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

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.

2020 ◽  
Vol 12 (1) ◽  
pp. 154-171
Author(s):  
Oluwatoyin M. ALADEJANA ◽  
Olaoluwa OYEDEJI ◽  
Olumide OMOBOYE ◽  
Mufutau BAKARE

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.


2018 ◽  
Vol 44 (2) ◽  
pp. 285-292
Author(s):  
Sereen Gul ◽  
Mujeeb Ur Rahman ◽  
Mohammad Ajmal ◽  
Abdul Kabir Khan Achakzai ◽  
Asim Iqbal

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.


2020 ◽  
Vol 71 (11) ◽  
pp. 1-9
Author(s):  
Noor Nihad Abdul Hussein ◽  
Aseel I. Ibrahim ◽  
Firas Hashim Kamar ◽  
Arelia Cristina Nechifor

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.


2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Neelima Deshpande ◽  
Prachi Choubey ◽  
Manasi Agashe

A species ofStreptomyces,Streptomyces ginsengisoli, a river isolate, was evaluated for production of an enzyme, L-asparaginase, with multiple functions mainly anticancer activity. The actinomycete was subjected to submerged fermentation by “shake flask” method. The quantity of L-asparaginase produced was estimated as 3.23 μmol/mL/min. The effect of various culture conditions on L-asparaginase production was studied by adopting a method of variation in one factor at a time. Of the various conditions tested, glucose (followed by starch) and peptone served as good carbon and nitrogen sources, respectively, for maximal production of enzyme at pH 8. The temperature of 30°C and an incubation period of 5 days with 0.05 g% asparagine concentration were found to be optimum for L-asparaginase production.


1997 ◽  
Vol 43 (6) ◽  
pp. 592-595 ◽  
Author(s):  
Rebecca Eagen ◽  
Anne Brisson ◽  
Colette Breuil

Sap-staining fungi, in particular Ophiostoma piceae, cause significant economic problems for Canada's lumber industry. For O. piceae and a few other staining fungi, the dark pigment is melanin. The choice of carbon and nitrogen sources added as supplements to a basic liquid mineral medium affected the colour and characteristics of the pigment produced by O. piceae. The mycelia became coloured early during the active growth phase and the colour reached a maximum intensity during the stationary growth phase.Key words: melanin, growth, Ophiostoma piceae, pigmentation, sap-staining fungus.


Author(s):  
Aastha Acharya ◽  
Amit Khanal ◽  
Mrishal Ratna Bajracharya ◽  
Aashish Timalsina ◽  
Arjun Bishwokarma ◽  
...  

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.


2018 ◽  
Vol 69 ◽  
pp. 1-11 ◽  
Author(s):  
Willian Daniel Hahn Schneider ◽  
Roselei Claudete Fontana ◽  
Simone Mendonça ◽  
Félix Gonçalves de Siqueira ◽  
Aldo José Pinheiro Dillon ◽  
...  

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