title: 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.
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.
authors.affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.
authors.forename: Noura El-Ahmady
authors.initials: Nel-A
authors.lastname: El-Naggar
authors.affiliation: Department of Agricultural Microbiology, National Research Center, Cairo, Egypt.
authors.forename: Hassan
authors.initials: H
authors.lastname: Moawad
authors.affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.
authors.forename: Nancy M
authors.initials: NM
authors.lastname: El-Shweihy
authors.affiliation: Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, Alexandria 21934, Egypt.
authors.forename: Sara M
authors.initials: SM
authors.lastname: El-Ewasy
mesh_terms: D001215:Asparaginase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D018929:Cell Culture Techniques; D005972:Glutaminase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D000960:Hypolipidemic Agents / Q000302:isolation & purification / Q000378:metabolism; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000254:growth & development
chemical_list: D000960:Hypolipidemic Agents; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D001215:Asparaginase; D005972:Glutaminase