ArticleScientific reports2018
Simultaneous cell disruption and semi-quantitative activity assays for high-throughput screening of thermostable L-asparaginases.
Article in Scientific reports, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Hyperthermophilic L-Asparaginase fromInternational journal of molecular sciences · 2025Article
- N-terminal truncated phospholipase A1 accessory protein PlaS from Serratia marcescens alleviates inhibitory on host cell growth and enhances PlaA1 enzymatic activity.Bioresources and bioprocessing · 2024Article
- Radiocarbon Flux Measurements Reveal Mechanistic Insight into Heat-Stress Induction of Nicotine Biosynthesis inInternational journal of molecular sciences · 2023Article
- Thermo-L-Asparaginases: From the Role in the Viability of Thermophiles and Hyperthermophiles at High Temperatures to a Molecular Understanding of Their Thermoactivity and Thermostability.International journal of molecular sciences · 2023Review
- Production, characterization and techno-economic evaluation of Aspergillus fusant L-asparaginase.AMB Express · 2023Article
- Current state of molecular and metabolic strategies for the improvement of L-asparaginase expression in heterologous systems.Frontiers in pharmacology · 2023Review
- Aliivibrio fischeri L-Asparaginase production by engineered Bacillus subtilis: a potential new biopharmaceutical.Bioprocess and biosystems engineering · 2022Article
- A Novel L-Asparaginase from Hyperthermophilic ArchaeonInternational journal of molecular sciences · 2021Article
- Microbial L-asparaginase for Application in Acrylamide Mitigation from Food: Current Research Status and Future Perspectives.Microorganisms · 2021Review
- L-asparaginase production review: bioprocess design and biochemical characteristics.Applied microbiology and biotechnology · 2021Review
- Bioprocess Optimization of Nutritional Parameters for Enhanced Anti-leukemic L-Asparaginase Production byInternational journal of peptide research and therapeutics · 2021Article
- Lys-Arg mutation improved the thermostability of Bacillus cereus neutral protease through increased residue interactions.World journal of microbiology & biotechnology · 2019Article
- Design of a high-efficiency synthetic system for l-asparaginase production inEngineering in life sciences · 2019Article
Corrections and comments
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Authors and funding
13 authors.
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No grant is acknowledged in the PubMed record.
Abstract
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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