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Background  Two sets of overlapping genes,  lacLMReu  and  lacLMAci , encoding heterodimeric β-galactosidases from  Lactobacillus reuteri  and  Lactobacillus acidophilus , respectively, have previously been cloned and expressed using the pSIP vector system and  Lactobacillus plantarum  WCSF1 as host. Despite the high similarity between these  lacLM  genes and the use of identical cloning and expression strategies, strains harboring  lacLMReu  produced about twenty-fold more β-galactosidase than strains containing  lacLMAci .    Results  In this study, the plasmid copy numbers (PCN) of expression vectors pEH9R ( lacLMReu ) and pEH9A ( lacLMAci ) as well as the transcription levels of both  lacLM  genes were compared using quantitative PCR methods. Analyses of parallel fermentations of  L. plantarum  harboring either pEH9R or pEH9A showed that the expression plasmids were present in similar copy numbers. However, transcript levels of  lacLM  from  L. reuteri  (pEH9R) were up to 18 times higher than those of  lacLM  from  L. acidophilus  (pEH9A). As a control, it was shown that the expression levels of regulatory genes involved in pheromone-induced promoter activation were similar in both strains.    Conclusion  The use of identical expression strategies for highly similar genes led to very different mRNA levels. The data indicate that this difference is primarily caused by translational effects that are likely to affect both mRNA synthesis rates and mRNA stability. These translational effects thus seem to be a dominant determinant for the success of gene expression efforts in lactobacilli.

microbial cell factories

Quantitative transcript analysis of the inducible expression system pSIP: comparison of the overexpression of Lactobacillus spp. β-galactosidases in Lactobacillus plantarum