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Characterization and application of a native lactic acid bacterium isolated from tannery fleshings for fermentative bioconversion of tannery fleshings

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Lactic acid bacteria (LAB) species isolated from limed and delimed tannery fleshings (TF) were evaluated for their fermentation efficiency and antibacterial property. The native LAB isolates efficiently fermented TF and resulted in a fermented mass with antioxidant properties, indicating their potential for effective eco-friendly bioconversion of TF. From among the LAB isolated, a proteolytic isolate showing better antimicrobial spectrum and reasonably good fermentation efficiency was identified as Enterococcus faecium HAB01 based on various biochemical and molecular tests. This isolate afforded a better degree of hydrolysis (81.36%) of TF than Pediococcus acidilactici (54.64%) that was previously reported by us. The bacteriocin produced by E. faecium was found to be antagonistic to several human pathogens including Listeria, Aeromonas, Staphylococcus and Salmonella. Further, E. faecium HAB01 bacteriocin was thermostable and had a molecular weight of around 5 kDa, apart from being stable at both acidic and alkaline conditions. The bacteriocin was unstable against proteases.

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  1. Ahmed J, Mahendrakar NS (1995) Effect of different levels of molasses and salt on acid production and volume of fermenting mass during ensiling of tropical freshwater fish viscera. J Food Sci Technol 32:115–118

  2. AOAC (1995) Official methods of analysis, Ch.12, 16th edn. Horowitz, Washington, DC, USA

  3. Arizcun C, Barcina Y, Torre P (1997) Identification and characterization of proteolytic activity of Enterococcus spp. isolated from milk and Roncal and Idiazábal cheese. Int J Food Microbiol 38:17–24

  4. Bhaskar N, Sakhare PZ, Suresh PV, Lalitha RG, Mahendrakar NS (2007) Biostabilization and preparation of protein hydrolysates from delimed leather fleshings. J Sci Ind Res 66:1054–1063

  5. Burianek LL, Yousef AE (2000) Solvent extraction of bacteriocins from liquid cultures. Lett Appl Microbiol 31:193–197

  6. Cotelle N, Bemier JL, Catteau JP, Pommery J, Wallet JC, Gaydou EM (1996) Antioxidant properties of hydroxyl flavones. Free Radic Biol Med 20:35–43

  7. Duan XJ, Zhang WW, Li XM, Wang BG (2006) Evaluation of antioxidant property of extract and fractions obtained from a red alga, Polysiphonia urceolata. Food Chem 95:37–43

  8. Edward AS, Boris VE, Vladimir VP, Evgini VM, Irina PM, Valery NB, Vladimir PL, Olga ES, Yuri NK, Yuri GS, Gregory RS, Bruce SS, Norman JS (2008) Diverse antimicrobial killing by Enterococcus faecium E 50-52 bacteriocin. J Agric Food Chem 56:1942–1948

  9. Franz CMAP, Stiles ME, Schleifer KH, Holzapfel WH (2003) Enterococci in food— a conundrum for food safety. Int J Food Microbiol 88:105–122

  10. Franz CMAP, Van Belkum MJ, Holzapfel WH, Abriovel H, Galvez A (2007) Diversity of enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev 31:293–310

  11. Ganesan P, Chandini SK, Bhaskar N (2008) Antioxidant properties of methanol extract and its fractions obtained from selected Indian red seaweeds. Bioresour Technol 99:2717–2723

  12. Geis AJ, Singh R, Teuber MJ (1983) Potential of lactic streptococci to produce bacteriocin. Appl Environ Microbiol 45:205–211

  13. Halami PM, Ramesh A, Chandrashekhar A (2000) Megaplasmid encoding novel sugar utilizing phenotypes, pediocin production and immunity in Pediococcus acidilactici C20. Food Microbiol 17:475–483

  14. Halami PM, Ramesh A, Chandrashekhar A (2005) Fermenting cucumber, a potential source for the isolation of pediocin-type bacteriocin producer. World J Microbiol Biotechnol 21:1351–1358

  15. He H, Chen X, Sun C, Zhang Y, Gao P (2006) Preparation and functional evaluation of oligopeptide rich hydrolysate from shrimp (Acetes chinensis) treated with crude protease from Bacillus sp. SM98011. Bioresour Technol 97:385–390

