou M, Han F, Li J, Zhao X (2008a) Isolation and identification of a novel alginate-degrading bacterium, Ochrobactrum sp. Songklanakarin J Sci Technol 30(2):135–140

    Google Scholar 

  16. Hu X, Jiang X, Hwang HM (2006) Purification and characterization of an alginate lyase from marine bacterium Vibrio sp. mutant strain 510-64. Curr Microbiol 53:135–140

    Article  Google Scholar 

  17. Zhou X, Wheeler MM, Oi FM, Scharf ME (2008b) RNA interference in the termite Reticulitermes flavipes through ingestion of double-stranded RNA. Insect Biochem Mol Biol 38:805–815

    Article  Google Scholar 

  18. Araki T, Hayakawa H, Lu Z, Karita S, Morishita T (1998) Purification and characterization of agarases from a marine bacterium, Vibrio sp.PO-303. J Mar Biotechnol 6:260–265

    Google Scholar 

  19. Li S, Wang L, Han F, Gong Q, Yu W (2016) Cloning and characterization of the first polysaccharide lyase family 6 oligoalginate lyase from marine Shewanella sp. Kz7. J Biochem 159(1):77–86

    Article  Google Scholar 

  20. Yanagisawa M, Kawai S, Murata K (2013) Strategies for the production of high concentrations of bioethanol from seaweeds: production of high concentrations of bioethanol from seaweeds. Bioengineered 4(4):224–235

    Article  Google Scholar 

  21. Hansen JB, Doubet RS, Ram J (1984) Alginase enzyme production by Bacillus circulans. Appl Environ Microbiol 47:704–709

    Google Scholar 

  22. Jang JS, Cho Y, Jeong GT, Kim SK (2012a) Optimization of saccharification and ethanol production by simultaneous saccharification and fermentation (SSF) from seaweed, Saccharina japonica. Bioprocess Biosyst Eng 35(1–2):11–18

    Article  Google Scholar 

  23. Jang SS, Shirai Y, Uchida M, Wakisaka M (2012b) Production of mono sugar from acid hydrolysis of seaweed. Afr J Biotechnol 11(8):1953–1963

    Google Scholar 

  24. Wong TY, Preston LA, Schiller NL (2000) Alginate lyase: review of major sources and enzyme characteristics, structure-function analysis, biological roles, and applications. Annu Rev Microbiol 54:289–340

    Article  Google Scholar 

  25. William AK, Eagon RG (1962) Studies on the alginase of Agarbacterium alginicums. Can J Microbiol 8:649–654

    Article  Google Scholar 

  26. Gacesa P (1992) Enzymic degradation of alginates. Int J Biochem 24:545–552

    Article  Google Scholar 

  27. Takeuchi T, Murata K, Isao K (1994) A method for depolymerization of alginate using the enzyme system of Flavobacterium multivolum. Nippon Suisan Gakkaishi 41:505–511

    Google Scholar 

  28. Nibu Y, Satoh T, Nishi Y, Takeuchi T, Murata K, Kusakabe I (1995) Purification and characterization of extracellular alginate lyase from Enterobacter cloacae M-1. Biosci Biotechnol Biochem 59:632–637

    Article  Google Scholar 

  29. Shimokawa T, Yoshida S, Kusakabe I, Takeuchi T, Murata K, Kobayashi H (1997) Some properties and action mode of “-L-guluronan lyase from Enterobacter cloacae M-1. Carbohydr Res 604:125–132

    Article  Google Scholar 

  30. Iwamoto Y, Araki R, Iriyama K, Oda T, Fukuda H, Hayashida S, Muramatsu T (2001) Purification and characterization of bifunctional alginate lyase from Alteromonas sp. strain no. 272 and its action on saturated oligomeric substrates. Biosci Biotechnol Biochem 65:133–142

    Article  Google Scholar 

  31. Li JB, Yu WG, Feng H, Han WJ, Kai S (2003) Purification and characterization of a novel alginate lyase from marine Vibrio sp. QY102. Acta Microbiol Sin 43:753–757

