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.

Noura El-Ahmady El-Naggar,Hassan Moawad,Sara M El-Ewasy,Nancy M El-Shweihy

doi:10.1155/2015/627031

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.

Highlights

L-asparaginase (L-asparagine aminohydrolase, EC 3.5.1.1) is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children, Hodgkin disease, acute myelocytic leukemia, acute myelomonocytic leukemia, chronic lymphocytic leukemia, lymphosarcoma treatment, reticulosarcoma, and melanosarcoma [1, 2]
The Lasparaginases of Erwinia chrysanthemi and E. coli have been employed for many years as effective drugs in the treatment
A statistical approach has been employed for which a Plackett-Burman design is used for identifying significant variables influencing glutaminase free L-asparaginase production by Streptomyces olivaceus NEAE-119

Summary

Introduction

L-asparaginase (L-asparagine aminohydrolase, EC 3.5.1.1) is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children, Hodgkin disease, acute myelocytic leukemia, acute myelomonocytic leukemia, chronic lymphocytic leukemia, lymphosarcoma treatment, reticulosarcoma, and melanosarcoma [1, 2]. L-asparaginase has an antioxidant property [3] It is used in food industry as a food processing aid; it can effectively reduce the level of acrylamide up to 90% in a range of starchy fried foods without changing the taste and appearance of the end product [4]. L-asparaginase administration has been limited by a high rate of hypersensitivity in the long-term use [5] and development of anti-asparaginase antibodies, which causes an anaphylactic shock or neutralization of the drug effect. To overcome these limitations, modified versions of Lasparaginase (such as L-asparaginase from other new sources, pegylated formulations, and L-asparaginase loaded into erythrocytes) have been recently proposed [6]. The Lasparaginases of Erwinia chrysanthemi and E. coli have been employed for many years as effective drugs in the treatment

Objectives

Methods

Results

Conclusion