Isolation of lactic acid bacteria from sugar cane juice and production of lactic acid from selected improved strains

Yoganand Sobrun, Archana Bhaw-Luximon, Dhanjay Jhurry, Daneshwar Puchooa

ANDI Centre of Excellence for Biomedical and Biomaterials Research, University of Mauritius, Réduit, MAURITIUS.
Faculty of Agriculture, University of Mauritius, Réduit, MAURITIUS.

DOI: 10.4236/abb.2012.34057                

Abstract

Lactic acid bacteria (LAB) were isolated from fresh sugar cane juice. It was found that several isolates exhibited a clear zone and growth on deMan, Rogosa, Sharpe (MRS) agar supplemented with sodium azide, bromocresol purple and sucrose. However, only 17 isolates which formed large yellow areas were selected for further investigations. These isolates were further identified according to their morphological and biochemical characteristics. It was found that 10 of these isolates were homofermenters. One of these 10 isolates was selected for mutagenesis using chemical (Ethidium bromide) and physical (UV-B) mutagens followed by biochemical characterisation. A total of 112 mutants were isolated and 9 homofermentative isolates were further investigated for their ability to produce lactic acid. 1H-NMR spectroscopy confirmed that all mutant isolates produced lactic acid as the sole fermentation product.

Keywords

Lactic Acid Bacteria; Sugar Cane Juice; Mutagenesis

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Oshiro M., Shinto H., Tashiro Y., Miwa N., Sekiguchi T., Okamoto M., Ishizaki A., Sonomoto K. 2009 Kinetic modeling and sensitivity analysis of xylose metabolism in Lactococcus lactis IO-1. J. Biosci. Bioeng. 108, 376–384.
[2]	Singhvi M., Joshi D., Adsul M., Varma A., Gokhale D. 2010 d-(-)-Lactic acid production from cellobiose and cellulose by Lactobacillus lactis mutant RM2-24. Green Chem. 12, 1106–1109.
[3]	Tashiro Y., Kaneko W., Sun Y., Shibata K., Inokuma K., Zendo T., Sonomoto K. 2011 Continuous d-lactic acid production by a novel thermotolerant Lactobacillus delbrueckii subsp lactis QU 41. Appl. Microbiol. Biotechnol. 89, 1741–1750.
[4]	Adnan A. F. M. and Tan I. K. P. (2007) Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresource Technology 98 (2007) 1380–1385.
[5]	Hofvendahl K. and Hahn-Hagerdal B. 2000 Factors affecting the fermentative lactic acid production from renewable resources. Enzyme Microb Technol. 26 (2-4), 87-107.
[6]	Ilmen M., Koivuranta K., Ruohonen L., Suominen P., Penttila M. 2007. Efficient production of l-lactic acid from xylose by Pichia stipitis. Appl. Environ. Microbiol. 73, 117–123.
[7]	Pandey A., Soccol C.R., Rodriguez-Leon J.A., Nigam, S. 2001 Production of organic acids by solid state fermentation. In: solid state fermentation in biotechnology: fundamentals and applications. Asiatech Publishers, New Delhi, p. 127.
[8]	Kharras G. B., Sanchez-Riera F., Severson D. K. 1993 Polymers of lactic acid. In: Molby, D.B. (Ed.), Plastics from microbes: Microbial synthesis of polymers and polymer precursors. Hanser Publ., pp. 93–137.
[9]	Tsai S. P., Coleman R. D., Moon S. H., Schneider K. A., Millard C. S. 1993 Strain screening and development for lactic acid fermentation. Appl. Biochem. Biotechnol. (39/40), 323–335.
[10]	Parekh S., Vinci V. A., Strobel R. J. 2000 Improvement of microbial strains and fermentation processes, Appl. Microbiol. Biotechnol. 54, 287–301.
