Characteristics and Multifarious Potential Applications of HAP Phytase of the Unconventional Yeast Pichia anomala

  • Swati Joshi
  • Tulasi SatyanarayanaEmail author


Most of cereal and legume seeds and their products contain 1–2% phytic acid that represents around 60% of the total phosphorus content. A large portion of phytic acid in seeds is in the form of salts known as phytates. The phytic acid-bound phosphorus (myoinositol 1,2,3,4,5, 6-hexakis dihydrogen phosphate) is poorly available to monogastrics. Therefore, inorganic phosphorus (Pi), a non-renewable mineral, is supplemented in diets for swine, poultry and fish to meet their Pi requirement. Furthermore, the unutilized phytate P from plant-based feeds is excreted, which becomes an environmental pollutant in the areas of intensive animal rearing. The excess P in soils flows into lakes and the sea that causes eutrophication, leading to water blooms and death of aquatic animals. The high negative charge on phytic acid results in the chelation of positively charged divalent metal ions (e.g. Fe2+, Ca2+, Zn2+, Cu2+, Mg2+, Mn2+) of nutritional significance, rendering a poor absorption and thus unavailable. This is partly attributed to the widespread human nutritional deficiencies of calcium, iron and zinc in developing countries where plant-based diets are predominantly consumed. The challenges in three areas of animal nutrition, environmental protection and human health justify research on phytases from different microbial sources for minimizing anti-nutritional effects of phytates and to enhance growth by improving phosphorus assimilation. This chapter reviews the developments on the production, characteristics and multifarious potential applications of phytase of the unconventional yeast Pichia anomala.


Pichia anomala Phytates Phytase Dephytinization Cell permeabilization Feed additive 


