scholarly journals Optimization Conditions of Extracellular Proteases Production from a Newly IsolatedStreptomyces PseudogrisiolusNRC-15

2012 ◽  
Vol 9 (2) ◽  
pp. 949-961 ◽  
Author(s):  
El-Sayed E. Mostafa ◽  
Moataza M. Saad ◽  
Hassan M. Awad ◽  
Mohsen H. Selim ◽  
Helmy M. Hassan

Microbial protease represents the most important industrial enzymes, which have an active role in biotechnological processes. The objective of this study was to isolate new strain ofStreptomycesthat produce proteolytic enzymes with novel properties and the development of the low-cost medium. An alkaline protease producer strain NRC-15 was isolated from Egyptian soil sample. The cultural, morphological, physiological characters and chemotaxonomic evidence strongly indicated that the NRC-15 strain represents a novel species of the genusStreptomyces, hence the nameStrptomyces pseudogrisiolusNRC-15. The culture conditions for higher protease production by NRC-15 were optimized with respect to carbon and nitrogen sources, metal ions, pH and temperature. Maximum protease production was obtained in the medium supplemented with 1% glucose, 1% yeast extract, 6% NaCl and 100 μmol/L of Tween 20, initial pH 9.0 at 50 °C for 96 h. The current results confirm that for this strain, a great ability to produce alkaline proteases, which supports the use of applications in industry.

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.


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.


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 ◽  
Author(s):  
Tayachew Desalegn ◽  
◽  
Ketema Bacha ◽  
Mesfin Tafesse ◽  
Chandran Masi ◽  
...  

Protease also called proteinase or peptidase is a digestive enzyme that is categorized under proteolytic enzymes and it has great potential in industrial application. Extracellular proteases are used in a variety of industries because they exhibit practically all of the characteristics needed for biotech applications such as detergent, bioremediation, food, and leather processing. In the synthesis of all three major types of acidic, neutral, and alkaline proteases, microbial sources have dominated an unbeatable area. Alkaline proteases are a large group of industrial enzymes formed by a wide variety of species, including animals, fungi, and bacteria. The fermentation method serves to make bacteria, fungi, and yeast alkaline proteases. Proteases are produced in large quantities by Gram-positive bacteria, especially those belonging to the Bacillus genus. Following standard procedures, the bacterial isolates PMOJ-01 and PMOJ-05 with the prominent zone of clearance and efficient enzyme development were further characterized to the genus level. Moreover, the growth conditions for the highest protease production were optimized with different pH, temperatures, and NaCl concentrations, in the results of PMOJ-01 and PMOJ- 05 pH (7 and 8), temperatures 45oC, and 1% NaCl concentrations both cases respectively. The proteases activities from PMOJ-01, Pseudomonas aeruginosa, and PMOJ-05, Bacillus subtilis were most active at pH 7.0 and pH 8.0 and temperature at 35oC and 45oC, respectively. The enzyme activity and the total solid protease sample of the crude enzyme of Pseudomonas aeruginosa and Bacillus subtilis were 0.299 U/ mL and 0.289 U/ mL, 1.37±0.14 U/mg, and 1.199 U/mg respectively. The effect on dehairing, distaining, and scum removal revealed that the purified protease enzyme of PMOJ-01 and PMOJ-05 can be used in detergent and leather industries.


2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Rayda Siala ◽  
Fakher Frikha ◽  
Samiha Mhamdi ◽  
Moncef Nasri ◽  
Alya Sellami Kamoun

Medium composition and culture conditions for the acid protease production byAspergillus nigerI1 were optimized by response surface methodology (RSM). A significant influence of temperature, KH2PO4, and initial pH on the protease production was evaluated by Plackett-Burman design (PBD). These factors were further optimized using Box-Behnken design and RSM. Under the proposed optimized conditions, the experimental protease production (183.13 U mL−1) closely matched the yield predicted by the statistical model (172.57 U mL−1) withR2=0.914. Compared with the initial M1 medium on which protease production was 43.13 U mL−1, a successful and significant improvement by 4.25 folds was achieved in the optimized medium containing (g/L): hulled grain of wheat (HGW) 5.0; KH2PO41.0; NaCl 0.3; MgSO4(7H2O) 0.5; CaCl2(7H2O) 0.4; ZnSO40.1; Na2HPO41.6; shrimp peptone (SP) 1.0. The pH was adjusted at 5 and the temperature at30°C. More interestingly, the optimization was accomplished using two cheap and local fermentation substrates, HGW and SP, which may result in a significant reduction in the cost of medium constituents.


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.


Polymers ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 287
Author(s):  
Thi Ngoc Tran ◽  
Chien Thang Doan ◽  
San-Lang Wang

Agro-byproducts can be utilized as effective and low-cost nutrient sources for microbial fermentation to produce a variety of usable products. In this study, wheat bran powder (WBP) was found to be the most effective carbon source for xylanase production by Streptomyces thermocarboxydus TKU045. The optimal media for xylanase production was 2% (w/v) WBP, 1.50% (w/v) KNO3, 0.05% (w/v) MgSO4, and 0.10% (w/v) K2HPO4, and the optimal culture conditions were 50 mL (in a 250 mL-volume Erlenmeyer flask), initial pH 9.0, 37 °C, 125 rpm, and 48 h. Accordingly, the highest xylanase activity was 6.393 ± 0.130 U/mL, 6.9-fold higher than that from un-optimized conditions. S. thermocarboxydus TKU045 secreted at least four xylanases with the molecular weights of >180, 36, 29, and 27 kDa when cultured on the WBP-containing medium. The enzyme cocktail produced by S. thermocarboxydus TKU045 was optimally active over a broad range of temperature and pH (40–70 °C and pH 5–8, respectively) and could hydrolyze birchwood xylan to produce xylobiose as the major product. The obtained xylose oligosaccharide (XOS) were investigated for 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and the growth effect of lactic acid bacteria. Finally, the solid waste from the WBP fermentation using S. thermocarboxydus TKU045 revealed the high adsorption of Congo red, Red 7, and Methyl blue. Thus, S. thermocarboxydus TKU045 could be a potential strain to utilize wheat bran to produce xylanases for XOS preparation and dye adsorbent.


2019 ◽  
Vol 19 (2) ◽  
pp. 470 ◽  
Author(s):  
Siti Nor Amira Rosli ◽  
Rohaida Che Man ◽  
Nasratun Masngut

Culture conditions including initial pH media, incubation period, inoculum size, type of carbon source, type of nitrogen source and its concentration, which affect xylanase production were screened via the one-factor-at-a-time approach. The bacteria used in the production of xylanase was isolated from the landfill site at Sg. Ikan, Kuala Terengganu, Malaysia. Three characterizations of the landfill soil were investigated for their moisture content, ash content, and pH. The culture conditions range used in the experimental work were between 6–30 h for the incubation period, with initial pH between 5–9, inoculum size between 1–20% v/v, carbon, nitrogen sources, and nitrogen source concentration between 1–5% w/v. Xylanase activity was estimated using dinitrosalicylic acid (DNS) based on the release of xylose under standard assay conditions. The landfill soil was observed to have pH between pH 3.4–7.2 with a moisture content between 12.4–33.7% and ash ranged between 3.5–4.3%. Results showed that the highest xylanase activity within studied ranges was recorded at 25.91±0.0641 U/mL with 10% (v/v) inoculum size, 1% (w/v) xylose as sole carbon source, mixture of 1% (w/v) peptone and 0.25% (w/v) ammonium sulphate as nitrogen sources, which was carried out at initial pH of 8.0 for 24 h incubation.


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