Abstract
Schizophyllum commune produces phytase through solid-state fermentation using different agroindustrial residues. After optimization of phytase production, a maximal level of phytase (113.7 Units/gram of dry substrate) was obtained in wheat bran based medium containing 5% sucrose, 50% humidity, 7.5% of biomass at 33 °C pH 7.0 during 72 h and a 285% improvement in enzyme titre was achieved. Analysis of fermentation parameters profile for phytase production showed the highest productivity (1.466 Units/gram of dry substrate/hour) in 66 h of fermentation. Phytase has an optimal pH of 5.0, an optimal temperature of 50 °C and K m and V max values of 0.16 mM and 1.85 μmol mL−1 min−1, respectively. Phytase activity was stimulated essentially in the presence of K+, Ca2+, Mg2+, Mn2+, Zn2+, Cu2+, Fe2+, Fe3+, Co2+, Ni2+, acetate and citrate at concentrations of 1 mM. Phytase had the best shelf life when stored at a cooling temperature, maintaining 38% of its initial activity after 112 days of storage, and still presenting enzymatic activity after 125 days of storage. Stability studies of phytase performed in aqueous enzyme extracts showed satisfactory results using polyethyleneglycol 3350, carboxymethylcellulose, methylparaben, mannitol and benzoic acid in concentrations of 0.25, 0.025, 0.025, 0.25, and 0.0025%, respectively. PEG 3350 was shown to be the best stabilizing agent, resulting in 109% of phytase activity from the initial crude extract remaining activity in after 90 days.
Similar content being viewed by others
References
Kumari M, Survase SA, Singhal RS (2008) Production of schizophyllan using Schizophyllum commune NRCM. Bioresour Technol 99:1036–1043
Collopy PD, Royse DJ (2004) Characterization of phytase activity from cultivated edible mushrooms and their production substrates. J Agric Food Chem 52:7518–7524
Spier MR, Greiner R, Rodriguez-León JA, Woiciechowski AL, Pandey A, Soccol VT, Soccol CR (2008) Phytase production using citric pulp and other residues of the agroindustry in SSF by fungal isolates. Food Technol Biotechnol 46:178–182
Sandberg AS, Andlid T (2002) Phytogenetic and microbial phytases in human nutrition. Int J Food Sci Technol 37:823–833
Pandey A, Soccol CR, Mitchell DA (2000) New developments in solid-state fermentation I—bioprocesses and products. Proc Biochem 35:1153–1169
Sharma A, Vivekanand V, Singh RP (2008) Solid-state fermentation for gluconic acid production from sugarcane molasses by Aspergillus niger ARNU-4 employing tea waste as the novel solid support. Bioresour Technol 99:3444–3450
Papagianni M, Nokes SE, Foler K (2000) Production of phytase by Aspergillus niger in submerged and solid-state fermentation. Process Biochem 35:397–402
Heinonen JK, Lahti RJ (1981) A new and convenient colorimetric determination of inorganic orthophosphate and its application to the assay of inorganic pyrophosphatase. Anal Biochem 113:313–317
Somogyi M (1945) A new reagent for the determination of sugars. J Biol Chem 160:61–68
Nelson NA (1944) A photometric adaptation of the Somogyi method for determination of glucose. J Biol Chem 153:375–380
Seitz LM, Sauer DB, Burroughs R, Mohr HE, Hubbard JD (1979) Ergosterol as a measure of fungal growth. Phytopathology 69:1202–1203
El-Gindy AA, Ibrahim ZM, Ali UF, El-Mahdy OM (2009) Extracellular phytase production by solid-state cultures of Malbranchea sulfurea and Aspergillus Niveus on cost-effective medium. Res J of Agric Biol Sci 5:42–62
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. Bioresour Technol 97:506–511
Chantasartrasamee K, Ayuthaya DI, Intarareugsorn S, Dharmsthiti S (2005) Phytase activity from Aspergillus oryzae AK9 cultivated on solid state soybean meal medium. Process Biochem 40:2285–2289
Chadha BS, Harmeet G, Mandeep M, Saini HS, Singh N (2004) Phytase production by the thermophilic fungus Rhizomucor pusillus. World J Microbiol Biotechnol 20:105–109
Singh B, Satyanarayana T (2008) Phytase production by a thermophilic mould Sporotrichum thermophile in cost-effective cane molasses medium and its application in bread. J Appl Microbiol 105:1858–1865