  1. Casey A, Walsh G (2004) Identification and characterization of a phytase of potential commercial interest. J Biotechnol 110:313–322PubMedGoogle Scholar
  2. Cho JS, Lee CW, Kang SH, Lee JC, Bok JD, Moon YS, Lee HG, Kim SC, Choi YJ (2003) Purification and characterization of a phytase from Pseudomonas syringae MOK1. Curr Microbiol 47:290–294PubMedGoogle Scholar
  3. Fredlund E, Broberg A, Boysen ME, Kenne L, Schnurer J (2004) Metabolite profiles of the biocontrol yeast Pichia anomala J121 grown under oxygen limitation. Appl Microbio lBiotechnol. 64:403–409Google Scholar
  4. Garcia-Estepa RM, Guerra-Hernandez E, Garcia-Villanova B (1999) Phytic acid content in milled cereal products and breads. Food Res Int 32:217–221Google Scholar
  5. Gibson D (1987) Production of extracellular phytase from Aspergillus ficuum on starch media. Biotechnol Lett 9:305–310Google Scholar
  6. Greiner R, Konietzny U, Jany KD (1993) Purification and characterization of two phytases from Escherichia coli. Arch Biochem Biophys 303:107–113PubMedGoogle Scholar
  7. Hara A, Ebina S, Kondo A, Funagua T (1985) A new type of phytase from Typha latifolia L. Agric Biol Chem 49:3539–3544Google Scholar
  8. Harland BF, Morris ER (1995) Phytate: a good or a bad food component. Nutr Res 15(5):733–754Google Scholar
  9. Hassan S, Altaff K, Satyanarayana T (2009) Use of soybean meal supplemented with cell bound phytase for replacement. Pak J Nutr. 8(4):341–344Google Scholar
  10. Hayakawa T, Toma Y, andIgaue I (1989) Purification and characterization of acid phosphatases with or without phytase activity from rice bran. Agric Biol Chem 53:1475–1483Google Scholar
  11. Howson SJ, Davis RP (1983) Production of phytate hydrolyzing enzymes by some fungi. Enzyme Microb Technol 5:377–389Google Scholar
  12. Huang WC, Tang IC (2007) Bacterial and yeast cultures-process characteristics, products, and applications. In: Yang ST (ed) Bioprocessing for value-added products from renewable resources: new technologies and applications. Elsevier, The Netherlands, pp 185–224Google Scholar
  13. Huebel F, Beck E (1996) Maize root phytase. Plant Physiol 112:1429–1436Google Scholar
  14. Ingvar S, Petter M (2011) Safety and regulation of yeasts used for biocontrol or biopreservation in the food or feed chain. Antonie Van Leeuwenhoek 99:113–119Google Scholar
  15. Joshi S, Satyanarayana T (2014) Optimization of heterologous expression of the phytase (PPHY) of Pichia anomala in P. pastoris and its applicability in fractionating allergenic glycinin from soy protein. J Ind Microbiol Biotechnol 41:977–987PubMedGoogle Scholar
  16. Joshi S, Satyanarayana T (2015a) Bioprocess for efficient production of recombinant Pichia anomala hytase and its applicability in dephytinization of chick feed and whole wheat flat Indian breads. J Ind Microbiol Biotechnol 42:1389–1400PubMedGoogle Scholar
  17. Joshi S, Satyanarayana T (2015b) Characteristics and applicability of phytase of the yeast Pichia anomala in synthesizing haloperoxidase. Appl Biochem Biotechnol 176:1351–1369PubMedGoogle Scholar
  18. Kaur P, Satyanarayana T (2005) Production of cell-bound phytase by Pichia anomalain an economical cane molasses medium: optimization using statistical tools. Process Biochem 40:3095–3102Google Scholar
  19. Kaur P, Satyanarayana T (2009) Improvement in cell-bound phytase activity of Pichia anomala by permeabilization and applicability of permeabilized cells in soymilk dephytinization. J Appl Microbiol 108:2041–2049PubMedGoogle Scholar
  20. Kaur P, Satyanarayana T (2010) Improvement in cell-bound phytase activity of P. anomala by permeabilization and applicability of permeabilized cells in soymilk dephytinization. J Appl Microbiol 108:2041–2049PubMedGoogle Scholar
  21. Kaur P, Singh B, Böer E, Straube N, Piontek M, Satyanarayana T, Kunze G (2010) Pphy—a cell-bound phytase from the yeast Pichia anomala: molecular cloning of the gene PPHY and characterization of the recombinant enzyme. J Biotechnol 149:8–15PubMedGoogle Scholar
