scholarly journals Production, Optimization, Purification, Kinetic analysis and applications of alkaline proteases produced from Bacillus subtilis through solid state fermentation of agricultural byproducts

2021 ◽  
Author(s):  
Roheena Abdullah ◽  
◽  
Maham Asim ◽  
Muhammad Nadeem ◽  
Kinza Nisar ◽  
...  

Proteases have gained more commercial value to date due to multiple applications in different industrial sectors. Current research was aimed to use the cheaper agricultural waste for optimal protease production. Maximum level of protease production was achieved at 37 °C, incubation period of 24 h, pH 9.0, inoculum size 3%, 1.5 g sucrose as a carbon source and 30% moisture content by using solid-state fermentation. Among the various inorganic and organic nitrogen sources, ammonium nitrate and yeast extract tremendously increased the production of protease. Among metal ions and surfactants tested, Ca2+ and Tween 40 showed the optimal protease production. The purification of protease was carried out by ammonium salt precipitation followed by sephadex G-100 gel filtration chromatography. The purification resulted in 1.3 fold of purified protease with a specific activity of 51.5 and a total yield of 37.5 %. Molecular weight of purified protease was predicted upon SDS-PAGE with a single band of ~36 kDa. The protease was quite stable over a temperature range of 35-45 °C and pH 7-9 with maximal activity at 40 °C and pH 9. The kinetic parameters Vmax (maximum rate) and Km (Michaelis-Menten constant) were calculated as 0.307 U/g and 11.2 mg/mL, respectively. The alkaline protease significantly de-hair the goat skin and successfully removed the animal blood stain from cotton cloth.

Engevista ◽  
2012 ◽  
Vol 15 (1) ◽  
pp. 61 ◽  
Author(s):  
Salah Din Mahmud Hasan ◽  
Citieli Giongo ◽  
Sonia Aparecida Reis Lopes-Shikida ◽  
Sérgio Luiz de Lucena ◽  
Mônica Lady Fiorese

The operation parameters used in solid-state fermentation (SSF) support the growth of filamentous fungi, which grow on solid substrates producing important metabolites such as proteases. The aim of this work is to obtain fungal proteases by SSF using the residues of a local brewery industry (barley bagasse and trub), that have high contents of proteins and soluble matter such as carbohydrates, vitamins, and mineral salts. The methodology includes the preparation of the residues, the screening of microorganisms, evaluation of the operation conditions for SSF using factorial design, purification and partial characterization of the protease. The results indicate that A. niger sp. UPT-03 isolated from the residue shows higher yields in terms of enzyme production (0.36 U gdm-1 h-1). The purification with DEAE-cellulose resulted in protease recovery with 30-fold of purification with a specific activity of 550 U mg protein-1. Higher proteolytic activity of purified enzyme was obtained at pH 5.5 and 55 ºC.


2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Abdilbar Usman ◽  
Said Mohammed ◽  
Jermen Mamo

Acid proteases represent an important group of enzymes, extensively used in food and beverage industries. There is an increased demand for acid proteases adapting to the industrial extreme environment, especially lower pH. Thus, this necessitates the search for a better acid protease from fungi that best performs in industrial conditions. The fungal isolates were isolated from grape and dairy farm soil using potato dextrose agar and further screened for protease production based on the hydrolysis of clear zone on skim milk agar. The potential fungi were then subjected to secondary screening under solid-state fermentation (SSF). After the secondary screening, the potential fungus was identified to the genus level by the macroscopic and microscopic methods. The growth conditions and media composition for the potential fungus were further optimized under SSF. The crude enzyme produced by the potential isolate was characterized after partial purification by acetone and ammonium sulfate precipitation. A total of 9 fungal isolates showed protease production in primary and secondary screening; however, one potential isolate (Z1BL1) was selected for further study based on its protease activity. The isolate was identified to the genus Aspergillus based on their morphological features. The maximum acid protease from the isolate Z1BL1 was obtained using fermentation media containing wheat bran as a solid substrate, 1 mL of 3.2 × 106 inoculum size, 50% moisture content, and pH 4.5 upon 120-h incubation at 30°C. The acetone-precipitated enzyme exhibited the maximum activity at 50°C and pH 5 with stability at pH 4–6 and temperature 40–60°C. Thus, the acid protease produced from Aspergillus showed suitable enzyme characteristics required in the industry and could be a candidate for application in the food industry after further purification.


