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Aspartic protease from Aspergillus niger: Molecular characterization and interaction with pepstatin A.

Kavya, P. and Sagar, K. Bhat and Sridevi Annapurna, Singh and Gopal, K. M. and Appu Rao, A. G. (2019) Aspartic protease from Aspergillus niger: Molecular characterization and interaction with pepstatin A. International Journal of Biological Macromolecules, 139. pp. 199-212. ISSN 0141-8130

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Abstract

In the pursuit of industrial aspartic proteases, aspergillopepsin A-like endopeptidase from the fungi Aspergillus niger, was identified and cultured by solid state fermentation. Conventional chromatographic techniques were employed to purify the extracellular aspartic protease to apparent homogeneity. The enzyme was found to have single polypeptide chain with a molecular mass of 50 ± 0.5 kDa. The optimum pH and temperature for the purified aspartic protease was found to be 3.5 and 60 °C respectively. The enzyme was stable for 60 min at 50 °C. The purified enzyme had specific activity of 40,000 ± 1800 U/mg. The enzyme had 85% homology with the reported aspergillopepsin A-like aspartic endopeptidase from Aspergillus niger CBS 513.88, based on tryptic digestion and peptide analysis. Pepstatin A reversibly inhibited the enzyme with a Ki value of 0.045 μM. Based on homology modeling and predicted secondary structure, it was inferred that the aspartic protease is rich in β-structures, which was also confirmed by CD measurements. Interaction of pepstatin A with the enzyme did not affect the conformation of the enzyme as evidenced by CD and fluorescence measurements. Degree of hydrolysis of commercial substrates indicated the order of cleaving ability of the enzyme to be hemoglobin N defatted soya flour N gluten N gelatin N skim milk powder. The enzyme also improved the functional characteristics of defatted soya flour. This aspartic protease was found to be an excellent candidate for genetic manipulation for biotechnological application in food and feed industries, due to its high catalytic turn over number and thermostability

Item Type: Article
Uncontrolled Keywords: Aspartic protease Structure and stability Pepstatin A interaction Commercial application
Subjects: 500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology > 04 Fungi
600 Technology > 08 Food technology > 16 Nutritive value > 05 Enzymes
Divisions: Protein Chemistry and Technology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 25 Nov 2020 05:39
Last Modified: 25 Nov 2020 05:39
URI: http://ir.cftri.com/id/eprint/14627

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