scholarly journals R unning Head: Novel Recombinant Phytate - d egrading Enzyme Production Statistical Optimization of the Induction of Phytase Production by Arabinose in a recombinant E. coli using Response Surface Methodology

2017 ◽  
Vol 26 (1) ◽  
Author(s):  
Abd - ElAziem Farouk ◽  
Anis Shobirin Meor Hussin ◽  
Ralf Greiner ◽  
Shareef Mohideen Ismail ◽  
Mohamed Faizal Batcha ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Incubation Temperature ◽  
Expression System ◽  
Phytase Production ◽  
E Coli ◽  
Significant Parameter ◽  
Central Composite Experimental Design ◽  
Full Factorial ◽  
Central Composite

The production of phytase in a recombinant E.coliusing thepBAD expression system wasoptimized using response surface methodology with full-factorial faced centered central composite design. The ampicilin and arabinose concentrationin the cultivation media and the incubation temperature were optimized in order to maximize phytase productionusing 23 central composite experimental design. With this design the number of actual experiment performed could be reduced while allowing eludidation of possible interactions among thesefactors. The most significant parameter was shown to be the linear and quadratic effect of the incubation temperature. OPtim al conditionsfor phytase production were determinedto be100 μg/ml ampicilin, 0.2 % arabinose and an incubation temperature of37ºC.The production of phytase in the recombinant E. coliwas scaledup to 100ml and 1000ml.

scholarly journals Statistical Optimization of the Induction of Phytase Production by Arabinose in a recombinant E. coli using Response Surface Methodology

2017 ◽  
Vol 26 (1) ◽  
pp. 57-64
Author(s):  
Abd-El Aziem Farouk ◽  
Anis Shobirin Meor Hussin ◽  
Ralf Greiner ◽  
Shareef Mohideen Ismail ◽  
Hamadah Mohd Nur Lubis
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Incubation Temperature ◽  
Expression System ◽  
Optimal Conditions ◽  
Phytase Production ◽  
E Coli ◽  
Central Composite Experimental Design ◽  
Full Factorial ◽  
Central Composite

The production of phytase in a recombinant E.coli using the pBAD expression  system was optimized using response surface methodology with full-factorial faced centered central composite design. The ampicilin and arabinose concentration in the cultivation media and the incubation temperature were optimized in order to maximize phytase production using 2 3  central composite experimental design. With this design the number of actual experiment performed could be reduced while allowing eludidation of possible interactions among these factors. The most significant parameter was shown to be the linear and quadratic effect of the incubation temperature.  Optimal conditions for phytase production were determined to be 100 µg/ml ampicilin, 0.2 % arabinose and an incubation temperature of 37ºC. The production of phytase in the recombinant E. coli was scaled up to 100 ml and 1000 ml.   

Optimization of Critical Medium Components for the Expression of Recombinant Human Transferrin in Insect Cells Baculovirus System

2012 ◽  
Author(s):  
Clarence M. Ongkudon ◽  
Badarulhisam Abdul Rahman ◽  
Azila Abd. Aziz
Keyword(s):  
Response Surface Methodology ◽  
Experimental Design ◽  
Central Composite Design ◽  
Response Surface ◽  
Insect Cells ◽  
Expression System ◽  
Proliferating Cells ◽  
Human Transferrin ◽  
Lipid Mixtures ◽  
Central Composite