  22. Kerovuo J, Rouvinen J, Hatzack F (2000) Analysis of myo-inositol hexakisphosphate hydrolysis by Bacillus phytase: indication of a novel reaction mechanism. Biochem J 352:623–628PubMedPubMedCentralGoogle Scholar
  23. Kim HW, Kim YO, Lee JH, Kim KK, Kim YJ (2003) Isolation and characterization of a phytase with improved properties from Citrobacter braakii. Biotechnol Lett 25:1231–1234PubMedGoogle Scholar
  24. Konietzny U, Greiner R (2002) Molecular and catalytic properties of phytate-degrading enzymes (phytases). Int J Food Sci Technol 37:91–812Google Scholar
  25. Kumar S, Tsai CJ, Nussinov R (2000) Factors enhancing protein thermostability. Protein Eng 13:179–191PubMedGoogle Scholar
  26. Kumar V, Yadav AN, Verma P, Sangwan P, Saxena A, Kumar K, Singh B (2017) β-Propeller phytases: diversity, catalytic attributes, current developments and potential biotechnological applications. Int J Biol Macromol 98:595–609PubMedGoogle Scholar
  27. Latiffi AA, Salleh AB, Rahman RN, Oslan SN, Basri M (2013) Secretary expression of thermostable alkaline protease from Bacillus stearothermophilus FI by using native signal peptide and α-factor secretion signal in Pichia pastoris. Genes Genet Syst 88:85–91PubMedGoogle Scholar
  28. McCollum EV, Hart EB (1908) On the occurrence of a phytin splitting enzyme in animal tissue. J Biol Chem 4:497–500Google Scholar
  29. Mullaney EJ, Daly CB, Ullah AH (2000) Advances in phytase research. Adv Appl Microbiol 47:157–199PubMedGoogle Scholar
  30. Nagai Y, Funahashi S (1962) Phytase (myo-inositol hexaphosphate phosphohydrolase) from wheat bran. Agric Biol Chem 26:794–803Google Scholar
  31. Nakamura Y, Fukuhara H, Sano K (2000) Secreted phytase activities of yeasts. Biosci Biotechnol Biochem 64:841–844PubMedGoogle Scholar
  32. Nakano T, Joh T, Tokumoto E, Hayakawa T (1999) Purification and characterization of phytase from bran of Triticum aestivum L.Cv. Nourin#61. Food Sci Technol Res 5:18–23Google Scholar
  33. Nelson TS, Sheih TR, Wodzinski RJ, Ware JH (1971) Effect of supplement phytase on the utilization of phytate phosphorus by chicks. J Nutr 101:1289–1294PubMedGoogle Scholar
  34. Oh BC, Choi WC, Park S, Kim YO, Oh TK (2004) Biochemical properties and substrate specificities of alkaline and histidine acid phytases. Appl Microbiol Biotechnol 63:362–372PubMedGoogle Scholar
  35. Pallauf J, Rimbach G (1997) Nutritional significance of phytic acid and phytase. Arch Anim Nutr 50:301–331Google Scholar
  36. Pasamontes L, Haiker M, Wyss M, Tessier M, Van Loon APGM (1997) Gene cloning, purification, and characterization of a heat stable phytase from the fungus Aspergillus fumigatus. Appl Environ Microbiol 63:1696–1700PubMedPubMedCentralGoogle Scholar
  37. Quan CS, Tian WJ, Fan SD, Kikuchi YI (2004) Purification and properties of a low-molecular-weight phytase from Cladosporium sp. FP-1. J Biosci Bioeng 97:260–266PubMedGoogle Scholar
  38. Raboy, V (1997) Accumulation and storage of phosphate and minerals. In: Larkins BA, Vasil IK (eds) Cellular and molecular biology of plant seed development, vol 4. Kluwer Academic Publishers, Dordrecht, pp 441–447Google Scholar
  39. Rao DECS, Rao KV, Reddy TP, Reddy VD (2009) Molecular characterization, physiochemical properties, Known and potential application of phytases: A review. Crit Rev Biotechnol 29(2):182–198PubMedGoogle Scholar
  40. Rapoport S, Leva E, Guest GM (1941) Phytase in plasma and erythrocytes of vertebrates. J Biol Chem 139:621–632Google Scholar
  41. Sajidan A, Farouk A, Greiner R, Jungblut P, Mueller EC, Borriss R (2004) Molecular and physiological characterisation of a 3- phytase from soil bacterium Klebsiella sp. ASR1. Appl Microbiol Biotechnol 65:110–118PubMedGoogle Scholar
  42. Sano K, Fukuhara H, Nakamura Y (1999) Phytase of the yeast Arxulaadeninivorans. Biotechnol Lett 21:33–38Google Scholar
  43. Satio T, Kohno M, Tsumura K, Kugimiya W, Kito M (2001) Novel method using phytase for separating soyabean β-conglycinin and glycinin. Biosci Biotechnol Biochem 65:884–887Google Scholar