  16. Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST (1994) Bergey's manual of determinative bacteriology, 9th edn. William and Wilkins, MD, pp 559–564

  17. Hoyle NT, Merritt JH (1994) Quality of fish protein hydrolysate from Herring (Clupea harengus). J Food Sci 59:76–79

  18. Kang JH, Lee MS (2004) Characterization of a bacteriocin produced by Enterococcus faecium GM-1 isolated from an infant. J Appl Microbiol 98:1169–1176

  19. Kimura M, Danno K, Yasui H (2006) Immunomodulatory function and probiotic properties of lactic acid bacteria isolated from Mongolian fermented milk. Biosci Microflora 25:147–155

  20. Kumar AG, Swarnalatha S, Sairam B, Sekaran G (2008a) Production of alkaline protease by Pseudomonous aeruginosa using proteinaceous solid waste generated from leather manufacturing industries. Bioresour Technol 99:1939–1935

  21. Kumar AG, Nagesh N, Prabhakar TG, Sekaran G (2008b) Purification of extracellular acid protease and analysis of fermentation metabolites by Synergistes sp. utilizing proteinaceous solid waste from tanneries. Bioresour Technol 99:2364–2372

  22. Nes IF, Diep DB, Holo H (2007) Bacteriocin diversity in Streptococcus and Enterococcus. J Bacteriol 189:1189–1198

  23. Oyedapo AF (1996) Preparation, properties and preservation of lactic acid fermented shrimp heads. Food Res Int 29:595–599

  24. Park SJ, Lim DS, Yoon SK, Baek YJ, Kim CH (1998) Isolation and identification of Enterococcus faecalis 2B4-1 containing antitumor substances. Korean J Appl Biotechnol 26:471–475

  25. Prieto P, Pineda M, Aguilar M (1999) Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 269:337–341

  26. Reuter G (1997) Present and future of probiotics in Germany and in Central Europe. Biosci Microflora 16:43–51

  27. Sachindra NM, Bhaskar N (2008) In-vitro antioxidant activity of liquor from fermented shrimp biowaste. Bioresour Technol 99:9013–9016

  28. Schagger H, Von Jagow G (1987) Tricine-sodium deodecyl sulphate polyacrylamide gel electrophoresis for the separation of proteins in the range from 1–100 kDa. Anal Biochem 166:368–379

  29. Shaw DM, Narasimha Rao D, Mahendrakar NS (1998) Rapid fermentation for ensiling of poultry intestines. Bioresour Technol 65:247–249

  30. Sambrook J, Russell DW (2001) Molecular cloning. A laboratory manual, vol I, 3rd edn. Cold Spring Harbor Laboratory Press, New York, USA

  31. Yashoda KP, Sachindra NM, Narasimha Rao D, Mahendrakar NS (2001) Changes in microbial population during fermentation of silkworm pupae. Sericologia 41:253–261

  32. Zakaria Z, Hall GM, Shama G (1998) Lactic acid fermentation of scampi waste in a rotating horizontal bioreactor for chitin recovery. Process Biochem 33:1–6

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This work is supported by funding from Council of Scientific and Industrial Research (CSIR) through the network programme (NWP-044) on Zero Emission Research Initiatives (ZERI). We thankfully acknowledge Mr. Masood Ahmed—Proprietor of M/s Mysore Super Reptiles Corporation, Bangalore (Karnataka, India)—for the gratis supply of limed TF. Authors thank Dr V Prakash, Director, CFTRI, for encouragement and permission to publish the work.

Conflict of interest statement

The authors declare that they have no conflict of interest.

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Correspondence to N. Bhaskar.

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Rai, A.K., Bhaskar, N., Halami, P.M. et al. Characterization and application of a native lactic acid bacterium isolated from tannery fleshings for fermentative bioconversion of tannery fleshings. Appl Microbiol Biotechnol 83, 757–766 (2009). https://doi.org/10.1007/s00253-009-1970-3

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  • Tannery fleshings
  • Lactic acid bacteria
  • Antioxidant activity
  • Bacteriocin
  • Fermentation