Transferin manusia (hTf) memainkan peranan yang penting dalam fungsi bakteriostatik dan pengangkutan ferum dari bahagian penyimpanan ke sel–sel yang membiak melalui proses endositosis janaan reseptor. Sistem ekspresi bakulovirus sel serangga telah dipakai secara meluas sebagai sistem alternatif dalam penghasilan Transferin manusia rekombinan (rhTf). Kajian ini ditumpukan ke atas pengoptimuman glutamina, glukosa dan campuran lipid 1000x yang dapat meningkatkan penghasilan rhTf. Reka bentuk eksperimen yang melibatkan 17 eksperimen reka bentuk komposit berpusat (CCD) telah digunakan dan hasil kajian dianalisis oleh Statistika (Statsoft v. 5.0). Metodologi permukaan tindak balas (RSM) telah mengenalpasti nilai optimum parameterparameter yang dikaji iaitu glutamina=2211.20 mg/L, glukosa=1291.95 mg/L, dan campuran lipid 1000x=0.64 %v/v. Hasil optimasi menunjukkan peningkatan hasil rhTf sebanyak tiga kali ganda, iaitu daripada 19.89 μg/ml kepada 65.12 μg/ml. Kata kunci: Transferin manusia; bakulovirus sel serangga; reka bentuk eksperimen; reka bentuk komposit berpusat; metodologi permukaan tindak balas Human Transferrin (hTf) plays a big role in providing bacteriostatic functions as well as to transport iron from the storage part to all proliferating cells by receptor mediated endocytosis. Insect cells baculovirus expression system has been widely used as an alternative expression system for the production of recombinant human Transferrin (rhTf). This work focused mainly on the optimization of glutamine, glucose and lipid mixtures 1000x to increase rhTf yield. An experimental design involving 17 central composite design (CCD) experiments was employed and results were analyzed by Statistica (Statsoft v. 5.0). The response surface methodology (RSM) had identified the optimum values where glutamine=2211.20 mg/L, glucose=1291.95 mg/L, and lipid mixtures 1000x=0.64 %v/v. Using the optimized parameters, the studies demonstrated an increase in the rhTf yield by three–fold from 19.89 μg/ml to 65.12 μg/ml. Key words: Human transferrin; insect cells baculovirus; experimental design; central composite design; response surface methodology

Research on Blanching Pretreatment of Quick Frozen Sword Bean (Canavalia gladiate)

2011 ◽  
Vol 140 ◽  
pp. 416-420
Author(s):  
Chen Wei Zhou ◽  
Juan Xu ◽  
Qing Qing Li ◽  
Rui Zhi Wang ◽  
Lu Yang ◽  
...  
Keyword(s):  
Response Surface Methodology ◽  
Statistical Model ◽  
Response Surface ◽  
Peroxidase Activity ◽  
Primary Determinant ◽  
Central Composite Experimental Design ◽  
Main Index ◽  
Temperature Time ◽  
Central Composite

Quick frozen could preserve the maximum micro-texture and taste quality of vegetables, and blanching pretreatment is primary determinant of quick frozen. In this study, the central composite experimental design and response surface methodology were adopted to derive a statistical model for the effect of blanching pretreatment on the quality of sword bean (Canavalia gladiate), with peroxidase activity as main index. The pretreatment process optimized with response surface methodology was blanching at 96 °C for 1.03 min. Through optimization, a statistical model was established for the relation between blanching temperature, time and peroxidase activity, which would facilitate the prediction of the effects of blanching on the quality of sword bean and could play a guidance role in the blanching pretreatment of many other vegetables.

scholarly journals Heterologous Ectoine Production inEscherichia coli: Optimization Using Response Surface Methodology

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
I Putu Parwata ◽  
Deana Wahyuningrum ◽  
Sony Suhandono ◽  
Rukman Hertadi
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Gene Cluster ◽  
Incubation Temperature ◽  
Halophilic Bacteria ◽  
Dry Weight ◽  
Halophilic Bacterium ◽  
E Coli ◽  
Halomonas Elongata ◽  
Recombinant Cell

Introduction. A halophilic bacterium of theHalomonas elongataBK-AG25 has successfully produced ectoine with high productivity. To overcome the drawbacks of high levels of salt in the production process, a nonhalophilic bacteria ofEscherichia coli(E. coli) was used to express the ectoine gene cluster of the halophilic bacteria, and the production of ectoine by the recombinant cell was optimized.Methods. The ectoine gene cluster from the halophilic bacterium was isolated and inserted into an expression plasmid of pET30(a) and subsequently transformed intoE. coliBL21 (DE3). Production of ectoine from the recombinantE. coliwas investigated and then maximized by optimizing the level of nutrients in the medium, as well as the bioprocess conditions using response surface methodology. The experimental designs were performed using a central composite design.Results. The recombinantE. colisuccessfully expressed the ectoine gene cluster ofHalomonas elongataBK-AG25 under the control of theT7promoter. The recombinant cell was able to produce ectoine, of which most were excreted into the medium. The optimization of ectoine production with the response surface methodology showed that the level of salt in the medium, the incubation temperature, the optical density of the bacteria before induction, and the final concentration of the inducer gave a significant effect on ectoine production by the recombinantE. coli. Interestingly, the level of salt in the medium and the incubation temperature showed an inverse effect on the production of intracellular and extracellular ectoine by the recombinant cell. At the optimum conditions, the production yield was about 418 mg ectoine/g cdw (cell dry weight) after 12 hours of incubation.Conclusion. This study is the first report on the expression of an ectoine gene cluster ofHalomonas elongataBK-AG25 inE. coliBL21, under the control of theT7promoter. Optimization of the level of nutrients in the medium, as well as the bioprocess condition using response surface methodology, has successfully increased the production of ectoine by the recombinant bacteria.