  44. Segueilha L, Lambrechts C, Boze H, Moulin G, Galzy P (1992) Purification and properties of the phytase from Schwanniomyces castellii. J Ferment Bioeng 74:7–11Google Scholar
  45. Singh B, Satyanarayana T (2008) Phytase production by a thermophilic mould Sporotrichum thermophile in solid state fermentation and its potential applications. Bioresourse Technol. 99:2824–2830Google Scholar
  46. Spencer JFT, Spencer DM (1997) Ecology: where yeasts live? Yeasts in Natural and Artificial Habitats. Springer, Berlin, pp 33–58Google Scholar
  47. Swick RA (2002) Soybean meal quality: assessing the characteristics of a major aquatic feed ingredient. Glob Aquacult Advocate. 5:46–49Google Scholar
  48. Tambe SM, Kaklij GS, Kelkar SM, Parekh LJ (1994) Two distinct molecular forms of phytase from Klebsiella aerogenes: evidence for unusually small active enzyme peptide. J Ferment Bioeng 77:23–27Google Scholar
  49. Van Eck JH, Prior BA, Brandt EV (1993) The water relations of growth and polyhydroxy alcohol production by ascomycetous yeasts. J Gen Microbiol 139:1047–1054Google Scholar
  50. Verma D, Satyanarayana T (2012) Phytase production by the unconventional yeast Pichia anomala in fed batch and cyclic fed batch fermentations. Afr J Biotechnol 11:13705–13709Google Scholar
  51. Vohra A (2002) Production, purification, characterization and application of phytase from Pichia anomala (Hansen) Kurtzman. Ph. D. thesisGoogle Scholar
  52. Vohra A, Satyanarayana T (2001) Phytase production by the yeast P. anomala. Biotechnol Lett 23:551–554Google Scholar
  53. Vohra A, Satyanarayana T (2002a) Statistical optimization of the medium components by response surface methodology to enhance phytase production by Pichia anomala. Process Biochem 37:999–1004Google Scholar
  54. Vohra A, Satyanarayana T (2002b) Purification and characterization of a thermostable and acid-stable phytase from P. anomala. World J Microbiol Biotechnol 18:687–691Google Scholar
  55. Vohra A, Satyanarayana T (2003) Phytases: microbial sources, production, purification, and potential biotechnological applications. Crit Rev Biotechnol 23:29–60PubMedGoogle Scholar
  56. Vohra A, Satyanarayana T (2004) A cost-effective cane molasses medium for enhanced cell-bound phytase production by Pichia anomala. J Appl Microbiol 97:471–476PubMedGoogle Scholar
  57. Vohra A, Rastogi SK, Satyanarayana T (2006) Amelioration in growth and phosphate assimilation of poultry birds using cell-bound phytase of P. anomala. World J Microbiol Biotechnol 22:553–558Google Scholar
  58. Wang Y, Wang Z, Du G, Hu Z, Liu L, Li J, Chen J (2009) Enhancement of alkaline polygalacturonate lyase production in recombinant Pichia pastoris according to the ratio of methanol to cell concentration. Biores Techol. 100:1343–1349Google Scholar
  59. Waterham HR, Digan ME, Koutz PJ, Lair SV, Cregg J (1997) Isolation of the Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase gene and regulation and use of its promoter. Gene 186:37–44PubMedGoogle Scholar
  60. Wyss M, Brugger R, Kronenberger A, Remy R, Fimbel R, Oesterhelt G, Lehmann M, Van Loon APGM (1999) Biochemical characterization of fungal phytases (myo-inositol hexakis phosphate phosphohydrolases): catalytic properties. Appl Environ Microbiol 65:367–373PubMedPubMedCentralGoogle Scholar
  61. Wyss M, Pasamontes L, Remy R, Kohler J, Kusznir E, Gadient M, Muller F, Van loon APGM (1998) Comparison of thermostability properties of three acid phosphatases from molds: Aspergillus fumigatus phytase, A. niger phytase and A. niger pH 2.5 acid phosphatase. Appl Environ Microbiol 64:4446–4451PubMedPubMedCentralGoogle Scholar

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© Springer Nature Singapore Pte Ltd. 2017

Authors and Affiliations

  1. 1.School of Life SciencesCentral University of GujaratGandhinagarIndia
  2. 2.Biological Sciences and EngineeringNetaji Subhas Institute of Technology (University of Delhi)DwarkaIndia

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