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Isolation and Biochemical Characterization of Acid Phytase from Aspergillus niger and Its Applications in Dephytinization of Phytic Acid in Poultry Feed Ingredients

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Abstract

Acidic phytase (PHY-B) was isolated from Aspergillus niger BIONCL8 strain, and assessed its application for the dephytinization of poultry feed ingredients. Aspergillus niger BIONCL8 strain was identified by precise molecular methods by targeting internal transcribed spacer (ITS) region of the fungi and molecular detection of PHY gene in the strain was confirmed by sequence analysis with respect to PHY production. The enzyme was isolated and purified by DEAE Sephadex A-50 and Bio-Gel P-60 Gel ion-exchange chromatography. The estimated molecular weight of the protein was 65 kDa on SDS-PAGE, it had a specific activity of 21.18 U/mg. The molecular weight of the protein was confirmed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The peptide sequence of PHY-B was identified using high resolution-mass spectrometry (LC-HR-MS), and its physicochemical parameters were studied. The PHY was stable at acidic pH 2.1, most active at 40°C and was stable up to 80°C, retaining 30% residual activity after 1 h of incubation. The PHY activity was enhanced in the presence of Mg2+ and EDTA, and activity was inhibited in the presence of Hg2+ and K+. The Km and Vmax of PHY were recorded as 3.35 mM and 1.27 U/mg, respectively. The PHY was shown to play a significant role in decreasing PA content in various poultry feed ingredients, ranging from 48.14 to 82.14%. The novel features of the enzyme can be used to decrease the PA content in feed ingredients and increase the bioavailability of nutrients to non-ruminant animals.

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ACKNOWLEDGMENTS

We would like to acknowledge CSIR-National Chemical Laboratory, Pune, India for providing necessary facilities.

Funding

This research has been supported by the Early Career Research (ECR) Award (file no. ECR/2018/001813), Science and Engineering Research Board (SERB), Department of Science and technology (DST), Delhi, India.

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Authors and Affiliations

  1. Biochemical Sciences Division, CSIR-National Chemical Laboratory, 411008, Pune, India

    Y. Bhandari, B. Sonwane & K. R. Vamkudoth

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, New Delhi, India

    B. Sonwane & K. R. Vamkudoth

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  1. Y. Bhandari
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  2. B. Sonwane
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Correspondence to K. R. Vamkudoth.

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Bhandari, Y., Sonwane, B. & Vamkudoth, K.R. Isolation and Biochemical Characterization of Acid Phytase from Aspergillus niger and Its Applications in Dephytinization of Phytic Acid in Poultry Feed Ingredients. Microbiology 92, 221–229 (2023). https://doi.org/10.1134/S0026261722601452

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