Response surface methodology for the optimization of keratinase production in culture medium containing feathers produced by Kocuria rosea

2006 ◽  
Vol 52 (5) ◽  
pp. 445-450 ◽  
Author(s):  
C Bernal ◽  
I Diaz ◽  
N Coello
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Batch Fermentation ◽  
Culture Medium ◽  
Medium Optimization ◽  
Fold Increase ◽  
Medium Composition ◽  
Central Composite Experimental Design ◽  
Kocuria Rosea ◽  
Central Composite

A 43-fold increase in keratinase production by Kocuria rosea was achieved in batch fermentation using response surface methodology. Factorial designs were used to select the components of a culture medium that showed a significant effect on keratinase production. An orthogonal–central composite experimental design was performed, with only two (feathers and magnesium) from nine initial compounds being further analyzed by response surface methodology. An optimum keratinase production of 14 886.9 U/mg was obtained with the following medium composition (per litre): NH4Cl, 0.3 g; NaCl, 0.3 g; K2HPO4, 3.2 g; KH2PO4, 4.0 g; MgSO4·6H2O, 0.5 g; yeast extract, 0.1 g; and finely milled feathers, 30 g. The medium was shaken at 400 r/min with an incubation period of 14 h at 40 °C.Key words: feathers, keratinases, Kocuria rosea, medium optimization, response surface methodology.

scholarly journals OPTIMASI FERMENTASI BAGAS TEBU OLEH Zymomonas mobilis CP4 (NRRL B-14023) UNTUK PRODUKSI BIOETANOL (Optimization of Sugarcane Bagasse Fermentation by Zymomonas mobilis CP4 (NRRL B-14023) for Bioethanol Production)

2014 ◽  
Vol 34 (03) ◽  
pp. 247 ◽  
Author(s):  
Atmiral Ernes ◽  
Lia Ratnawati ◽  
Agustin Krisna Wardani ◽  
Joni Kusnadi
Keyword(s):  
Response Surface Methodology ◽  
Experimental Design ◽  
Response Surface ◽  
Sugarcane Bagasse ◽  
Zymomonas Mobilis ◽  
Urea Concentration ◽  
Fermentation Time ◽  
Inoculum Concentration ◽  
Central Composite Experimental Design ◽  
Central Composite

Second generation bioethanol can be produced from fermentation of natural renewable materials, such as agricultural crops, as well as from industrial and domestic waste. The present study was aimed to optimize the fermentation process (inoculum concentration, urea concentration, and fermentation time) for ethanol production from sugarcane bagasse byZymomonas mobilis CP4 using response surface methodology (RSM) central composite experimental design (CCD). The RSM model predicted the optimum value of ethanol content was 1.257% (v/v) at inoculum concentration 15% (v/v), urea concentration 0.3% (w/v), and fermentation time 45 h. Based on the experiment, the ethanol concentrationwas 1.213% (v/v), which was in close agreement with the predicted value. Ethanol yield of this experiment was 0.479 with fermentation effi ciency of 93.9%. The results presented here proved a signifi cant contribution of Z. mobilis CP4 to the production of bioethanol from sugarcane bagasse.Keywords: Bioethanol, sugarcane bagasse, Zymomonas mobilis CP4, fermentation optimization ABSTRAKBioetanol generasi kedua dapat diproduksi dari fermentasi bahan terbarukan, seperti produk hasil pertanian, dan limbah atau hasil samping pengolahan industri dan rumah tangga. Tujuan penelitian ini adalah optimasi parameter fermentasi yang meliputi konsentrasi inokulum, konsentrasi urea, dan lama fermentasi untuk produksi etanol dari bagas tebu oleh Zymomonas mobilis CP4 dengan menggunakan response surface methodology (RSM) central composite experimental design (CCD). Kondisi respon yang optimal berdasarkan prediksi model diperoleh pada konsentrasi inokulum 15% (v/v), konsentrasi urea 0,3% (b/v), dan lama fermentasi 45 jam, dengan prediksi respon kadar etanol sebesar 1,257%(v/v). Berdasarkan hasil penelitian, kadar etanol optimal diperoleh sebesar 1,213% (v/v), yang menunjukkan hasil yang tidak berbeda jauh dengan prediksi model. Yield etanol yang diperoleh sebesar 0,479 dengan efi siensi fermentasi 93,9%. Hasil penelitian ini membuktikan bahwa strain bakteri Zymomonas mobilis CP4 memiliki potensi yang cukup menjanjikan sebagai mikroba penghasil etanol.Kata kunci: Bioetanol, bagas tebu, Zymomonas mobilis CP4, optimasi fermentasi

