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Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology.

El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM - Biomed Res Int (2015)

Bottom Line: The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out.Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design.The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.

ABSTRACT
Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rRNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product (1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

No MeSH data available.


Related in: MedlinePlus

The main effects of the fermentation conditions on L-asparaginase production according to the Packett-Burman experimental results.
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Related In: Results  -  Collection


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fig4: The main effects of the fermentation conditions on L-asparaginase production according to the Packett-Burman experimental results.

Mentions: Sixteen different independent (assigned) variables including temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O and four unassigned variables (commonly referred to as dummy variables) were screened in Plackett-Burman experimental design of 20 trials to study the effect of the selected variables on the production of L-asparaginase. Four dummy variables are used to estimate experimental errors in data analysis (Table 3). Table 3 represents the results of the screening of significant variables for L-asparaginase production and the corresponding response (Y) using Plackett-Burman design. The maximum L-asparaginase activity (49.874 U/mL) was achieved in the run number 15, while the minimum L-asparaginase activity (5.181 U/mL) was observed in the run number 5. Statistical analysis of the L-asparaginase activity was performed and represented in Table 4. With respect to the main effect of each variable (Figure 4), we can see that seven variables from the fifteen named including temperature, inoculum size, inoculum age, agitation speed, dextrose, starch and L-asparagine positively affect L-asparaginase production where the other eight variables named pH, incubation time, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O negatively affect L-asparaginase production. The Pareto chart illustrates the order of significance of the variables affecting L-asparaginase production in Plackett-Burman experimental design (Figure 5). Among the fifteen variables, agitation speed showed the highest positive effect by 15.87%, followed by inoculum age (9.96%), and then temperature by 7.55%. Among the 15 variables, NaCl showed the highest negative significance by 13.03%. Next to NaCl, incubation time showed negative effect by 10.75% followed by FeSO4·7H2O and MgSO4·7H2O by 9.87% and 7.95%, respectively.


Optimization of Culture Conditions for Production of the Anti-Leukemic Glutaminase Free L-Asparaginase by Newly Isolated Streptomyces olivaceus NEAE-119 Using Response Surface Methodology.

El-Naggar Nel-A, Moawad H, El-Shweihy NM, El-Ewasy SM - Biomed Res Int (2015)

The main effects of the fermentation conditions on L-asparaginase production according to the Packett-Burman experimental results.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477217&req=5

fig4: The main effects of the fermentation conditions on L-asparaginase production according to the Packett-Burman experimental results.
Mentions: Sixteen different independent (assigned) variables including temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O and four unassigned variables (commonly referred to as dummy variables) were screened in Plackett-Burman experimental design of 20 trials to study the effect of the selected variables on the production of L-asparaginase. Four dummy variables are used to estimate experimental errors in data analysis (Table 3). Table 3 represents the results of the screening of significant variables for L-asparaginase production and the corresponding response (Y) using Plackett-Burman design. The maximum L-asparaginase activity (49.874 U/mL) was achieved in the run number 15, while the minimum L-asparaginase activity (5.181 U/mL) was observed in the run number 5. Statistical analysis of the L-asparaginase activity was performed and represented in Table 4. With respect to the main effect of each variable (Figure 4), we can see that seven variables from the fifteen named including temperature, inoculum size, inoculum age, agitation speed, dextrose, starch and L-asparagine positively affect L-asparaginase production where the other eight variables named pH, incubation time, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O negatively affect L-asparaginase production. The Pareto chart illustrates the order of significance of the variables affecting L-asparaginase production in Plackett-Burman experimental design (Figure 5). Among the fifteen variables, agitation speed showed the highest positive effect by 15.87%, followed by inoculum age (9.96%), and then temperature by 7.55%. Among the 15 variables, NaCl showed the highest negative significance by 13.03%. Next to NaCl, incubation time showed negative effect by 10.75% followed by FeSO4·7H2O and MgSO4·7H2O by 9.87% and 7.95%, respectively.

Bottom Line: The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out.Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design.The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

View Article: PubMed Central - PubMed

Affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.

ABSTRACT
Among the antitumor drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological, and biochemical properties together with 16S rRNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product (1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett-Burman experimental design and response surface methodology was carried out. Fifteen variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4·7H2O, NaCl, and FeSO4·7H2O) were screened using Plackett-Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age, and agitation speed) were further optimized by the face-centered central composite design-response surface methodology.

No MeSH data available.


Related in: MedlinePlus