Nigham PS, Pandey A (2009) Biotechnology for agro-industrial residues utilization. In: Pre-treatment of agro-industrial residues, 1st edn. Netherlands, Springer, p 25
Bhavsar K, Ravi Kumar V, Khire JM (2010) High level phytase production by Aspergillus niger NCIM 563 in solid state culture: response surface optimization, up-scaling, and its partial characterization. J Ind Microbiol Biotechnol. doi:10.1007/s10295-010-0926-z
Spier MR, Scheidt G, Portella AC, Rodríguez-León JA, Woiciechowski AL, Greiner R, Soccol CR (2011) Increase in phytase synthesis during citric pulp fermentation. Chem Eng Commun 198:286–297
Bogar B, Szakacs G, Pandey A, Abdulhameed S, Linden JC, Tengerdy RP (2003) Production of phytase by Mucor racemosus in solid-state fermentation. Biotechnol Prog 19:312–319
Bogar B, Szakacs G, Linden JC, Pandey A, Tengerdy RP (2003) Optimization of phytase production by solid substrate fermentation. J Ind Microbiol Biotechnol 30:183–189
Spier MR, Woiciechowski AL, Letti LAJ, Scheidt GN, Sturm W, Rodriguez-León JA, Carvalho JC, Dergint DEA, Soccol CR (2010) Monitoring fermentation parameters during phytase production in column-type bioreactor using a new data acquisition system. Bioprocess Biosyst Eng 33:1033–1041
Esakkiraj P, Sandoval G, Sankaralingam S, Immanuel G, Palavesam A (2010) Preliminary optimization of solid-state phytase production by moderately halophilic Pseudomonas AP-MSU 2 isolated from fish intestine. Ann Microbiol 60:461–468
Costa M, Lerchundi G, Villarroel F (2009) Phytase production by Aspergillus ficuum in submerged and solid state fermentation using agroindustrial waste as support. Rev Colomb Biotechnol 11:73–83
Vassilev N, Vassileva M, Bravo V, Fernández-Serrano M, Nikolaeva I (2007) Simultaneous phytase production and rock phosphate solubilization by Aspergillus niger grown on dry olive wastes. Ind Crop Prod 26:332–336
Sabu A, Sarita S, Pandey A, Bogar B, Szakacs G, Soccol CR (2002) Solid-state fermentation for production of phytase by Rhizopus oligosporus. Appl Biochem Biotechnol 102–103:251–260
Spier MR, Letti LA, Woiciechowski AL, Soccol CR (2009) A simplified model for A. niger FS3 growth during phytase formation in solid state fermentation. Braz Arch Biol Technol 52:151–158
Kaur O, Satyanarayana T (2005) Production of cell-bound phytase by Pichia anomala in an economical cane molasses medium: optimization using statistical tools. Process Biochem 40:3095–3102
Vats P, Banerjee UC (2002) Studies on the production of phytase by a newly isolated strain of Aspergillus niger var teigham obtained from rotten wood-logs. Process Biochem 38:211–217
Casey A, Walsh G (2003) Purification and characterization of extracellular phytase from Aspergillus niger ATCC 9142. Bioresour Technol 86:183–188
Spier MR, Fendrich R, Almeida P, Noseda M, Greiner R, Konietzny U, Woiciechowski A, Soccol VT, Soccol CR (2010) Phytase produced on citric byproducts: purification and characterization. World J Microbiol Biotechnol 27:267–274
Vohra A, Satyanarayana T (2003) Phytases: microbial sources, production, purification, and potential biotechnological applications. Crit Rev Biotechnol 23:29–60
Casey A, Walsh G (2004) Identification and characterization of a phytase of potential commercial interest. J Biotechnol 110:313–322
Konietzny U, Greiner R (2002) Molecular and catalytic properties of phytate-degrading enzymes (phytases). Int J Food Sci Technol 37:791–812
Boyce A, Walsh G (2007) Purification and characterisation of an acid phosphatase with phytase activity from Mucor hiemalis Wehmer. J Biotechnol 132:82–87
Woodzinski RJ, Ullah AHJ (1996) Biochemical characterization of cloned Aspergillus fumigatus phytase (phyA). Adv Appl Microbiol 42:263–302
Shimizu M (1993) Purification and characterization of phytase and acid phosphatase produced by Aspergillus oryzae K1. Biosci Biotechnol Biochem 57:1354–1365
