Cloning of 1,3-Propanediol dehydrogenase-encoding gene from Lactobacillus buchneri
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Abstract
Nowadays, Biofuel industry is growing rapidly, resulting in an increasing level of glycerol which is a byproduct from biofuel production. Glycerol can be converted to 1,3-Propanediol (1,3-PD) which is a chemical precursor presenting in many industrial applications. In microbial metabolic pathway, dhaT encodes 1,3-Propanediol dehydrogenase (1,3-PDH), which leads to synthesis of 1,3-PD. In this study, dhaT gene from Lactobacillus buchneri was amplified by polymerase chain reaction (PCR), then inserted into expression vector pET-28a(+) and subsequently introduced into Escherichia coli BL21. The correct transformants were screened by colony PCR and recombinant plasmid was isolated and sequenced. The result showed that the dhaT gene from L. buchneri was successfully cloned. The activity of 1,3-PDH was 154.228 mU/ml in recombinant E. coli while the activity of the same enzyme was 208.793 mU/ml in L. buchneri. The results suggest that the dhaT gene was successfully expressed in this work and might provide further knowledge to develop genetically engineered Escherichia coli strain for producing 1,3-PD from glycerol in the future.
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