Immobilization of chymotrypsin (E.C.3.4.21.1), subtilisin (E.C.3.4.21.14) and neutral proteinase from Bacillus subtilis by covalent binding to benzoquinone-activated pearl cellulose

1983 ◽  
Vol 48 (10) ◽  
pp. 2874-2878 ◽  
Author(s):  
Helena Škachová ◽  
Jiří Kučera
Keyword(s):  
Activation Energy ◽  
Enzyme Activity ◽  
Bacillus Subtilis ◽  
Substrate Concentration ◽  
Covalent Binding ◽  
Neutral Proteinase

Chymotrypsin (E.C.3.4.21.1), subtilisin (E.C.3.4.21.14), and a neutral proteinase from Bacillus subtilis were immobilized by covalent binding to benzoquinone-activated pearl cellulose. The yield of the immobilized protein was 55% in the case of chymotrypsin and 50% in the case of subtilisin and neutral proteinase from B. subtilis. The determination of activation energy at a low substrate concentration showed that the enzyme activity is limited by diffusion under these conditions. The activity yields are generally very good yet the activity of the enzymes immobilized is relatively low, most likely because of the presence of benzoquinone, as shown in experiments with the immobilization of chymotrypsin by the same technique on supports with different hydrophilicity.

Colorimetric Determination of Serum Guanase Activity

1966 ◽  
Vol 12 (4) ◽  
pp. 187-193 ◽  
Author(s):  
Wendell T Caraway
Keyword(s):  
Enzyme Activity ◽  
Differential Diagnosis ◽  
Substrate Concentration ◽  
Colorimetric Method ◽  
Colorimetric Determination ◽  
Concentration Time

Abstract A colorimetric method for the determination of serum guanase has been developed whereby ammonia, formed by the deteramination of guanine, is measured by the phenatehypochlorite reaction. Variations in substrate concentration, time of incubation, pH, temperature, and concentration of enzyme have been investigated with respect to enzyme activity. The determination of serum guanase appears to be useful in the differential diagnosis of jaundice.

scholarly journals Determination of Optimal Fermentation Condition for N-acetylglucosamine Production Using Mucor circinelloides Extracellular Chitinase

2019 ◽  
Vol 21 (2) ◽  
pp. 105
Author(s):  
Yuniwaty Halim ◽  
Hardoko Hardoko ◽  
Reinald Febryanto Pengalila
Keyword(s):  
Enzyme Activity ◽  
Experimental Method ◽  
Substrate Concentration ◽  
Chitinase Activity ◽  
Mucor Circinelloides ◽  
Fermentation Condition ◽  
Optimal Temperature ◽  
Fermentation Time ◽  
Optimal Ph

This research aimed to determine the best fermentation condition, consists of pH, temperature, fermentation time and substrate concentration, in N-acetylglucosamine production from shrimp shells using crude extracellular chitinase obtained from Mucor circinelloides mould. The method used was experimental method with fermentation treatment of different pH (5, 6, 7, 8 and 9) and temperature (30, 40, 50, 60, 70 and 80°C). The optimal pH and temperature of fermentation obtained was used to determine the maximum substrate concentration (0.5, 1, 1.5 and 2%) and fermentation time (2, 4, 6 and 24 hours) to produce the highest concentration of N-acetylglucosamine. The optimal pH for fermentation was 8, with chitinase activity of 4.38±0.06 U/ml, while the optimal temperature was 50°C with enzyme activity of 5.42±0.06 U/ml. Substrate concentration and fermentation time affected the N-acetylglucosamine production. The optimal fermentation condition was obtained with substrate concentration of 1.5% and fermentation time of 2 hours resulted to N-acetyl Glucosamine concentration of 2195.83±15.14 ppm.

scholarly journals PRODUKSI DAN PENENTUAN KONDISI OPTIMUM ENZIM XILANASE Bacillus amyloliquefaciens FUKUMOTO PADA SUBSTRAT XILAN JERAMI

2015 ◽  
Vol 2 (1) ◽  
pp. 74
Author(s):  
Widiyanti Sekatresna ◽  
Abdi Dharma ◽  
Periadnadi
Keyword(s):  
Enzyme Activity ◽  
Rice Straw ◽  
Incubation Time ◽  
Substrate Concentration ◽  
Bacillus Amyloliquefaciens ◽  
Specific Activity ◽  
Enzymatic Reaction ◽  
Optimal Conditions ◽  
Enzyme Specific Activity

