Electrotransformation of Lactobacillus manihotivorans LMG 18010~T and LMG 18011

摘要

An electroporation procedure was developed for the genetic transformation of intact cells of Lactobacillus manihotivorans, a new Lactobacillus species isolated from Cassava sour starch fermentation in Colombia. Transformation efficiency of Lact. manihotivorans strains LMG 18010~T and LMG 18011 was measured and compared with electroporability of Lact. plantarum strains NCIMB 8299 and LMG 9211, by investigating the effect of changes in various parameters. For Lact. manihotivorans strain LMG 18010~T, the composition of the culture medium, such as the type of peptone and the presence of Tween-80, was found to be the most critical parameter, as well as the aeration conditions of the culture; better electroporation was obtained without air. The presence of MgCl_2 in the recovery medium was favourable to regeneration of electroporated cells. Plasmid-curing of the cells did not improve their electroporability. Transformants were obtained with Lact. manihotivorans strain LMG 18010~T and the plasmids pLZ12 and pGK13, whereas Lact. manihotrorans strain LMG 18011 was transformable with plasmids pLP825 and pLZ12, with different electroporation conditions.