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Joers, A.

Publications and source records attributed to Joers, A..

3 recordsLinked to original sources

Decoupling growth and production by removing the origin of replication from a bacterial chromosome

Efficient production of biochemicals and proteins in cell factories frequently benefits from a two-stage bioprocess in which growth and production phases are decoupled. Here we describe a novel growth switch based on the permanent removal of the origin of replication (oriC) from the Escherichia coli chromosome. Without oriC, cells cannot initiate a new round of replication and they stop growing while their metabolism remains active. Our system relies on a serine recombinase from bacteriophage phiC31 whose expression is controlled by the temperature-sensitive cI857 repressor from phage lambda. Reporter protein expression in switched cells continues after cessation of growth, leading to protein levels up to five times higher compared to non-switching cells. Switching induces a unique physiological state that is different from both normal exponential and stationary phases. Switched cells remain in this state even when not growing, retain their protein synthesis capacity, and do not induce proteins associated with the stationary phase. Our switcher technology is potentially useful for a range of products and applicable in many bacterial species for decoupling growth and production.

synthetic biology↗

Dividing subpopulation of Escherichia coli in stationary phase

The bacterial growth cycle contains different phases: after the growth substrate is exhausted of the toxic waste products accumulate the growth stops. In this non-growing culture the number of colony forming bacteria remains constant or starts to decrease. It has been shown that during prolonged incubation there is constant growth and death of bacteria and certain mutant populations take over the culture. Here we show that the dynamic cell division and death balance can be obtained even before mutants take over the culture.

microbiology↗

Muropeptides Stimulate Growth Resumption from Stationary Phase in Escherichia coli

When nutrients run out, bacteria enter a dormant metabolic state. This low or undetectable metabolic activity helps bacteria to preserve their scant reserves for future, but also diminishes their ability to trace the environment for new growth-promoting substrates. However, neighboring microbial growth is a sure indicator of favorable environment and thus, can serve as a cue for exiting the dormancy. Here we report that for Escherichia coli this cue is the basic peptidoglycan unit (i.e. muropeptide). We show that several forms of muropeptides can stimulate growth resumption of dormant E. coli cells, but the sugar - peptide bond is crucial for activity. We also demonstrate that muropeptides from several different species can induce growth resumption of E. coli and also Pseudomonas aeruginosa. These results, together with the previous identification of muropeptides as germination signal for bacterial spores, makes muropeptides rather universal cue for bacterial growth.

microbiology↗