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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

Three-dimensional response surface plots showing the effect of temperature (X1), inoculum age (X5) and agitation speed (X6) and their mutual effect on the production of L-asparaginase.
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fig7: Three-dimensional response surface plots showing the effect of temperature (X1), inoculum age (X5) and agitation speed (X6) and their mutual effect on the production of L-asparaginase.

Mentions: The interaction effects and optimal levels of the variables were determined by plotting the three-dimensional response surface curves (Figures 7(a)–7(c)) when one of the variables is fixed at optimum value and the other two are allowed to vary. Figure 7(a) represents the L-asparaginase activity as a function of temperature (X1) and inoculum age (X5) by keeping agitation speed (X6) at optimum value. It showed that lower and higher levels of temperature support relatively low levels of L-asparaginase activity; the highest value of L-asparaginase activity was obtained with middle level of temperature and inoculum age. Further increase of inoculum age did not result in higher L-asparaginase activity. Figure 7(b) represents the L-asparaginase activity as a function of temperature (X1) and agitation speed (X6) by keeping inoculum age (X5) at optimum value; the maximum L-asparaginase activity was attained at moderate to high levels of agitation speed and moderate levels of temperature and further increase in the temperature resulted in a gradual decrease in the L-asparaginase activity. Figure 7(c) showed that the maximum L-asparaginase production was attained beyond middle levels of inoculum age and lower and higher levels of inoculum age resulted in a gradual decrease in L-asparaginase production. Highest value of L-asparaginase production was obtained beyond high agitation speed.


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)

Three-dimensional response surface plots showing the effect of temperature (X1), inoculum age (X5) and agitation speed (X6) and their mutual effect on the production of L-asparaginase.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig7: Three-dimensional response surface plots showing the effect of temperature (X1), inoculum age (X5) and agitation speed (X6) and their mutual effect on the production of L-asparaginase.
Mentions: The interaction effects and optimal levels of the variables were determined by plotting the three-dimensional response surface curves (Figures 7(a)–7(c)) when one of the variables is fixed at optimum value and the other two are allowed to vary. Figure 7(a) represents the L-asparaginase activity as a function of temperature (X1) and inoculum age (X5) by keeping agitation speed (X6) at optimum value. It showed that lower and higher levels of temperature support relatively low levels of L-asparaginase activity; the highest value of L-asparaginase activity was obtained with middle level of temperature and inoculum age. Further increase of inoculum age did not result in higher L-asparaginase activity. Figure 7(b) represents the L-asparaginase activity as a function of temperature (X1) and agitation speed (X6) by keeping inoculum age (X5) at optimum value; the maximum L-asparaginase activity was attained at moderate to high levels of agitation speed and moderate levels of temperature and further increase in the temperature resulted in a gradual decrease in the L-asparaginase activity. Figure 7(c) showed that the maximum L-asparaginase production was attained beyond middle levels of inoculum age and lower and higher levels of inoculum age resulted in a gradual decrease in L-asparaginase production. Highest value of L-asparaginase production was obtained beyond high agitation speed.

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