Isolation and identification of phytase producing fungi for extraction and use of enzymes in animal feed and poultry

Document Type : Research Paper

Authors

1 Member of scientific board of ASRI

2 Razi Vaccine and Serum Research Institute.Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

Abstract

In the present study, native fungi producing phytase from soil and other environmental samples (Includes: soil around the root of leguminous plants, Cows and goats excrement, Chicken floor, Vermicompost, Mineral phosphorus mine, Sewage water, Plants and fruits infected with fungi and Samples prepared from research centers of the country)were isolated and incubated for 5 days at 28 ° C on SDA(Saburad Dextrose Agar). Then a phytase screening medium (PSM) was used to identify and isolate fungi capable of producing phytase enzyme. Morphological characteristics of fungal strains were studied using optical microscopy.Samples which produced clear halo were chosen for enzymatic activity assay based on the release of a phosphorus nanomol in one minute. Based on the amount of enzyme and amount of biomass production, were evaluated by the respective isolates.The optimum pH and temperature for maximum production of enzyme through the selected isolates were 5.5 and 30 °C, respectively. Finally, the isolates were identified as Fusariumoxysporum and Aspergillus Niger using Genetic analyzer system. The maximum amount of enzyme production was recorded as 130.11 and 125.27 enzymatic units for Fusarium oxysporum and Aspergillus Niger, respectively. The results showed that from five to seven days during enzyme culture are the most suitable time for enzyme extraction and purification.The results of morphological studies were completely consistent with the molecular identification results

Keywords

References

آهنگی، ز.شجاع ساداتی، س.ع. نیکوپور، ه. (1385).تولید پروتئین قارچی با استفاده از فوزاریوم اگزیسپوروم PTCC 5115. مجله علوم تغذیه و صنایع غذایی ایران. دوره 1، شماره 2.
 سجادی، گ. شجاعی، ا. فاضلی، م . امینی، ج جمالی فر، ح.(1388) . تولید برون سلولی نانوذرات نقره به وسیله قارچ فوزاریوم اگزیسپوروم در مقیاس آزمایشگاهی. نشریه دنیای میکروب ها. دوره 2، شماره 1.
Anna, l., Tania, C., Okcha, L.and  Keiyh, S. ( 2008). Colony Multiplex-Tandem PCR for Rapid, Accurate Identification of [10] Fungal Cultures.J Clin Microbiol.( 46): 4058-4060.
Bae , H.D., Yanke, L.J., Cheng , K. J. and SELINGER, L.B.( 1999). A novel staining method for detecting phytase activity. Journal of Microbiological Methods.( 39): 17-22.
Bijender, S.( 2012). Production of phytate-hydrolyzing enzymes by thermophilic moulds. African Journal of Biotechnology. 11, 59. DOI= http://dx.doi.org/10.5897/ajb12.695.
Brinch-Pedersen, H., Sørensen, L. and Holm, P.( 2002). Engineering crop plants getting a handle on phosphate. Trend in Plant Science-Cell. 7, 3, 118-125.
 
Bu, Y., Huang, H., Zhou, G.( 2008). Direct polymerase chain reaction (PCR) from human whole blood and filter-paper-dried blood by using a PCR buffer with a higher pH. Analytical Biochemistry (Mosc.) 375, 370-372.
 
Costello, A., Glonek, J.R. and Myers, T.C). 1976. (Phosphorus-31 nuclear magneticresonance – pH titration of hexaphosphate (phytic acid). In Proceedings of the Inositol Phosphates: Their Chemistry, Biochemistry and Physiology (Elsevier, New York), 156-171.
 
Chung,T. K.) 2002). How to get the best out of phytase. Feed Mix. 10, 5, 27-29.
 
De angelis, M.( 2003). Phytase activity in sourdough lactic acid bacteria: purification and characterization of a phytase from Lactobacillus sanfranciscensis CB1. International Journal of Food Microbiology. 87, 3, 259-270. DOI= http://dx.doi.org/10.1016/s0168-1605(03)00072-2.
Gargova, S., Roshkova, Z.  and Vancheva, G.( 1997). Screening of fungi for phytase production. Biotechnology Techniques. 11, 4 (April), 221–224.
Gonita-Mishra, I., Deshmukh, D., Tripathi, N., Bardiya-Bhurat, K., Taniwai, K.  and Tiwari, S.( 2013). Isolation, morphological and molecular characterization of phytate-hydrolysing fungi by 18S rDNA sequence analysis. Brazilian Journal of Microbiology. 44, 1, 317-323.
Greiner, R. and  Konietzny, U.( 2006). Phytase for Food Application. Food Technol. Biotechnol. 44, 2 (March), 124-140.
Gunashree, B.S.( 2006). Studies on the production and characterization of phytate degrading enzymes in Aspergillus Niger. In Food Microbiology Department. MYSORE, 243.
Haefner, S., Knietsch, A., Scholten, E., Braun. J., Lohscheidt, M. and O, Z.( 2005). Biotechnological production and applications of phytases. Applied Microbiology and Biotechnology Advances. 68, 588–597.
Howson, S.J. and Davis, R.P.( 1983). Production of phytate-hydrolysing enzyme by some fungi. Enzyme and Microbial Technology. 5, 5, 377-382. DOI= http://dx.doi.org/10.1016_0141-0229(83)90012-1.