2020 ◽  
Author(s):  
Abdilbar Usman ◽  
Said Mohammed ◽  
Jermen Mamo

Abstract 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.


2008 ◽  
Vol 83 (7) ◽  
pp. 1012-1018 ◽  
Author(s):  
Jarun Chutmanop ◽  
Sinsupha Chuichulcherm ◽  
Yusuf Chisti ◽  
Penjit Srinophakun

2018 ◽  
Vol 81 (1) ◽  
Author(s):  
Halifah Pagarra ◽  
Roshanida A. Rahman ◽  
Nur Izyan Wan Azelee ◽  
Rosli Md Illias

Polygalacturonases represent an important member of pectinases group of enzymes with immense industrial applications. The activity of exo-polygalacturonase produced by Aspergillus niger was studied in solid state fermentation (SSF) using Nephrolepis biserrata leaves as substrate. Central composite design (CCD) was used to optimize four significant variables resulted from the screening process that has been initially analyzed for the production of exo-polygalacturonase which are incubation time, temperature, concentration of pectin and moisture content. The optimum exo-polygalacturonase production obtained was 54.64 U/g at 120 hours of incubation time, temperature at 340C, 5.0 g/L of pectin concentration and 75.26% of moisture content. For partial characterization of exo-polygalacturonase, the optimum temperature and pH were obtained at 50°C and pH 4.0, respectively. SDS-PAGE analysis showed that molecular weight of exo-polygalacturonase were 35 and 71 kDa. This study has revealed a significant production of exo-polygalacturonase by A. niger under SSF using cheap and easily available substrate and thus could found immense potential application in industrial sectors and biotechnology


2012 ◽  
Vol 47 (1) ◽  
pp. 99-104 ◽  
Author(s):  
G Dharani ◽  
NS Kumaran

The purpose of this work is to study the optimized cultural conditions for the production of amylase by Aspergillus niger in solid state and submerged liquid fermentation. Four solid substrates banana peel, corn, potato and tapioca with different moisture conditions were taken for solid state fermentation (SSF). Basal medium was used for submerged liquid fermentation (SLF) with different pH (3 to 8), temperature (25, 30, 35 and 40ºC), carbon concentration (1, 2 and 3 g) and nitrogen source (0.1, 0.2 and 0.3 g). In SSF, tapioca yielded highest amylase activity and specific activity (4.43U/ml and 4.58U/mg) at 50% moisture content. In SLF, 2 g starch and 0.3 g peptone concentration showed 0.78 and 1.23 U/ml amylase activities under the optimum pH (5) and temperature (30ºC) the amylase activities reached to 0.86 U/ml and 0.76 U/ml respectively. In SSF using tapioca as substrate the enzyme yield is about five times higher than that achieved with submerged liquid culture. DOI: http://dx.doi.org/10.3329/bjsir.v47i1.7310 Bangladesh J. Sci. Ind. Res. 47(1), 99-104, 2012


2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Ponnuswamy Vijayaraghavan ◽  
Sreekumar Saranya ◽  
Samuel Gnana Prakash Vincent

Cow dung and agroresidues were used as the substrates for the production of alkaline proteases by Pseudomonas putida strain AT in solid-state fermentation. Among the various substrates evaluated, cow dung supported maximum (1351±217 U/g) protease production. The optimum conditions for the production of alkaline proteases were a fermentation period of 48 h, 120% (v/w) moisture, pH 9, and the addition of 6% (v/w) inoculum, 1.5% (w/w) trehalose, and 2.0% (w/w) yeast extract to the cow dung substrate. The enzyme was active over a range of temperatures (50–70°C) and pHs (8–10), with maximum activity at 60°C and pH 9. These enzymes showed stability towards surfactants, detergents, and solvent and digested various natural proteins.


2016 ◽  
Vol 182 (2) ◽  
pp. 511-528 ◽  
Author(s):  
R. Renganath Rao ◽  
M. Vimudha ◽  
N. R. Kamini ◽  
M. K. Gowthaman ◽  
B. Chandrasekran ◽  
...  

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