scholarly journals Production, purification, and characterization of alkaline protease from Aspergillus flavus and its compatibility with commercial detergents

BioResources ◽  
2020 ◽  
Vol 16 (1) ◽  
pp. 291-301
Author(s):  
Amtul W. Wajeeha ◽  
M. Javaid Asad ◽  
Raja Tahir Mahmood ◽  
Tayyaba Zainab ◽  
Sidrah Nazir ◽  
...  

Aspergillus flavus was used to produce alkaline protease. Solid state fermentation (SSF) strategy was adopted to explore the most favorable physical and nutritional conditions for enzyme production. Maximum production was achieved at pH 6.0 and a temperature of 30 °C after 84 h of growth period. For the optimization of the chemical parameters, different carbon and nitrogen sources were used including glucose, fructose, sucrose, ammonium sulphate, and urea. Maximum production was observed with 0.3% concentration of all the compounds. Ammonium sulphate salting out and gel chromatography was used to purify the enzyme. The enzyme was completely precipitated out at 80% saturation. The value of Vmax was 3.9 U/mL, while the value of Km was 1.9 mg/mL. The enzyme was tested for its compatibility with a few famous detergents available on the market. With the alkaline protease under study, the enzyme displayed a maximum retention of its activity i.e. 80.8% in the presence of commercial detergent Surf excel. The activity dropped down to 61.5% when the enzyme was allowed to work in the presence of another locally used detergent, i.e., Bonus. Protease production from A. flavus was carried out on rice bran and wheat bran and the wheat bran gave better results.

2019 ◽  
pp. 1-8
Author(s):  
Tapasi Polley ◽  
Uma Ghosh

Aim: The current study aimed at studying the optimum fermentation conditions and nutritional conditions for alkaline protease production by submerge fermentation using Alternaria alternata TUSGF1, isolated from poultry farm soil. Study Design: The results of environmental and nutritional parameters for protease production by OVAT method was analyzed by origin 6.1 software. Place and Duration of Study: Department of Food technology and Biochemical Engineering, Jadavpur University, Kolkata, West Bengal, India between March 2017 and May 2017. Methodology: A protease producing microorganism was isolated from a poultry farm soil and identified as Alternaria alternata TUSGF1. Various environmental and nutritional process parameters such as volume of medium, fermentation time, temperature, age of inoculums, agitation and carbon sources and nitrogen sources were standardized for the maximum yield of alkaline protease. Results: The optimum conditions of protease activity was 30°C at volume of medium 60 ml with 7 days age of inoculum in the medium containing 168 h of incubation and 120 rpm agitation rate. Peptone, casien, skimmed milk, urea and yeast extract were good nitrogen sources whilst maltose, fructose, starch, and sucrose were appropriate for enzyme production by submerge fermentation. Conclusion: Alkaline protease production by a newly isolated Alternaria alternata TUSGF1 from poultry farm soil was studied in shake flask conditions by submerge fermentation. It was established that the optimum protease production was recorded at 30°C, 60 ml volume of medium leaves and incubation time of 168 h. The best carbon and nitrogen sources for protease production were fructose and casein, respectively.


2021 ◽  
Vol 18 (1) ◽  
pp. 185-195
Author(s):  
Gousiya Begum ◽  
Srinivas Munjam

Pectinases are the commercial enzymes that are abundantly employed in various industries like fruit juice industries for clarification, wine indutsry and paper industry for bleaching up pulp. The present work was done on culture conditions optimization for production of pectinases under submerged fermentation using wheat bran as a substrate. Fungal strains were isolated from vegetable waste dump yard soils of Warangal district of Telangana state and screened for their activity on pectin agar medium. Among 30 isolates, two fungal strains showed good activity and identified them as A. niger and A. flavus. The effects of the different carbon and nitrogen sources on pectinases viz. exo-PG, endo-PG, endo-PL and PME by A. niger with 1% wheat bran was carried out in submerged fermentation. These studies revealed that carbon and nitrogen sources have shown considerable influence on enzyme production. Among all the carbon sources tried, sucrose at 1% was shown to be efficient carbon source for all four types of pectinases under investigation. For endo-PG, endo-PL and PME maximum enzyme production were recorded on 8th day of incubation period but for exo-PG enhanced production was observed on 12th day. A. niger could not produce PME on 12th day from 2.50% to subsequent concentrations. Among nine different nitrogen sources were screened, maximum pectinase production was recorded in sodium nitrate at 0.2 % for A. niger. Endo-PG, endo-PL and PME maximum production were recorded on 8th day of incubation and for exo-PG maximum production was observed on 12th day. No PME production was observed in A. niger on 12th day.