Enzyme-assisted Extraction for Optimized Recovery of Phenolic Bioactives from Peganum hermala Leaves Using Response Surface Methodology

2018 ◽  
Vol 17 (4) ◽  
pp. 349-354
Author(s):  
Qadir Rahman ◽  
Anwar Farooq ◽  
Amjad Gilani Mazhar ◽  
Nadeem Yaqoob Muhammad ◽  
Ahmad Mukhtar
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Enzyme Concentration ◽  
Coumaric Acid ◽  
Assisted Extraction ◽  
Pre Treatment ◽  
Optimized Conditions ◽  
Enzyme Complexes ◽  
Hydrolysis Of ◽  
Central Composite

This study investigates the effect of enzyme formulations (Zympex-014, Kemzyme dry-plus and Natuzyme) on recovery of phenolics from Peganum hermala (harmal) leaves, under optimized conditions using response surface methodology. As compared to the other enzyme complexes, the yield (34 g/100g) obtained through Zympex-014-assisted extraction was higher under optimized conditions such as time (75 min), temperature (70°C), pH (6.5) and enzyme concentration (5 g/100 g) using central composite design (CCD). Effectiveness of Zympex-014 towards hydrolysis of P. hermala leaves cell wall was examined by analyzing the control and enzyme-treated leave residues using scanning electron microscope (SEM). GC/MS characterization authenticated the presence of quercetin (1.44), gallic acid (0.23), caffeic acid (0.04), cinnamic acid (0.05), m-coumaric acid (0.23) and p-coumaric acid (0.37 μg/g) as the potent phenolics in Zympex-014 based extract. It can be concluded from the findings of the current work that pre-treatment of P. hermala leaves with Zympex-014 significantly enhanced the recovery of phenolics that supports its potential uses in the nutra-pharamaceutical industry.

Response Surface Methodology towards Optimization of Calotropis Procera Essential oil Extraction by using Supercritical CO2.

2019 ◽  
Vol 09 ◽  
Author(s):  
Hossein Zaeri ◽  
Bahareh Kamyab Moghadas ◽  
Bijan Honarvar ◽  
Ali Shokuhi Rad
Keyword(s):  
Response Surface Methodology ◽  
Essential Oil ◽  
Response Surface ◽  
Extraction Time ◽  
Calotropis Procera ◽  
Retrieval Rate ◽  
The Family ◽  
Effects Of Temperature ◽  
Carbon Dioxide Co2 ◽  
Central Composite

: In this research, the extraction of essential oil from Calotropis Procera with the family name of Asclepiadaceae, by supercritical carbon dioxide (CO2) solvent has been investigated in detail, and the yield and chemical profile of the extracts achieved by this method were compared with those resulted by the conventional Hydro distillation method. To optimize the process parameters of CO2 supercritical extraction (SCE) of the Calotropis Procera, the Response Surface Methodology (RSM) with central composite design (CCD) was employed. The effects of temperature, pressure, and extraction time on the oil yield are considered for investigation. Results showed that the data were sufficiently fitted into the second-order polynomial model. The extraction conditions, including pressure, temperature, and extraction time, were studied between 150-200 bar, 40-50 ºC, and 50-100 min, respectively. The optimal conditions are achieved as the temperature of 47.19ºC, the pressure of 172.2 bar, and time of 86 minutes with the retrieval rate of 31.39%.

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