scholarly journals Isolation of Thermostable Protease Producing Bacteria from the Microbial Mats

2021 ◽  
Vol 23 (10) ◽  
pp. 222-235
Author(s):  
Malathi, M ◽  
◽  
Latha, D ◽  
Keyword(s):  
Enzyme Production ◽  
Microbial Mats ◽  
Skim Milk ◽  
Inoculum Size ◽  
Protease Production ◽  
Carbon And Nitrogen ◽  
Test Culture ◽  
Higher Temperature ◽  
Optimized Conditions ◽  
Near Future

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.

scholarly journals Novel Application of Mahua (Madhuca sp.) Flowers for Augmented Protease Production from Aeromonas sp. S1

2012 ◽  
Vol 7 (10) ◽  
pp. 1934578X1200701 ◽  
Author(s):  
Amrik Bhattacharya ◽  
Vandana Saini ◽  
Anshu Gupta
Keyword(s):  
Enzyme Production ◽  
Low Cost ◽  
Inoculum Size ◽  
Protease Production ◽  
Agitation Rate ◽  
Value Addition ◽  
New Approach ◽  
Flower Extract ◽  
Optimized Conditions ◽  
Optimum Production

The present study explored the utilization of Mahua ( Madhuca sp.) flowers, a major non-timber forest product (NTFP) of India, as a low-cost, natural substrate for protease production under submerged fermentation. Bacterial strain Aeromonas sp. S1, previously reported by us, was used as the protease producer. Using Mahua flower extract (MFE) as the medium additive, the protease production could successfully be enhanced by 5.6-fold (564.5 UmL−1) after 24 h of fermentation under optimized conditions compared with initial production of 99.9 UmL−1 in the absence of MFE. The cultural parameters for optimum production of protease were determined to be: incubation time-24 h; pH-7.0; MFE concentration-5% (v/v); inoculum size-0.3% (v/v) and agitation rate-200 rpm. The results obtained demonstrate the potential of cheaper and abundantly available Mahua flowers for induction of proteases, and thus offer a new approach for value addition to this biomass through industrial enzyme production.

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

2021 ◽  
Author(s):  
Cyr Jonas Morabandza ◽  
Valentin Dibangou ◽  
Faly Armel Soloka Mabika ◽  
Elgie Viennechie Gatse ◽  
Tarcisse Baloki Ngoulou ◽  
...  
Keyword(s):  
Enzyme Production ◽  
Skim Milk ◽  
Nitrogen Sources ◽  
Petri Dish ◽  
Culture Conditions ◽  
Luria Bertani ◽  
Protease Production ◽  
Carbon And Nitrogen ◽  
Cassava Leaves ◽  
Milk Casein

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.

Statistical Optimization of the production of protease from Bacillus sp. MSK-01 in submerged fermentation

2018 ◽  
Vol 23 (4) ◽  
Author(s):  
Kulwant Kaur ◽  
Sonica Sondhi ◽  
Palki Sahib Kaur
Keyword(s):  
Submerged Fermentation ◽  
Milk Powder ◽  
Skim Milk ◽  
Fold Increase ◽  
Skim Milk Powder ◽  
Protease Production ◽  
High Yield ◽  
Bacillus Sp ◽  
Increasing Demand ◽  
Optimized Conditions

Protease has been in increasing demand in industries due to its hydrolytic nature. In industries, high yield of enzyme is required to meet the industrial need at a relatively cheaper cost. In the present study, the protease from Bacillus sp. MSK-01 was produced in large quantity by submerged fermentation. Statistical techniques including Plackett-Burman and Response surface methodology are useful tools for optimizing many parameters at a time and are used for increasing the protease production from Bacillus sp. MSK-01. 19 different parameters were chosen, out of which 15 factors had positive effect on protease yield. Four maximum influencing factors were peptone, magnesium sulphate, skim milk powder and casein were chosen to further increase the protease yield. 397.3 IU ml-1 of enzyme yield was obtained under optimized conditions which lead to 198 fold increase in the yield of protease from unoptimized condition. 

scholarly journals Rice bran as a substrate for proteolytic enzyme production

2006 ◽  
Vol 49 (5) ◽  
pp. 843-851 ◽  
Author(s):  
Alagarsamy Sumantha ◽  
Paul Deepa ◽  
Chandran Sandhya ◽  
George Szakacs ◽  
Carlos Ricardo Soccol ◽  
...  
Keyword(s):  
Rice Bran ◽  
Enzyme Production ◽  
Proteolytic Enzyme ◽  
Initial Moisture Content ◽  
Fold Increase ◽  
Maximal Activity ◽  
Protease Production ◽  
Rhizopus Microsporus ◽  
Rhizopus Sp ◽  
Optimized Conditions

