Enrichment And Characterization Of Limonin Degrading  Microorganisms Isolated From Kinnow Orchard Soil,  Peel And Waste Site

Kusum Dua; Gurvinder Singh Kocher

doi:10.48165/

Authors

Kusum Dua Department of Microbiology, Punjab Agricultural University, Ludhiana – 140 000, Punjab (India)
Gurvinder Singh Kocher Department of Microbiology, Punjab Agricultural University, Ludhiana – 140 000, Punjab (India)

DOI:

https://doi.org/10.48165/

Keywords:

Bacillus megaterium, degradation, immobilized cells, limonin, Meyerozyma caribbica

Abstract

The present research was aimed to screen and characterize limonin degrading microorganisms and assess their efficiency in limonin reduction. ight bacterial isolates from kinnow orchard soil and two yeast isolates from soft kinnow peel and kinnow waste site (Abohar, Punjab) were isolated. All the isolates were evaluated for their limonin utilizing ability by swarm plate assay. Two bacterial isolates (S1 and S2) and both yeast isolates (PY1 and AY1) showed considerable zone of TTC reduction. All the four selected isolates were studied for their growth profile and rate of limonin degradation in minimal media broth with limonin (28 ppm) as a sole carbon source. Bacterial isolate S1 showed maximum limonin degradation (52.9%), followed by yeast isolates PY1 (48.5%), AY1 (31.1%) and bacterial isolate S2 (26.4%). Ca-alginate immobilized cells of PY1 yeast and S1 bacteria were also studied for their limonin degrading ability which respectively showed 94.8 and 98% limonin reduction. The two microbial isolates were molecularly identified as Bacillus megaterium strain KU570370.1. on 16SrDNA basis and Meyerozyma caribbica isolate KP860076.1. on 18S rDNA basis These microbial isolates could be exploited for the production of enzyme limonin dehydrogenase for possible use in debittering kinnow juice.

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References

Barnett, J.A., Payne, R.W. and Yarrow, D. 2000. Yeasts: Characteristics and Identification. Cambridge University Press, Cambridge, England.

Chang, W.C., Chen, M.W. and Lee, T.M. 1999. 2,3,5-triphenyltetrazolium reductions in the viability assay of Ulva fasciata (Chlorophyta) in response to salinity stress. Botanical Bsssulletin of Academia Sinica. 40(3): 207-212.

Chibata, I. and Tosa, T. 1977. Transformation of organic compounds by immobilized microbial cells. Advances in Applied Microbiology, 22: 1-27.

Dua, K. and Kocher, G.S. 2017. Fermentative processing of kinnow juice and extraction of limonin from kinnow waste. Current Trends in Biomedical Engineering and Biosciences, 4(3): 1-4. Hasegawa, S., Bennett, R.D., Maier, V.P. and King, A.D. 1972. Limonoate dehydrogenase from Arthrobacter globiformis. Journal of Agricultural and Food Chemistry, 20: 1031-1034. Hasegawa, S., Maier, V.P. and King, A.D. 1974. Isolation of new limonoate dehydrogenase from Pseudomonas. Journal of Agricultural and Food Chemistry, 22: 523-526.

Hasegawa, S., Pelton, V.A. and Bennet, R.D. 1983. Metabolism of limonoids by Arthrobacter globiformis. II. Basis for practical means of reducing the limonin content of orange juice by immoblized cells. Journal of Agricultural Food Chemistry, 31: 102-107.

Hasegawa, S., Vandercook, C.E., Choi G.Y., Herman, Z. and Ou, P. 1985. Limonoid debittering of citrus juice sera by immobilized cells of Corynebacterium fascians. Journal of Food Science, 50: 330-332.

Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J T. and Williams, S.T. 1994. Bergey's Manual of Determinative Bacteriology. Williams & Wilkins, Baltimore, USA.

Humanes, L., López-Ruiz, A., Merino, M.T., Roldán, J.M. and Diez, J. 1997. Purification and characterization of limonoate dehydrogenase from Rhodococcus fascians. Applied and Environmental Microbiology, 63: 3385-3389.

Iborra, J.L., Manjo´n, A., Ca´novas, M., Lozano, P. and Martı´nez, C. 1994. Continuous limonin degradation by immobilized Rhodococcus fascians cells in K. carragenaan. Applied Microbiology and Biotechnology, 41: 487-493.

Jain, S. 2003. Molecular and Physiological Studies on Pseudomonas putida (Thesis). Department of Biotechnology and Environmental Sciences, Thapar Institute of Engineering and Technology, Patiala, India.

Kimura, M. 1980. A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16: 111-120. Kreger-Van Rij, N.J.W. 1984. The Yeast a Taxonomic Study. Elsevier Science Publ., New York, USA.

Kurtzman, C.P., Fell, J.W. and Boekhout, T. 2011. The Yeasts: A Taxonomic Study (5th edn.). Elsevier Science Publ., Amsterdam, Netherland.

Lazarova, V. and Manem, J. 1995. Biofilm characterization and activity analysis in water and wastewater treatment. Water Research, 29: 2227-2245.

Malik, M and Ghosh, M. 2012. Immobilization parameters statistically optimized for whole cells of P. putida G7 to enhance limonin biotransformation. Journal of Advanced Laboratory Research in Biology, 3: 266-275.

Maniatis, T., Fritsch, E F. and Sambrook, J. 1982. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.

Kusum Dua and G.S. Kocher

Moat, A.G., Foster, J.W. and Spector, M.P. 2002. Microbial Physiology (4th edn), John Wiley, New York, USA.

Nagra, R.P. and Puri, M. 2010. Screening and characterization of limonin hydrolyzing microorganism for production of limonin dehydrogenase. pp. 170. In: National Conference on Recent Advances in Chemical and Environmental Sciences. Multani Mal Modi College, Patiala, Punjab, India.

Punjab Horticulture Mission. 2015. Punjab Horticulture Mission. 2014-2015. (http://punjab horticulture.com/wp-content/themes/twentyeleven/document/fruits/Fruits%2020 14 -15.pdf). Puri, M., Kaur, L. and Marwaha, S.S. 2002. Partial purification and characterization of limonoate dehydrogenase from Rhodococcus fascians for the degradation of limonin. Applied Microbiology and Biotechnology, 12: 669-673.

Tamura, K., Dudley, J., Nei, M. and Kumar, S. 2007. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 24: 1596-1599. Vaks, B. and Lifshitz, A. 1981. Debittering of orange juice by bacteria which degrade limonin. Journal of Agricultural and Food Chemistry, 29: 1258-1261.

Van der Walt, J.P and Yarrow, D. 1984. Methods for the isolation, maintenance, classification and identification of yeasts. pp. 45-105. In: The Yeasts: A Taxonomic Study (ed. N.J. Kreger-van Rij). Elsevier Science Publ., Amsterdam, Netherland.

Verma, J.P., Singh, S., Shrivastava, P.K. and Ghosh, M. 2010. Identification and characterization of cellular locus of limonin biotransforming enzyme in Pseudomonas putida. International Journal of Food Science and Technology, 45: 319-326

Yarrow, D. 2000. Methods for isolation, maintenance and identification of yeasts. pp. 77-100. In: The Yeasts: A Taxonomic Study (eds. C.P. Kurtzman and J.W. Fell). Elsevier Science Publ., Amsterdam. Netherland.