scholarly journals Production, Purification and Optimisation of Amylase by Submerged Fermentation Using Bacillus Subtilis

2019 ◽  
pp. 265-275
Author(s):  
Aastha Acharya ◽  
Amit Khanal ◽  
Mrishal Ratna Bajracharya ◽  
Aashish Timalsina ◽  
Arjun Bishwokarma ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Maximum Yield ◽  
Cell Immobilization ◽  
Nitrogen Sources ◽  
Biochemical Tests ◽  
Carbon And Nitrogen ◽  
Nutrient Agar Medium ◽  
Amylase Production ◽  
Bacillus Spp ◽  
Almost All

Amylase is abundantly present in nature. The main source of this enzyme is the microbial origin. It is known that about two-third of the industrial enzymes (amylase, protease, cellulose, penicillinase, chitinase, etc,) are produced by the Bacillus spp. The present work comprised in the identification of amylase producing Bacillus spp and exposure of the producers to various parameters for the maximum yield of the enzyme. To isolate and identify the amylase and protease producing strain, soil samples were collected from different vegetation from the altitude at 4367.35 feet above sea level. The isolates were screened and various biochemical tests and morphological observations were done to identify the isolates. The enzymes were produced by the submerged state fermentation (SmF) from the isolates and purified by dialysis. Effects of temperature, pH, and different carbon and nitrogen sources of the medium using SmF were optimized. Among 95 isolates, 36 were identified. Among the identified isolates, Bacillus subtilis produced the maximum yield and thus, it was optimized for the amylase production. The maximum amylase production was found at 42 C temperature, in fructose as a carbon sugar, peptone as a nitrogen source and at pH 7. Almost all the enzyme producers inhabited the roots of leguminous plants. In the present study, starch is used with the nutrient agar medium to help in cell immobilization for maximum production of amylase by strains of Bacillus. More sophisticated process of purification such as chromatography and electrophoresis will yield more enzyme as compared to the dialysis.

scholarly journals Caseinase Production and Media Optimization from Bacillus subtilis

2020 ◽  
Vol 71 (11) ◽  
pp. 1-9
Author(s):  
Noor Nihad Abdul Hussein ◽  
Aseel I. Ibrahim ◽  
Firas Hashim Kamar ◽  
Arelia Cristina Nechifor
Keyword(s):  
Bacillus Subtilis ◽  
Enzyme Production ◽  
Paper Industry ◽  
Nitrogen Sources ◽  
The Body ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Sequencing Result

Caseinase is involved in the breakdown of milk protein casein and converts casein into smaller simple sugars which can be easily utilized by the body for the production of ATP and Fat. Casein can be an instant energy source to the body and involves in muscle building. Caseinase enzyme can be extensively used at the industrial scale for Milk, Textile, Dairy, Paper industry and several other medical purposes. In view of the importance of caseinase, the current research deals with the isolation and identification of caseinase producing bacteria from soil. This is followed by the production of enzyme and its purification. The study also includes its kinetic characterization using the parameters Temperature, pH as well as Carbon and Nitrogen Sources. The organism which was isolated from soil and capable of producing the caseinase enzyme was identified to be Bacillus subtilis based on the Biochemical tests and 16S rRNA sequencing result. The optimal carbon and nitrogen sources were identified to be Glucose and casein respectively. Regarding the optimal conditions, the suitable temperature for maximum enzyme production was found to be 40 0C and pH was 9. When the organism was cultured under the optimal condition using casein as a nitrogen source and glucose as the carbon source, at 40 0C and pH 9, 1590 ng/mL of enzyme production was estimated.

scholarly journals Production, purification and characterization of thermostable alpha amylase from Bacillus subtilis Y25 isolated from decaying yam (Dioscorea rotundata) tuber

2020 ◽  
Vol 12 (1) ◽  
pp. 154-171
Author(s):  
Oluwatoyin M. ALADEJANA ◽  
Olaoluwa OYEDEJI ◽  
Olumide OMOBOYE ◽  
Mufutau BAKARE
Keyword(s):  
Bacillus Subtilis ◽  
Metal Ion ◽  
Specific Activity ◽  
Nitrogen Sources ◽  
16S Rrna Gene Sequencing ◽  
Exchange Chromatography ◽  
Rrna Gene ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Amylase Production

