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Novel isoforms of proteinaceous α-amylase inhibitor (α-AI) from seed extract of Albizia lebbeck

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Abstract

Proteinaceous inhibitors of digestive α-amylase occur naturally in leguminous seeds and find applications in agriculture and clinical studies. We have detected and isolated eight novel α-amylase inhibitor isoforms in the seed extract of Albizia lebbeck. They are designated as AL-αAI-1 to AL-αAI-8. These isoforms specifically inhibit human salivary α-amylase and porcine pancreatic α-amylase. The occurrence and profile of α-amylase inhibitor isoforms were revealed by 7 % native-PAGE containing 0.1 % starch. The apparent molecular weights of native bands of AL-αAIs were 97.4, 68.6, 61.0, 57.2, 56.0, 54.7, 51.1, and 47.7 kDa, respectively. Partial purification of potent α-amylase inhibitor was achieved using ammonium sulfate fractionation and gel filtration chromatography on G-100 Sephadex column followed by preparative gel electrophoresis. SDS-PAGE analysis of partially purified AL-αAI showed two polypeptide bands of ~35.8 and ~32.6 kDa. All these isoforms showed effective resistance to in vitro proteolysis by pepsin, trypsin, and chymotrypsin. These inhibitors are stable over a wide range of pH and temperature and have optimum activity at pH 7 and at 37 °C. The finding and information obtained in the present investigation about novel isoforms of α-amylase inhibitors from A. lebbeck could be important and may find applications in clinical studies to modulate starch digestion and glycemic index.

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Abbreviations

α-AI:

α-Amylase inhibitor

AL-αAI:

Albizia lebbeck α-amylase inhibitor

BPB:

Bromophenol blue

BSA:

Bovine serum albumin

DNSA:

3,5-Dinitrosalicylic acid

HSA:

Human salivary α-amylase

PAGE:

Polyacrylamide gel electrophoresis

PPA:

Porcine pancreatic α-amylase

PVP:

Polyvinylpyrrolidone

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Acknowledgments

First author of the manuscript is grateful to University Grant Commission (UGC), Government of India, New Delhi for providing financial assistance in the form of Maulana Azad national fellowship (MANF).

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Authors and Affiliations

  1. Department of Biochemistry, Dr. Babasaheb Ambedkar Marathwada University, Aurangabad, India

    Faiyaz K. Shaikh, Prafull P. Gadge, Ashok A. Shinde, Bhimrao V. Jaiwal, Keshav D. Shinde, Monohar V. Padul & Manvendra S. Kachole

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  1. Faiyaz K. Shaikh
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  2. Prafull P. Gadge
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  3. Ashok A. Shinde
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  4. Bhimrao V. Jaiwal
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  7. Manvendra S. Kachole
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Correspondence to Faiyaz K. Shaikh.

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Communicated by M. Stobiecki.

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Shaikh, F.K., Gadge, P.P., Shinde, A.A. et al. Novel isoforms of proteinaceous α-amylase inhibitor (α-AI) from seed extract of Albizia lebbeck . Acta Physiol Plant 35, 901–909 (2013). https://doi.org/10.1007/s11738-012-1133-5

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