Optimization of Milk-Clotting Protease Production by a Local Isolate of Aspergillus Niger Ffb1 in Solid-State Fermentation

Bensmail, Souhila; Mechakra, Aicha; Fazouane–Naimi, Fethia

doi:10.15414/jmbfs.2015.4.5.467-472

Cited by 10 publications

(27 citation statements)

References 31 publications

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“…This could be due to lamentous fungi preferred SSF for production of enzymes (18). Similar protease activities were recorded by Aspergillus species (26)(27)(28)(29) and Mucor species under SSF.…”

Section: Discussionsupporting

confidence: 65%

Production, Optimization and Characterization of an Acid Protease From Filamentous Fungi by Solid-state Fermentation

Usman

Mohammed

Mamo

2020

Preprint

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Acid proteases represent an important group of enzymes, extensively used in food and beverage industries. There are a diversification of food industries and thus an increasing demand for biocatalysts capable of adapting the industrial extreme environments. These demands can be covered by a plant and animal proteases; however there is a shortage to meet the present industrial demands. This necessitates the search for an alternative acid protease sources from fungi. The fungal isolates were recovered from grape and dairy farm soil using potato-dextrose Agar. The fungi were further screened for protease production based on the hydrolysis of clear-zone on skim-milk agar media. The potential fungi were then subjected to secondary screening under solid-state fermentation. After primary and secondary screening, the potential fungus (isolate Z1BL1) was identified to the genus level by combination of macroscopic and microscopic morphological study. The growth condition and media composition for potential fungal isolate (Z1BL1) was further optimized under solid-state fermentation. The crude enzyme produced from isolate Z1BL1 was characterized after partial purification by acetone and ammonium sulphate precipitation. A total of 9 fungal isolates were showed protease production in primary and secondary screening, however 1 potential isolate (Z1BL1) was selected for further study based on its protease activity. The potential fungus, isolate Z1BL1 was identified to the genus Aspergillus based on their morphological features. The optimization of media composition and growth conditions for acid protease production from Z1BL1 were slightly increased the protease activity. The acetone precipitated enzyme exhibited the maximum activity at 50 0C and pH 5 with stability at pH 4-6 and temperature 40-60 0C. Thus based on the above findings, the acid protease produced from Aspergillus was shown suitable enzyme characteristics required in industry and could be a candidate to be applicable in food industry after further purification by high resolution techniques.

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Section: Discussionsupporting

confidence: 65%

Production, Optimization and Characterization of an Acid Protease From Filamentous Fungi by Solid-state Fermentation

Usman

Mohammed

Mamo

2020

Preprint

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“…Although the particle size of each substrate was not determined, the difference may be due to differences in particle size between wheat bran and the other three substrates which is partly related to porosity [25]. Wheat bran is considered as the best substrate for the production of acid protease from A. oryzae MTCC 5341 [4], other Aspergillus species [20,26], and Mucor sp. [27].In the present study, both wheat bran moistened with a mineral solution and wheat bran moistened with HCL were used for milk-clotting protease production; however, the result showed that wheat bran moistened with HCL was better for MCP production (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…The effect of initial moisture content on enzyme production was tested by moistening substrate using distilled water in different percentages of moisture content, 45, 50, 55, 60, 65, and 70%, to find out the best percentage for enzyme production [20]. The milk-clotting and protease activities were assayed according to [6].…”

Section: Effect Of Moisture Contentmentioning

confidence: 99%

Optimization of media composition and growth conditions for production of milk-clotting protease (MCP) from Aspergillus oryzae DRDFS13 under solid-state fermentation

et al. 2020

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This study reports the optimization of milk-clotting protease production from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) in both one-variable-at-a-time and response surface methodology (RSM). The production and optimization of milk-clotting protease obtained from Aspergillus oryzae DRDFS13 under solid-state fermentation (SSF) using different agroindustrial wastes as solid substrates were studied. The agro-industrial wastes used included wheat bran, rice bran, pea bran, and grass pea bran. The chemical composition of the best solid substrate was tested using standard methods. Others cultivation parameters were studied, and the results showed that the optimum fermentation medium composed of wheat bran, casein (1% w/w), and glucose (0.5% w/w) and the conditions for maximum milk-clotting protease production were at the moisture content of 55.0%, inoculum of 0.5*10 6 spores/mL, incubation temperature of 30°C, pH of 6.0, and fermentation time of 5 days. The highest milk-clotting activity was obtained from the crude enzyme extracted using 0.1 M NaCl and partial purification of the crude enzyme using chilled acetone, and 80% (NH 4) 2 SO 4 increased the ratio of MCA/PA from 0.56 to 1.30 and 0.65, respectively. Moreover, the highest MCA (137.58 U/mL) was obtained at a casein concentration of 0.5%, pH 4.0, and 25°C, using RSM. Thus, results from the present study showed that the optimization of milk-clotting protease production from A. oryzae DRDFS 13 under SSF by both one-variable-at-a-time and RSM significantly increased the milk-clotting activity. This is the first report from a fungus in the Ethiopian setting and a modest contribution to highlight the potential of harnessing microbial protease enzymes for industrial applications.