Amylases have wide biotechnological potentials for applications in various industries. An α-amylase-producing bacterium was isolated from deteriorating yam tubers. Molecular characterization using the 16S rRNA gene sequencing was used to confirm the identity of the bacterium as Bacillus subtilis Y25. The effect of some cultural and nutritional factors such as pH, temperature, carbon and nitrogen sources on α-amylase production from the bacterium was determined. Maximum α-amylase production was observed using starch and peptone as carbon and nitrogen sources, respectively, with an initial medium pH of 8.0 and incubation at 45 °C for 36 h. The enzyme was purified by ion exchange chromatography on CM Sepharose CL-6B. The kinetic parameters Km and Vmax of the enzyme, as well as the effect of pH, temperature, metal ions and ethylenediaminetetra acetic acid (EDTA) on the activity of the purified enzyme were studied. The specific activity of the partially purified enzyme was determined to be 15.21 Units/mg protein with a purification fold of 3.80. The molecular weight of the purified enzyme was estimated to be 58.0 kDa. The Vmax and Km values obtained with soluble starch for Bacillus subtilis Y25 α-amylase were 314.10 ± 23.30 Units/mg protein and 53.98 ± 12.03 mg/ml, respectively. The enzyme exhibited optimum activity at a temperature of 60 °C and pH 8.0. The metal ion Ca2+ had no effect on the enzyme at 20 mM concentration, whereas Na+ and Mg2+, as well as EDTA inhibited the enzyme at the same concentration. The characteristics of the α-amylase from Bacillus subtilis Y25 revealed it to be a thermostable and an alkaline metalloenzyme with potential for applications in the detergent and saccharification industries.

scholarly journals Optimization of the growth of and α-amylase production by Bacillus subtilis IP 5832 in shake flask and laboratory fermenter batch cultures

2011 ◽  
Vol 76 (7) ◽  
pp. 965-972 ◽  
Author(s):  
Natasa Bozic ◽  
Jordi Ruiz ◽  
Josep López-Santín ◽  
Zoran Vujcic
Keyword(s):  
Bacillus Subtilis ◽  
Shake Flask ◽  
Nitrogen Sources ◽  
Stationary Growth Phase ◽  
Stationary Growth ◽  
Carbon And Nitrogen Sources ◽  
Productivity Level ◽  
Batch Cultures ◽  
Carbon And Nitrogen ◽  
Amylase Production

Cell growth and the level of ?-amylase in response to the carbon and nitrogen sources used for the growth of the strain Bacillus subtilis IP 5832 were examined. Based on the amylase productivity level in shake flask cultures after 24 hours of growth, the growth medium containing starch and peptone was selected as the best medium. Amylase production was greatly reduced when glutamate or citrate as sources of carbon were used. Experiments performed at different initial concentrations of starch showed that although the strain grew well with all the starch concentration used, 0.5 % starch was necessary for maximum ?-amylase production, inducing 1.55 IU mL-1 of amylase to be secreted after 8 h of cultivation in shaking flasks. During the batch fermentation of B. subtilis IP 5832 strain in 2 L laboratory fermenter, a 60 % higher activity (2.5 IU mL-1) was obtained. The production of the enzyme was directly related to the growth of the strain. Maximum enzyme activity was obtained at the beginning of the stationary growth phase.

scholarly journals Effects of carbon and nitrogen sources on production of proteases by Bacillus subtilis IC-5

2018 ◽  
Vol 44 (2) ◽  
pp. 285-292
Author(s):  
Sereen Gul ◽  
Mujeeb Ur Rahman ◽  
Mohammad Ajmal ◽  
Abdul Kabir Khan Achakzai ◽  
Asim Iqbal
Keyword(s):  
Bacillus Subtilis ◽  
Sodium Nitrate ◽  
Inorganic Nitrogen ◽  
Carbon Sources ◽  
Basal Medium ◽  
Nitrogen Sources ◽  
Neutral Protease ◽  
Carbon And Nitrogen Sources ◽  
Inorganic Nitrogen Source ◽  
Carbon And Nitrogen

