scholarly journals Immobilization of α - amylase from locale bacteria isolate Bacillus subtilis ITBCCB148 with diethylaminoethyl cellulose (DEAE-Cellulose)

2010 ◽  
Vol 7 (1) ◽  
pp. 123-128 ◽  
Author(s):  
Yandri. Yandri ◽  
Tati Suhartati ◽  
Sutopo Hadipo Hadi
Keyword(s):  
Thermal Stability ◽  
Bacillus Subtilis ◽  
Immobilized Enzyme ◽  
Storage Temperature ◽  
Residual Activity ◽  
Deae Cellulose ◽  
Ionic Exchange ◽  
Optimum Temperature ◽  
Diethylaminoethyl Cellulose ◽  
Exchange Matrix

The thermal stability increase of a-amylase obtained from locale bacteria isolate Bacillus subtilis ITBCCB148 was achieved by immobilization process using an ionic exchange matrix of DEAE-Cellulose. The result showed that the immobilized enzyme has an optimum temperature of 60°C; KM 14.8 mL substrate and Vmax 42.4 U/mL. The thermal stability storage temperature of 60°C, pH 9.0 and 60 minutes demonstrated the immobilized enzyme has residual activity of 28.1%; ki = 0.0224 min.-1; and ΔGi = 103.7 kJ mol-1. Although the immobilized enzyme’s thermal stability was only increased 1.5 times, at higher temperatures, it was much more stable than the native enzyme.

scholarly journals Increasing Stability of a-amylase Obtained from Bacillus subtilis ITBCCB148 by Immobilization with Chitosan

2020 ◽  
Vol 10 (2) ◽  
pp. 155-161
Author(s):  
Yandri Yandri ◽  
Tati Suhartati ◽  
Heri Satria ◽  
Arum Widyasmara ◽  
Sutopo Hadi
Keyword(s):  
Thermal Stability ◽  
Bacillus Subtilis ◽  
Half Life ◽  
Thermodynamic Data ◽  
Immobilized Enzyme ◽  
Residual Activity ◽  
Optimum Temperature ◽  
Chitosan Matrix ◽  
T Values ◽  
Thermal Stability Of

In this research, the immobilization of α-amylase from Bacillus subtilis ITBCCB148 by crosslinking method on chitosan matrix has been performed. This research aims to know the effect of immobilization on the thermal stability of α-amylase. The results showed that the native α-amylase has an optimum temperature of  65oC, KM = 1.6 mg mL-1 substrate, and Vmax = 39.7 µmol mL-1 min-1. The immobilized α-amylase has optimum temperature of 75oC, KM = 3.5 mg mL-1 substrate, and Vmax = 7.05 µmol mL-1 min-1. The residual activity of the native and immobilized enzyme on thermal stability test at 65oC for 80 minutes was 58% and 86.15%, respectively. The immobilized enzyme can be reused up to six repeated cycles.The thermodynamic data of native enzyme was t½ = 113.6 min, ki = 6.1x10-3 min-1, and ΔGi = 107.3 kJ mol-1, while the immobilized enzyme was t½ = 433.1 min, ki= 1.6x10-3 min-1, and ΔGi 111.1 kJ mol-1. Based on the decrease of ki, and the increase of ΔGi and half-life(t½) values, the immobilization of α-amylase with chitosan can increase the thermal stability of this enzyme.

Peningkatan Stabilitas Termal Enzim α-Amilase dari Bacillus subtilis ITBCCB148 dengan Amobilisasi Menggunakan Zeolit

2019 ◽  
Vol 2 (2) ◽  
Author(s):  
Yandri ◽  
Fathaniah Sejati ◽  
Tati Suhartati
Keyword(s):  
Thermal Stability ◽  
Bacillus Subtilis ◽  
Immobilized Enzyme ◽  
Specific Activity ◽  
Residual Activity ◽  
Stability Test ◽  
Optimum Temperature ◽  
Before And After ◽  
Cation Exchange Column ◽  
Thermal Stability Of

The objective of the research is to increase the thermal stability of -amylase from Bacillus subtilis ITBCCB148 by immobilization using zeolite. For that reason, firstly we need to produce, isolate, and purify the enzyme. The purification of the enzyme was conducted by the following steps: fractionation with ammonium sulphate, dialysis, and CM-cellulose cation exchange column chromatography. The purified enzyme was immobilized using zeolite. The success in immobilization of the enzyme was evaluated by comparing the thermal stability of the enzyme before and after immobilization. Activity of α-amylase was determined by the Mandels and Fuwa method. The protein content was determined based on the method by Lowry. The results showed that the specific activity of purified enzyme was 2473.7 U / mg, increased 19 times compared to crude extract of enzyme having specific activity of 1285.9 U / mg. The purified enzyme has the optimum temperature at 65ºC, while the immobilized enzyme has the optimum temperature at 75ºC. The thermal stability test of the purified enzyme at 65ºC for 100 minutes showed the purified enzyme having residual activity of 20%; t 1 / 2 = 30 min, k i = 0.023 min -1 and ΔGi = 103.65 kJ mol -1 . The thermal stability test of the immobilized enzyme at 65ºC for 100 minutes showed that the immobilized enzyme had residual activity of 40%; t 1/ 2 = 49 min, k i = 0.014 min -1 and ΔGi = 105.03 kJ mol -1 . Immobilization using zeolite has succeeded in increasing the thermal stability of enzyme by 1.64 times compared to the purified enzyme, which is indicated by the decreasing of k i value, the increase of half-life and denaturation energy change (ΔGi).

