Factors Affecting Protease Production by Bacillus stearothermophilus RM-67

1983 ◽  
Vol 46 (12) ◽  
pp. 1020-1025 ◽  
Author(s):  
A. K. CHOPRA ◽  
D. K. MATHUR

Amongst the nitrogen sources, tryptone and yeast extract at 0.5% and 0.15% level, respectively, caused maximum enzyme production by Bacillus stearothermophilus RM-67. Addition of sodium chloride (0.5%) to the basal medium enhanced the enzyme production by 63%. Various sugars incorporated into the standardized basal medium proved inhibitory to enzyme elaboration. Maximum enzyme production was observed in the early decline growth phase of the organism in tryptone-yeast extract-salt medium (pH 6.5) when inoculated at 4% level and incubated on a rotary shaker at 55°C for 8 h and subsequently at 45°C up to 24 h.

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.


1979 ◽  
Vol 9 (1) ◽  
pp. 65-71
Author(s):  
Robert K. Bailey ◽  
Jack L. Voss ◽  
Rodney F. Smith

The rate of isolation of organisms resembling Haemophilus vaginalis (Corynebacterium vaginale) from vaginal specimens was not significantly affected by anaerobic versus carbon dioxide incubation atmospheres or whether specimens were inoculated on isolation media immediately after collection or after a delay of 6 h. Forty-one clinically isolated strains were provisionally divided into 30 H. vaginalis strains and 11 H. vaginalis -like (HVL) strains based on morphological and growth characteristics. The H. vaginalis strains were less reactive in API-20A identification test strips, (Analytab Products, Inc.) using Lombard-Dowell broth, than in a modified basal medium that contained proteose peptone no. 3 (Difco). The numbers and kinds of substrates fermented by 30 clinical and 2 reference strains of H. vaginalis varied among conventional, API, Minitek (Baltimore Biological Laboratory), and rapid buffered substrate fermentation systems. A greater number and variety of carbohydrates were fermented by the 11 HVL strains more consistently in all four test systems. Analysis of volatile and nonvolatile fermentation end products by gas-liquid chromatography did not reveal significant differences between the H. vaginalis and HVL strains. However, the latter group grew in peptone-yeast extract-glucose broth, whereas the H. vaginalis strains did not grow without the addition of starch to peptone-yeast extract-glucose. All of the reference and clinical strains were similar in their susceptibilities to a variety of antimicrobial compounds except sulfonamides, which inhibited the HVL strains and bifidobacteria but not the H. vaginalis strains. Sulfonamide susceptibility or resistance corresponded in part to the H. vaginalis and HVL-bifidobacteria strain reactions on selected conventional fermentation substrates. Susceptibility or resistance to sulfonamides and metronidazole in conjunction with fermentation tests is described to aid in the separation of H. vaginalis from other possibly unrecognized biotypes of H. vaginalis or other vaginal bacteria that presumptively resemble the organism. A human blood medium known as V agar was also of considerable value in distinguishing H. vaginalis from HVL strains, because only the H. vaginalis strains produced diffuse beta-hemolysis on V agar.


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.


1950 ◽  
Vol 28c (6) ◽  
pp. 586-599 ◽  
Author(s):  
W. M. Dion

The main factors that influence the production of proteolytic enzymes by a few selected cultures have been studied. The time taken to reach the maximum yield of proteases is dependent upon the growth rate of each organism, and varies from two to five days. The fungi tested require the presence of an easily available carbohydrate source in addition to a protein substrate in order to produce high yields of proteolytic enzymes. The Streptomyces cultures will produce proteases in the absence of a carbohydrate source, but yields are generally low. The fungi studied will not produce significant amounts of proteases when grown on predominately inorganic nitrogen sources in contrast with the Streptomyces cultures, one of which produced almost as high yields of proteolytic enzymes when grown with sodium nitrate as when grown with Klim. Of a number of protein sources Klim and malt sprouts provided the best media for protease production. The temperature of incubation and pH of the culture medium are also important factors affecting the yield of proteolytic enzymes.


1981 ◽  
Vol 44 (9) ◽  
pp. 661-664 ◽  
Author(s):  
A. K. CHOPRA ◽  
HARISH CHANDER ◽  
V. K. BATISH ◽  
B. RANGANATHAN

Mucor racemosus isolated from butter exhibited maximum lipase production at 22 C in 3 days at pH 5.0, when grown as a static culture. Supplementation of the medium with glucose and lactose at 1% level resulted in maximum (18.0 μmoles FFA) and minimum (10.5 μmoles FFA) enzyme production. Of the five different nitrogen sources tested, peptone at the 2% level supported highest production of lipase. Calcium, potassium and sodium citrate (0.1 %), when incorporated in the medium, stimulated production of enzyme to the extent of 80, 60 and 47%, respectively.