Dvoràkovà O, Volfovà J, Kopecky J (1997) Characterization of phytase produced by Aspergillus niger. Folia Microbiol 42:349–352
Quan C, Zhang L, Wang Y, Ohta Y (2001) Production of phytase in a low phosphate medium by a novel yeast Candida krusei. J Biosci Bioeng 92:154–160
Van Staden J, Denhaan R, Van Zyl WH, Botha A, Viljoen-Bloom M (2007) Phytase activity in Cryptococcus laurentii ABO 510. FEMS Yeast Res 7:442–448
Beal L, Mehta T (2006) Zinc and phytate distribution in peas. Influence of heat treatment, germination, pH, substrate, and phosphorus on pea phytate and phytase. J Food Sci 50:96–100
Yang W-J, Kim K-W (1994) Kinetic properties of rat intestinal phytase/alkaline phosphatase. Korean Biochem J 27:342–345
Ullah AHJ (1988) Production, rapid purification and catalytic characterization of extracellular phytase from Aspergillus ficuum. Prep Biochem 18:443–458
Greiner R (2004) Purification and properties of a phytate-degrading enzyme from Pantoea agglomerans. Protein J. 23:567–576
Greiner R, Konietzny U, Jany KD (1997) Purification and properties of a phytase from rye. J Food Biochem 22:143–161
Tadashi N, Satoshi K, Tadanori Y, Hideharu A, Yoko K, Seiji S, Keiichi Y (2001) Phytase having a low michaelis constant for phytic acid from Monascus. US Patent 6261592. http://www.freepatentsonline.com/6261592.html. Accessed 06 Jun 2010
Sri-Akkharin W (2004) Production and characterization of extracellular phytase from Pseudomonas sp. Thesis, degree of master of science biotechnology, Faculty of Graduate Studies of Mahidol University, Bangkok
Soni SK, Magdum A, Khire JM (2010) Purification and characterization of two distinct acidic phytases with broad pH stability from Aspergillus niger NCIM 563. World J Microbiol Biotechnol 26:2009–9018
Fu D, Huang H, Luo H, Wang Y, Yang P, Meng K, Bay Y, Wu N, Yao B (2008) A highly pH stable phytase from Yersinia kristensenii: cloning, expression, and characterization. Enzyme Microb Technol 42:499–505
Nair VC, Laflamme J, Duvnjak Z (2006) Production of phytase by Aspergillus ficuum and reduction of phytic acid content in canola meal. J Sci Food Agric 54:355–365
Brugger R, Nunes CS, Hug D, Vogel K, Guggenbhul P, Mascarello F, Augem S, Wyss M, Van Loon APGM, Pasamontes L (2004) Characteristics of fungal phytases from Aspergillus fumigatus and Sartorya fumigata. Appl Microbiol Biotechnol 63:383–389
Health and Consumer Protection Directorate-General, Directorate B—Scientific Health Opinions, Unit B3—Management of scientific committees II Opinion of the SCAN on the use of enzymatic product Natuphos® 5000 (3-phytase; EC 3.1.3.8) as feed additive http://ec.europa.eu/food/fs/sc/scan/out44_en.pdf. Accessed 20 Jun 2010
Costa SA, Tzanova T, Carneiro AF, Paar A, Gübitz GM, Cavavo-Paulo A (2002) Studies of stabilization of native catalase using additives. Enzyme Microb Technol 30:387–391
Michiaki M, Koji K, Kazuo K (1997) Effects of polyols and organic solvents on thermostability of lipase. J Chem Technol Biotechnol 70:188–192
Longo MA, Combes D (1999) Thermostability of modified enzymes: a detailed study. J Chem Technol Biotechnol 74:25–32
Wang W (1999) Instability, stabilization, and formulation of liquid protein pharmaceuticals Int. J. Pharm 185:129–188
Iyer PV, Ananthanarayan L (2008) Enzyme stability and stabilization—aqueous and nonaqueous environment. Process Biochem 43:1019–1032
Rowe RC, Sheskey PJ, Owen SC (2005) Handbook of pharmaceutical excipients, 5th edn. APhA Publications, Washington
Acknowledgments
The authors would like to thank the National Council of Technological and Scientific Development (CNPq) for financial resources.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Salmon, D.N.X., Piva, L.C., Binati, R.L. et al. A bioprocess for the production of phytase from Schizophyllum commune: studies of its optimization, profile of fermentation parameters, characterization and stability. Bioprocess Biosyst Eng 35, 1067–1079 (2012). https://doi.org/10.1007/s00449-012-0692-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00449-012-0692-6