Abstract
dTDP-l-Rhamnose biosynthetic gene cluster was cloned from Thermus caldophilus. A cluster of four open reading frames, strmlA, B, C and D, responsible for the production of dTDP-l-rhmanose, was screened from the genomic library. Thermophilic glucose-1-phosphate thymidylyltransferase, encoding 356 amino acids with a calculated molecular weight 38 kDa, was expressed under the control of the tac promoter in E. coli. The expressed enzyme, stRmlA is thermostable up to 70 °C and apparently retained its activity even up to 90 °C. It shares 73% sequence identity to glucose-1-phosphate thymidylyltransferase from Streptomyces argillaceus. Amino acid sequence comparison of stRmlA with ten glucose-1-phosphate thymidylyltransferases indicated higher number of unusual hydrophobic residues (A10, A58, A69, A252, V225, V257, V265, I242 and I246) and charged residues (D43, D160, D248, D249, D315, H124, H201, H283 and H354) in stRmlA.
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Parajuli, N., Lee, DS., Lee, H.C. et al. Cloning, expression and characterization of glucose-1-phosphate thymidylyltransferase (strmlA) from Thermus caldophilus . Biotechnology Letters 26, 437–442 (2004). https://doi.org/10.1023/B:BILE.0000018264.35237.86
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DOI: https://doi.org/10.1023/B:BILE.0000018264.35237.86