2020 ◽  
Author(s):  
Sangeeta Negi ◽  
Sapna Jain ◽  
Anand Kumar

Abstract In order to achieve high yield of fungal protease in a very cost effective way and to meet its increased market demand, current study deals with the screening of various agro-wastes as carbon source for the production of protease from Rhizopus oryzae (SN5) / NCIM-1447 under solid state fermentation. Substrates and culture parameters such as wheat bran, soybean meal, black-gram husk, rice husk, mixture of wheat bran, soybean meal, nitrogen sources, pH, temperature and incubation time were first optimized with one factor at time strategy and then EVOP factorial and yield of alkaline protease was achieved 412.8 U/gds at 28 o C and pH=6 after 72 hours of fermentation taking wheat bran and soybean as a substrate in 4:1 ratio. Further Artificial Neural Networks (ANN), was trained with data of EVOP and yield of protease was enhanced up to 422.6 U/gds with wheat bran: soyabean in ratio of 70:30, pH 6.2 at 30°C. The evolved process and Rhizopus oryzae (SN5)/ NCIM-1447 strain would be promising for protease production at industrial scale at low cost.


Molecules ◽  
2021 ◽  
Vol 26 (4) ◽  
pp. 1139
Author(s):  
Aida Karray ◽  
Mona Alonazi ◽  
Habib Horchani ◽  
Abir Ben Bacha

This study was conducted to identify a new alkaline and thermophilic protease (Ba.St.Pr) produced from Bacillus stearothermophilus isolated from olive oil mill sols and to evaluate its culture conditions, including temperature, pH, carbon and nitrogen sources, and incubation time. The optimum culture conditions for cell growth (10 g/L) and protease production (5050 U/mL) were as follows: temperature 55 °C, pH 10, inoculation density 8 × 108 CFU/mL, and incubation time 24 h. The use of 3% yeast extract as the nitrogen sources and galactose (7.5 g/L) as the carbon sources enhanced both cell growth and protease production. Using reversed-phase analytical HPLC on C-8 column, the new protease was purified with a molecular mass of approximately 28 kDa. The N-terminal sequence of Ba.St.Pr exhibited a high level of identity of approximately 95% with those of Bacillus strains. Characterization under extreme conditions revealed a novel thermostable and alkaline protease with a half-life time of 187 min when incubated with combined Ca2+/mannitol. Ba.St.Pr demonstrated a higher stability in the presence of surfactant, solvent, and Ca2+ ions. Consequently, all the evaluated activity parameters highlighted the promising properties of this bacterium for industrial and biotechnological applications.


2018 ◽  
Vol 10 (4) ◽  
pp. 1210-1215 ◽  
Author(s):  
Raghu Ram M ◽  
Suman Kumar Yepuru

Aspergillus oryzae isolatedon  Potato dextrose agar  from soil samples of kottakoduru seashore of Bay of Bengal, Andhra Pradesh, India seashore of Bay of Bengal by spread plate method and was screened for alkaline protease production on Skim milk containing agar plates and identified by clear zones of protein hydrolysis around colonies. Different physical and chemical parameters such as pH, temperature, substrate concentration and incubation time were optimized for the better production of alkaline protease. The maximum protease activity was found at pH of 8 containing 10% wheat bran at 300C, after 72 hours of fermentation.ZnSO4was effective activator for protease activity and sodium dsulphate had shownmore than 50% inhibition of enzyme activity. Among the different oil cakes used for the production of enzyme the Sesame  oil cake proved to be suitable substrate after wheat bran for the production of protease by Aspergillus oryzae.