    Google Scholar 

  32. Oh Y, Xu X, Kim JY, Park JM (2015) Maximizing the utilization of Laminaria japonica as biomass via improvement of alginate lyase activity in a two-phase fermentation system. Biotechnol J 10(8):1281–1288

    Article  Google Scholar 

  33. Hisano T, Yonemoto Y, Yamashita T, Fukuda Y, Kimura A, Murata K (1995) Direct uptake of alginate molecules through a pit on the bacterial cell surface: a novel mechanism for the uptake of macromolecules. J Ferment Bioeng 79:538–544

    Article  Google Scholar 

  34. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Anal Chem 28:350–356

    Article  Google Scholar 

  35. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem 31:426

    Article  Google Scholar 

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

    Google Scholar 

  37. Garrity GM, Bell JA, Lilburn T (2005) The revised road map to the manual. In Brenner, Krieg, Staley and Garrity (Eds.), Bergey’s manual of systematic bacteriology. The Proteobacteria, part A, introductory essays. Springer, 2 (2) pp 159–220

  38. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.

    Google Scholar 

  39. Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from microorganisms. J Mol Biol 3:208–218

    Article  Google Scholar 

  40. Babu TG, Nithyanand P, Babu NKC, Pandian SK (2009) Evaluation of cetyltrimethylammonium bromide as a potential short-term preservative agent for stripped goat skin. World J Microbiol Biotechnol 25:901–907

    Article  Google Scholar 

  41. Kanagasabhapathy M, Sasaki H, Nagata S (2008) Phylogenetic identification of epibiotic bacteria possessing antimicrobial activities isolated from red algal species of Japan. World J Microbiol Biotechnol 24:2315–2321

    Article  Google Scholar 

  42. Wei N, Quartreman J, Jin YS (2013) Marine macroalgae: an untapped resource for producing fuels and chemicals. Trends Biotechnol 31(2):70–77

    Article  Google Scholar 

  43. Ohta Y, Hatada Y, Nogi Y, Miyazaki M, Li Z, Akita M, Hidaka Y, Goda S, Ito S, Horikoshi K (2004) Enzymatic properties and nucleotide and amino acid sequences of a thermostable β-agarase from a novel species of deep-sea Microbulbifer. Appl Microbiol Biotechnol 64:505–514

    Article  Google Scholar 

  44. Uchimura K, Miyazaki M, Nogi Y, Kobayashi T, Horikoshi K (2010) Cloning and sequencing of alginate lyase genes from deep-sea strains of Vibrio and Agarivorans and characterization of a new Vibrio enzyme. Mar Biotechnol 12:526–533

    Article  Google Scholar 

  45. Kawamoto H, Horibe A, Miki Y, Kimura T, Tanaka K, Nakagawa T, Kawamukai M, Matsuda H (2006) Cloning and sequencing analysis of alginate lyase genes from the marine bacterium Vibrio sp. O2. J Mar Biotechnol 8:481–490

    Article  Google Scholar 

  46. Aoki T, Araki T, Kitamikado M (1990) Purification and characterization of a novel beta-agarase from Vibrio sp. AP-2. Eur J Biochem 187:61–465

    Article  Google Scholar 

  47. Kim HS, Lee C-G, Lee EY (2008) Alginate lyase: structure, property, and application. Biotechnol Bioprocess Eng 16:843–851

    Article  Google Scholar 

  48. Jonnadula RC, Verma P, Shouche YS, Ghadi SC (2009) Characterization of Microbulbifer strain CMC-5, a new biochemical variant of Microbubifer elongatus type strain DSM6810Tisolation from decomposing seaweeds. Curr Microbiol 59:600–607

    Article  Google Scholar 

  49. Tang JC, Taniguchi H, Chu H, Zhou Q, Nagata S (2009) Isolation and characterization of alginate-degrading bacteria for disposal of seaweed wastes. Lett Appl Microbiol 48:38–43

    Article  Google Scholar 

  50. Alexeeva YV, Ivanova EP, Zvaygintseva TN, Mikhailov VV (2002) Optimization of glycosidases production by Pseudoalteromonas issachenkonii KMM 3549. Lett Appl Microbiol 35:343–346