Rice bran was used as the substrate for screening nine strains of Rhizopus sp. for neutral protease production by solid-state fermentation. The best producer, Rhizopus microsporus NRRL 3671, was used for optimizing the process parameters for enzyme production. Fermentation carried out with 44.44 % initial moisture content at a temperature of 30 C for 72 h was found to be the optimum for enzyme secretion by the fermenting organism. While most of the carbon supplements favored enzyme production, addition of casein resulted in a marginal increase in protease yield. Fermentation was then carried out under optimized conditions to obtain the crude extract of the enzyme, which was partially purified by precipitation and dialysis. A 3-fold increase in the enzyme purity was achieved in this manner. The enzyme was found to be a metalloprotease, being activated by Mn2+, with maximal activity at a temperature of 60 C and pH 7.0.

Cytotoxic Effect of Pseudomonas aurogenosa Protease on Cancer Cells

2019 ◽  
Vol 10 (03) ◽  
Author(s):  
Ghanyia J. Shanyoor ◽  
Fatima R. Abdul ◽  
Nehad A. Taher ◽  
Ihsan A. Raheem
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Cytotoxic Effect ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) and the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method and it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column and the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , and one normal cell line Ref ( Rat embryonic fibroblast ) . The cancer and normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8and 0.16 mg/ml) then incubated for additional 48h at 37C0 and the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andslight toxicity ( 37.12% ) on normal cell line (Ref) in a concentration (0.8mg/ml).

Constraints in Adoption ofModern Farm Machines by Tribal Farmers in Ramgarh District of Jharkhand

2019 ◽  
Vol 6 (03) ◽  
Author(s):  
PK SUNDARAM ◽  
BIKASH SARKAR ◽  
UJJWAL KUMAR ◽  
AP ANURAG ◽  
DK RAGHAV ◽  
...  
Keyword(s):  
Cell Line ◽  
Normal Cell ◽  
Human Cancer ◽  
Skim Milk ◽  
Exchange Chromatography ◽  
Protease Production ◽  
Normal Cell Line ◽  
Protease Enzyme ◽  
Tribal Farmers ◽  
Embryonic Fibroblast

About (20) Pseudomonas rogenosa isolate were experienced for their ability of protease production by calculating the diameter of lysis area after developing on skim milk agar medium (qualitatively ) andamp; the results exhibited that only isolate no (5), was higher isolate for protease making of (26mm) of lysis area. Then, the protein concentration also identified by Bradford method andamp; it was found of 0.16 mg/ ml , then purification was done by using an ion- exchange chromatography with DEAE sephadex G- 100 column andamp; the results showed the presence of 1 peak of enzyme with 50 Kd of molecular weight 2 peaks of other proteins . we tried to investigate the invitro Cytotoxic effect of purified enzyme against two human cancer lines, HeP2 (Human larynx epidermed carcinoma ) , RD ( Rabdo- Sarcoma ) , andamp; one normal cell line Ref ( Rat embryonic fibroblast ). The cancer andamp; normal cells were treated with different concentrations of protease enzyme ranging from ( 0.05, 0.1, 0.2, 0.4,0.8andamp; 0.16 mg/ml) then incubated for additional 48h at 37C 0 andamp; the results showed highest toxicity ( 80.28%) of protease enzyme on RD , moderate cytotoxicity (45.52%) on Hep andamp;slight toxicity ( 37.12% )on normal cell line (Ref) in a concentration (0.8mg/ml).

scholarly journals Optimal Conditions For Production Acidic D-Xylanase By Aspergillus niger in Submerged Fermentation

2011 ◽  
Vol 5 (3) ◽  
pp. 14-21
Author(s):  
Muhamed Omar Abdulatif ◽  
Hyder H. Assmaeel ◽  
Raghad kadhim Obeid ◽  
Ayat Adnan Abbas
Keyword(s):  
Aspergillus Niger ◽  
Rice Husk ◽  
Enzyme Production ◽  
Inoculum Size ◽  
Optimal Conditions ◽  
Optimum Conditions ◽  
Xylanase Production ◽  
Primary Isolation ◽  
Optimum Ph ◽  
Optimum Period

he Xylanase producing strain Aspergillus niger was isolated from soil on potato dextrose agar in the presence of xylan as its first substrate for primary isolation, and then grown under liquid medium fermentation in the presence of crude xylan (rice husk) to produce D-Xylanase. the optimum conditions were determined as follows: the Optimum pH for xylanase production was found pH 5.0, xylanase was induced by xylan (rice husk) 0.1% and the production was (61.221 U/ml) and nitrogen source Yeast extract recorded highest enzyme production( 89.71 U/ml), and repressed by carbon source xylose the highest enzyme production (88.69 U/ml). The optimum temperature was 40°с for xylanase production was (35.15 U/ml), the optimum period after 7 days of incubation was (52.33 U/ml) ,the optimum substrate concentration 0.1% was (45.95 U/ml), and the optimum inoculum size was 1 x 106 (spore /ml) recorded (57.19 U/ml ).

