Abstract

L-asparaginase (L-AsnA) is widely distributed among microorganisms and has important applications in medicine and in food technology sectors. Therefore, the ability of the production, purification, and characterization of AsnA from Spirulina maxima (SM) were tested. SM cultures grown in Zarrouk medium containing different N2 (in NaNO3 form) concentrations (1.25, 2.50, and 5.0 g/L) for 18 days contained a significant various quantity of dry biomass yields and AsnA enzyme levels. MS L-AsnA activity was found to be directly proportional to the N2 concentration. The cultures of SM at large scales (300 L medium, 5 g/L N2) showed a high AsnA enzyme activity (898 IU), total protein (405 mg/g), specific enzyme activity (2.21 IU/mg protein), and enzyme yield (51.28 IU/L) compared with those in low N2 cultures. The partial purification of crude MS AsnA enzyme achieved by 80% ammonium sulfate AS precipitated and CM-Sephadex C-200 gel filtration led to increases in the purification of enzyme with 5.28 and 10.91 times as great as that in SM crude enzymes. Optimum pH and temperature of purified AsnA for the hydrolyzate were 8.5 and 37 ± 0.2°C, respectively. To the best of our knowledge, this is the first report on L-asparaginase production in S. maxima.

Highlights

  • Enzymes are proteins that catalyze biochemical reactions

  • These AsnA enzymes are produced by a large number of microorganisms that include bacteria (Streptomyces gulbargensis, Enterobacter cloacae, and Serratia marcescens), fungi, and yeast

  • Nitrogen is required for all biosynthesis leading to reproduction, product formation, and cell maintenance

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Summary

Introduction

Enzymes are proteins that catalyze biochemical reactions. They are more important than vitamins and minerals for general health [1]. L-asparaginase (L-asparagine amidohydrolase EC, 3.5.1.1) is an enzyme of high intensive chemotherapeutic application due to its use in wide variety of cancer therapy mainly in acute lymphoblastic leukemia [2] It is used for treatment of various diseases such as childhood acute lymphoblastic leukemia, myelomonocytic leukemia, reticulum sarcoma, melanoma sarcoma, non-Hodgkin’s lymphoma, pancreatic carcinoma, and bovine lymphoma sarcoma [3, 4]. Since the tumor cells require the high quantity of asparagine from the diet and from other cells for their rapid proliferation and depend on an external supply of L-asparagine for their growth, by continuous administering of Lasparaginase, the tumor cells are made to stave off that particular amino acid, which leads to the death of lymphoblasts by apoptosis [6] These AsnA enzymes are produced by a large number of microorganisms that include bacteria (Streptomyces gulbargensis, Enterobacter cloacae, and Serratia marcescens), fungi, and yeast. Considering the importance of asparaginase, this study was carried out to produce optimized and characterized Lasparaginase from SM and determine the purification process and characterization of its properties

Material and Methods
Cultivation of Algae
Results and Dissection
Conclusion
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