scholarly journals The Stability Improvement of α-Amylase Enzyme from Aspergillus fumigatus by Immobilization on a Bentonite Matrix

2022 ◽  
Vol 2022 ◽  
pp. 1-7
Author(s):  
Yandri Yandri ◽  
Ezra Rheinsky Tiarsa ◽  
Tati Suhartati ◽  
Heri Satria ◽  
Bambang Irawan ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Half Life ◽  
Thermal Inactivation ◽  
Immobilized Enzyme ◽  
Residual Activity ◽  
Maximum Velocity ◽  
Partial Purification ◽  
Soluble Enzyme ◽  
Optimum Temperature ◽  
The Stability

The stability of the α-amylase enzyme has been improved from Aspergillus fumigatus using the immobilization method on a bentonite matrix. Therefore, this study aims to obtain the higher stability of α-amylase enzyme from A. fumigatus; hence, it is used repeatedly to reduce industrial costs. The procedures involved enzyme production, isolation, partial purification, immobilization, and characterization. Furthermore, the soluble enzyme was immobilized using 0.1 M phosphate buffer of pH 7.5 on a bentonite matrix, after which it was characterized with the following parameters such as optimum temperature, Michaelis constant (KM), maximum velocity V max , thermal inactivation rate constant (ki), half-life (t1/2), and the change of energy due to denaturation (ΔGi). The results showed that the soluble enzyme has an optimum temperature of 55°C, KM of 3.04 mg mL−1 substrate, V max of 10.90 μmole mL−1 min−1, ki of 0.0171 min−1, t1/2 of 40.53 min, and ΔGi of 104.47 kJ mole−1, while the immobilized enzyme has an optimum temperature of 70°C, KM of 8.31 mg mL−1 substrate, V max of 1.44 μmole mL−1 min−1, ki of 0.0060 min−1, t1/2 of 115.50 min, and ΔGi of 107.37 kJ mole−1. Considering the results, the immobilized enzyme retained 42% of its residual activity after six reuse cycles. Additionally, the stability improvement of the α-amylase enzyme by immobilization on a bentonite matrix, based on the increase in half-life, was three times greater than the soluble enzyme.

scholarly journals Preparation and studies on immobilized α-glucosidase from baker’s yeast Saccharomyces cerevisiae

2007 ◽  
Vol 72 (12) ◽  
pp. 1255-1263 ◽  
Author(s):  
Khaled Ahmed ◽  
Nenad Milosavic ◽  
Milica Popovic ◽  
Radivoje Prodanovic ◽  
Zorica Knezevic ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Thermal Stability ◽  
Immobilized Enzyme ◽  
Immobilized Enzymes ◽  
Optimum Temperature ◽  
Activity Profile ◽  
Original Activity ◽  
Yeast Saccharomyces Cerevisiae ◽  
Optimum Ph ◽  
Promising Solution

?-Glucosidase from S. cerevisiae was covalently immobilized onto Sepabeads EC-EA by the glutaraldehyde method. An analysis of the variables controlling the immobilization process is first presented and it is shown that the highest coupling of ?-glucosidase occurred within 24 h. Also, a loading of 30 mg/g support proved to be effective, resulting in a rather high activity of around 45 U g-1 with a satisfactory degree of enzyme fixed. Both free and immobilized enzymes were then characterized by determining the activity profile as a function of pH, temperature and thermal stability. The obtained immobilized preparation showed the same optimum pH, but a higher optimum temperature compared with the soluble one. In addition, the immobilized enzyme treated at 45 ?C for 1 h still retained an activity of around 20 %, whereas the free enzyme completely lost its original activity under this condition. In conclusion, the developed immobilization procedure is quite simple, easily reproducible and provides a promising solution for the application of immobilized ?-glucosidase.

scholarly journals Enzyme activity and stability of lactase immobilized on two different supports: Calcium alginate and magnetic chitosan