Jorquera, MA., Gabler, S., Inostroza, NG., Acuña, JJ., Campos, MA., Menezes-Blackburn D., Greiner, R(2018). Screening and Characterization of Phytases from Bacteria Isolated from Chilean Hydrothermal Environments.Microb Ecol. 75(2):387-399.

 

Khan, A. and Ghosh, K.( 2012). Characterization and Identification of Gut-Associated Phytase-Producing Bacteria in Some Freshwater Fish Cultured in Ponds. Acta Ichthyologica et Piscatoria 42, 1, 37-45. DOI= http://dx.doi.org/10.3750/aip2011.42.1.05.
Kerovuo, J., Ruovinen, J. and Hatzack, F. (2000). Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase: indication of a novel reaction mechanism. Biochemical Journal 352, 623-628.
Layton, A., Mckayl , L., Williams, D., Garrett, V.,Gentry, R. and Sayler, G. (2006). Development of Bacteroides 16S rRNA Gene TaqMan-Based Real- Time PCR Assays for Estimation of Total, Human  and Bovine Fecal Pollution in Water. applied and environmental Microbiology Reports. 72, 4214-4224.
Lee, D. H., Choi , S. U. and  Hwng, Y. I.( 2005). Culture Conditions and Characterizations of a New Phytase-Producing Fungal Isolate, Aspergillus sp. L117. Mycobiology .33, 4 (November), 223-229.
Lei, G. and Poress, J.( 2003). Phytase enzyology, applications, and biotechnology. Biotechnology Letters. 25, 1787-1794
Michaud, V., Gil, P., Kwiatek, O., Prome, S., Dixon, L. and Romero,L.(2007). Long-term storage at tropical temperature of dried-blood filter papers for detection and genotyping of RNA and DNA viruses by direct PCR. Journal of Virological Methods (Orlando). 146, 257-265.
Mirhendi, H., Diba, K., Rezaei, A., Jalalizand, N., Hosseinpur, L. and  Khodadadi, H.( 2007). Colony PCR Is a Rapid and Sensitive Method for DNA Amplification in Yeasts. Iranian Journal of Public Health Monograph. 36, 40-44.
Murugesan, G., Angayarkanni, J. and Swaminathan, K. ( 2002). Effect of tea fungal enzymes on the quality of black  tea. Food Chemistry. 79, 407-411.
Nelson, T.S., Shieh, T.R., Wodzinski, R.J. and Ware, J.H.(1983.( The availability of phytate phosphorus in soybean meal before and after treatment with a mold phytase. Poultry Science. 47, 1842-1848.

Neira-Vielma, AA., Aguilar, CN., Ilyina, A., Contreras-Esquivel, JC., Carneiro-da-Cunha, MDG, Michelena-Álvarez G, Martínez-Hernández, JL.(2017). Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate. Biotechnol Rep (Amst).  15;17:49-54

Ocampo, M.B., Patiño, L.F.C., Marin, M.M., Salazar, M.Y. and Gutierrez , P.A.S. (2012). Isolation and Characterization of Potential Phytase-Producing Fungi from Environmental Samples of Antioquia (Colombia). Revista Facultad Nacional de Agronomía, Medellín. 65, 1, 6291-6303.
Oh, B., Choi, W., Park, S., Kim , Y. and Oh, T.( 2004). Biochemical properies and substrate specificities of alkaline and histidine acid phytases. Applied Microbiology and Biotechnology Advances. 63, 362-372
Powar , V.K. and Jagannathan, V.(1982). Purification and Properties of Phytate-Specific Phosphatase from Bacillus subtilist. Journal of Bacteriology. 151, 3 (September), 1102-1108.
Sasirekha, B., Bedashree, T. and Champa, K.L. (2012). Optimization and partial purification of extracellular phytase from Pseudomonas aeruginosa p6. European Journal of Experimental Biology. 2, 95-104.
Shankaranand, V.S. and Lonsane, B.K.( 1994). Coffee husk: an inexpensive substrate for production of citric acid by Aspergillus niger in a solid state fermentation system. World Journal of Microbiology and Biotechnology. 10, 165-168.
Singh, N.K., Joshi, D.K. and Gupta, R.K.( 2013). Isolation of Phytase Producing Bacteria and Optimization of Phytase Production Parameters. Jundishapur Journal of Microbiology. 6, 5. DOI= http://dx.doi.org/10.5812/jjm.6419.
Sreedevi, S. and Reddy, B.N.( 2013). Purification and Biochemical Characterization of Phytase from newly isolated Bacillus subtilis C43. Advanced BioTech. 12, 8 (February), 1-6.
Vatsa, P. and Banerjee, U.C.( 2004). Production studies and catalytic properties of phytases (myo-inositolhexakisphosphate phosphohydrolases). Enzyme and Microbial Technology. 35, 1 (6 July), 3-14. DOI= http://dx.doi.org/10.1016.
Vohra, A. and Satyanarayana, T .( 2003). Phytases: Microbial Sources, Production, Purification, and Potential Biotechnological Applications. Critical Reviews in Biotechnology. 23, 29-60.
Yanke, L.J., Selinger, L.B. and Cheng, K.J. (1999). Phytase activity of Selenomonas ruminantium: a preliminary characterization. Letters in Applied Microbiology. 29(April), 20-25.
Animal Sciences Journal
Volume 32, Issue 125
March 2020
Pages 185-204

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