2017 ◽  
Vol 22 (1) ◽  
pp. 31 ◽  
Author(s):  
Eris Septiana ◽  
Partomuan Simanjuntak

Antioxidant is an interesting topic due to their capability to inhibit free radical and prevent damage because of oxidative processes. Endophyt fungi is one of antioxidant compound resources in nature. The low yield to gain antioxidant compound from fungi challenges to look for the composition of media and optimalization of growth conditions. This research aimed to know the effect of medium condition in different carbon and nitrogen sources as well as initial pH towards antioxidant activity of endophyt fungi Bo.Ci.Cl.A3. Shaker fermentation was used on 120 rpm at room temperature for 14 days. The carbon sources were glucose, sucrose, and starch and nitrogen sources were NaNO3, NH4NO3, and yeast extract with initial pH at 5, 7, and 9. Ethyl acetate was used as extractor. The results showed that endophyt fungi can produce secondary metabolite as antioxidant at all variation of fermented media. The nitrogen source of yeast extract could increase antioxidant activity of endophyt fungi Bo.Ci.Cl.A3, while other sources such as nitrogen source, carbon sources, and different initial pH on the basal medium that were used did not give increasing antioxidant activity. The conclusion of this research was the substitution of nitrogen source with yeast extract (3 g/L) on the basal medium Czapek Dox’s Broth could increase antioxidant activity of endophyt fungi Bo.Ci.Cl.A3.


Author(s):  
Malika Benkahoul ◽  
Amina Bramki ◽  
Aicha Belmessikh ◽  
Aicha Mechakra-Maza

The production of an acid protease by liquid fermentation is performed on a medium based on decommissioned dates by a local mold isolated from an extreme environment. The used mold is isolated from the thermal soil in Hammam Safsaf of Teleghma (Algeria). Phenotypic and molecular identification has shown that it is Aspergillus niger. This fungi strain exhibited exo-protease activity on milk agar. The fermentation medium based on decommissioned dates is enriched by other factors according to a statistical method; the plan of Plackett and Burman. This method (N = 8 experiments and N-1 factors) allowed the optimization of the enzyme production and the growth of the mold. The statistical analysis of the obtained results shows an increased enzyme activity (650.20U), in the presence of yeast extract and salts with probabilities p = 0.239 for the first and p = 0.190 for the second. Furthermore, the corn-steep-liquor (p = 0.229) and yeast extract (p = 0.053) present a significant effect on mold growth. In conclusion, the culture of A. niger on optimized medium gives good yields of biomass and proteolytic activity compared to the basal medium. These results are encouraging. In fact, the use of a cheap and available substrate such as decommissioned dates saves the cost price of fermentation on an industrial scale.


1971 ◽  
Vol 24 ◽  
Author(s):  
W. H. Verstraete

Some  factors affecting the L-asparaginase activity of E.  aroideae were investigated. Increasing  concentrations of glucose in the culture medium had an inhibiting effect on  the production of L-asparaginase by this microorganism. Buffering of the  culture medium in order to stabilize the pH during growth resulted in a decrease  of the L-asparaginase activity. From the different nitrogen sources examined,  tryptone, proteose peptone nr 2 and nr 3 stimulated the L-asparaginase  production. Toluene treatment of the cells practically destroyed the  L-asparaginase. Acetone dried cells showed an L-asparaginase activity  comparable with the activity of living cells.


1952 ◽  
Vol 30 (4) ◽  
pp. 360-370 ◽  
Author(s):  
Egon Stark ◽  
P. A. Tetrault

Thirty-five cultures of Bacillus stearothermophilus hydrolyzed five starches under various cultural conditions. Hydrolysis occurred regardless of the type, brand, or batch of starch; regardless of the initial pH or of the subsequent pH changes of the medium. Starch in broth was better attacked than in agar media. Some cultures hydrolyzed 0.5%, but not 1% starch; others hydrolyzed easily 10% soluble starch. Length of incubation was important. Certain cultures never formed acid or sugar from starch. Dextrinization was a more reliable indication of starch hydrolysis than was the formation of acid or sugar. Soluble starch gave more consistent results in repeated experiments than did nonsoluble starches. The type of protein medium determines strongly the formation of amylase. Trypticase was the best commercial medium, yeast extract came second. The other 10 media yielded fewer amylolytic cultures. Yeast extract added to media enhanced amylase formation, except with trypticase. Tryptose, proteose-peptone, and neopeptone inhibited the growth of most cultures.


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