Yeast Isolates From Diverse Sources For Cold-Active Polygalacturonase And Amylase Production
K. Divya, P. Naga Padma
Keywords: Amylase, cold-active enzyme, Geotrichum sps, polygalacturonase, ruthenium red, screening, yeasts.
Abstract: Cold–active polygalacturonase and amylase producers were screened using enrichment culture technique. The diverse sources screened were cold stored spoilt fruits and vegetables from different local super markets, market waste dumped soils, fruit waste dumped soils, mountain soils and Himalayan soils. About sixty yeasts showing pectinolytic activity were isolated by ruthenium red plate assay. Eight yeasts with higher zones of pectin hydrolysis were selected and tested for cold-active polygalacturonase and amylase production. The cultures were tested for cold active pectinase and amylase enzyme activity by dinitrosalicylic acid (DNS) method. The cultures were grown at both room temperature (20-25 °C) and cold temperatures (5°C) but the cold active enzyme activity was tested at 5°C. Highest cold-active pectinase producing yeast culture with good cold-active amylase activity was selected for further study. Thus the present cold-active polygalacturonase producer with amylase activity could have better application in fruit juice clarification and so could be a potential isolate.
 I. Alkorta, C. Garbisu, M.J. Llama and J.L. Serra, “Industrial Applications of Pectic Enzymes: A Review”. Process Biochemistry. 33, 21-28, 1998.
 K. Anuradha, P. Naga Padma, S. Venkateshwar and Gopal Reddy, “Fungal Isolates from Natural Pectic Substrates for Polygalaturonase and Multienzyme production”. Indian Journal of Microbiology. 50, 339-344, 2010.
 H. Birgisson, O. Delgado, L. Garcia Arroyo, R. Hatti-kaul, & B. Mattiasson, “Cold-adapted yeasts as producers of cold-active polygalacturonases”. Extremophiles. 7, 185−193, 2003.
 P. Blanco, C. Sieiro, A. Diaz, & T.G. Villa, “Production and partial characterization of an endo-polygalacturonase from Saccharomyces cerevisiae”. Canadian Journal of Microbiology. 40, 974−977, 1994.
 L. Ceci, & J. Lozano, “Determination of enzymatic activities of commercial pectinases for the clarification of apple juice”. Food Chemistry. 61, 237-241, 1998.
 A. Collmer, J.L. Reid, & M.S. Mount, “Assay procedures for pectic enzymes”. Methods in Enzymology. 161, 329-335, 1988.
 A. Gainvors, N. Karam, C. Lequart and A. Berarbi, “Use of Saccharomyces cerevisiae for the clarification of fruit juices”. Biotechnology Letters. 16, 1329-1334, 1994.
 A.S. Ismail, “Utilization of orange peels for the production of multienzyme complexes by some fungal strains”. Proc. Biochem. 31: 645-650, 1996.
 R.S. Jayani, S. Saxena, and R. Gupta, “Microbial pectinolytic enzymes: A review”. Process Biochemistry. 40, 2931-2944, 2005.
 J.L. Loo, O.M. Lai, K. Long. K and H.M. Ghazali, “Identification and Characterisation of a Locally Isolated Lipolytic Microfungus - Geotrichum candidum”. Malaysian Journal of Microbiology, Vol 2(1), pp.22-29, 2006.
 R. Margesin, & F. Schinner, “Properties of cold-adapted microorganisms and their potential role in biotechnology”. Journal of Biotechnology. 33, 1-14, 1994.
 G.L. Miller, “Use of dinitrosalicylic acid reagent for determination of reducing sugar”. Anal Chem 31: 426-428, 1959.
 T. Nakagawa, T. Nagaoka, S. Tanuguchi, T. Miyaji and N. Tomizuka, “Cold-active Pectinases from psycrophilic and pectinolytic Yeasts: Isolation, enzymatic properties and applications”. (pp. 1-16): Research Signpost, 2005.
 T. Nakagawa, T. Nagaoka, T. Miyaji and N. Tomizuka, (2005). “Cold-Active Polygalacturonase from Psychrophilic-Basidiomycetous Yeast Cystofilobasidium strain PYY-1”. Bioscience Biotechnolology Biochemistry. 69 (2), 419-421, 2005.
 P. Naga Padma, K. Anuradha, and Gopal reddy, “Pecticnolytic yeast isolates for cold active polygalactouronase production”. Innovative food Science and emerging technologies. 12, 178-181, 2011.
 Natalia Lorena Rojas, Sebastián Fernando, Cavalitto Carlos Fernando Mignone, Roque Alberto Hours., (2008). “Role of PPase-SE in Geotrichum klebahnii, a yeast-like fungus able to solubilize pectin”. Electronic Journal of Biotechnology ISSN: 0717-3458 Vol 11 No. 1, 2008.
 D.B. Pedrolli, E. Gomes, R. Monti, E.C. Carmona, “Studies on productivity and characterization of polygalacturonase from Aspergillus giganteus submerged culture using citrus pectin and orange waste”. Appl Biochem Biotechnol 144: 191-200, 2008.
 M. Roohi.M, I. Kuddus, Z. Ahmad and J.M. Arif, “Production of cold –active extracellular α- amylase by newly isolated Microbacterium foliorum”. Asian journal of biotechnology, 3:449-459, 2011.
 Y. Sudheer Kumar, S.P. Reddy and O.V.S. Reddy, “Optimization of fermentation conditions for Mango (Mangifera indica L) wine production by employing response surface methodology”. International Journal of Food Science and Technology, 44: 2320-2327, 2009.
 Sandeep Singh and Reena Gupta,“Apple juice clarification using fungal pectinolytic enzyme and gelatin”. Indian Journal of Biotechnology, 3: 573-576, 2004.
 SF. Cavalitto, RA. Hours, and CF. Mignone, (2000). “Growth and protopectinase production of Geotrichum klebahnii in batch and continuous cultures with synthetic media”. Journal of Industrial Microbiology & Biotechnology. 25, 260-265, 2000.
 Wen-Chi Hou, Wei-Hsien Chang and Chii-Ming Jiang, “Qualitative distinction of carbozyl group distributions in pectin with ruthenium red”. Bot Bull Acad Sin 40: 115-119, 1999.
 J.R. Whitaker, (1984). “Pectic substances, pectic enzymes and haze formation in fruit juices”. Enzyme Microbial Technology. 6, 341-349, 1984.
 Y.D. Hang, E.E. Woodams, (1992) “Production and characterization of polygalacturonase from Geotrichum candidum” World Journal of Microbiology and Biotechnology, 1992.