l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization

Kotzia, Georgia A.; Labrou, Nikolaos E.

doi:10.1016/j.jbiotec.2006.07.037

Cited by 146 publications

(98 citation statements)

References 51 publications

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“…These results are in the range with those obtained by many investigators [43,58] reported that the molecular weight of L-asparaginase from P. sativumseeds was 68 KDa using Sephadex LH-20 and 69 KDa using Sephacryl S-200 chromatography. Other studies found the molecular weight of L-asparaginase produced from P. sativum was 58 KDa [12,13,68] However, the molecular weight of L-asparaginase by Sephacryl S-200 yielded a value of 79 kDa. Vignaunguiculata with a molecular weight of 70000Dalton using Sephacryl S-200 chromatography [7,10,21].…”

Section: Molecular Weight Of L-asparaginasementioning

confidence: 98%

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Anticancer Activity of L-Asparaginase Produced from Vigna Unguiculata

Moharib

2018

World Scientific Research

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The present study aimed to production of purified L-asparaginase from Vignaunguiculata. Different physiological parameters, such as pH, temperature and incubation period, were optimized for growth and maximum L-asparaginase production. The optimum parameters were 37°C, 30 min and pH 8.5. Maximum L-asparaginase was 886.4U/ml with a specific activity of 1140.7 U/ml (31 fold purification with 28 %yield) were obtained at optimum conditions. The purified L-asparaginase produced from Vignaunguiculata was used for characterization and general properties. The effect of pH and temperature on L-asparaginase activity as well as stability at different pH and temperature were determined. The optimum pH 8.5 and 37ºC temperature on L-asparaginase showed 100% residual activity. Stability of pH around 8.5 and temperature 70ºC showed 90 and 78 % residual activity at 30 and 60 min respectively. The Lasparaginase showed high stability at alkaline pH (pH 8.5) when incubated for up to 60h.The molecular weight of the produced L-asparaginase was close to 68.5 kDa. Cytotoxic activity of Lasparaginase was examined in vitro using four carcinoma cell lines. L-aspargenase has higher effective in growth inhibition against HEPG2 and HCT-116 but lower against HELLA and MCF7 carcinoma cell lines. The data show that L-aspargenase has a higher cytotoxic activity against HEPG2 and HCT116, revealed higher percentage of cell death, indicating antitumor properties, and demonstrate direct effect on cancer cell proliferation of HEPG2 and HCT116. Therefore, Vignaunguiculata was considered to be a suitable source for production of Lasparaginase has higher activity and good stability. Purified L-asparaginase obtained from Vignaunguiculata could be employed in drug chemotherapy and treatment of cancer.

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Section: Molecular Weight Of L-asparaginasementioning

confidence: 98%

“…These results are in agreement with those reported by other investigators [9] found the optimum activity of L-asparaginase at pH 8.5. E.coli and S. marsconshave an optimum activity at pH8.6 [2,68]. While the optimum pH for L-asparaginase activity of Bacillus spp.…”

Section: Effect Of Ph On L-asparaginase Activitymentioning

confidence: 99%

Anticancer Activity of L-Asparaginase Produced from Vigna Unguiculata

Moharib

2018

World Scientific Research

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“…The use of L-asparaginase in anti-cancer therapy is based on its ability to catalyze the conversion of L-Asparagine to L-Aspartate and ammonia in human body fluid. On the whole lymphoblasts are unable to produce endogenous L-asparagine, starvation for this amino acid leads to death of these cells [2]. Enormous numbers of researches have been accomplished on the biosynthesis of this therapeutically important L-Asparaginase, since Mashburn and Wriston demonstrated anti tumor activity [3].…”

Section: Introductionmentioning

confidence: 99%

Isolation and Purification of High Efficiency L-asparaginase by Quantitative Preparative Continuous-elution SDS PAGE Electrophoresis

M¹,

Selvam²

2011

J Microbial Biochem Technol

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An Unique extracelluar glutaminase free L-asparaginase from novel marine Actinomycetes was isolated to perceptible homogeneity in agro industrial wastes. Quantitative Preparative Continuous-Elution SDS PAGE Electrophoresis is a high-resolution method for the preparative isolation of L-Asparaginase in biological samples. The enzyme was purified 248.68-fold and showed a final specific activity of 5035.28 IU/mg with an 80.71% yield. The homotetramer enzyme has a molecular mass of 133.25 kDa and an isoelectric point of approximately 5.4.Kinetic parameters, Km and Vmax of purified L-asparaginase from Streptomyces radiopugnans MS1 were found to be 0.0598, 3.5478 IU µg -1 respectively. The de novo sequencing strategy presented here provides a rapid and reliable means to identify proteins in Streptomyces radiopugnans MS1. The purified L-asparaginase has no glutaminase activity, which can diminish the leeway of side effects during the itinerary of anti-malignancy therapy.

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“…These values are lower as compared to Krasotkina et al (2004), and cannot be compared to a measurement using the same method because this value was not provided (Kotzia and Labrou, 2005). On the other hand, when compared to E. chrysanthemi L-asparaginase (Kotzia and Labrou, 2007), these values are higher, and, surprisingly, our values were also higher than E. coli L-asparaginase. According to Duval et al (2002), it would be expected that E. coli L-asparaginase would be more effective in depletion L-Asn than Erwinia L-asparaginase during the childhood lymphoid malignancies treatment.…”

Section: Discussionmentioning

confidence: 56%

Comparison between Two Erwinia carotovora L-Asparaginase II Constructions: cloning, Heterologous Expression, Purification, and Kinetic Characterization

Wink¹,

Bogdawa²,

Renard³

et al. 2010

J Microbial Biochem Technol

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L-Asparaginase II from Erwinia carotovora may represent an important alternative therapy in the treatment of acute childhood lymphoblastic leukaemia, despite its promising lower glutaminase activity than Escherichia coli and Erwinia chrysanthemi L-asparaginases II, currently used in treatment of this disease. Here we describe cloning, expression, purification and determination of steady-state kinetic parameters for E. carotovora L-asparaginase II: with (AspSP) and without the signal peptide (AspMP). AspMP was purified to homogeneity by a single-step protocol with 91% yield, and AspSP by a two-step protocol with 28% yield. In addition, both enzymes presented similar high specific activities: 208.1 and 237.6 U mg -1 , respectively. The K m and k cat values showed that AspMP has lower glutaminase activity than AspSP. Moreover AspMP is produced by a simpler purification protocol, and at higher yield. This process can be amenable to large scale production and be of interest to researchers and biopharmaceutical companies.

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l-Asparaginase from Erwinia Chrysanthemi 3937: Cloning, expression and characterization

Cited by 146 publications

References 51 publications

Anticancer Activity of L-Asparaginase Produced from Vigna Unguiculata

Anticancer Activity of L-Asparaginase Produced from Vigna Unguiculata

Isolation and Purification of High Efficiency L-asparaginase by Quantitative Preparative Continuous-elution SDS PAGE Electrophoresis

Comparison between Two Erwinia carotovora L-Asparaginase II Constructions: cloning, Heterologous Expression, Purification, and Kinetic Characterization

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