Journal of Food, Agriculture and Environment

Screening of phytate degrading microorganisms inhabiting fresh sprouts: Bacillus subtilis detected with high phytase activity

Author(s):

Savita P. D’Souza*, B. Kanchanashri, M. Yallappa, S. B. Niveditha, V. C. Suvarna, A. N. Balakrishna

Recieved Date: 2017-02-03, Accepted Date: 2017-03-27

Abstract:

Selective isolation on modified MRS medium and improved screening assays aided in investigating the population, diversity and efficacy of phytate degrading/solubilizing microorganisms dwelling fresh sprouts of cereals, pulses, condiments and oilseeds. Morphological and biochemical characterization indicated that isolates belonged to the group of lactic acid bacteria (Lactobacillus, Leuconostoc and Streptococcus/Lactococcus) and Gram-positive rods (Bacillus). The above phytate solubilizing bacterial strains showed microaerophilic nature, produced acids from lactose and hydrolyzed starch and casein. Budding (Saccharomyces and Meyerozyma) and non-budding yeasts (Geotrichum) were also identified developing clear zones of phytate solubilization. All the isolates were non-haemolytic. However, in this study only bacteria were selected for further screening. The screening approaches used in the experiment allowed comparative analysis of bacterial efficacy in phytate solubilization. It resolved doubts of hydrolysis due to microbial acids. Although isolates produced acids, they failed to solubilize tricalcium phosphate indicating that phytate degradation in this case was enzyme specific. Screening by agar-well plate assay on phytase screening medium (PSM) revealed horse grain sprout isolate, Bacillus with high extracellular phytase activity and the area of hydrolysis was up to 29.3 mm² in 24 h. In lactic acid bacteria area of hydrolysis ranged from 1.73 to10.47 mm² in 24 h. Screening by PSM broth assay (organic phytate as the only source of P) categorized Bacillus from horse grain as an excellent phytase producer. Amount of inorganic phosphates (Pi) released was up to 167.8 µg/100 mL broth in 48 h. The 16SrRNA analysis showed 98% homology with Bacillus subtilis. SDS-PAGE analysis of the extracellular phytase produced by B. subtilis recorded molecular weight of approximately 44 kDa.

Keywords:

Sprouts, isolation, screening, phytate degradation, Gram positive bacteria, phytase activity

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Journal: Journal of Food, Agriculture and Environment
Year: 2017
Volume: 15
Issue: 2
Category: Food and Health
Pages: 47-51

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