Screening for efficient phytase producing bacterial strains from different soils

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Research Paper 01/01/2013
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Screening for efficient phytase producing bacterial strains from different soils

S. Sreedevi, B.N. Reddy
Int. J. Biosci.3( 1), 76-85, January 2013.
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Phytic acid (myo-inositol hexakisphosphate) is the main form of phosphate constituting about 80% of the total phosphorus in cereals, legumes and oilseeds used as animal feeds. Phytases are the enzymes which hydrolyze phytic acid to less phosphorylated myo-inositol derivatives releasing inorganic phosphate. The supplementation of animal feed with phytases reduces the cost of diets by removing or reducing the need for supplemental inorganic phosphate and increases the bioavailability of phosphorous in monogastric animals. Rhizosphere soil, Cattle shed soil and poultry farm soil collected from various regions of Hyderabad, Andhra Pradesh, India were used as source material for isolation and screening of phytase producing bacteria. A total of 162 colonies were obtained on wheat extract agar media plates. Fifty four colonies showed clear zones of hydrolysis around them which were replated and hydrolysis efficiency calculated .Twenty one isolates showed more than 50% efficiency which were investigated for phytase production in submerged fermentation. C43 isolate was found to produce significantly high phytase activity than other isolates.


Adeola O. 1999. Nutrient management procedures to enhance environmental conditions: An introduction. Journal of Animal Science 77, 427-429.

Angelis MD, Gallo G, Corbo MR, McSweeney PLH, Faccia 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, 259-270.

Anis Shobirin MH, Farouk A, Greiner R. 2009. Potential phytate-degrading enzyme producing bacteria isolated from Malaysian maize plantation. African Journal of Biotechnology 8 (15), 3540-3546.

Arpana mittall, Gulab Singh, Varsha Goyal, Anita Yadav, Kamal Rai Aneja, Sanjeev Kumar Gautam, Neeraj Kumar Aggarwal. 2011. Isolation and biochemical characterization of acido-thermophilic extracellular phytase producing bacterial strain for potential application in poultry feed.Asian journal of experimental biological sciences 2(4), 663-666.

Baharak Hosseinkhani, Giti Emtiazi, Iraj Nahvi. 2009. Analysis of phytase producing bacteria (Pseudomonas sp.) from poultry faeces and optimization of this enzyme production. African Journal of Biotechnology 8 (17), 4229-4232.

Baruah K, Sahu NP, Pal AK, Debnath. 2004. Dietary phytase: An ideal approach for a cost effective and low polluting aquafeed. Naga World fish center quaterly 27 (3&4), 15-19.

Bushra Uzair, Nuzhat Ahmed 2007. Screening of phytate hydrolysing marine bacteria isolated from Baluchistan Coast. Journal of Basic & Applied Sciences 3(1), 19-24.

Casey A, Walsh G. 2003. Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142. Bioresource Technology 86, 183-188.

Choi WC, Oh BC, Kim HK, Kang SC, Oh TK. 2002. Characterization and cloning of a phytase from Escherichia coli WC7. Korean Journal of Microbiology and Biotechnology 30, 1-7.

Chunshan Q, Linghua Z, Yunji W, Yoshiyuki O. 2001.Production of phytase in slow phosphate medium by a novel yeast candida krusei. Journal of Bioscience and Bioengineering 92,154-160.

Common FH. 1989. Biological availability of phosphorus for pigs. Nature 143, 370-380.

Dost K, Tokul O. 2006. Determination of phytic acid in wheat and wheat products by reverse phase high performance liquid chromatography. Analytica Chimica Acta, 558, 22-27.

Ebune A, Al-Asheh S, Duvnjak Z. 1995. Effects of phosphate, surfactants and glucose on phytase production and hydrolysis of phytic acid in canola meal by Aspergillus ficuum during solid-state fermentation. Bioresource Technology 54, 241-247.

Gibson DM, Ullah ABJ. 1990. Inositol metabolism in plants: phytases and their action on phytic acid. Wiley-Liss, Inc., Plant Biology 9, 77-92.

Graf E. 1983. Calcium binding to phytic acid. Journal of agricultural and food chemistry 31, 851-855.

Greiner R, Haller E, Konietzny U, Jany KD. 1997. Purification and characterization of a phytase from Klebsiella terrigena. Archives of Biochemistry and Biophysics 341, 201-206.

