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 Extruction of keratinase from the local isolate Bacillus licheniformis and partially purified and characterized

2009 ◽  
Vol 3 (2) ◽  
pp. 41-52
Author(s):  
Rasha T. Abdullah ◽  
Abdulkareem J. Hashim ◽  
JASIM M. Karhout
Keyword(s):  
Enzyme Activity ◽  
Bovine Serum Albumin ◽  
Ion Exchange ◽  
Bacillus Licheniformis ◽  
Enzyme Characterization ◽  
Optimal Temperature ◽  
Chicken Feathers ◽  
Local Isolate ◽  
Cellulose Column ◽  
Optimal Ph

The keratinase produced from local isolate Bacillus licheniformis was purified by two steps included precipitation by ammonium sulphate with 40% saturation; followed by ion exchange using CM-Cellulose column. The enzyme was purified to 12.6 times in the last step with an enzyme yield of 17%. Enzyme characterization results indicated that: The optimal pH for enzyme activity was 7.5 and it was stable at 7-9.5. The optimal temperature for enzyme activity was 50°C and it was stable for 30 min at 25-45 °C. Substrate specifity was tested using casein, Bovine serum albumin, gelatin, hooves, human hair, chicken feathers and wool; higher specifity was recorded using casein gave 0.6 unit /ml. The enzyme was inhibited by PMSF and metal ions like Hg+2, Fe+2, Cu+2 and Mn+2, and activated by Ca+2, Mg+2, Zn+2and Al+3.

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.

scholarly journals Utilization of Crude Intracellular Chitinase Enzyme from Providencia stuartii for Glucosamine Production from Shrimp Shells

REAKTOR ◽  
2019 ◽  
Vol 19 (2) ◽  
pp. 62-67
Author(s):  
Hardoko Hardoko ◽  
Titri Siratantri Mastuti ◽  
Desy Puspasari ◽  
Yuniwaty Halim
Keyword(s):  
Optimum Condition ◽  
Substrate Concentration ◽  
Intracellular Enzyme ◽  
Fermentation Time ◽  
Shrimp Shells ◽  
Optimum Ph ◽  
Chitin Hydrolysis ◽  
Providencia Stuartii ◽  
Optimum Ph And Temperature

Chitin hydrolysis using enzyme is one of the methods to produce glucosamine in shorter time compared to using microbial cells, but the ability to produce glucosamine at enzyme’s optimum condition is influenced by substrate concentration and fermentation time. The objective of this research was to determine the optimum substrate concentration and fermentation time of shrimp shells’ chitin to produce glucosamine at the optimum pH and temperature of crude intracellular chitinase enzyme from Providencia stuartii. Method used was experimental method, started by extraction of intracellular enzyme from P. stuartii, followed by determination of optimum pH and temperature of enzyme. The optimum condition was used for experiment of shrimp shells’ chitin fermentation with treatments of chitin substrate concentration (0.5; 1.0; 1.5; 2.0%) and fermentation time (2, 4, 6 and 24 hours). Results showed that optimum enzyme activity occurred at pH of 5.0 and temperature of 40oC, which was about 6.03 U/ml. Concentration of chitin substrate and fermentation time influenced the amount of glucosamine obtained. Fermentation of shrimp shells’ chitin using crude intracellular enzyme was optimum at 1.0% substrate concentration and 6 hours fermentation time, which produced glucosamine about 1680.06±58.49 ppm. Keywords: intracellular chitinase enzyme, glucosamine, shrimp shells’ chitin, P. stuartii

Production and purification of an extracellular β-galactosidase from the Dutch elm disease fungus Ophiostoma novo-ulmi

1997 ◽  
Vol 43 (11) ◽  
pp. 1011-1016 ◽  
Author(s):  
Thomas Binz ◽  
Colette Gremaud ◽  
Giorgio Canevascini
Keyword(s):  
Enzyme Activity ◽  
Culture Filtrate ◽  
Galacturonic Acid ◽  
Gel Filtration ◽  
Total Activity ◽  
Dutch Elm Disease ◽  
Optimal Temperature ◽  
Ophiostoma Ulmi ◽  
Sds Page ◽  
Optimal Ph

