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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

L-asparaginase activity of Streptomyces sp. NEAE-119 detected by plate assay. (a, b) Production of the enzyme indicated by color change in the medium from yellow to pink zone surrounding the colony after two and five days, respectively. (c) Control plates were prepared as inoculated medium without dye. (d) Uninoculated medium with dye.
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fig1: L-asparaginase activity of Streptomyces sp. NEAE-119 detected by plate assay. (a, b) Production of the enzyme indicated by color change in the medium from yellow to pink zone surrounding the colony after two and five days, respectively. (c) Control plates were prepared as inoculated medium without dye. (d) Uninoculated medium with dye.

Mentions: L-asparaginase activity of Streptomyces sp. NEAE-119 was detected by plate assay. Production of the enzyme was indicated by color change in the medium from yellow to pink zone surrounding the colony (Figure 1). L-asparaginase activity was confirmed by agar-well diffusion technique. The potential culture, strain NEAE-119, was identified on the basis of morphological, cultural, physiological, and chemotaxonomic properties, together with 16S rRNA sequence as Streptomyces olivaceus strain NEAE-119.


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)

L-asparaginase activity of Streptomyces sp. NEAE-119 detected by plate assay. (a, b) Production of the enzyme indicated by color change in the medium from yellow to pink zone surrounding the colony after two and five days, respectively. (c) Control plates were prepared as inoculated medium without dye. (d) Uninoculated medium with dye.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig1: L-asparaginase activity of Streptomyces sp. NEAE-119 detected by plate assay. (a, b) Production of the enzyme indicated by color change in the medium from yellow to pink zone surrounding the colony after two and five days, respectively. (c) Control plates were prepared as inoculated medium without dye. (d) Uninoculated medium with dye.
Mentions: L-asparaginase activity of Streptomyces sp. NEAE-119 was detected by plate assay. Production of the enzyme was indicated by color change in the medium from yellow to pink zone surrounding the colony (Figure 1). L-asparaginase activity was confirmed by agar-well diffusion technique. The potential culture, strain NEAE-119, was identified on the basis of morphological, cultural, physiological, and chemotaxonomic properties, together with 16S rRNA sequence as Streptomyces olivaceus strain NEAE-119.

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