scholarly journals Optimization of Culture Conditions for Protease Production using Three Strains of Bacillus

Author(s):  
Cyr Jonas Morabandza ◽  
Valentin Dibangou ◽  
Faly Armel Soloka Mabika ◽  
Elgie Viennechie Gatse ◽  
Tarcisse Baloki Ngoulou ◽  
...  

The aim of this work was to determine the effect of a few external factors on bacterial growth and the production of enzymes with a proteolytic effect in three strains of Bacillus: CMS5 (Bacillus subtilis), CMS4 (Bacillus sp.) and SPo5 14′ (Bacillus velenzensis) isolated from squashes packed in traditionally prepared cassava leaves, but also to determine the best source of carbon and nitrogen. All three strains have the ability to actively degrade milk casein. The strains were grown in Luria Bertani medium and the suspension from the cell culture was used to measure optical density and demonstrate enzyme activity on a petri dish containing skim milk. Several parameters were verified including the influence of temperature, pH, and carbon and nitrogen source on growth and enzyme production. Growth was possible from 25 to 60°C with an optimal temperature of 30°C after 24 hours. Enzyme production was observed from 25 to 55°C with an optimum at 37°C. For pH, growth and enzyme production was possible from pH 5.7 and 9 with an optimum of 7 in all three strains. Among the sources of carbons used, galactose is the best source for growth after 24 h in all three strains, and starch for production. Among nitrogen sources, Bacto-peptone is best for growth as well as production.

2021 ◽  
Vol 23 (10) ◽  
pp. 222-235
Author(s):  
Malathi, M ◽  
◽  
Latha, D ◽  

As thermostable protease has more commercial value in different industries, the aim of this study was to search for such an enzyme producing bacteria from the microbial mats. Investigation was continued on the isolate for its ability to produce mass amount of enzyme and its activity under suitable optimized conditions. Different parameters including cheap carbon and nitrogen substrates, inoculum size and temperature was selected to optimize the enzyme production conditions. Initially five different isolates from two microbial mats collected from different sources were analyzed for its ability to produce thermostable protease after exposing to higher temperature incubation conditions. Test culture tentatively named as 1F from microbial mat-1 was selected as more enzyme producer among the ten isolates. The organism was selected based on the zone of clearance on skim milk agar by the isolate, that indicating more protease production. Under each optimization parameter, each type of carbon (Lactose), and nitrogen (yeast extract) source showed more enzyme production and activity respectively. About 1% inoculum size and a thermostable temperature of 45°C produced significant amount of enzyme and its activity. The obtained results emphasized the need for thermostable protease for different commercial industries in the existing and near future.


2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Abbas Akhavan Sepahy ◽  
Leila Jabalameli

Soil samples of Tehran jungle parks were screened for proteolytic Bacilli. Among eighteen protease producers one of the isolates obtained from Lavizan park, in north east of Tehran, was selected for further experimental studies. This isolate was identified as Bacillus sp. strain CR-179 based on partial sequencing of 16S rRNA. Various nutritional and environmental parameters affected protease production by Bacillus sp. strain CR-179. Protease production by this Bacillus cultivated in liquid cultures reached a maximum at 24 h, with levels of 340.908 U/mL. Starch and maltose were the best substrates for enzyme production while some pure sugars such as fructose, glucose, and sucrose could not influence production of protease. Among various organic nitrogen sources corn steep liquor, which is commercial, was found as the best substrate followed by yeast extract, whey protein, and beef extract. The optimal pH and optimal temperature of enzyme production were 8.0 and 45°C, respectively. Studies on enzymatic characterization revealed that crude protease showed maximum activity at pH 9.0 and 60°C, which is indicating the enzyme to be thermoalkaline protease.


2020 ◽  
Vol 19 (1) ◽  
pp. 11-20
Author(s):  
Aysun Pekşen ◽  
Beyhan Kibar

Macrolepiota procera, commonly called the Parasol Mushroom, is a delicious mushroom collected from the nature and commonly consumed by the public in many regions of Turkey. This study was conducted to determine the optimum culture conditions (pH, temperature, carbon and nitrogen sources) for mycelial growth of M. procera. Three pH values (pH 5.0, 5.5 and 6.0), four incubation temperatures (15, 20, 25 and 30°C), seven carbon (C) sources (dextrose, glucose, lactose, maltose, mannitol, sucrose and xylose) and six nitrogen (N) sources ((NH4)2HPO4, NH4NO3 and Ca(NO3)2, malt extract, peptone and yeast extract) were investigated. In the second step of the study, the effect of seven pH values (4.0, 4.5, 5.0, 5.5, 6.0, 6.5 and 7.0) on the mycelial colony diameter was examined at 20 and 25°C since these temperatures gave the best mycelial growth in the previously conducted temperature experiment. The best mycelial growth was determined at pH 6.0. The optimum temperature for mycelial growth of M. procera was found as 25°C. The use of glucose as carbon source and yeast extract and peptone as nitrogen source in the culture medium gave the best results for mycelial growth. Determining of optimum culture conditions for mycelial growth of M. procera will provide important contributions to the fortcoming studies on it’s commercially cultivation in Turkey.


Biomolecules ◽  
2019 ◽  
Vol 9 (9) ◽  
pp. 461 ◽  
Author(s):  
Soo Kweon Lee ◽  
Ju Hun Lee ◽  
Hyeong Ryeol Kim ◽  
Youngsang Chun ◽  
Ja Hyun Lee ◽  
...  

