Increasing loop flexibility affords low-temperature adaptation of a moderate thermophilic malate dehydrogenase from Geobacillus stearothermophilus
| العنوان: | Increasing loop flexibility affords low-temperature adaptation of a moderate thermophilic malate dehydrogenase from Geobacillus stearothermophilus |
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| المؤلفون: | Yuya Shimozawa, Yoshiaki Nishiya, Tsutomu Nakamura, Tomoki Himiyama |
| المصدر: | Protein Engineering, Design and Selection. 34 |
| بيانات النشر: | Oxford University Press (OUP), 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | L-Lactate Dehydrogenase, biology, Stereochemistry, Thermophile, Mutant, Temperature, Wild type, Bioengineering, Nicotinamide adenine dinucleotide, Biochemistry, Malate dehydrogenase, Geobacillus stearothermophilus, Kinetics, chemistry.chemical_compound, chemistry, Malate Dehydrogenase, Lactate dehydrogenase, biology.protein, Citrate synthase, Molecular Biology, Biotechnology |
| الوصف: | Malate dehydrogenase (MDH) catalyzes the reversible reduction of nicotinamide adenine dinucleotide from oxaloacetate to L-malate. MDH from moderate thermophilic Geobacillus stearothermophilus (gs-MDH) has high thermal stability and substrate specificity and is used as a diagnostic reagent. In this study, gs-MDH was engineered to increase its catalytic activity at low temperatures. Based on sequential and structural comparison with lactate dehydrogenase from G. stearothermophilus, we selected G218 as a mutation site to increase the loop flexibility pivotal for MDH catalysis. The G218 mutants showed significantly higher specific activities than the wild type at low temperatures and maintained thermal stability. The crystal structure of the G218Y mutant, which had the highest catalytic efficiency among all the G218 mutants, suggested that the flexibility of the mobile loop was successfully increased by the bulky side chain. Therefore, this study demonstrated the low-temperature adaptation of MDH by facilitating conformational changes during catalysis. |
| تدمد: | 1741-0134 1741-0126 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2116ca7729c9dc0dcd725a73e8cd0dc6Test https://doi.org/10.1093/protein/gzab026Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....2116ca7729c9dc0dcd725a73e8cd0dc6 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 17410134 17410126 |
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