Cloning, expression and characterization of the recombinant Yersinia pseudotuberculosis l-asparaginase

Pokrovskaya, M. V.; Александрова, С. С.; Pokrovsky, Vadim S.; Omeljanjuk, N.M.; Borisova, Anna A.; Anisimova, Natalia; Соколов, Н. Н.

doi:10.1016/j.pep.2011.12.005

Cited by 44 publications

(22 citation statements)

References 14 publications

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“…The optimum pH was different from those of asparaginases from Yersinia pseudotuberculosis and Helicobacter pylori where optimum pH valueswere8 and 10, respectively. However, this pH was exactly the same as that of asparaginase found in Thermococcus kodakaraensis 7,9,10 .…”

Section: Characterization Of Asparaginasessupporting

confidence: 76%

Purification and Characterization of Pseudomonas aeruginosa PAO1 Asparaginase

Kuwabara

Prihanto

Wakayama

et al. 2015

Procedia Environmental Sciences

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The prevalence of bacterial infection has been gradually increased for the last decades. This phenomenon is predicted as one of serious problems to human security in the next decades. Helicobactor pylori has been clarified to be related to gastric inflammation and a variety of diseases including gastric cancer. Recently, it has been known that H. pylori asparaginase is necessary for the growth in host infection and might be involved in inhibition of lymphocyte function at gastric niche. On the other hand, Pseudomonas aeruginosa is known to cause a fence-sitting infectious disease. However, no studies on a physiological role of asparaginase in relation to pathogenicity of P. aeruginosa have been found. Also, there are no studies on its physiological role in relation to pathogenicity of P. aeruginosa have been found. Therefore, we focused on purification and characterization of P. aeruginosa PAO1 asparaginase in order to clarify its physiological roles in this study. Intracellular asparaginase was highly purified and characterized from P. aeruginosa PAO1. The profile of DEAE-cellufine A 500 column chromatography suggested that two types of asparaginases were produced in P. aeruginosa PAO1. The optimum pH and temperature of the one enzyme was 9.5 and 40°C, respectively while those of the another was 9.5 and 65°C, respectively. Those results indicated that P. aeruginosa PAO1 had two different types of asparaginases.

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Section: Characterization Of Asparaginasessupporting

confidence: 76%

Purification and Characterization of Pseudomonas aeruginosa PAO1 Asparaginase

Kuwabara

Prihanto

Wakayama

et al. 2015

Procedia Environmental Sciences

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“…Unlike bacterial L-asparaginases, which are found as tetramers of identical subunits with molecular masses in the range of 140 to 160 kDa, the purified RmAsnase homodimer's molecular mass was higher than that of some other fungal L-asparaginases (1,5). The specific activity of RmAsnase was similar to that of L-asparaginase from Pectobacterium carotovorum (2,020.9 U/mg) (11) but much higher than the specific activities of most bacterial and fungal L-asparaginases (14,(30)(31)(32). The optimum pH and temperature of RmAsnase are similar to those reported for many L-asparaginases (2,31,33).…”

Section: Discussionmentioning

confidence: 99%

“…RmAsnase, the fungal L-asparaginase from R. miehei, showed antiproliferative activity against cells from several human leukemia cell lines and displayed very low L-glutaminase activity. Thus, RmAsnase may also be useful as an alternative to bacterial L-asparaginases in leukemia treatment (2,6,11,32,35).…”

Section: Discussionmentioning

confidence: 99%

Biochemical Characterization of a Novel l -Asparaginase with Low Glutaminase Activity from Rhizomucor miehei and Its Application in Food Safety and Leukemia Treatment

Huang

Liu

Sun

et al. 2014

Appl Environ Microbiol

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A novel fungal gene encoding the Rhizomucor miehei L-asparaginase (RmAsnase) was cloned and expressed in Escherichia coli. Its deduced amino acid sequence shared only 57% identity with the amino acid sequences of other reported L-asparaginases. The purified L-asparaginase homodimer had a molecular mass of 133.7 kDa, a high specific activity of 1,985 U/mg, and very low glutaminase activity. RmAsnase was optimally active at pH 7.0 and 45°C and was stable at this temperature for 30 min. The final level of acrylamide in biscuits and bread was decreased by about 81.6% and 94.2%, respectively, upon treatment with 10 U RmAsnase per mg flour. Moreover, this L-asparaginase was found to potentiate a lectin's induction of leukemic K562 cell apoptosis, allowing lowering of the drug dosage and shortening of the incubation time. Overall, our findings suggest that RmAsnase possesses a remarkable potential for the food industry and in chemotherapeutics for leukemia.

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“…Reported two-step purification procedure was more efficient and rapid in comparison with the four-step procedure described by Vanelly (2007), which produced an overall yield of 6%. The efficiency of expression of PAL seems to be extremely low, if compared with other recombinant enzymes, such as L-asparaginases, where overall yield more than 40% is quite typical (Khushoo, 2004;Pokrovskaya, 2012).…”

Section: Resultsmentioning

confidence: 99%

Efficient Expression of Recombinant L-phenylalanine Ammonia-lyase From Rhodosporidium toruloides using Escherichia coli

Prosekov¹,

Babich²,

Pokrovsky³

et al. 2014

Journal of Applied Biotechnology

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We explored simple and cost effective method to obtain recombinant PAL of Rhodosporidium toruloides using E. coli expression system. E. coli strain BL21 (DE3) was transformed by pET28 plasmid containing L-phenylalanine ammonia-lyase (PAL) gene from Rhodosporidium toruloides and cultured in different conditions. To assess the physiological activity of microbial cells, we studied the growth kinetics in a series of batch fermentation and changes in its activity after the induction. The kinetics of the process of cultivation of recombinant E. coli strain was studied by the Ludeking-Paire model. A growth and induction regimen was established for use in bioreactor which results in the accumulation of 1,9 g l -1 of recombinant PAL. The point of maximum activity of the PAL produced was determined at 38.0-38.5 h after induction. The optimum concentration of inducer (IPTG) was 2 mM, we recommend to add the inducer at OD540=1.3. This simple and cost effective production process could be recruited for large scale production of PAL.

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Cloning, expression and characterization of the recombinant Yersinia pseudotuberculosis l-asparaginase

Cited by 44 publications

References 14 publications

Purification and Characterization of Pseudomonas aeruginosa PAO1 Asparaginase

Purification and Characterization of Pseudomonas aeruginosa PAO1 Asparaginase

Biochemical Characterization of a Novel l -Asparaginase with Low Glutaminase Activity from Rhizomucor miehei and Its Application in Food Safety and Leukemia Treatment

Efficient Expression of Recombinant L-phenylalanine Ammonia-lyase From Rhodosporidium toruloides using Escherichia coli

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