2020 ◽  
Vol 16 (4) ◽  
pp. 413-417
Author(s):  
Farah Syafiqah Mohd Zawawi ◽  
Latiffah Karim ◽  
Siti Radhiah Omar ◽  
Asma’ Ali
Keyword(s):  
Thermal Stability ◽  
Enzyme Activity ◽  
Enzymatic Activity ◽  
Calcium Alginate ◽  
Lactose Intolerance ◽  
Immobilized Enzyme ◽  
Soluble Form ◽  
Optimum Temperature ◽  
Magnetic Chitosan ◽  
Important Enzyme

Lactase is a very important enzyme to cure lactose intolerance problem. However, it is naturally existing in soluble form and cannot be reused. The current study was performed to compare the productivity and stability of lactase immobilized on calcium alginate and magnetic chitosan. The reusability of immobilized enzyme was measured for 28 days. Thermal stability was measured at 27, 37, 50 and 70 ºC. Lactase immobilized on calcium alginate showed a better stability after 21 days where it retained up to 62% of enzyme activity. However, lactase on magnetic chitosan expresses a better thermal stability as it produced 6% more sugar than lactase on calcium alginate at the optimum temperature 50 ºC. Lactase immobilized on calcium alginate and magnetic chitosan showed significantly different enzymatic activity, stability, and reusability.

Immobilization and Characterization of Tannase and its Haze-removing

2009 ◽  
Vol 15 (6) ◽  
pp. 545-552 ◽  
Author(s):  
Erzheng Su ◽  
Tao Xia ◽  
Liping Gao ◽  
Qianying Dai ◽  
Zhengzhu Zhang
Keyword(s):  
Kinetic Parameter ◽  
High Activity ◽  
Green Tea ◽  
Storage Stability ◽  
Residual Activity ◽  
Black Tea ◽  
Optimum Temperature ◽  
Oolong Tea ◽  
Optimum Ph

Tannase was effectively immobilized on alginate by the method of crosslinking-entrapment-crosslinking with a high activity recovery of 76.6%. The properties of immobilized tannase were investigated. Its optimum temperature was determined to be 35 ° C, decreasing 10 °C compared with that of free enzyme, whereas the optimum pH of 5.0 did not change. The thermal and pH stabilities of immobilized tannase increased to some degree. The kinetic parameter, Km, for immobilized tannase was estimated to be 11.6 × 10-4 mol/L. Fe2+ and Mn2+ could activate the activity of immobilized tannase. The immobilized tannase was also applied to treat the tea beverage to investigate its haze-removing effect. The content of non-estern catechins in green tea, black tea and oolong tea increased by 52.17%, 12.94% and 8.83%, respectively. The content of estern catechins in green tea, oolong tea and black tea decreased by 20.0%, 16.68% and 5.04%, respectively. The anti-sediment effect of green tea infusion treated with immobilized tannase was significantly increased. The storage stability and reusability of the immobilized tannase were improved greatly, with 72.5% activity retention after stored for 42 days and 86.9% residual activity after repeatedly used for 30 times.

Milk-Coagulating Extract Produced from Solanum aethiopicum Shum Fruits: Multivariate Techniques of Preparation, Thermal Stability and Effect on Milk Solids Recovery in Curd

2014 ◽  
Vol 10 (2) ◽  
pp. 211-222 ◽  
Author(s):  
Valentin Désiré Guiama ◽  
Robert Germain Beka ◽  
Esther Ngah ◽  
David Gabriel Libouga ◽  
Dominique Vercaigne-Marko ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Multivariate Techniques ◽  
Extraction Temperature ◽  
Optimum Temperature ◽  
Solanum Aethiopicum ◽  
Fruit Maturity ◽  
Nacl Concentration ◽  
Actual Yield ◽  
Milk Solids ◽  
Thermal Stability Of

Abstract This study investigated a novel procedure of Solanum aethiopicum Shum fruits extract (SASFE) preparation using multivariate experimental designs as factorial and Box–Behnken. The thermal stability of optimized extract as well as its influence on the milk solids in curd was determined. The results showed that extraction time, fruit maturity and pH did not affect significantly SASFE preparation, while the amount of fruits, extraction temperature and NaCl concentration of extractant had a significant effect (p < 0.05). The greatest coagulant index was obtained under the following conditions: 12.5% of fruits, 25°C of extraction temperature and 4% NaCl concentration of extractant. It was thermosensitive and exhibited optimum temperature at 50°C. There was no statistical difference between SASFE and calf rennet in terms of solids yield in curd, estimated yield and actual yield. On the basis of these results, SASFE can be used as a vegetable alternative to calf rennet.