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“…Aspergillus tamari and Penicillium pinophillum also demonstrated great milk-clotting protease production in liquid medium containing whey as one of the main substrates (Benlounissi et al, 2014). In the study of Bensmail et al (2015), A. niger produced proteolytic enzymes with wheat bran as substrate in submerged and solid-state fermentation. Castro et al (2014) also studied milk-clotting enzymes secreted by A. niger.…”

Section: Protease Production and Milk-clotting Activitymentioning

confidence: 99%

“…Submerged fermentation (SmF) is a well-known process to produce enzymes by the growing of microorganisms in liquid substrate or in the presence of excess water while solid-state fermentation (SSF) involves the growth of microorganisms on solid substrates in the absence or near absence of free water in the space between particles (Bensmail et al, 2015;Singhania et al, 2010). Because of need for new microbial milk-clotting proteases sources due the shortage of rennet and the increase demand for cheese, the aim of this study was to investigate the protease production by A. flavo furcatis DPUA 1608 in submerged and solid state fermentation and also certify the nonproduction of aflatoxin by this species.…”

Section: Introductionmentioning

confidence: 99%

Aspergillus flavo furcatis: Aflatoxin test and milk-clotting protease production in submerged and solid state fermentation

Alecrim¹,

Martim²,

Souza³

et al. 2017

Afr. J. Microbiol. Res.

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Proteolytic enzymes are metabolites that can be produced by microbial sources and develop important functions in food industry as in cheese manufacturing. However, it is necessary to ensure the final product safety by testing toxins production by microorganisms. As a result of this, the aim of this study was to investigate the production of proteases by Aspergillus flavo furcatis DPUA 1608 in submerged and solid state fermentation and also certify the non-production of aflatoxin by this species. The aflatoxin test was carried out using the method of ammonia vapor. In this test, A. flavo furcatis DPUA 1608 was inoculated in seven different media cultures (COA, YES, CZ, CYA, GMS, PMS and PDA) and the production of toxins was confirmed by the color change of culture reverse after adding a 25% (w/v) ammonia solution. The protease production was conducted using four inoculums (SAB+GLI, SAB+SAC, BDA+GLI and BDA+SAC) in three media cultures of submerged fermentation obtained by a base mineral solution (MA01, MAGli and MASac) and in solid state fermentation using açai seeds and rice bran as substrate (SAFA). According to ammonia vapor test, A. flavo furcatis is not an aflatoxin producer. There was no color change in the colonies reverse of any culture media. All crude extracts obtained in both fermentations were tested for protease production. The best protease activity was observed in the medium MA01 (inoculums SAB+SAC, BDA+GLI and BDA+SAC). Milk clotting activity was determined in all crude extracts of submerged and solid fermentation. However, the clot formed was considered as strong milk coagulation only in the media MA01 of submerged fermentation (inoculums SAB+SAC, BDA+GLI and BDA+SAC) and in all inoculums of solid state fermentation (SAFA). A. flavo furcatis DPUA 1608 showed potential milk-clotting protease production in both fermentations media used.

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Optimization of Milk-Clotting Protease Production by a Local Isolate of Aspergillus Niger Ffb1 in Solid-State Fermentation

Cited by 10 publications

References 31 publications

Production, Optimization and Characterization of an Acid Protease From Filamentous Fungi by Solid-state Fermentation

Production, Optimization and Characterization of an Acid Protease From Filamentous Fungi by Solid-state Fermentation

Optimization of media composition and growth conditions for production of milk-clotting protease (MCP) from Aspergillus oryzae DRDFS13 under solid-state fermentation

Aspergillus flavo furcatis: Aflatoxin test and milk-clotting protease production in submerged and solid state fermentation

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