 ABSTRACT The production and determination of  optimal condition of xylanase produced by Bacillus amyloliquefaciens on rice straw xylan were investigated in this study. The parameters to be observed were optimal conditions of pH, temperature, substrate concentration and incubation time. Xilanase activity was determined by measuring the amount of reducing sugar formed in the enzymatic reaction based on Somogyi Nelson method. Optimal conditions needed for the production of xylanase were at pH 7, temperature 27°C and six days of incubation time. While optimal conditions of xylanase action were reached at pH 8.2, temperature 45°C, substrate concentration 3.5%(w/w) and 15 minutes of incubation time with enzyme activity and enzyme specific activity of 1.285 U/mL and 0.738 U/mg respectively. As a comparison, xylanase was also produced on pure xylan  (birchwood), enzyme activity and enzyme specific activity obtained were 2.701 U/mL and 1.658 U/mg respectively. Cellulase content in enzyme produced on rice straw xilan showed the enzyme activity of 0.094 U/mL.  Keywords : xylanase, Bacillus amyloliquefaciens, rice straw xilan

Respirometry for determination of the influents SS-concentration

1996 ◽  
Vol 33 (1) ◽  
pp. 311-323 ◽  
Author(s):  
A. Witteborg ◽  
A. van der Last ◽  
R. Hamming ◽  
I. Hemmers
Keyword(s):  
Experimental Design ◽  
Activated Sludge ◽  
Substrate Concentration ◽  
Full Scale ◽  
Activated Sludge Process ◽  
Respiration Rates ◽  
On Line ◽  
Large Measurement ◽  
Set Up

A method is presented for determining influent readily biodegradable substrate concentration (SS). The method is based on three different respiration rates, which can be measured with a continuous respiration meter which is operated in a cyclic way. Within the respiration meter nitrification is inhibited through the addition of ATU. Simulations were used to develop the respirometry set-up and decide upon the experimental design. The method was tested as part of a large measurement programme executed at a full-scale plant. The proposed respirometry set-up has been shown to be suitable for a semi-on-line determination of an influent SS which is fully based on the IAWQ #1 vision of the activated sludge process. The YH and the KS play a major role in the principle, and should be measured directly from the process.

Development of HPLC Method for the Purity Test by Design of Experiments and Determination of Activation Energy of Hydrolytic Degradation Reactions of Sofosbuvir

2020 ◽  
Vol 16 (7) ◽  
pp. 976-987
Author(s):  
Jakub Petřík ◽  
Jakub Heřt ◽  
Pavel Řezanka ◽  
Filip Vymyslický ◽  
Michal Douša
Keyword(s):  
Activation Energy ◽  
Design Of Experiments ◽  
Mobile Phase ◽  
Hydrolytic Degradation ◽  
Isoconversional Method ◽  
Hplc Method ◽  
Organic Modifier ◽  
Calculation Methods ◽  
Degradation Reactions

Background: The present study was focused on the development of HPLC method for purity testing of sofosbuvir by the Design of Experiments and determination of the activation energy of hydrolytic degradation reactions of sofosbuvir using HPLC based on the kinetics of sofosbuvir degradation. Methods: Following four factors for the Design of Experiments were selected, stationary phase, an organic modifier of the mobile phase, column temperature and pH of the mobile phase. These factors were examined in two or three level experimental design using Modde 11.0 (Umetrics) software. The chromatographic parameters like resolution, USP tailing and discrimination factor were calculated and analysed by partial least squares. The chromatography was performed based on Design of Experiments results with the mobile phase containing ammonium phosphate buffer pH 2.5 and methanol as an organic modifier. Separation was achieved using gradient elution on XBridge BEH C8 at 50 °C and a flow rate of 0.8 mL/min. UV detection was performed at 220 nm. The activation energy of hydrolytic degradation reactions of sofosbuvir was evaluated using two different calculation methods. The first method is based on the slope of dependence of natural logarithm of the rate constant on inverted thermodynamic temperature and the second approach is the isoconversional method. Results and Conclusion: Calculated activation energies were 77.9 ± 1.1 kJ/mol for the first method and 79.5 ± 3.2 kJ/mol for the isoconversional method. The results can be considered to be identical, therefore both calculation methods are suitable for the determination of the activation energy of degradation reactions.

scholarly journals Determination of cleavage sites of APendonuclease from Bacillus subtilis.

1980 ◽  
Vol 44 (7) ◽  
pp. 1713-1715
Author(s):  
Yoshihiko SAKO ◽  
Aritsune UCHIDA ◽  
Hajime KADOTA
Keyword(s):  
Bacillus Subtilis ◽  
Cleavage Sites
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