Protein phosphorylation on serine, threonine, and tyrosine (Ser/Thr/Tyr) is well established as a key regulatory posttranslational modification in eukaryotes, but little known about its extent function prokaryotes. Although protein kinases phosphatases have been predicted identified variety of bacterial species, classical biochemical approaches so far revealed only few substrate proteins even fewer sites. Bacillus subtilis model Gram-positive bacterium which two-dimensional...

ABSTRACT We constructed a library of synthetic promoters for Lactococcus lactis in which the known consensus sequences were kept constant while separating spacers randomized. The consists 38 differ strength from 0.3 up to more than 2,000 relative units, latter among strongest this organism. ranking promoter activities was somewhat different when assayed Escherichia coli , but are efficient modulating gene expression bacterium as well. DNA sequencing revealed that weaker (which had below 5...

ABSTRACT The nature of the control glycolytic flux is one central, as-yet-uncharacterized issues in cellular metabolism. We developed a molecular genetic tool that specifically induces ATP hydrolysis living cells without interfering with other aspects Genes encoding F 1 part membrane-bound (F 0 ) H + -ATP synthase were expressed steadily growing Escherichia coli cells, which lowered intracellular [ATP]/[ADP] ratio. This resulted strong stimulation specific concomitant smaller decrease growth...

A minimal growth medium containing glucose, acetate, vitamins, and eight amino acids allowed for of Lactococcus lactis subsp. , with a specific rate in batch culture μ = 0.3 h -1 . With 19 added, the increased to 0.7 exponential phase proceeded until high cell concentrations were reached. We show that morpholinepropanesulfonic acid (MOPS) is suitable buffer L. may be applied concentrations.

Most metabolic reactions are connected through either their utilization of nucleotides or regulation by these metabolites. In this review the biosynthetic pathways for pyrimidine and purine metabolism in lactic acid bacteria described including interconversion pathways, formation deoxyribonucleotides salvage use exogenous precursors. The data enzymatic genetic reviewed, as well gene organizations different bacteria. Mutant phenotypes methods manipulation nucleotide pools also discussed. Our...

Abstract Recent phosphoproteomics studies of several bacterial species have firmly established protein phosphorylation on Ser/Thr/Tyr residues as a PTM in bacteria. In particular, our recent reports the phosphoproteomes model organisms Bacillus subtilis and Escherichia coli detected over 100 events each species. Here we extend analyses to Lactococcus lactis , lactic acid bacterium widely employed by food industry, which at was barely studied all. Despite lack almost any prior evidence L....

Metabolic cofactors such as NADH and ATP play important roles in a large number of cellular reactions, it is great interest to dissect the role these different aspects metabolism. Toward this goal, we overexpressed oxidase soluble F1-ATPase Escherichia coli lower level ATP, respectively. We used global interaction network, comprising protein interactions, transcriptional regulation, metabolic networks, integrate data from transcription profiles, fluxes, metabolite levels. identified...

Sensing and responding to environmental cues is a fundamental characteristic of bacterial physiology virulence. Here we identify polyamines as novel signals essential for virulence Salmonella enterica serovar Typhimurium, major intracellular pathogen model organism studying typhoid fever. Central its are two loci Pathogenicity Island 1 2 (SPI1 SPI2). SPI1 promotes invasion epithelial cells, whereas SPI2 enables S. Typhimurium survive proliferate within specialized compartments inside host...

Persister cells constitute a small subpopulation of bacteria that display remarkably high antibiotic tolerance and for pathogens such as Staphylococcus aureus are suspected culprits chronic recurrent infections. Persisters formed during exponential growth characterized by low ATP levels but less is known in stationary phase. By enrichment from transposon mutant library S. we identified mutants this phase displayed enhanced persister cell formation. We found inactivation either sucA or sucB,...

In this article, we review some of the expression systems that are available for Metabolic Control Analysis and Engineering, examine their advantages disadvantages in different contexts. a recent approach, artificial promoters modulating gene micro-organisms were constructed using synthetic degenerated oligonucleotides. From work, promoter library was obtained Lactococcus lactis, containing numerous individual covering wide range activities. Importantly, activities covered small steps...

The membrane-bound H(+)-ATPase plays a key role in free-energy transduction of biological systems. We report how the carbon and energy metabolism Escherichia coli changes response to deletion atp operon that encodes this enzyme. Compared with isogenic wild-type strain, growth rate yield were decreased less than expected for shift from oxidative phosphorylation glycolysis alone as source ATP. Moreover, respiration strain was increased by 40% compared strain. This result is surprising, since...

A synthetic promoter library (SPL) for Lactobacillus plantarum has been developed, which generalizes the approach obtaining promoters. The consensus sequence, derived from rRNA promoters extracted L. WCFS1 genome, was kept constant, and non-consensus sequences were randomized. Construction of SPL performed in a vector (pSIP409) previously developed high-level, inducible gene expression sakei. wide range strengths obtained with approach, covering 3-4 logs levels small increments activity....

The use of fossil carbon sources for fuels and petrochemicals has serious impacts on our environment is unable to meet the demand in future. A promising sustainable alternative substitute with microbial cell factories converting lignocellulosic biomass into desirable value added products. However, such bioprocesses require availability suitable efficient biocatalysts, capable utilizing C5 sugars tolerant inhibitory compounds generated during pretreatment biomass. In this study, performance a...

Abstract Lactococcus lactis is essential for most cheese making and this mesophilic bacterium has its growth optimum around 30 °C. We have, through adaptive evolution, isolated a mutant TM29 that grows well up to 39 °C continuous at 40 possible if pre-incubated slightly lower temperature. At the maximal permissive temperature wild-type, 38 °C, 33% faster 12% higher specific lactate production rate than parent MG1363, which results in fast accumulation. Genome sequencing was used reveal...

Sustainable and economically feasible ways to produce ethanol or other liquid fuels are becoming increasingly relevant due the limited supply of fossil environmental consequences associated with their consumption. Microbial production fuel compounds has gained a lot attention focus mostly been on developing bio-processes involving non-food plant biomass feedstocks. The high cost enzymes needed degrade such feedstocks into its constituent sugars as well problems various inhibitors generated...

Abstract The potential that lies in harnessing the chemical synthesis capabilities inherent living organisms is immense. Here we demonstrate how biosynthetic machinery of Lactococcus lactis , can be diverted to make (3 R )-acetoin and derived 2,3-butanediol isomers meso -(2,3)-butanediol (m-BDO) (2 ,3 )-butanediol (R-BDO). Efficient production was accomplished using a strain where competing lactate, acetate ethanol forming pathways had been blocked. By introducing different alcohol...

ABSTRACT A new approach for modulating gene expression, based on randomization of promoter (spacer) sequences, was developed. The method applied to chromosomal genes in Lactococcus lactis and shown generate libraries clones with broad ranges expression levels target genes. In one example, overexpression achieved by introducing an additional copy into a phage attachment site the chromosome. This resulted series strains phosphofructokinase activities from 1.4 11 times wild-type activity level....

The C and D cell cycle periods of seven Escherichia coli K-12 strains three E. B/r were determined by computer simulation DNA histograms obtained flow cytometry batch cultures grown at several different generation times. To obtain longer times two the cultivated dilution rates in glucose-limited chemostats. replication period (C period) was found to be similar At below 60 min constant; above it increased linearly with increasing time. from termination division (D more variable. It much...