l-Asparaginase from Streptomyces griseus NIOT-VKMA29: optimization of process variables using factorial designs and molecular characterization of l-asparaginase gene

Meena, Balakrishnan; Anburajan, Lawrance; Sathish, T.; Rangamaran, Vijaya Raghavan; Dharani, G.; Vinithkumar, Nambali Valsalan; Kirubagaran, R.

doi:10.1038/srep12404

Cited by 40 publications

(16 citation statements)

References 22 publications

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“…L-asparaginase (EC 3.5.1.1), which catalyses the conversion of L-asparagine to L-aspartate and ammonia, has been identified in various organisms, including animals, plants, yeast, fungi, bacteria, and archaea 1 – 3 . L-asparaginase has high commercial value due to its vast applications in the food and pharmaceutical industries 2 , 4 .…”

Section: Introductionmentioning

confidence: 99%

Simultaneous cell disruption and semi-quantitative activity assays for high-throughput screening of thermostable L-asparaginases

Zhang

et al. 2018

Sci Rep

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L-asparaginase, which catalyses the hydrolysis of L-asparagine to L-aspartate, has attracted the attention of researchers due to its expanded applications in medicine and the food industry. In this study, a novel thermostable L-asparaginase from Pyrococcus yayanosii CH1 was cloned and over-expressed in Bacillus subtilis 168. To obtain thermostable L-asparaginase mutants with higher activity, a robust high-throughput screening process was developed specifically for thermophilic enzymes. In this process, cell disruption and enzyme activity assays are simultaneously performed in 96-deep well plates. By combining error-prone PCR and screening, six brilliant positive variants and four key amino acid residue mutations were identified. Combined mutation of the four residues showed relatively high specific activity (3108 U/mg) that was 2.1 times greater than that of the wild-type enzyme. Fermentation with the mutant strain in a 5-L fermenter yielded L-asparaginase activity of 2168 U/mL.

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Section: Introductionmentioning

confidence: 99%

Simultaneous cell disruption and semi-quantitative activity assays for high-throughput screening of thermostable L-asparaginases

Zhang

et al. 2018

Sci Rep

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“…The optimal level of carbon source for L-asparaginase production by S. longsporusflavus F-15 was 1.5% starch, 45 while it was 1% starch for Streptomyces griseus NIOT-VKMA 29 41 . The final pH values of the fermentation broths of Streptomyces labedae amended with starch, glycerol, maltose and galactose independently developed alkaline conditions where as the media supplemented with other carbon sources developed acidic, which may be the reason for low productivity of the enzyme.…”

Section: Selection Of Best Carbon and Nitrogen Sources For Maximum L-mentioning

confidence: 99%

Extracellular L-Asparaginase from Streptomyces labedae VSM-6: Isolation, Production and Optimization of Culture Conditions Using RSM

Mangamuri¹,

Vijayalakshmi²,

Ganduri³

et al. 2017

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Objective: The present study was intended to isolate actinomycetes VSM-6 from deep sea sediment samples of Bay of Bengal that is potent to produce L -asparaginase. Materials and Methods: The identification of the isolate was executed by polyphasic taxonomy.Optimization was carried out using one factor at a time (O-F-A-T) for the production of the L-asparaginase. RSM was pledged to optimize the L -asparaginase production by S.labedae VSM-6. Central composite design was applied to study the influence of the variables and their interactive effects on the production of L -asparaginase. Unstructured Kinetic modelling for L -asparaginase production was adopted using Leudeking-Piret (LILP) and Logistic Incorporated Modified Leudeking-Piret (LIMLP) models. Results: Optimization using One-Factor-At-A-time registered a turnout of 8.92 IU/ml of L -asparaginase production. But results obtained from the statistical design are in agreement with the experimental results. The model followed the second order polynomial equation and the model adequacy was determined by the P value (<0.0001), Coefficient determination (R 2 ) with a value of 0.9942 and the adjusted R 2 = 0.9087 which determines that the model was significant. The experimental values are in compliance with the model anticipated values and catalogued an escalation in yield of L -asparaginase (10.17 IU/ml) by RSM. Unstructured Kinetic modelling for L -asparaginase production adopting Leudeking-Piret (LILP) and Logistic Incorporated Modified Leudeking-Piret (LIMLP) models showed L -asparaginase production of (10.17 IU/ml), closer to model anticipated value (10.23 IU/ml) so unstructured models provided a better approximation for L -asparaginase production by S.labedae VSM-6. Conclusion: From our study we have reported for the first time the production of L -asparaginase from S.labedae VSM-6 using central composite design and kinetic modelling.

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“…However, the low yield of L-asparaginase has restricted the application of this enzyme in the food and pharmaceutical industries. A series of strategies have been used to improve L-asparaginase production, such as optimization of fermentation medium and process [9][10][11], signal peptide screening, promoter mutation, and N-terminal deletion [12]. To date, the highest yield of L-asparaginase obtained was 2168 U/mL, which was reported in our previous work [13].…”

Section: Introductionmentioning

confidence: 69%

Design of a high‐efficiency synthetic system for l‐asparaginase production in Bacillus subtilis

Zhang

et al. 2019

Engineering in Life Sciences

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l‐asparaginase has high application value in medicine and food industry, but the low yield limits its application. In this study, we designed a synthetic system in Bacillus subtilis to produce l‐asparaginase by improving gene expression and optimizing the fermentation agitation speed. Gene expression was improved by respectively increasing transcription levels and translation speeds through screening promoters and RBS sequences. With the optimal promoter, P43, and the synthetic RBS sequence, the yield obtained in a shake flask was 371.87 U/mL, which was 2.09 times that with the original strain. To further enhance production in a 5‐L fermenter, a multistage agitation speed control strategy was adopted, involving agitation at 600 rpm for the first 12 h, followed by a gradual increase in speed to 900 rpm, which resulted in the highest yield of l‐asparaginase, 5321 U/mL, after 42 h of fermentation.

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l-Asparaginase from Streptomyces griseus NIOT-VKMA29: optimization of process variables using factorial designs and molecular characterization of l-asparaginase gene

Cited by 40 publications

References 22 publications

Simultaneous cell disruption and semi-quantitative activity assays for high-throughput screening of thermostable L-asparaginases

Simultaneous cell disruption and semi-quantitative activity assays for high-throughput screening of thermostable L-asparaginases

Extracellular L-Asparaginase from Streptomyces labedae VSM-6: Isolation, Production and Optimization of Culture Conditions Using RSM

Design of a high‐efficiency synthetic system for l‐asparaginase production in Bacillus subtilis

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