The causal agents of Dutch elm disease, Ophiostoma ulmi (isolate H200) and Ophiostoma novo-ulmi (isolate CKT-11), secreted similar amounts of β-galactosidase in liquid shake cultures when grown on galacturonic acid or sodium pectate (1.45 ± 0.16 and 1.03 ± 0.24 nkat∙mL−1 for O. ulmi, respectively, and 1.30 ± 0.08 and 1.28 ± 0.26 nkat∙mL−1 for O. novo-ulmi, respectively). Rhamnose and pectin also stimulated secretion but to a lesser extent, whereas on glucose, enzyme activity was barely detectable (≤0.01 nkat∙mL−1). Ophiostoma novo-ulmi was shown by Q-Sepharose chromatography to form two β-galactosidases, named β-galactosidases I and II. In cultures grown on galacturonic acid β-galactosidase I accounted for approximately 75% of the total activity in the culture filtrate. β-Galactosidase I was further purified to apparent electrophoretic homogeneity by means of Sephacryl gel filtration chromatography, chromatofocusing, and Superdex75 gel filtration. The molecular mass of the enzyme was 135 kDa by SDS–PAGE and 123 kDa by gel filtration. Its isoelectric point, determined by chromatofocusing, was 4.9. The optimal pH for enzyme activity was 5.8 and the optimal temperature was 50 °C. The Km values for p-nitrophenyl β-D-galactopyranoside and lactose were 7.52 and 14.23 mM, respectively, and the maximum velocities for these substrates were 1733 and 355 nkat∙mg protein−1, respectively. The Ki value for D(−)-galactonic acid γ-lactone was 2.29 mM.Key words: Dutch elm disease, β-galactosidase, Ophiostoma ulmi, Ophiostoma novo-ulmi.

scholarly journals Produksi Crude Selulase dari Bahan Baku Ampas Tebu Menggunakan Kapang Phanerochaete chrysosporium

2017 ◽  
Vol 1 (1) ◽  
pp. 17
Author(s):  
Sri Rulianah ◽  
Zakijah Irfin ◽  
Mufid Mufid ◽  
Prayitno Prayitno
Keyword(s):  
Enzyme Activity ◽  
Salicylic Acid ◽  
Phanerochaete Chrysosporium ◽  
Substrate Concentration ◽  
Lignin Content ◽  
Raw Material ◽  
Cellulase Enzymes ◽  
Fermentation Time ◽  
Acid Reagent ◽  
High Cellulose

Bagasse mengandung selulosa yang cukup tinggi sehingga berpotensi sebagai bahan baku produksi crude selulase menggunakan kapang Phanerochaete chrysosporium. Kapang ini memiliki kemampuan untuk memproduksi enzim selulase dari substrat yang mengandung selulosa dan juga menghasilkan enzim yang dapat memecah lignin sehingga tidak perlu dilakukan proses delignifikasi. Tujuan dari penelitian ini adalah untuk memanfaatkan limbah ampas tebu sebagai bahan baku pembuatan crude selulase menggunakan kapang Phanerochaete chrysosporium dan mengetahui pengaruh penambahan konsentrasi substrat dan waktu fermentasi terhadap aktivitas crude selulase yang dihasilkan. Penelitian ini dilakukan dengan cara mengeringkan dan memperkecil ukuran ampas tebu, meremajakan kapang Phanerocheate chrysoporium, membuat inokulum dalam media cair, memfermentasi ampas tebu sesuai dengan variabel, dengan media Nitrogen Limited Media (NLM) menggunakan kapang Phanerocheate chrysoporium. Hasil fermentasi disaring, dan filtratnya dianalisa aktivitasnya sebagai crude selulase. Variabel dalam penelitian ini adalah waktu fermentasi 9, 11, 13, 15 dan 17 hari dan konsentrasi ampas tebu sebagai media: 5, 6, dan 7 % b/v. Ekstrak kasar selulase (crude) yang dihasilkan disaring menggunakan filter vakum, dan aktivitas filtrat (crude cellulase) diuji dengan pereaksi DNS (dinitro salicylic acid) dengan menggunakan spektrofotometer UV-Vis. Hasil penelitian menunjukkan bahwa aktivitas selulase tertinggi diperoleh pada variabel konsentrasi ampas tebu sebesar 7% b/v dan waktu inkubasi selama 17 hari yaitu sebesar 91.304 U/mL.Bagasse contain high cellulose which potentially to be used to raw material for producing cellulase enzyme using fungi Phanerochaete chrysosporium. This fungus has ability to produce cellulase enzymes from substrates which contain cellulose and also produce enzymes that can degrade lignin content so it didn’t need the delignification process. The objective of this study was to convert cellulose in bagasse to be crude cellulase enzymes by using Phanerochaete chrysosporium and determine the effect of substrate concentration and fermentation time to the enzyme activity. This research was conducted by drying and reducing the bagasse particle size, rejuvenating mold Phanerocheate chrysoporium, making inoculum in liquid medium, fermenting bagasse in accordance with the variable, with media NLM (nitrogen limited media) using Phanerocheate chrysoporium. Fermentation results were filtered, and it was analyzed the activity of crude cellulase. The variable in this study was the time of fermentation 9, 11, 13, 15, and 17 days and substrate concentration: 5, 6, and 7 % b/v. Crude cellulose was filtered and was analyzed the enzyme activity by DNS (dinitro salicylic acid) reagent, using UV-Vis spectrophotometer. The best result of this study was the crude cellulase with highest activity 91,304 U/mL for 7 % substrate concentration with fermentation time 17 days.