Greiner R, Konietzny U, Jany KD. 1993. Purification and characterization of two phytases from Escherichia coli. Archives of Biochemistry and Biophysics 303, 107-113.

Gulati HK, Chadha BS, Saini HS. 2007. Production and characterization of thermostable alkaline phytase from Bacillus laevolacticus isolated from rhizosphere soil. Journal of Industrial Microbiology and Biotechnology 34(1), 91-98.

Harland BF, Morris ER. 1995. Phytate: A good or bad food component. Nutition Research 15(5), 733-754.

Hegeman CE, Grabau EA. 2001. A novel phytase with sequence similarity to purple acid phosphatases is expressed in cotyledons of germinating soybean seedlings. Plant Physiology. 126, 1598-1608.

Heinonen JK, Lahti RJ. 1981. A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase. Analytical Biochemistry 113, 313-317.

Howson SJ, Davis RJ. 1983. Production of phytate-hydrolysing enzyme by fungi. Enzyme and Microbial Technology 5, 377-382.

In NJ, Jang ES, Kim YJ, Oh NS. 2004. Purification and properties of an extracellular acid phytase from Pseudomonas fragi Y9451. Journal of Microbiology and Biotechnology 14, 1004-1008.

Kaur P, Satyanarayana T. 2005. Production of cell-bound phytase by Pichia anomala in an economical cane molasses medium: Optimization using statistical tools. Process Biochemistry 40, 3095-3102.

Kim B H, Namkung H, Paik I K. 2002. Utilization of plant phytase to improve phosphorus availability. Korean Journal of Animal Science 44(4), 407-418.

Kim HW, Kim YO, Lee JH, Kim KK, Kim YJ. 2003. Isolation and characterization of a phytase with improved properties from Citrobacter braakii. Biotechnology Letters 25, 1231-1234.

Kim YO, Kim HK, Bae KS, Yu JH, Oh TK. 1998 Purification and properties of a thermostable phytase from Bacillus sp. DS11. Enzyme and Microbial Technology 22, 2-7.

Lambrechts C, Boze H, Moulin G, Galzy P. 1992. Utilization of phytate by some yeast. Biotechnology Letters 14, 61-66.

Lantzsch HJ, Wist S, Drochner W. 1995. The effect of dietary calcium on the efficacy of microbial phytase in rations for growing pigs. Journal of Animal Physiology and Animal Nutrition 73, 19-26.

Lee D, Schroeder J, Gordon D T. 1988. Enhancement of Cu bioavailability in the rat by phytic acid. Journal of Nutrition 118, 712–717.

Lei X, Pao K, Elwyn R M, Ullrey D E, Yokoyama M T. 1993. Supplemental microbial phytase improves bioavailability of dietary zinc to weanling pigs. Journal of Nutrition 123, 1117–1123.

Lestienne I, Icard-Verniere C, Mouquet C, Picq C, Treche S. 2005. Effects of soaking whole cereal and legume seeds on iron, zinc and phytate contents. Food Chemistry 89, 421-425.

Mukesh Kumar DJ, Balakumaran MD, Kalaichelvan PT, Pandey A, Singh A, Raja RB. 2011.  Isolation,  Production  &  Application  of Extracellular   Phytase   by   Serratia  Marcescens. Jundishapur journal of microbiology 4(4), 273-282.

Mukesh P, Suma S, Singaracharya MA, Lakshmipathi V. 2004. Isolation of phytate hydrolyzing microbial strains from traditional waste water of rice fermentation and liquid cattle feeds. World Journal of Microbiology and Biotechnology 20, 531-534.

Mullaney EJ, Daly CB, Ullah AHJ. 2000. Advances in Phytase Research. Advances in Applied Microbiology 47,157-199.

Nam-Soon Oh and Man-Jin In 2009. Phytate degradation by Leuconostoc mesenteroides KC51 cultivation in soymilk. African Journal of Biotechnology 8 (13), 3023-3026.

Nasi M. 1990. Microbial phytase supplementation for improving bioavailability of plant phosphorus in the diet of growing pigs. Journal of Agricultural Science 62, 435-442.

Nayini NR, Markakis P. 1983. Effects of inositol phosphates on mineral utilization. Federation proceedings 45, 819-826.

Nayini NR, Markakis P. 1984. The phytase of yeast. Food Science and Technology 17, 126-132.