ESTIMATION OF GROWTH – TIME TEMPERATURE CHARACTERISTICS FOR ISOLATES OF FOMES PINI, A HEART ROT FUNGUS

1964 ◽  
Vol 42 (12) ◽  
pp. 1635-1652 ◽  
Author(s):  
Lee A. Paine ◽  
Gideon Schwarzbart ◽  
William G. O'Regan
Keyword(s):  
Regression Analysis ◽  
Cross Section ◽  
Growth Pattern ◽  
Storage Temperature ◽  
Three Dimensional ◽  
Douglas Fir ◽  
Growth Time ◽  
Optimum Temperature ◽  
Analysis Techniques ◽  
White Fir

Regression analysis techniques were applied to an estimation of three-dimensional surfaces representing the growth of Fomes pini as a function of time and temperature. These methods were judged to be valuable in their economy of data and in their provision of readily available plotting points for any desired cross section of the surface.The growth pattern of F. pini taken from Douglas fir was distinct from that of the form of F. pini found on nearby white fir. Growth of isolates from Douglas fir was more than twice that of white fir isolates after 18 days at near-optimum temperatures on malt agar. Estimates of growth trends and optimum temperatures were examined both for individual isolates of F. pini and for averages of isolates from the two host species, Douglas fir and white fir. Results suggest that chronological changes in the optimum temperature may be affected by the relation between the storage temperature preceding initial measurements and the terminal optimum temperature.

scholarly journals Production of Aspartame by Immobilized Thermolysin

2019 ◽  
pp. 1232-1239
Author(s):  
Mohammed A Alsoufi ◽  
Raghad A. Aziz
Keyword(s):  
Reaction Time ◽  
Immobilized Enzyme ◽  
Bentonite Clay ◽  
Optimum Temperature ◽  
Initial Activity ◽  
Reaction Conditions ◽  
Original Activity ◽  
Ph Profile ◽  
Full Activity ◽  
Main Activity

The aim of this study was the production of aspartame by using immobilized thermolysin in bentonite clay. The yield of immobilized thermolysin in bentonite was 92% of the original enzyme amount. pH profile of free and immobilized enzyme was 7.0 and 7.5 respectively which was stable at 6.5-9.0 for 30min. The optimum temperature of both enzymes was 50°C, while they were stable at 65°C for 30min. however, they lost 52.73 and 61.72% from its main activity at 80°C respectively. Immobilized thermolysin has retained all activity within 27 days, but it kept 68.27% of initial activity when stored for 60 days at 4°C whereas, it retained a full activity after 20 continue usage. In addition, it retained 86.53% of its original activity after 30 continuing usages. The yield of produced aspartame was increased with reaction time; it was 9% after 1h and increased gradually to 100% after 10h at reaction conditions.

scholarly journals ENHANCED ENZYMATIC ACTIVITY OF STREPTOMYCES GRISEOPLANUS L-ASPARGINASE VIA ITS INCORPORATION IN AN OIL-BASED NANOCARRIER

2020 ◽  
pp. 203-210
Author(s):  
ABEER A. EL-HADI ◽  
HANAN MOSTAFA AHMED ◽  
RANIA A. ZAKI ◽  
AMIRA MOHAMED MOHSEN
Keyword(s):  
Thermal Stability ◽  
Oleic Acid ◽  
Enzymatic Activity ◽  
Specific Activity ◽  
Residual Activity ◽  
Tween 80 ◽  
Biochemical Properties ◽  
Lymphocytic Leukemia ◽  
Tween 20 ◽  
Ph Range

Objective: L-asparaginase (L-asp) is a vital enzyme used as a therapeutic agent in combination with other drugs in the treatment of acute lymphoma, melanosarcoma and lymphocytic leukemia. Immobilization of enzymes through loading on nanoemulsion (NE) results in some advantages such as enhancing their stability and increasing their resistance to proteases. Aim of the present study is to formulate L-asp loaded nanoemulsion to enhance its efficiency and thermal stability. Methods: Nanoemulsion loaded with L-asp crude extract (specific activity 13.23U/mg protein) was prepared employing oleic acid as oil, tween 20/tween 80 as surfactants and propylene glycol (PG) as co-surfactant. L-asp loaded NE underwent several thermodynamic stability studies and the optimized formulae were further examined for their biochemical properties and thermal stability. Results The developed formulations were spherical in shape and their sizes were in the nanometric dimensions with negatively charged zeta potential values. Upon comparing the enzyme activity of L-asp loaded NE employing tween 20 (F1) or tween80 (F4) at different concentrations, the results revealed that F4 NE showed higher enzymatic activity [323 U/ml] compared to F1 NE [197 U/ml] at the same concentration. The nanosized immobilized L-asp was more stable in the pH range from 8 to 8.5 as compared to free L-asp. The immobilized enzyme preserved about 59.11% of its residual activity at 50 °C; while free L-asp preserved about 33.84%. Conclusion: In the view of these results, NE composed of oleic acid, tween 80 and PG represents a promising dosage form for enhancing the activity and stability of Streptomyces griseoplanus L-asp.

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