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 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

scholarly journals Production of N-acetylglucosamine from shrimp shells’ chitin using intracellular chitinase from Mucor circinelloides

Food Research ◽  
2020 ◽  
Vol 4 (5) ◽  
pp. 1582-1587
Author(s):  
Yuniwaty Halim ◽  
Fransiska ◽  
Hardoko ◽  
R. Handayani
Keyword(s):  
Incubation Period ◽  
Substrate Concentration ◽  
Chitinase Activity ◽  
Mucor Circinelloides ◽  
Optimum Temperature ◽  
Shrimp Shells ◽  
Incubation Periods ◽  
Optimum Ph ◽  
Substrate Concentrations ◽  
Optimum Ph And Temperature

Chitin is a natural biopolymer found in shrimp shells and can be processed into Nacetylglucosamine which is extensively used as a dietary supplement to treat osteoarthritis, back pain and knee pain. This research was conducted to determine the optimum pH, temperature, substrate concentration and incubation period to produce Nacetylglucosamine using crude and semi pure intracellular chitinase extracted from Mucor circinelloides. Chitinase activity was measured to determine optimum pH and temperature by using various pHs (3, 4, 5, 6, 7, 8 and 9) and temperatures (30oC, 40oC, 50oC, 60oC, 70oC and 80oC). Different substrate concentrations (0.5%, 1.0%, 1.5% and 2.0%) and incubation periods (2, 4, 6 and 24 hrs) were used to determine the optimum condition to produce N-acetylglucosamine. Results showed that crude intracellular chitinase had an optimum pH of 5 with chitinase activity of 4.16±0.07 U/mL and optimum temperature of 60oC with chitinase activity of 4.22±0.07 U/mL. The optimum substrate concentration obtained was 0.5% and the optimum incubation period obtained was 6 hrs with about 961.67±9.13 ppm N-acetylglucosamine produced. Semi pure intracellular chitinase had an optimum pH of 4 with chitinase activity of 4.75±0.09 U/mL and optimum temperature of 50oC with chitinase activity of 5.03±0.08 U/mL. The optimum substrate concentration obtained was 1.5% and the optimum incubation period obtained was 4 hrs with about 1150.56±12.55 ppm N-acetylglucosamine produced.

scholarly journals Characteristics of Curds With Milk Clotting Enzyme from Indonesian Local Isolate of Lactic Acid Bacteria

2020 ◽  
Vol 1 (2) ◽  
pp. 79-86
Author(s):  
Wendry Putranto ◽  
Apon Mustopa ◽  
Jendri Mamangkey ◽  
Netty Aritonang
Keyword(s):  
Enzyme Activity ◽  
Ammonium Sulfate ◽  
Observational Research ◽  
Ammonium Sulfate Precipitation ◽  
Fermentation Time ◽  
Milk Clotting ◽  
Milk Clotting Enzyme ◽  
Qualitative Observation ◽  
Milk Clotting Activity

To get the potential of lalcat acid bacteria isolate to produce Milk Clotting Enzyme (MCE), it is necessary to screen milk clotting activity both quantitatively and qualitatively. Through qualitative observation, the characteristics of the curd resulting from enzyme activity can be obtained. MCE is a protease that has the characteristics of milking. Based on the results of this observational research, the curd characteristic produced can be used as a benchmark to determine the length of time of fermentation and optimization of the determination of ammonium sulfate precipitation concentration. Isolate BAL shows the results of a compact curd at a fermentation time of 25 hours at 37 ℃ and the optimization results of the deposition of ammonium sulfate which shows the characteristics of a compact curd by 45% ammonium sulfate.

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.

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