    Article  Google Scholar 

  51. Li JW, Dong S, Song J, Li CB, Chen XL, Xie BB, Zhang YZ (2011) Purification and characterization of a bifunctional alginate lyase from Pseudoalteromonas sp. SM0524. Mar Drugs 9(1):109–123

  52. Moen E, Horn S, Østgaard K (1997) Alginate degradation during anaerobic digestion of Laminaria hyperborea stipes. J Appl Phycol 9:157–166

    Article  Google Scholar 

  53. Doubet RS, Quatrano RS (1984) Properties of alginate lyases from marine bacteria. Appl Environ Microbiol 47(4):699–703

    Google Scholar 

  54. Turovskiy Y, Kashtanov D, Paskhover B, Chikindas ML (2007) Quorum sensing: fact, fiction, and everything in between. Adv Appl Microbiol 62:191–234

    Article  Google Scholar 

  55. Thomas F, Lundqvist LC, Jam M, Jeudy A, Barbeyron T, Sandström C, Czjzek M (2013) Comparative characterization of two marine alginate lyases from Zobellia galactanivorans reveals distinct modes of action and exquisite adaptation to their natural substrate. J Biol Chem 288(32):23021–23037

    Article  Google Scholar 

  56. Moen E, Østgaard K (1997) Aerobic digestion of Ca-alginate gels studied as a model system of seaweed tissue degradation. J Appl Phycol 9:261–267

    Article  Google Scholar 

  57. Xu Y, Tian XP, Liu YJ, Li J, Kim CJ, Yin H, Li WJ, Zhang S (2013) Sinomicrobium oceani gen. nov., sp. nov., a member of the family Flavobacteriaceae isolated from marine sediment. Int J Syst Evol Microbiol 63(Pt 3):1045–1050

    Article  Google Scholar 

  58. Cho H, Kim H, Kim Y (2012) Isolation and characterization of novel alginate-degrading Pseudoalteromonas sp. Y-4. Fish Aquat Sci 15(3):259–263

    Google Scholar 

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Acknowledgments

The authors thank the Department of Science and Technology, New Delhi for financial support (ref. no. DST/TSG/AF/2010/12) and Shri AMM Murugappa Chettiar Research Centre and Alagappa Government Arts and Science College (Alagappa University) for providing the laboratory facilities.

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Authors and Affiliations

  1. Marine Algae Research Division, Post Graduate and Research Department of Botany, Alagappa Government Arts and Science College (Alagappa University), Karaikudi, Tamil Nadu, 630003, India

    A. Jegatheesan & C. Poonam

  2. Shri AMM Murugappa Chettiar Research Centre (MCRC), Taramani, Chennai, Tamil Nadu, 600113, India

    M. P. Sudhakar & K. Perumal

  3. Department of Plant Science, Central University of Kerala, Riverside Transit Campus, Padanakkad, Nileshwar, Kerala, 671314, India

    K. Arunkumar

Authors
  1. A. Jegatheesan
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  2. M. P. Sudhakar
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  3. C. Poonam
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  4. K. Perumal
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  5. K. Arunkumar
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Corresponding author

Correspondence to K. Arunkumar.

Electronic supplementary material

Table 1

Alginolytic bacteria isolated from brown seaweeds, sediments and industrial wastes. (DOCX 12 kb)

Table 2

Reagents for DNA extraction. (DOCX 11 kb)

Plate 1

Bacterial isolates AAl02 and AAl04 growing on basal medium supplemented with alginate (1 %) forming lesions as a result of alginate degradation: (a) isolation of alginate-degrading bacteria by spread plating method, (b) isolation of pure culture (bacteria) degrading alginate (c)AAl02 and (d)AAl04 activity on alginate containing plates. (DOCX 115 kb)

Figure 1

Phyllogenetic analysis of 16S rRNA sequence of promising alginolytic bacteria (a) AAl 02 (b) AAl 04 isolated from the Sargassum longifolium industrial waste. (DOCX 77 kb)

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Jegatheesan, A., Sudhakar, M.P., Poonam, C. et al. Isolation and characterization of alginate-degrading bacteria Sinomicrobium oceani . Biomass Conv. Bioref. 7, 51–58 (2017). https://doi.org/10.1007/s13399-016-0212-z

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