1963 ◽  
Vol 9 (2) ◽  
pp. 211-220 ◽  
Author(s):  
D. W. S. Westlake

A number of molds and bacteria were screened for their ability to degrade quercitrin. The molds, but not the bacteria, were particularly active and produced carbon monoxide. The degradation of quercitrin is dependent upon the synthesis of an inducible glycosidase (quercitrinase). This enzyme is synthesized by only a few members of the Aspergillus flavus group. Two of these strains synthesized quercitrinase and excreted it and other enzymes into the culture medium. Maximum production of quercitrinase was obtained with organic nitrogen sources such as yeast extract or phytone. Quercitrinase is induced by readily metabolized flavonols and flavonol-glycosides. The glycosidase is quite specific, liberating the rhamnose from the 3-position of quercitrin and myricitrin and the 7-position of robinin. The aglycone, quercetin, is subsequently metabolized to carbon monoxide and the depside of phloroglucinol-carboxylic acid and proto-catechuic acid. Evidence is also presented for an alternative pathway for the metabolism of the flavonol nucleus.


2020 ◽  
Author(s):  
Sangeeta Negi ◽  
Kumari Vibha

Abstract In order to achieve high yield of fungal protease in a very cost effective way and to meet its increased market demand, current study deals with the screening of various agro-wastes as carbon source for the production of protease from Rhizopus oryzae (SN5) / NCIM-1447 under solid state fermentation. Substrates and culture parameters such as wheat bran, soybean meal, black-gram husk, rice husk, mixture of wheat bran, soybean meal, nitrogen sources, pH, temperature and incubation time were first optimized with one factor at time strategy and then EVOP factorial and yield of alkaline protease was achieved 412.8 U/gds at 28oC and pH=6 after 72 hours of fermentation taking wheat bran and soybean as a substrate in 4:1 ratio. Further Artificial Neural Networks (ANN), was trained with data of EVOP and yield of protease was enhanced up to 422.6 U/gds with wheat bran: soyabean in ratio of 70:30, pH 6.2 at 30°C. The evolved process and Rhizopus oryzae (SN5)/NCIM-1447 strain would be promising for protease production at industrial scale at low cost.


2018 ◽  
Vol 7 (3) ◽  
pp. 48
Author(s):  
Hind A. A. Al-Zahrani

A potent alkaline protease producing strain characterized and identified as Streptomyces sp. H1 was isolated from soil around red sea shore. The enzyme was produced extracellulary in submerged culture revealed maximum level during early stationary phase. Alkaline protease showed the highest activity at incubation time, pH and inoculum size of 3 days, 9 and 8% respectively. Among different carbon sources beet molasses gave a maximum production followed by starch, sucrose and fructose. High yield of protease production was noticed with casein followed by peptone, yeast extract and ammonium sulphate. Furthermore, it was optimized with 7g/l NaCl resulted in higher level of protease. Optimization of the process parameters resulted in about 3.4 fold increase in the alkaline protease. Partial purification of the crude enzyme was achieved by fractional precipitation using ammonium sulfate at 50% saturation. Due to the maximum production of protease in the presence of cheaper substrate as beet molasses, stability at alkaline pH 9 and temperature up to 70 oC besides salt tolerance make the strain and its enzyme useful in different industrial applications.  


2020 ◽  
Author(s):  
Sangeeta Negi ◽  
Sapna Jain ◽  
Anand Kumar

Abstract In order to achieve high yield of fungal protease in a very cost effective way and to meet its increased market demand, current study deals with the screening of various agro-wastes as carbon source for the production of protease from Rhizopus oryzae (SN5) / NCIM-1447 under solid state fermentation. Substrates and culture parameters such as wheat bran, soybean meal, black-gram husk, rice husk, mixture of wheat bran, soybean meal, nitrogen sources, pH, temperature and incubation time were first optimized with one factor at time strategy and then EVOP factorial and yield of alkaline protease was achieved 412.8 U/gds at 28 o C and pH=6 after 72 hours of fermentation taking wheat bran and soybean as a substrate in 4:1 ratio. Further Artificial Neural Networks (ANN), was trained with data of EVOP and yield of protease was enhanced up to 422.6 U/gds with wheat bran: soyabean in ratio of 70:30, pH 6.2 at 30°C. The evolved process and Rhizopus oryzae (SN5)/ NCIM-1447 strain would be promising for protease production at industrial scale at low cost.


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