Production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation

Alagarsamy Sumantha, Chandran Sandhya, G. Szakács, Carlos R. Soccol, Ashok Pandey

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Five strains of fungi belonging to Aspergillus sp. were evaluated by casein agar plate assay and a wheat bran-based solid-state fermentation for selecting a neutral protease-producing culture. Based on the results, A. oryzae NRRL 2217 was selected for further studies. Sixteen different agro-industrial residues were evaluated for their potential to serve as a substrate for neutral protease production by this fungal strain. Results showed that a combination of coconut oil cake and wheat bran in the mass ratio of 1:3 was the best substrate for enzyme production. Various process parameters influencing protease production including fermentation time, initial moisture content, and fermentation temperature were optimised. The medium was supplemented with different nutrients in the form of organic and inorganic nitrogen and carbon sources. Supplementation of chitin increased the enzyme production significantly. Ammonium nitrate as inorganic nitrogen supplement slightly enhanced enzyme production. No organic nitrogen supplement was effective enhancer of enzyme production. Fermentation was performed under optimised conditions (initial moisture content V/m = 50 %, temperature 30°C, 48 h). Partial purification of the enzyme resulted in a 3-fold increase in the specific activity of the enzyme. The partially purified enzyme was characterised by various features that govern the enzyme activity such as assay temperature, assay pH and substrate concentration. The effect of various metal ions and known protease inhibitors on the enzyme activity was also studied. The enzyme was found to be stable in pH range 7.0-7.5, and at temperature of 50°C for 35 min. By the activating effect of divalent cations (Mg 2+, Ca2+, Fe2+) and inhibiting effect of certain chelating agents (EGTA, EDTA), the enzyme was found to be a metalloprotease.

Original languageEnglish
Pages (from-to)313-319
Number of pages7
JournalFood Technology and Biotechnology
Volume43
Issue number4
Publication statusPublished - Oct 2005

Fingerprint

Metalloproteases
metalloproteinases
Fermentation
Purification
Enzymes
fermentation
Substrates
enzymes
proteinases
wheat bran
enzyme activity
Assays
Peptide Hydrolases
Nitrogen
coconut oil cake
ethylene glycol tetraacetic acid
Enzyme activity
temperature
assays
nitrogen

Keywords

  • Agro-industrial residues
  • Aspergillus
  • Neutral protease
  • Solid-state fermentation

ASJC Scopus subject areas

  • Food Science
  • Biotechnology

Cite this

Production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation. / Sumantha, Alagarsamy; Sandhya, Chandran; Szakács, G.; Soccol, Carlos R.; Pandey, Ashok.

In: Food Technology and Biotechnology, Vol. 43, No. 4, 10.2005, p. 313-319.

Research output: Contribution to journalArticle

Sumantha, Alagarsamy ; Sandhya, Chandran ; Szakács, G. ; Soccol, Carlos R. ; Pandey, Ashok. / Production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation. In: Food Technology and Biotechnology. 2005 ; Vol. 43, No. 4. pp. 313-319.
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