The effects of various carbon and nitrogen sources were evaluated on production of proteases by Bacillus subtilis IC-5. Both type and concentration of carbon and nitrogen sources influenced the production of proteases. Among the carbon sources glucose was found to be the most effective. It gave maximum production at 2% w/v concentration i.e., 1875 and 950 U/ml, alkaline and neutral protease, respectively. The response of Bacillus subtilis IC-5 towards synthesis and excretion of enzymes varied with the type of nitrogen sources. The addition of organic nitrogen sources to basal medium repressed the synthesis of proteases while the addition of inorganic nitrogen source such as sodium nitrate was found to be the best stimulating for alkaline and neutral protease synthesis. Sodium nitrate enhanced the production up to 62.40 and 10.52% of alkaline and neutral protease, respectively against w.r.t. control.

scholarly journals Optimization of Cultural Conditions for Production of Antibacterial Metabolites from Streptomyces coelicoflavus BC 01

2015 ◽  
Vol 7 (2) ◽  
pp. 151-159
Author(s):  
Kothagorla Venkata RAGHAVA RAO ◽  
Dadi BHASKARA RAO ◽  
Botcha SATYANARAYANA ◽  
Tamanam RAGHAVA RAO
Keyword(s):  
Andhra Pradesh ◽  
Maximum Yield ◽  
Nitrogen Sources ◽  
Fermentation Medium ◽  
Carbon And Nitrogen ◽  
Mangrove Soil ◽  
Cultural Conditions ◽  
Bean Meal ◽  
Metabolites Production ◽  
Soya Bean Meal

The aim of the present study was to optimize various cultural conditions for the production of antibacterial metabolites by Streptomyces coelicoflavus BC 01 isolated from mangrove soil, Visakhapatnam, Andhra Pradesh, India. The effect of various factors such as carbon and nitrogen sources, different concentrations of NaCl and K2HPO4, different temperature, pH, incubation time and agitation on antibacterial metabolites production were studied. The production of antibacterial metabolites by the isolate Streptomyces coelicoflavus BC 01 was greatly influenced by the cultural conditions. Glucose (1.2%) and soya bean meal (1%) seemed to be the best carbon and nitrogen source respectively, followed by NaCl (1%) and K2HPO4 (0.25%). Maximum production of antibacterial metabolites was observed at a temperature of 30 °C, with pH 7.2, at 160 rpm for 96 hrs. These optimized parameters can be further useful to design a fermentation medium to achieve maximum yield of antibacterial metabolites from Streptomyces coelicoflavus BC 01.

scholarly journals Screening and Optimization of Thermo-Tolerant Bacillus Sp. For Amylase Production and Antifungal Activity

2019 ◽  
Vol 24 (1) ◽  
pp. 47-56 ◽  
Author(s):  
Susmita Sapkota ◽  
Sujan Khadka ◽  
Aava Gautam ◽  
Rojina Maharjan ◽  
Ruby Shah ◽  
...  
Keyword(s):  
Aspergillus Flavus ◽  
Industrial Applications ◽  
Antagonistic Activity ◽  
Soil Samples ◽  
Bacillus Sp ◽  
Biochemical Tests ◽  
Wide Range ◽  
Amylase Production ◽  
Bacillus Spp ◽  
Two Parameters

Amylases are starch degrading enzymes which are produced by plants, animals and microorganisms. Amylases produced by microorganisms have a wide range of industrial applications such as in pharmaceutical, food, textile and paper industries. However, there are still limitations in the isolation of amylase producing microorganisms. The objective of this study was to isolate the potent amylase producing Bacillus sp. from soil samples and evaluate their abilities for inhibiting the aflatoxin producing Aspergillus flavus. In this study, 30 soil samples were used. For the screening and identification of Bacillus strain, morphological and biochemical tests were performed. Iodine assay was done to screen the potent amylase producers. Two parameters (pH and temperature) were used to optimize the cultural conditions for the production of amylase. To determine the total reducing sugar, dinitrosalicylic acid (DNS) assay was used. Altogether 29 colonies were selected and identified as Bacillus spp out of which 16 were selected to determine enzyme activity by cup plate method. Four isolates (DK9, DK10, IM4 and KD7) showing highest amylolytic activities (16 mm, 12 mm, 14 mm and 14 mm zone of hydrolysis) were subjected for further study. Isolate KD7 showed the highest amylolytic activity (0.19 U/mL) compared to other isolates. Maximum amylase production was found at pH 6 and temperature 50° C (0.19 U/mL). Among these 4 isolates, DK9 and KD9 showed strong antagonistic activity against Aspergillus flavus while DK10 and IM4 showed moderate antifungal activities. Thus, the bacterial isolate KD7 was identified as the most potent strain for maximum amylase production.