Cordycepin, a beneficial bioactive product specifically found in Cordyceps, has received attention in various bioindustrial applications such as in pharmaceuticals, functional foods, and cosmetics, due to its significant functions. However, low productivity of cordycepin is a barrier to commercialization. In this study, Cordyceps militaris was mutated by UV irradiation to improve the cordycepin production. The highest producer KYL05 strain was finally selected and its cordycepin production was increased about 1.5-fold compared to wild type. In addition, the effects of culture conditions were fundamentally investigated. Optimal conditions were as follows: pH 6, temperature of 25 °C, shaking speed of 150 rpm, and culture time of 6 days. Effects of medium component on cordycepin production were also investigated by using various carbon and nitrogen sources. It was found that glucose and casein hydrolysate (CH) were most effective as carbon and nitrogen sources in cordycepin production (2.3-fold improvement) with maximum cordycepin production of about 445 mg/L. In particular, production was significantly affected by CH. These results should be of value in improving the efficiency of mass production of cordycepin.


2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Gustavo Carvalho do Nascimento ◽  
Ryhára Dias Batista ◽  
Claudia Cristina Auler do Amaral Santos ◽  
Ezequiel Marcelino da Silva ◽  
Fabrício Coutinho de Paula ◽  
...  

β-fructofuranosidase (invertase) andβ-D-fructosyltransferase (FTase) are enzymes used in industrial processes to hydrolyze sucrose aiming to produce inverted sugar syrup or fructooligosaccharides. In this work, a blackAspergillussp. PC-4 was selected among six filamentous fungi isolated from canned peach syrup which were initially screened for invertase production. Cultivations with pure carbon sources showed that invertase and FTase were produced from glucose and sucrose, but high levels were also obtained from raffinose and inulin. Pineapple crown was the best complex carbon source for invertase (6.71 U/mL after 3 days of cultivation) and FTase production (14.60 U/mL after 5 days of cultivation). Yeast extract and ammonium chloride nitrogen sources provided higher production of invertase (6.80 U/mL and 6.30 U/mL, respectively), whereas ammonium nitrate and soybean protein were the best nitrogen sources for FTase production (24.00 U/mL and 24.90 U/mL, respectively). Fermentation parameters for invertase using yeast extract wereYP/S= 536.85 U/g andPP= 1.49 U/g/h. FTase production showed values ofYP/S= 2,627.93 U/g andPP= 4.4 U/h using soybean protein. The screening for best culture conditions showed an increase of invertase production values by 5.10-fold after 96 h cultivation compared to initial experiments (fungi bioprospection), while FTase production increased by 14.60-fold (44.40 U/mL) after 168 h cultivation.A. carbonariusPC-4 is a new promising strain for invertase and FTase production from low cost carbon sources, whose synthesized enzymes are suitable for the production of inverted sugar, fructose syrups, and fructooligosaccharides.


2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
S. P. Gautam ◽  
P. S. Bundela ◽  
A. K. Pandey ◽  
Jamaluddin Khan ◽  
M. K. Awasthi ◽  
...  

The main purpose of this study is to reduce the production cost of cellulase by optimizing the production medium and using an alternative carbon source such as municipal solid waste residue. In the present investigation, we aim to isolate the two novel cellulase producing fungi (Aspergillus niger and Trichoderma sp.) from municipal solid waste. Municipal solid waste residue (4-5% (w/v)) and peptone and yeast extract (1.0% (w/v)) were found to be the best combination of carbon and nitrogen sources for the production of cellulase by A. niger and Trichoderma sp. Optimum temperature and pH of the medium for the cellulase production by A. niger were 40°C and 6-7, whereas those for the production of cellulase by Trichoderma sp. were 45°C and 6.5. Cellulase production from A. niger and Trichoderma sp. can be an advantage as the enzyme production rate is normally higher as compared to other fungi.


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 ◽  
Vol 10 (24) ◽  
pp. 8867
Author(s):  
Osama M. Darwesh ◽  
Ibrahim A. Matter ◽  
Hesham S. Almoallim ◽  
Sulaiman A. Alharbi ◽  
You-Kwan Oh

The color of food is a critical factor influencing its general acceptance. Owing to the effects of chemical colorants on health, current research is directly aimed at producing natural and healthy food colorants from microbial sources. A pigment-producing fungal isolate, obtained from soil samples and selected based on its rapidity and efficiency in producing red pigments, was identified as Monascus ruber OMNRC45. The culture conditions were optimized to enhance pigment production under submerged fermentation. The optimal temperature and pH for the highest red pigment yield were 30 °C and 6.5, respectively. The optimum carbon and nitrogen sources were rice and peptone, respectively. The usefulness of the pigment produced as a food colorant was evaluated by testing for contamination by the harmful mycotoxin citrinin and assessing its biosafety in mice. In addition, sensory evaluation tests were performed to evaluate the overall acceptance of the pigment as a food colorant. The results showed that M. ruber OMNRC45 was able to rapidly and effectively produce dense natural red pigment under the conditions of submerged fermentation without citrinin production. The findings of the sensory and biosafety assessments indicated the biosafety and applicability of the red Monascus pigment as a food colorant.


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