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. 2016:2016:1785938.
doi: 10.1155/2016/1785938. Epub 2016 Jul 25.

Optimization of Growth Conditions for Purification and Production of L-Asparaginase by Spirulina maxima

Hanaa H Abd El Baky  1 , Gamal S El Baroty  2
Affiliations

Optimization of Growth Conditions for Purification and Production of L-Asparaginase by Spirulina maxima

Hanaa H Abd El Baky et al. Evid Based Complement Alternat Med. 2016.

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.

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Figures

Figure 1
Figure 1
Effect of nitrogen concentration on growth of Spirulina maxima.
Figure 2
Figure 2
Detection of L-asparaginase (L-AsnA) activity of macroalgae and microalgae assayed by Gulati et al. [19] method using phenol red indicator: (a) L-AsnA production from microalgae, (b) L-ASase production from macroalgae, and (c) control without L-AsnA extract.
Figure 3
Figure 3
Effect of pH on Spirulina maxima L-asparaginase activity. Enzyme samples (containing 1 mg/mL of protein) were incubated at pH ranges from 5 to 10.
Figure 4
Figure 4
The effect of temperature on the stability of L-asparaginase activity. Enzyme samples (containing 1 mg/mL of protein) were incubated at temperature ranges from 20 to 45°C.
See this image and copyright information in PMC

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