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Production of feed enzymes (phytase and plant cell wall hydrolyzing enzymes) byMucor indicus MTCC 6333: Purification and characterization of phytase

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Abstract

The production of phytase and associated feed enzymes (phosphatase, xylanase, CMCase, α-amylase and β-glucosidase) was determined in a thermotolerant fungusMucor indicus MTCC 6333, isolated from composting soil. Solid-substrate culturing on wheat bran and optimizing other culture conditions (C and N sources, level of N, temperature, pH, culture age, inoculum level), increased the yield of phytase from 266±0.2 to 513±0.4 nkat/g substrate dry mass. The culture extract also contained 112, 194, 171, 396, and 333 nkat/g substrate of phosphatase, xylanase, CMCase, β-glucosidase and α-amylase activities, respectively. Simple 2-step purification employing anion exchange and gel filtration chromatography resulted in 21.9-fold purified phytase. The optimum pH and temperature were pH 6.0 and 70 °C, respectively. The phytase was thermostable under acidic conditions, showing 82 % residual activity after exposure to 60 °C at pH 3.0 and 5.0 for 2 h, and displayed broad substrate specificity. TheK m was 200 nmol/L andv lim of 113 nmol/s per mg protein with dodecasodium phytate as substrate.In vitro feed trial with feed enzyme resulted in the release of 1.68 g inorganic P/kg of feed after 6 h of incubation at 37 °C.

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Abbreviations

DMS:

dry mass substrate

Pi :

inorganic phosphorus

NSP:

non-starch polysaccharides

DSP:

dodecasodium phytate

PSM:

phytase screening medium

MTCC:

Microbial Type Culture Collection

SSF:

solid state fermentation

PHY:

phytase(s); 3-phytase (EC 3.1.3.8) 4-phytase (EC 3.1.3.26)

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Authors and Affiliations

  1. Department of Microbiology, Guru Nanak Dev University, 143005, Amritsar, India

    H. K. Gulati, B. S. Chadha & H. S. Saini

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  1. H. K. Gulati
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  2. B. S. Chadha
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  3. H. S. Saini
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Correspondence to B. S. Chadha.

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Gulati, H.K., Chadha, B.S. & Saini, H.S. Production of feed enzymes (phytase and plant cell wall hydrolyzing enzymes) byMucor indicus MTCC 6333: Purification and characterization of phytase. Folia Microbiol 52, 491–497 (2007). https://doi.org/10.1007/BF02932109

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  • DOI: https://doi.org/10.1007/BF02932109

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