Isolation of phytase-producing fungi from the soil poultry house
Subject Areas : microbiology
Omid Nikbakht Chemazketi
1
,
Amir Hossain Esmaili
2
,
Issa Gholampour Azizi
3
1 - Faculty of Veterinary Medicine, Bob.C. Islamic Azad University, Babol, Iran.
2 - Department of Paramedicine, Bob.C. Islamic Azad University, Babol, Iran.
3 - Department of Mycology, Bab.C., Islamic Azad University, Babol, Iran
Keywords: fungus, PSM, phytase, Aspergillus, soil,
Abstract :
Objective: Phosphorus is an essential micronutrient of plants and plays a role in important biological functions. In nature, phosphorus is mostly found in immobile minerals and in fixed organic forms, including phytic acid and phosphoesteric compounds. However, the bioavailability of limited phosphorus can be increased by using phosphate solubilizing microorganisms such as bacteria and fungi.
Materials and methods: Samples were taken from the depth of 3-5 cm of the soil where the poultry live and after preparing 10-1 to 10-5 dilutions and pore plate culture in Potato dextrose agar (PDA) medium containing antibiotics, after one-week storage at temperature 28 degrees, molds were isolated and identified. Then these molds were inoculated in PSM (Phytase Screening Medium) culture medium and the mushrooms that had the clearest aura were selected and the enzyme index (EI) and phosphate solubility efficiency (SE) were determined and finally the phytase enzyme activity was determined with a spectrophotometer was measured.
Findings: In this study, the fungi that were able to form a clear halo around the colony in the specific culture medium of phytase screening (PSM) included Aspergillus niger and Aspergillus flavus. Aspergillus flavus was more effective than Niger by 2.17%. Their average enzyme activity was 5.07±0.17 and 4.94±0.15 Iu/L, respectively.
Conclusion: This research can provide effective solutions for the development of an ideal phytase to improve animal nutrition, human health and environmental protection in the near future.
1. Balwani I, Chakravarty K, Gaur S. Role of phytase producing microorganisms towards agricultural sustainability. Biocatalysis and Agricultural Biotechnology. 2017 Oct 1;12:23-9.
2. Sadaf N, Haider MZ, Iqbal N, Abualreesh MH, Alatawi A. Harnessing the Phytase production potential of soil-borne fungi from wastewater irrigated fields based on eco-cultural optimization under shake flask method. Agriculture. 2022 Jan 12;12(1):103.
3. Manghwar H, Hussain A, Ali Q, Saleem MH, Abualreesh MH, Alatawi A, Ali S, Munis MF. Disease severity, resistance analysis, and expression profiling of pathogenesis-related protein genes after the inoculation of Fusarium equiseti in wheat. Agronomy. 2021 Oct 23;11(11):2124.
4-. Huang SJ, Chen CH, Tsai SY. Phytase production by Grifola frondosa and its application in inositol-enriched solid-state fermentation brown rice. Int. J. Food Eng. 2018 Dec;4:263-7.
5. Jatuwong K, Suwannarach N, Kumla J, Penkhrue W, Kakumyan P, Lumyong S. Bioprocess for production, characteristics, and biotechnological applications of fungal phytases. Frontiers in Microbiology. 2020 Feb 14;11:188.
6. Hosseinkhani B, Hosseinkhani G. Analysis of phytase producing bacteria (Pseudomonas sp.) from poultry faeces and optimization of this enzyme production. African Journal of Biotechnology. 2009;8(17).
7. Wang Y, Ye X, Ding G, Xu F. Overexpression of phyA and appA genes improves soil organic phosphorus utilisation and seed phytase activity in Brassica napus. PloS one. 2013 Apr 3;8(4):e60801.
8. Kim T, Mullaney EJ, Porres JM, Roneker KR, Crowe S, Rice S, Ko T, Ullah AH, Daly CB, Welch R, Lei XG. Shifting the pH profile of Aspergillus niger PhyA phytase to match the stomach pH enhances its effectiveness as an animal feed additive. Applied and environmental microbiology. 2006 Jun;72(6):4397-403.
10. Greiner R. Purification and properties of a phytate-degrading enzyme from Pantoea agglomerans. The protein journal. 2004 Nov;23:567-76.
11. Haefner S, Knietsch A, Scholten E, Braun J, Lohscheidt M, Zelder O. Biotechnological production and applications of phytases. Applied microbiology and biotechnology. 2005 Sep;68:588-97.
12. Lei XG, Porres JM. Phytase enzymology, applications, and biotechnology. Biotechnology letters. 2003 Nov;25:1787-94.
13. Nassiri M, Ariannejad H. Comparative analysis of peripheral alkaline phytase protein structures expressed in E. coli. Reports of Biochemistry & Molecular Biology. 2015 Oct;4(1):10.
14. Kumar A, Chanderman A, Makolomakwa M, Perumal K, Singh S. Microbial production of phytases for combating environmental phosphate pollution and other diverse applications. Critical Reviews in Environmental Science and Technology. 2016 Mar 18;46(6):556-91.
15. Agu KC, Umeoduagu ND, Victor-Aduloju AT, Uwanta LI, Adepeju DM, Udenweze EC, Awari VG, Chidubem-Nwachinemere NO, Nwosu JC, Udeh KC. Isolation and Characterization of Proteolytic Enzyme Produced from Fungi. 16. Gaind S, Nain L. Soil–phosphorus mobilization potential of phytate mineralizing fungi. Journal of plant nutrition. 2015 Dec 6;38(14):2159-75.
17. Qasim Ss, Shakir Ka, Al-Shaibani Ab. Isolation, Screening And Production Of Phytate Degrading Enzyme (Phytase) From Local Fungi Isolate. Iraqi Journal Of Agricultural Sciences. 2016 Jan 1;47(1).
18. Bae HD, Yanke LJ, Cheng KJ, Selinger LB. A novel staining method for detecting phytase activity. Journal of microbiological methods. 1999 Dec 1;39(1):17-22.
19. Sasirekha B, Bedashree T, Champa KL. Optimization and partial purification of extracellular phytase from Pseudomonas aeruginosa p6. Eur J Exp Biol. 2012;2(1):95-104.
20. Singh B, Satyanarayana T. Fungal phytases: characteristics and amelioration of nutritional quality and growth of non‐ruminants. Journal of Animal Physiology and Animal Nutrition. 2015 Aug;99(4):646-60.