Purification and Characterization of Phytase Produced by Aspergillus niger Isolated from Poultry Farm Soil Using Submerged Fermentation
doi.org/10.26538/tjnpr/v5i6.17
Keywords:
Phytase, Aspergillus niger, Industry, Submerged fermentationAbstract
Phytase is used in feed processing as an exogenous supplement in monogastric animal feed. This study was aimed at determining phytase production capacity of Aspergillus niger isolated from poultry farm soil. A. niger was isolated from poultry farm soil using phytase screening medium (PSM) containing sodium phytate as substrate. Quantitative estimation of phytase production was carried out by submerged fermentation using 3 x 105 spores/100 mL PSM for 5 days at 30oC and 150 rpm. The obtained crude phytase was purified using a combined purification process of ammonium sulphate precipitation and gel filtration chromatography using a 50 x 1.5 cm Sephadex G-75 column. The activity of the partially purified enzyme was assessed by subjecting it to varying pH, temperature, and nutrient conditions. The optimum temperature and pH were 50oC and 6 respectively. The enzyme remained active for over 30 min at 70oC and completely lost activity after 10 min at 80oC. No activity was observed at 100oC. The phytase produced with glucose as carbon source had higher activity (0.087 U/mL) than that produced with sucrose (0.018 U/mL). The A. niger isolate used in this study was unable to utilize ammonium sulphate as a source of nitrogen (no activity). This study establishes the potential of indigenous fungi for industrial phytase production.
References
Muslim SN, Ali AN, AL-Kadmy IM, Khazaal SS, Ibrahim SA, Al-Saryi NA, Al-Saadi LG, Muslim SN, Salman BK, Aziz SN. Screening, nutritional optimization and purification for phytase produced by Enterobacter aerogenes and its role in enhancement of hydrocarbons degradation and biofilm inhibition. Microb Pathogen. 2018; 115:159-167.
Sorour MA, Abdel-Gawad AS, Mehanni AE, Oraby RE. The use of phytate- degrading enzyme extracted from germinated legumes in food processing. Nutr Food Toxicol.2017; 2(1):274-282.
Dersjant-Li Y, Awati A, Schulze H, Partridge G. Phytase in non-ruminant animal nutrition: a critical review on phytase activities in the gastrointestinal tract and influencing factors. J Sci Food Agric. 2014; 95:878-896.
Obafemi YD, Ajayi AA, Taiwo OS, Olorunsola SJ, Isibor PO. Isolation of polygalacturonase-producing bacterial strain from tomatoes (Lycopersicon esculentum Mill.). Int J Microbiol. 2019; Article ID 7505606.
Jain J, Sapna, Singh B. Characteristics and biotechnological applications of Bacterial phytases. Proc Biochem. 2016; 51:159-169.
Ajayi AA, Lawal B, Salubi AE, Onibokun AE, Oniha MI, Ajayi OM. Pectinase Production by Aspergillus niger using Pineapple Peel Pectin and Its Application in Coconut Oil Extraction. IOP Conference Series: Earth Environ Sci. 2021; 655(1):012014.
Betancur OM. Cervantes PLF, Montoya MM, Yepes SM, Sánchez GPA. Isolation and characterization of potential phytase-producing fungi from environmental samples of Antioquia (Colombia). 2012; Revista Facultad Nacional de Agronomía Medellín, 65(1):6291-6303.
Sethi BK, Jana A, Nanda PK, Dasmohapatra PK, Sahoo SL, Kumar J. Production of α-Amylase by Aspergillus terreusNCFT 4269.10 Using Pearl Millet and Its Structural Characterization. Front Plant Sci. 2016; 1:639.
Saithi S and Tongta A Phytase Production of Aspergillus niger on Soybean Meal by Solid-State Fermentation Using a Rotating Drum Bioreactor. Agric Agric Sci. 2016; 11:25-30.
Neira-Vielma AA, Aguilar CN, Ilyina A, ContrerasEsquivel JC, Carneiro-da-Cunha M, Michelena-Álvarez G and Martínez-Hernández J. Purification and biochemical characterization of an Aspergillus niger phytase produced by solid-state fermentation using triticale residues as substrate. Biotech Rep. 2018; 17:49-54.
Qasim SS, Shakir KA, Al-Shaibani AB. Isolation, screening and production of phytate degrading enzyme (phytase) from local fungi isolate. Iraqi J Agric Sci. 2016; 74(Special Issue):121-128.
Lee D, Choi S, Hwan Y. Culture conditions and characterizations of a new phytase-producing fungal isolate, Aspergillus sp. L117. Mycobiol. 2005; 33(4):223-229.
Domsch KH, Gam W, Anderson TH. Compendium of Soil Fungi. (Volume 1). London: Academic Press; 1980. 860 p.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Bio Chem. 1951; 193(1):265-275.
Sanni DM, Lawal OT, Enujiugha VN. Purification and characterization of phytase from Aspergillus fumigatusisolated from African Giant Snail (Achatina fulica) Biocat Agric Biotech. 2019; 17:225-232.
Kalsi HK, Singh R, Dhaliwal HS, Kumar V. Phytase from Enterobacter and Serratia species with desirable characteristics for food and feed applications. Biotech. 2016; 6(1):1-13.
Thyagarajan R, Namasivayam SKR, Narendrakumar G. Partial purification of phytase from Hypocrea lixii surt01, a poultry isolate. Int J Pharm Bio Sci. 2014; 5(4):680-687.
Ajith S, Ghosh J, Shet D, ShreeVidhya S, Punith BD, Elangovan AV. Partial purification and characterization of phytase from Aspergillus foetidus MTCC 11682. AMB Express. 2019; 9(1):1-11.
Bhavsar K, Kumar VR, Khire JM. 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 Biotech. 2011; 38(9):1407-1417.
Shah PC, Kumar VR, Dastager SG, Khire JM. Phytase production by Aspergillus niger NCIM 563 for a novel application to degrade organophosphorus pesticides AMB Express. 2017; 7(1):66.
Meletiadis J, Meis JF, Mouton JW, Verweij PE. Analysis of growth characteristics of filamentous fungi in different nutrient media. J Clin Microbiol. 2001; 39(2):478-484.
Tariq M, Nawaz M, Aftab AA, Sana S, Hafeez MA, Nazir J, Shahzad W, Najeeb MI 2017 Production and characterization of phytase from indigenous Aspergillus niger isolates. Pak J Agric Sci. 2017; 54(4):799-806.
Pedri ZC, Lozano LMS, Hermann KL, Helm CV, Peralta RM and Tavares LBB. Influence of nitrogen sources on the enzymatic activity and grown by Lentinula edodes in biomass Eucalyptus benthamii. Braz. J Bio. 2015; 75(4):940-947.
Bala A, Jain J, Kumari A, Singh B. Production of an extracellular phytase from a thermophilic mould Humicola nigrescens in solid state fermentation and its application in dephytinization. Biocatal. Agric. Biotechnol. 2014; 3(4):259-64.
Shahryari Z, Fazaelipoor MH, Setoodeh P, Nair RB, Taherzadeh MJ, Ghasemi Y. Utilization of wheat straw for fungal phytase production. Int J Recycl Org Waste Agric. 2018; 27(4):345-355.
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