Hyperthermophilic aldolases as biocatalyst for C-C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima
| العنوان: | Hyperthermophilic aldolases as biocatalyst for C-C bond formation: rhamnulose 1-phosphate aldolase from Thermotoga maritima |
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| المؤلفون: | Isabel Oroz-Guinea, Israel Sánchez-Moreno, Eduardo García-Junceda, Montaña Mena |
| المصدر: | Digital.CSIC. Repositorio Institucional del CSIC instname |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Stereochemistry, Aldolases, Molecular Sequence Data, medicine.disease_cause, Applied Microbiology and Biotechnology, Cofactor, Catalysis, Enzyme catalysis, Enzyme Stability, medicine, Thermozyme, Escherichia coli, Hyperthermophilic enzymes, Thermotoga maritima, Amino Acid Sequence, Cloning, Molecular, Thermostability, Aldehyde-Lyases, chemistry.chemical_classification, biology, Aldolase A, Temperature, General Medicine, Hydrogen-Ion Concentration, biology.organism_classification, Recombinant Proteins, Kinetics, Enzyme, chemistry, Biochemistry, Biocatalysis, biology.protein, Biotechnology, Half-Life |
| الوصف: | The TM1072 gene from Thermotoga maritima codifies for a putative form of a rhamnulose-1-phosphate aldolase (Rha-1PATm). To investigate this enzyme further, its gene was cloned and expressed in Escherichia coli. The purified enzyme was activated by Co2+ as a divalentmetal ion cofactor, instead of Zn2+ as its E. coli homologue, and exhibited a maximum of activity at 95 °C. Furthermore, the enzyme displayed a high stability against extreme reaction conditions, retaining 90 % of its activity in the presence of 40 % of acetonitrile and showing a half-life greater than 3 h at 115 °C. The kinetic parameters at room temperature (R/T) were also studied; the KM was calculated to be 3.6±0.33 mM, while kcat/KM was found to be 0.7× 103 s−1 M−1. Given these characteristics, Rha-1PA Tm is an attractive enzyme for use as a biocatalyst for industrial applications, offering intriguing possibilities for practical biocatalysis. We thank the Spanish Ministerio de Economía y Competitividad (Grant PI11/01436) and Comunidad de Madrid (Grant S2009/PPQ-1752) for financial support. I. O.-G. is a JAEPredoc fellow from CSIC. |
| تدمد: | 1432-0614 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::bf6dce67380d59585a3e67687d1d9180 https://pubmed.ncbi.nlm.nih.gov/25324130 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....bf6dce67380d59585a3e67687d1d9180 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14320614 |
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