scholarly journals Process optimization for amylase production of Bacillus subtilis M4 mutant strain

2019 ◽  
Vol 27 (02) ◽  
pp. 8-19
Author(s):  
Naramchimeg B ◽  
Altantsetseg Kh ◽  
Urantulkhuur B
Keyword(s):  
Bacillus Subtilis ◽  
Mutant Strain ◽  
Inorganic Nitrogen ◽  
Nitrogen Sources ◽  
Fold Increase ◽  
Medium Composition ◽  
Optimization Techniques ◽  
Amylase Production ◽  
Ph Range ◽  
Inoculum Volume

There are many factors that influence the character of bacterial metabolism and enzyme production.For themaximum production of the desired products, the media components and fermentation conditions should beoptimized. In our investigation, we improved the amylase production of Bacillus subtilis M4 mutant strain bythe combination of two optimization techniques. The cultural conditions (time period, temperature, pH,inoculum volume) and medium ingredients (various carbon, organic and inorganic nitrogen sources, chlorides, sulfates, phosphates, carbonates) were optimized by one factor at a time methodology (OFAT) and response surface methodology (RSM) to increase the amylase production. The optimum conditions for amylase production were found be the following: 35ºC, pH range 7 and incubation time 72h, inoculum volume 8% (v/v). Optimum medium composition for amylase production was the following: starch 12.9 g, peptone 9.75g, calcium carbonate 0.439 g, magnesium sulfate 0.464 g and potassium chloride 0.464 g per liter. Whenapplied to our optimized medium in the fermentation process, the enzyme activity increased from 0.741 to1.58 U/ml, which means a 2.1-fold increase compared to the original medium.

scholarly journals Lab-Scale Optimization of Polyhydroxyalkanoate Production by Bacterial Strain cmg1415 on Local Cheap Substrates Using One Variable at a Time Approach

2021 ◽  
Vol 1 (1) ◽  
pp. 25-31
Author(s):  
Muhammadi Muhammadi ◽  
Shabina Shafiq
Keyword(s):  
Sugar Beet ◽  
Minimal Medium ◽  
Maximum Yield ◽  
Nitrogen Sources ◽  
Sole Source ◽  
Labor Cost ◽  
Culture Conditions ◽  
Carbon And Nitrogen Sources ◽  
Carbon And Nitrogen ◽  
Scale Optimization

Production of polyhydroxyalkanoate (PHA) under optimum culture conditions using local cheap feedstocks is indispensable to overcome the current cost of PHA-based plastics. For this purpose, optimum culture conditions and cheap feedstocks were investigated to produce maximum yield of PHA in CMG1415. Maximum yield was obtained with sucrose or sugar beet as sole source of precursors for PHA in 8 days of incubation at 35 °C in a minimal medium adjusted at pH 7. Further, for maximum yield no mechanical shaking was needed. Local cheap feedstock such as sugar beet and molasses were found to play as significant carbon and nitrogen sources for maximum PHA yield.  Bacterial plastic produced under these low-labor-cost culture conditions may to reduce the present cost of degradable bioplastic and be much effective alternate of nondegradable varieties of synthetic plastic.

scholarly journals Bacillus megaterium Biodegradation Glyphosate

2021 ◽  
Author(s):  
Nibal Khaleel Mousa ◽  
Abdul-Jabbar Ali ◽  
Maha Hussein
Keyword(s):  
Incubation Period ◽  
Bacillus Megaterium ◽  
Organophosphorus Pesticides ◽  
Nitrogen Sources ◽  
Mineral Salt ◽  
Morphological Identification ◽  
Biochemical Tests ◽  
Carbon And Nitrogen ◽  
Degradation Ratio ◽  
Glyphosate Degradation

The Bacillus megaterium ability was evaluated in this paper to degrade the Glyphosate. organophosphorus pesticides, The bacteria re-cultured that isolated from other researches of Baghdad soils and morphological identification and biochemical tests besides by selectivity media. The (5 and 25) ppm showed the highest growth results were within two days to two months on mineral salt media. The highest glyphosate degradation ratio % were (70) % per 25 ppm/two months. Incubation period Increasing led to highest glyphosate degradation ratio% at (25) ppm led to conclusion that bacteria digestive the pesticides as carbon and nitrogen sources and will be well harvest it form contaminated areas.

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