Purification and characterization of phytase from Bacillus subtilis P6: Evaluation for probiotic potential for possible application in animal feed

Abstract

The antinutritional property of phytate is the main problem associated with food animals, but phytase-producing probiotic strain could be the best alternative for resolving this problem. In the present study, phytase from probiotic strain Bacillus subtilis P6 was purified to homogeneity, which showed a relative molecular mass of 40 kDa on native and SDS-PAGE. The purification fold of purified phytase was 86.99, with a specific activity of 104.39 U/mg protein. The purified enzyme showed optimum activity at 40 • C and pH 6.0. Semi-metal ions, such as K + and Ca 2+ , slightly enhance the enzyme activity while Cu 2+ , Co 2+ , and Zn 2+ prominently inhibited phytase activity. Purified phytase showed the highest substrate specificity toward its natural substrate phytate and did not hydrolyze other substrates having a phosphate bond. The purified enzyme exhibited strong proteolytic tolerance against trypsin and pepsin. Further, we evaluated the probiotic potential of B. subtilis and noted that it showed excellent resistance toward high acidic and bile concentrations. Subsequently, it revealed exceptional antimicrobial activity, good hydrophobicity, autoaggregation ability, nonhemolytic, and nonlecithinase activity. Additionally, B. subtilis P6 displayed susceptibility against major antibiotics. Owing to the excellent biochemical properties of phytase with notable probiotic characteristics, B. subtilis P6 may offer the possibility of being a promising candidate for the application in animal feed and food supplements. However, the in vivo use of strain secreting phytase needs to be studied in animals for its intended benefits

Characterization and in vitro assessment of probiotic potential of Lactiplantibacillus plantarum BXM2 from fermented honey passion fruit beverage

“…Screening of microorganisms from traditional fermented foods has been a powerful tool to obtain new strains for industrial use, providing better fermentation properties and/or desirable health benefits (M'hamed et al., 2022; Trivedi et al., 2022). From a naturally fermented fruit beverage, this work isolated and identified a Lactiplantibacillus plantarum strain BXM2, which could be beneficial and genetically stable for industrial food fermentation.…”

Insight into phytase-producing microorganisms for phytate solubilization and soil sustainability

“…Phytases have been reported in bacteria such as Aerobacter aerogenes, Bacillus sp. (Bacillus licheniformis, Bacillus subtilis P6, and B. subtilis) (Zhao et al, 2021;Trivedi et al, 2022), Enterobacterium, anaerobic rumen bacteria (Megasphaera elsdenii and Selenomonas ruminantium), Escherichia coli, Lactobacillus amylovorus, and Pseudomonas sp.…”

“…Phytases have been reported in bacteria such as Aerobacter aerogenes , Bacillus sp. ( Bacillus licheniformis , Bacillus subtilis P6, and B. subtilis ) ( Zhao et al, 2021 ; Trivedi et al, 2022 ), Enterobacterium , anaerobic rumen bacteria ( Megasphaera elsdenii and Selenomonas ruminantium ), Escherichia coli , Lactobacillus amylovorus, and Pseudomonas sp. Lactobacillus sanfranciscensis , which were discovered as the highest phytase producers among the lactic acid bacterial strains recovered from sour doughs ( Konietzny and Greiner, 2004 ).…”

Unraveling the potential of bacterial phytases for sustainable management of phosphorous

“…Cyberlindnera jadinii CJ2 Cell bound Capusoni et al 96 Finarome sp. Cell bound Klosowski et al 97 Pichia kudriavzevii Cell bound Hellstrom et al 98 Pseudomonas strains Cell bound Zhuang et al 99 Bacillus subtilis P6 Intracellular Trivedi et al 100 Talaromyces purpureogenus NSA20 Intracellular Ahmed et al 101 Pediococcus acidilactici SMVDUDB2 Intracellular Bhagat et al 102 Pseudomonas mandelii GS10-1 Extracellular Li et al 103 Bacillus licheniformis Extracellular Dan et al 104 Pediococcus pentosaceus KTU05-8, KTU05-9 Extracellular Cizeikiene et al 105 Lactic acid bacteria Extracellular Nuobariene et al 106 Lactobacillus fermentum spp. KA1 Extracellular Sharma et al 107 purification, and potential biotechnological applications.…”

“…In this respect, there are various phytase purification techniques, the selection of which further depends on the source of extracted crude form of phytase type under study. 122 Trivedi et al 123 reported the purification and characterization of extracellular phytase from B. subtilis P6, a probiotic strain. In this study, crude form of extracted bacterial phytase was purified to homogeneity by the combination of three steps including ammonium sulfate precipitation, DEAE-sepharose, and Sephadex G-100.…”