Transformation of Clostridium beijerinckii Br21 for insertion of the gene dhaB, to encode the enzyme glycerol dehydratase

Abstract

Sustainable development is one of the pillars of bioeconomy, which aims to conserve natural resources and to reuse industrial waste. One of the bioprocesses in line with these objectives is the fermentation of glycerol, the main residue of biodiesel production, by bacteria of the genus Clostridium. Specifically, Clostridium beijerinckii Br21 can produce 1,3-propanediol from glycerol, but it does not have the gene dhaB, responsible for the first step of the reaction. This microorganism possibly produces 1,3-propanediol via an alternative route. Thus, C. beijerinckii Br21 will be transformed for insertion of the gene dhaB, which should increase the reductive pathway of glycerol metabolism. In addition, we seek to learn about and to apply the Clostridium transformation methodologies, which are not trivial, used by the German research group in our laboratory. We will use pure glycerol as substrate and will characterize fermentation by measuring OD600, pH, and concentrations of substrates and products. In addition, this procedure will help to understand the actual biochemical pathway that C. beijerinckii is making use to form 1,3-propanediol, a totally non-explored task in current literature.