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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 phylogenetic tree was constructed via the bootstrap test of neighbor-joining algorithm based on the 16S rRNA gene sequences of strain NEAE-119 and related species of the genus Streptomyces. Only bootstrap values above 50%, expressed as percentages of 1000 replications, are shown at the branch points. GenBank sequence accession numbers are indicated in parentheses after the strain names. Phylogenetic analyses were conducted in the software package MEGA4. Bar: 0.5 substitution per nucleotide position.
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Related In: Results  -  Collection


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fig3: The phylogenetic tree was constructed via the bootstrap test of neighbor-joining algorithm based on the 16S rRNA gene sequences of strain NEAE-119 and related species of the genus Streptomyces. Only bootstrap values above 50%, expressed as percentages of 1000 replications, are shown at the branch points. GenBank sequence accession numbers are indicated in parentheses after the strain names. Phylogenetic analyses were conducted in the software package MEGA4. Bar: 0.5 substitution per nucleotide position.

Mentions: The 16S rRNA gene sequence (1509 bp) was determined for strain NEAE-119. A BLAST search [33] of the GenBank database using this sequence showed its similarity to that of many species of the genus Streptomyces. A phylogenetic tree (Figure 3) based on 16S rRNA gene sequences of members of the genus Streptomyces was constructed according to the bootstrap test of neighbor-joining algorithm method of Saitou and Nei [35] with MEGA4 [34]. This tree shows the close phylogenetic association of strain NEAE-119 with certain other Streptomyces species. Phylogenetic analysis indicated that the strain NEAE-119 consistently falls into a clade together with Streptomyces enissocaesilis strain ACCA1 (GenBank/EMBL/DDBJ accession number JX042471.1, 99% sequence similarity), Streptomyces plicatus strain RT-57 (GenBank/EMBL/DDBJ accession number HQ909761.1, 99% sequence similarity), and Streptomyces olivaceus strain RT-54 (GenBank/EMBL/DDBJ accession number HQ909759.1, 99% sequence similarity). On the basis of the collected data and in view of the comparative study of the recorded properties of isolate number NEAE-119 in relation to the closest related species of the genus Streptomyces, it is most closely related to the type strains of Streptomyces olivaceus strain RT-54 (GenBank/EMBL/DDBJ accession number HQ909759.1) (99% sequence similarity). Therefore, this strain was identified as Streptomyces olivaceus strain NEAE-119 and its sequencing product was deposited in the GenBank database under accession number KJ200342.


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 phylogenetic tree was constructed via the bootstrap test of neighbor-joining algorithm based on the 16S rRNA gene sequences of strain NEAE-119 and related species of the genus Streptomyces. Only bootstrap values above 50%, expressed as percentages of 1000 replications, are shown at the branch points. GenBank sequence accession numbers are indicated in parentheses after the strain names. Phylogenetic analyses were conducted in the software package MEGA4. Bar: 0.5 substitution per nucleotide position.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

fig3: The phylogenetic tree was constructed via the bootstrap test of neighbor-joining algorithm based on the 16S rRNA gene sequences of strain NEAE-119 and related species of the genus Streptomyces. Only bootstrap values above 50%, expressed as percentages of 1000 replications, are shown at the branch points. GenBank sequence accession numbers are indicated in parentheses after the strain names. Phylogenetic analyses were conducted in the software package MEGA4. Bar: 0.5 substitution per nucleotide position.
Mentions: The 16S rRNA gene sequence (1509 bp) was determined for strain NEAE-119. A BLAST search [33] of the GenBank database using this sequence showed its similarity to that of many species of the genus Streptomyces. A phylogenetic tree (Figure 3) based on 16S rRNA gene sequences of members of the genus Streptomyces was constructed according to the bootstrap test of neighbor-joining algorithm method of Saitou and Nei [35] with MEGA4 [34]. This tree shows the close phylogenetic association of strain NEAE-119 with certain other Streptomyces species. Phylogenetic analysis indicated that the strain NEAE-119 consistently falls into a clade together with Streptomyces enissocaesilis strain ACCA1 (GenBank/EMBL/DDBJ accession number JX042471.1, 99% sequence similarity), Streptomyces plicatus strain RT-57 (GenBank/EMBL/DDBJ accession number HQ909761.1, 99% sequence similarity), and Streptomyces olivaceus strain RT-54 (GenBank/EMBL/DDBJ accession number HQ909759.1, 99% sequence similarity). On the basis of the collected data and in view of the comparative study of the recorded properties of isolate number NEAE-119 in relation to the closest related species of the genus Streptomyces, it is most closely related to the type strains of Streptomyces olivaceus strain RT-54 (GenBank/EMBL/DDBJ accession number HQ909759.1) (99% sequence similarity). Therefore, this strain was identified as Streptomyces olivaceus strain NEAE-119 and its sequencing product was deposited in the GenBank database under accession number KJ200342.

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