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

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.

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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

ASEAN Journal on Science and Technology for Development ◽

10.29037/ajstd.179 ◽

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.

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

Jurnal Teknologi ◽

10.11113/jt.v45.338 ◽

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

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Supercritical CO2 Extraction of Plumula nelumbinis Oil Rich in γ-Sitosterol Using Response Surface Methodology

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.864-867.213 ◽

2013 ◽

Vol 864-867 ◽

pp. 213-216

Author(s):

Dong Dong Jia ◽

Rong Lan ◽

Yong Yue Sun

Keyword(s):

Response Surface Methodology ◽

Factorial Design ◽

Response Surface ◽

Surface Analysis ◽

Full Factorial Design ◽

Response Surface Analysis ◽

Optimal Conditions ◽

Full Factorial

Supercritical CO2extraction ofPlumula nelumbinisoil rich in γ-sitosterol was investigated with a 42full factorial design and response surface analysis. At optimal conditions (P=35 MPa,T=55 C,dp=0.22 mm,Q=2.0 L/min), the yield of the extracted oil was up to 12.2%, in which the concentration of γ-sitosterol was 7.38%, indicating that the γ-sitosterol contents inPlumula nelumbinisand its oil were much higher than that in other vegetables.

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The Brazilian Way to Consume acai: Do guarana Extract and Sugar Concentrations Influence on Acceptance?

Journal of Food Research ◽

10.5539/jfr.v3n1p39 ◽

2013 ◽

Vol 3 (1) ◽

pp. 39

Author(s):

Gabriela Montenegro ◽

Karen S. Pereira ◽

Lauro Melo

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Critical Value ◽

Consumer Acceptance ◽

Central Composite Rotatable Design ◽

Optimal Conditions ◽

F Test ◽

Sensory Acceptance ◽

Amazon Estuary ◽

Central Composite

Acai is a palm widely distributed in the Amazon estuary floodplains. The edible pulp of acai fruits is commonly macerated with water to produce a thick, purple beverage of creamy texture, oily appearance, and characteristic flavor. Depending on the Brazilian region, the acai based-product is prepared adding sugar and guarana extract, but their ideal proportions have never been determined in order to optimize consumers’ sensory acceptance. This research investigated these concentrations using response surface methodology (RSM) based on a five-level, two variable central composite rotatable design (CCRD). Dependent variable was consumer acceptance (flavor, texture and overall liking) and results were analyzed by multivariate regressions. Analyses of Variance (ANOVAs) showed significant models – F-test values (29.3 for flavor, for texture 37.8 and 30.4 for overall liking) higher than the critical value of 4.35 (d.f. = 3; p < 0.05; R2 of 0.926 for flavor, 0.942 for texture and 0.929 for overall liking). Acceptance models are presented (significant parameters). Results showed that guarana extract has a stronger influence (negative) on acceptance compared to sugar (positive), both not on optimal conditions yet. Therefore, more studies are needed in order to optimize acai acceptance.

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

Applied Mechanics and Materials ◽

10.4028/www.scientific.net/amm.140.416 ◽

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.

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Phytoremediation of Soil Contaminated with Nickel by Lepidium sativum; Optimization by Response Surface methodology

Global NEST Journal ◽

10.30955/gnj.000996 ◽

2013 ◽

Vol 15 (1) ◽

pp. 69-75 ◽

Cited By ~ 9

Keyword(s):

Response Surface Methodology ◽

Response Surface ◽

Soil Samples ◽

Lepidium Sativum ◽

Nickel Concentration ◽

Optimal Conditions ◽

Polluted Soils ◽

Independent Variables ◽

Central Composite ◽

Accumulator Plant

Phytoremediation is an alternative to traditional chemical and ways of treating polluted soils. The current study was carried out to investigate the phytoremediation of soil contaminated with nickel (Ni) by Lepidium sativum. Soil samples from 0 to 10 cm depth were collected. Lepidium sativum was transplanted in pots containing 5 kg of the collected soils. Central composite design and response surface methodology were employed in order to illustrate the nature of the response surface in the experimental design and explain the optimal conditions of the independent variables. Different concentrations for Ni (1 to 20 mg kg-1) and times for collecting samples (10 to 40 days) were used. The results showed the amount of Ni removed was ranged from 8.62 mg kg-1 (Ni concentration of 20 mg kg-1 and time for taking samples of 10 days) to 7.066 mg kg-1 (Ni concentration of 10.50 mg kg-1 and time for taking samples of 40days). Additionally, the findings explained that the Lepidium sativum is an effective accumulator plant for phytoremediation of Ni polluted soils. Optimum conditions for nickel concentration and time for taking samples were 19.66 mg kg-1 and 39.28 days, respectively. For the optimum condition, the amount of Ni removed was 10.8095 mg kg-1.

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Optimization of baker`s yeast production on grape juice using response surface methodology

Acta periodica technologica ◽

10.2298/apt2152089s ◽

2021 ◽

pp. 89-110

Author(s):

Mahmood Sawsan ◽

Ali Ali ◽

Darwesh Ayhem ◽

Zam Wissam

Keyword(s):

Saccharomyces Cerevisiae ◽

Response Surface Methodology ◽

Response Surface ◽

Ammonium Sulfate ◽

Grape Juice ◽

Optimal Conditions ◽

Fermentation Time ◽

Fermentation Conditions ◽

Central Composite Experimental Design ◽

Kinematic Models

There is an increasing interest in improving biological processes, including fermentation processes, improving fermentation conditions is difficult, as it requires the use of an appropriate improvement method that allows operating the biological fermenter under optimal conditions in order to obtain the largest possible amount of the final product. The aim of this work was to succeed in examples of fermentation conditions to produce the largest possible quantity of dry yeast biomass Saccharomyces cerevisiae using grape juice as the sole carbon source. The optimum values of five factors that have an effect on the production of dry biomass from baker`s yeast were determined. The design of the experiments was carried out using the central composite experimental design (CCD) and the number of experiments according to the design was (54) experiments, the response surface methodology method was used to determine the best possible amount of production of yeast, and has reached (41.44 g/L) after 12 hours of fermentation, under the following optimal conditions (temperature (30.11??), pH (4.75), sugar concentration (158.36 g/L), the ratio of carbon to nitrogen (an essential nutrient for yeast growth ) is (11.9), initial concentration of yeasts (2.5 g/L), the amount of urea was 6.65 g/L and the amount of ammonium sulfate used was 6.65 g/L, so that the concentration of added urea and ammonium sulfate was (50-50)% and the required C/N ratio was achieved, and the used agitation speed was equal to 200 r.p.m during the fermentation process. The fermenter power of the obtained yeast was 470 ml. Three kinematic models (Monod, Verhulst, and Tessier) were also selected for the purpose of studying the kinetic performance of Saccharomyces cerevisiae yeast. Monod and Tessier`s models did not give satisfactory results, while the best results were according to the Verhulst model. Also, the Leudeking Piret model has also been successfully used to predict substrate consumption during fermentation time.

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Heterologous Ectoine Production inEscherichia coli: Optimization Using Response Surface Methodology

International Journal of Microbiology ◽

10.1155/2019/5475361 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 4

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.

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

Canadian Journal of Microbiology ◽

10.1139/w05-139 ◽

2006 ◽

Vol 52 (5) ◽

pp. 445-450 ◽

Cited By ~ 35

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.

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