Pallauf J, Rimbach G. 1996. Nutritional significance of phytic acid and phytase. Archives of Animal Nutrition 50, 301-319.

Pandey A, Szakacs G, Soccol CR, Rodriguez-Leon JA, Soccol VT. 2001. Production, purification and properties of microbial phytases. Bioresource. Technology 77, 203-214.

Powar VK, Jagannathan V. 1982. Purification and properties of phytate-specific phosphatase from Bacillus subtilis. Journal of. Bacteriology 151, 1102-1108.

Raun A, Cheng E, Burroughs W. 1956. Phytate phosphorus hydrolysis and availability to rumen microorganisms. Journal of Agricultural and Food Chemistry 4, 869-871.

Reddy NR, Sathe SK, Salunkhe DK. 1982. Phytate in legumes and cereals. Advances in Food Research 28, 1-92.

Rodrigues H, Fraga R. 1999. Phosphate solubilizing bacteria and their role in plant growth promotion. Biotechnology Advances 17, 319-339.

Roopesh K, Ramachandran S, Nampoothiri KM, Szakacs G, Pandey A. 2006. Comparison of phytase production on wheat bran and oilcakes in solid-state fermentation by Mucor racemosus. Bioresource Technology 97, 506-511.

Sasirekha B, Bedashree T , Champa KL. 2012. Optimization and partial purification of extracellular phytase from Pseudomonas aeruginosa p6. European Journal of Experimental Biology 2 (1), 95-104.

Scott JJ, Loewus FA. 1986. A calcium activated phytase from pollen of Lilium longiflorum. Plant Physiology 82, 333-335.

Scott JJ. 1991. Alkaline phytase activity in nonionic detergent extracts of legume seeds. Plant Physiology 95, 1298-1301.

Shieh TR, Ware JH. 1968. Survey of microorganisms for the production of extracellular phytase. Applied Microbiology 16, 1348-1351.

Shieh TR, Wodzinski RJ, Ware JH. 1969. Regulation of the formation of acid phosphatase by inorganic phosphate in Aspergillus ficuum. Journal of Bacteriology 100, 1161-1165.

Shimizu M. 1992. Purification and characterization of phytase from Bacillus subtilis (natto) N-77. Bioscience, Biotechnology, and Biochemistry 56, 1266–1269.

Shimizu M. 1993. Purification and characterization of phytase and acid phosphatase by Aspergillus oryzae K1. Bioscience, Biotechnology and Biochemistry 57, 1364-1365.

Spier MR, Greiner R, Rodriguez-Leon JA, Woiciechowski AL, Pandey A, et al. 2008. Phytase production using citric pulp and other residues of the agro-industry in SSF by fungal isolates. Food Technology and Biotechnology 46, 178-182.

Stephen Joseph, Jisha MS. 2008. Phosphate Solubilisation efficiency. Journal of Agricultural & Environmental Sciences 4(1),110-112.

Sutardi M, Buckle K A. 1986. The characteristics of soybean phytase. Journal of Food Biochemistry 10, 197-216.

Sutardi M, Buckle KA. 1988. Characterization of extra- and intracellular phytase from Rhizopus oligosporus used in tempeh production. International Journal of Food Microbiology 6, 67-69.

Thomson LU, Yoon JH. 1984. Starch digestibility as effected by polyphenols and phytic acid. Journal of Food Science 49, 1228-1229.

Tseng YH, Fang TJ, Tseng SM. 2000. Isolation and characterization of a novel phytase from Penicillium simplicissimum. Folia Microbiologica 45, 121–127.

Waldroup PW. 1999. Nutritional approaches to reducing phosphorus excretion in poultry. Poultry Science.78, 683-691.

Wang HL, Swain EW, Hasseltine CW. 1980. Phytase of molds used in oriental food fermentation. Journal of Food Science 45, 1264-1266.

Wodzinski RJ, Ullah AHJ. 1996. Phytase. Advances in Applied Microbiology 42, 263-302.

Yanke LJ, Bae HD, Selinger LB, Cheng KJ. 1998. Phytase activity of anaerobic ruminal bacteria. Microbiology 144, 1565-1573.

Yoon SJ, Choi YJ, Min HK, Cho K K, Kim JW. et al. 1996. Isolation and identification of phytase-producing bacterium, Enterobacter sp.4 and enzymatic properties of phytase enzyme. Enzyme and Microbial Technology 18(6), 449-45