[11]	de Man J. C., Rogosa M., Sharpe M. E.1960 Medium for the cultivation of lactobacilli. J. Appl. Bacteriol. 23, 130–135. J. Appl. Bacteriol. 23, 130–135.
[12]	Sperber W. H. and Swan J. 1976 Hot-Loop Test for the Determination of Carbon Dioxide Production from Glucose by Lactic Acid Bacteria. Applied & Environmental Microbiology, Vol. 31 (6), 990-991.
[13]	Gawel, D.; Maliszewska-Tkacyk, M.; Jonczyk, P.; Schaaper, R. M. and Fijalkowska, I.J. (2002). Lack of Strand Bias in UV-Induced Mutagenesis in Escherichia coli. Bacteriology. 184 (16), 4449-4454.
[14]	Ramanjooloo A., Bhaw-Luximon A., Jhurry D and Cadet F. (2009) 1H NMR Quantitative of Lactic Acid Produced by Biofermentation of Cane Sugar Juice. Spectroscopy Letters, 42, 296–304.
[15]	Pullman A. and Pullman B. 1963 The mechanism of ultraviolet-induced mutations. Biochimica et Biophysica Acta 75, 269–271.
[16]	Rohwerder T., Gehrke T., Kinzler K., Sand W. 2003 Bioleaching review part A progress in bioleaching: fundamentals and mechanisms of bacterial metal sulfide oxidation. Applied Microbiology and Biotechnology 63 (3), 239–248.
[17]	Yang Y., Zhang S., Xu A. L., Zou L. H., Li L., Qiu G. Z. 2010 UV-induced mutagenesis and bioleaching of Acidiphilium cryptum and Acidithiobacillus ferrooxidans. Journal of Central South University (Science and Technology) 41 (2), 393–399.
[18]	Beggs C. B. 2002 A quantitative method for evaluating the photoreactivation of ultraviolet damaged microorganisms. Photochemistry and Photobiology Science 1 (6), 431–437.
[19]	Slonimski P. P., Perrodin G, and Croft J. H. 1968 Ethidium bromide induced mutation of yeast mitochondria: complete transformation of cells into respiratory deficient non-chromosomal "petites." Biochem. Biophys. Res. Commun. 30, 232-239.
[20]	Rubin S. J. and Rosenblum E. D. 1971. Effects of ethidium bromide on growth and on loss of the penicillinase plasmid of Staphylococcus aureus. J. Bacteriol. 108, 1200-1204.
[21]	Tomchick R. and Mandel H. G. 1964 Biochemical effects of ethidium bromide in microorganisms. J. Gen. Microbiol. 36, 225-236.
[22]	Maillet S. 1969 Action mutagene specifique de trois clorants, sur la mutation reverse d'un gene ProA- chez E. coli K12. C.R. Acad. Sci. 269, 1708-1711.
[23]	Waring M. J. 1966. Structural requirements for the binding of ethidium to nucleic acids. Biochim. Biophys. Acta 114, 234-244.
[24]	Gitler, C., Rubalcava B. and Caswell A. 1969 Fluorescence changes of ethidium bromide on binding to erthrocyte and mitochondrial membranes. Biochim. Biophys. Acta 193, 479-481.
[25]	Yu M, Yurong S, Wei X, Yang Y, Zhou Y, Hao X, Zhang N and Niu R (2007). Depletion of mitochondrial DNA by ethidium bromide treatment inhibits the proliferation and tumorigenesis of T47D human breast cancer cells. Toxicology Letters 170, 83-93
[26]	Goncalves, L.M.D.; Ramos, A.; Almeida, J.C.; Xavier, A.M.R.B. and Carrondo, M.J.T. (1997). Elucidation of the mechanism of lactic acid growth inhibition and production in batch cultures of Lactobacillus rhamnosus. Applied Microbiology and Biotechnology 48, 346-350.
[27]	Kotzamanidis, Ch.; Roukas, T. and Skaracis, G. (2002). Optimization of lactic acid production from beet molasses by Lactobacillus delbrueckii NCIMB 8130. World journal of Microbiology and Biotechnology 18, 441-448.