Non-redundant functionality of lactiplantibacillus plantarum phospho-β-glucosidases revealed by carbohydrate utilization signatures associated to pbg2 and pbg4 gene mutants.

Abstract

Aim: To increase our knowledge on the functionality of 6-phospho-β-glucosidases linked to phosphoenolpyruvate-dependent phosphotransferase systems (PTS) that are encountered in high redundancy in the Lactiplantibacillus plantarum WCFS1 genome.

Methods and results: Two L. plantarum WCFS1 gene mutants that lacked one of the 6-phospho-β-glucosidases, ∆pbg2 (or ∆lp_0906) or ∆pbg4 (or ∆lp_2777) were constructed and the metabolic impact of these mutations assessed by high-throughput phenotyping (Omnilog). The ∆pbg2 mutant displayed a reduced metabolic performance, having lost the capacity to utilize 20 out of 57 carbon (C)-sources used by the wild-type strain. Conversely, the ∆pbg4 mutant conserved the capacity to metabolize most of the C-sources preferred by the wild type strain. This mutant utilized 56 C-sources albeit the range of substrates used and hence its metabolic profiling differed from that of the WCFS1 strain. The ∆pbg2 mutant notably reduced or abolished the capacity to metabolize substrates related to pentose and glucoronate interconversions and was unable to assimilate fatty acids or nucleosides as sole C-sources for growth. The ∆pbg4 mutant acquired the capacity to utilize efficiently glycogen, indicating an efficient supply of glucose from this source.

Conclusion: Lactiplantibacillus plantarum gene mutants that lack individual 6-phospho-β-glucosidases display very different carbohydrate utilization signatures showing that these enzymes can be crucial to determine the capacity of L. plantarum to consume different C-sources and hence for the nutrition and physiology of this microorganism.

Keywords: knock-out mutants; lactiplantibacillus plantarum; carbohydrate utilization; high-throughput phenotyping; metabolic adaptation; phospho-β-glucosidases.