Production and characterisation of thermostable alkaline protease from Bacillus subtilis isolated from LA hot spring, Terengganu

2021 ◽  
Vol 16 (7) ◽  
pp. 84-91
Author(s):  
Maslinda Alias ◽  
Hakim Che Harun Mohammad ◽  
Ashraf Razali Nurul ◽  
Jasnizat Saidin ◽  
Nazaitulshila Rasit ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Alkaline Protease ◽  
Protease Activity ◽  
Hot Spring ◽  
Skim Milk ◽  
Industrial Applications ◽  
Protease Production ◽  
Significant Activity ◽  
Original Activity ◽  
Optimum Ph

This research aims to produce thermostable alkaline protease from Bacillus subtilis isolated from La Hot Spring, Terengganu, Malaysia. The study was also conducted to determine the optimum conditions for protease production and stability by considering several parameters including pH, temperature and salt concentration. All seven bacteria were screened on skim milk agar overnight at 37 °C. Three strains with the highest proteolytic activity were identified in protease specific medium. The thermostable alkaline protease had an optimum temperature of 60 °C which achieved 85.73, 82.90 and 83.05 U/mL of protease activity for the three strains respectively. Furthermore, the strains exhibited significant activity of more than 90% from their original activity. Meanwhile, the optimum pH for protease production was pH 9 with the protease activity of 76.76, 79.71 and 88.39 U/mL for TB4, TB6 and TB9 strains, respectively. Proteases were found stable at pH 9 where the loss did not exceed 30% of its original activity. Collectively, all of the data emphasised that proteases from B. subtilis were alkaline thermostable proteases in accordance with a recent report. The finding highlights the viability of the proteases for biotechnological and industrial applications.

Studies on the manufacture of sweet cream starter butter: II. Effect of variation in proportion of starter added to cream

1960 ◽  
Vol 27 (1) ◽  
pp. 91-102 ◽  
Author(s):  
F. H. McDowall ◽  
J. A. Singleton ◽  
B. S. Le Heron
Keyword(s):  
Lactic Acid ◽  
Low Temperatures ◽  
Starter Culture ◽  
Skim Milk ◽  
Ph Values ◽  
Commercial Scale ◽  
Increase With Increase ◽  
Keeping Quality ◽  
Higher Temperature

SummaryProduction of diacetyl and acetoin by starters in cold skim-milk and cream was shown to increase with increase in the proportion of starter culture added, with some limitations at the higher rates of starter addition.With Streptococcus diacetilactis starter in skim-milk at 50°F the relation between proportion of starter added and production of diacetyl was linear up to addition at the 4% level, whereas at 43°F it was approximately linear up to the 10% level. At both 50 and 43°F the relation between the proportion of starter added and the production of acetoin was linear up to the 10% level.With Camb starter in skim-milk at both 50 and 43°F there were regular increases in production of diacetyl up to the 4% level of addition, but only minor changes thereafter with increase in the proportion of starter added up to 10%. At both temperatures the maximum production of acetoin was reached with the 7% rate of addition.Production of diacetyl and acetoin in skim-milk was greater at 50°F than at 43°F with both starters for all proportions up to 10%, and it was greater for Str. diacetilactis than for the mixed cultures.Except at the higher rates of addition of starter and at the higher temperature there were no concomitant increases in the acidity of the milk or lowering of the pH values. It appears that at low temperatures production of diacetyl by starters in sweet milk and cream proceeds independently of production of lactic acid.Similar results were obtained in a series of experimental buttermaking trials and some small commercial-scale trials, in which varying proportions of starter were added to creams after pasteurizing and before holding overnight for churning. With the cream-holding temperatures used, mainly 40–50°F, the pH values of the butters were not appreciably lowered by the starter additions to the cream. At all the rates of addition there were with Str. diacetilactis starter higher contents of diacetyl in the butter than with Camb starter. There was no indication of any relationship between the proportion of starter added and the keeping quality of the butter.

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