Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation
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-industri...
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University of Zagreb Faculty of Food Technology and Biotechnology
2005-01-01
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| Series: | Food Technology and Biotechnology |
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| Online Access: | http://hrcak.srce.hr/file/162917 |
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| author | Alagarsamy Sumantha Chandran Sandhya George Szakacs Carlos R. Soccol Ashok Pandey |
| author_facet | Alagarsamy Sumantha Chandran Sandhya George Szakacs Carlos R. Soccol Ashok Pandey |
| author_sort | Alagarsamy Sumantha |
| collection | DOAJ |
| description | 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 (Mg2+, Ca2+, Fe2+) and inhibiting effect of certain chelating agents (EGTA, EDTA), the enzyme was found to be a metalloprotease. |
| first_indexed | 2024-03-09T08:10:03Z |
| format | Article |
| id | doaj.art-f4def07f3e9d4a3bb8e520260ec25c93 |
| institution | Directory Open Access Journal |
| issn | 1330-9862 1334-2606 |
| language | English |
| last_indexed | 2024-03-09T08:10:03Z |
| publishDate | 2005-01-01 |
| publisher | University of Zagreb Faculty of Food Technology and Biotechnology |
| record_format | Article |
| series | Food Technology and Biotechnology |
| spelling | doaj.art-f4def07f3e9d4a3bb8e520260ec25c932023-12-02T23:29:52ZengUniversity of Zagreb Faculty of Food Technology and BiotechnologyFood Technology and Biotechnology1330-98621334-26062005-01-01434313319Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate FermentationAlagarsamy Sumantha0Chandran Sandhya1George Szakacs2Carlos R. Soccol3Ashok Pandey4Biotechnology Division, Regional Research Laboratory (CSIR), Trivandrum, IndiaBiotechnology Division, Regional Research Laboratory (CSIR), Trivandrum, IndiaDepartment of Agricultural Chemical Technology, Technical University of Budapest, H-1111 Budapest, HungaryLaboratory of Process Biotechnology, Federal University of Parana, Curitiba, BrazilBiotechnology Division, Regional Research Laboratory (CSIR), Trivandrum, IndiaFive 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 (Mg2+, Ca2+, Fe2+) and inhibiting effect of certain chelating agents (EGTA, EDTA), the enzyme was found to be a metalloprotease.http://hrcak.srce.hr/file/162917neutral proteasesolid-state fermentationAspergillusagro-industrial residues |
| spellingShingle | Alagarsamy Sumantha Chandran Sandhya George Szakacs Carlos R. Soccol Ashok Pandey Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation Food Technology and Biotechnology neutral protease solid-state fermentation Aspergillus agro-industrial residues |
| title | Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation |
| title_full | Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation |
| title_fullStr | Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation |
| title_full_unstemmed | Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation |
| title_short | Production and Partial Purification of a Neutral Metalloprotease by Fungal Mixed Substrate Fermentation |
| title_sort | production and partial purification of a neutral metalloprotease by fungal mixed substrate fermentation |
| topic | neutral protease solid-state fermentation Aspergillus agro-industrial residues |
| url | http://hrcak.srce.hr/file/162917 |
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