Pcal_0970: an extremely thermostable l-asparaginase from Pyrobaculum calidifontis with no detectable glutaminase activity

Chohan, Shahid Mahmood; Rashid, Naeem; Sajed, Muhammad; Imanaka, Tadayuki

doi:10.1007/s12223-018-0656-6

Cited by 31 publications

(9 citation statements)

References 35 publications

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“…A higher specific activity (1.08 U mg −1 ) was obtained in the present work. Chohan et al (2018), obtained a volumetric activity of 20.13 U L −1 working with L-ASNase gene from Pyrobaculum calidifontis expressed in E. coli, lower than that found in the present study [42]. However, in a work published by Khushoo and collaborators (2005), the production of extracellular L-ASNase achieved the volumetric activity of 850,000 U L −1 in the extracellular medium, higher than that found in the present study [72].…”

Section: Fermentationcontrasting

confidence: 74%

Development of Processes for Recombinant L-Asparaginase II Production by Escherichia coli Bl21 (De3): From Shaker to Bioreactors

et al. 2020

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Since 1961, L-asparaginase has been used to treat patients with acute lymphocytic leukemia. It rapidly depletes the plasma asparagine and deprives the blood cells of this circulating amino acid, essential for the metabolic cycles of cells. In the search for viable alternatives to produce L-asparaginase, this work aimed to produce this enzyme from Escherichia coli in a shaker and in a 3 L bioreactor. Three culture media were tested: defined, semi-defined and complex medium. L-asparaginase activity was quantified using the β-hydroxamate aspartic acid method. The defined medium provided the highest L-asparaginase activity. In induction studies, two inducers, lactose and its analog IPTG, were compared. Lactose was chosen as an inducer for the experiments conducted in the bioreactor due to its natural source, lower cost and lower toxicity. Batch and fed-batch cultures were carried out to reach high cell density and then start the induction. Batch cultivation provided a final cell concentration of 11 g L−1 and fed-batch cultivation produced 69.90 g L−1 of cells, which produced a volumetric activity of 43,954.79 U L−1 after lactose induction. L-asparaginase was produced in a shaker and scaled up to a bioreactor, increasing 23-fold the cell concentration and thus, the enzyme productivity.

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Section: Fermentationcontrasting

confidence: 74%

Development of Processes for Recombinant L-Asparaginase II Production by Escherichia coli Bl21 (De3): From Shaker to Bioreactors

et al. 2020

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“…fuscus NBRC 14270 [58], Staphylococcus sp. OJ82 [65], термофильных архей P. horikoshii OT3 [54], P. furiosus DSM 3638 [52], T. kodakaraensis KOD1 [47], T. gammatolerans EJ3 [48], T. zilligii [66], P. calidifontis [56]. Как и у L-АСПаз мезофильных микроорганизмов, каждый мономер, размером в 35 кДа, состоит из N-и C-концевого доменов, объединённых междоменным линкером [47,48,66,70].…”

Section: олигомеризация L-аспарагиназ экстремофильных микроорганизмовunclassified

“…Психрофильные грибные L-аспарагиназы Trichosporon asahii IBBLA1 30,0 7,0 20,57 [61] Ионы Mg 2+ , Mn 2+ , Ca 2+ (1 мМ) не оказывали влияния на активность бактериальной L-АСПазы T. thermophilus, в то время как Zn 2+ в концентрациях 0,5 мМ, либо 1 мМ снижал активность на 40% и 60% соответственно [59]. Аналогичные результаты были получены для L-АСПазы P. calidifontis [56].…”

Section: влияние ионов металлов на активность экстремофильных L-аспарагиназunclassified

“…Устойчивость к экстремальным условиям внешней среды -отличительная особенность представителей архей. К настоящему времени достаточно подробно изучены L-АСПазы эвриархеот родов Thermococcus [46][47][48], Pyrococcus [49][50][51][52][53][54], Archaeoglobus [55]; кренархеот -рода Pyrobaculum [56].…”

Section: Introductionunclassified

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L-asparaginases of extremophilic microorganisms in biomedicine

Думина¹,

Eldarov²,

Zdanov

et al. 2020

BIOMED KHIM

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L-asparaginase is extensively used in the treatment of acute lymphoblastic leukemia and several other lymphoproliferative diseases. In addition to its biomedical application, L-asparaginase is also of prospective use in food industry to reduce the formation of acrylamide, which is classified as probably neurotoxic and carcinogenic to human, and in biosensors for determination of L-asparagine level in medicine and food chemistry. The importance of L-asparaginases in different fields, disadvantages of commercial ferments, and the fact that they are widespread in nature stimuli the search for biobetter L-asparaginases from new producing microorganisms. In this regard, extremofile microorganisms exhibit unique physiological properties such as thermal stability, adaptability to extreme cold conditions, salt and pH tolerance and so provide one of the most valuable sources for novel L-asparaginases. The present review summarizes the recent results on studying the structural, functional, physicochemical and kinetic properties, stability of extremophilic L-asparaginases in comparison with their mesophilic homologues.

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“…L-ASNases of extremophilic origin have been already reported from Thermococcus kodakarensis [ 24 , 34 , 42 ], Thermococcus zilligii [ 43 ], Thermococcus gammatolerans [ 44 ], Pyrococcus yayanosii CH1 [ 45 ], Pyrococcus furiosus [ 33 , 46 , 47 , 48 ], Pyrococcus horikoshii [ 49 ], Archaeoglobus fulgidus [ 23 ] and Pyrobaculum calidifontis [ 50 ], living under extremely high temperatures.…”

Section: Introductionmentioning

confidence: 99%

A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application

Думина

Zhgun

Pokrovskaya

et al. 2021

IJMS

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L-asparaginase (L-ASNase) is a vital enzyme with a broad range of applications in medicine and food industry. Drawbacks of current commercial L-ASNases stimulate the search for better-producing sources of the enzyme, and extremophiles are especially attractive in this view. In this study, a novel L-asparaginase originating from the hyperthermophilic archaeon Thermococcus sibiricus (TsA) was expressed in Escherichia coli, purified and characterized. The enzyme is optimally active at 90 °C and pH 9.0 with a specific activity of 2164 U/mg towards L-asparagine. Kinetic parameters KM and Vmax for the enzyme are 2.8 mM and 1200 µM/min, respectively. TsA is stable in urea solutions 0–6 M and displays no significant changes of the activity in the presence of metal ions Ni2+, Cu2+, Mg2+, Zn2+ and Ca2+ and EDTA added in concentrations 1 and 10 mmol/L except for Fe3+. The enzyme retains 86% of its initial activity after 20 min incubation at 90 °C, which should be enough to reduce acrylamide formation in foods processed at elevated temperatures. TsA displays strong cytotoxic activity toward cancer cell lines K562, A549 and Sk-Br-3, while normal human fibroblasts WI-38 are almost unsensitive to it. The enzyme seems to be a promising candidate for further investigation and biotechnology application.

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Pcal_0970: an extremely thermostable l-asparaginase from Pyrobaculum calidifontis with no detectable glutaminase activity

Cited by 31 publications

References 35 publications

Development of Processes for Recombinant L-Asparaginase II Production by Escherichia coli Bl21 (De3): From Shaker to Bioreactors

Development of Processes for Recombinant L-Asparaginase II Production by Escherichia coli Bl21 (De3): From Shaker to Bioreactors

L-asparaginases of extremophilic microorganisms in biomedicine

A Novel L-Asparaginase from Hyperthermophilic Archaeon Thermococcus sibiricus: Heterologous Expression and Characterization for Biotechnology Application

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