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Citrate, low pH and amino acid limitation induce citrate utilization in Lactococcus lactis biovar diacetylactis.

ABSTRACT: In Lactococcus lactis subsp. lactis biovar diacetylactis, citrate transport is facilitated by the plasmid-encoded citrate permease (CitP). In this work, we analysed regulation of citrate utilization by pH, nutrient limitation and the presence of citrate at four different levels: (i) plasmid copy number, (ii) citP transcription, (iii) citP mRNA processing and (iv) citrate utilization capacity. Citrate was supplied as cosubstrate together with lactose. The citP gene is known to be induced in cells grown at low pH. However, we demonstrated that transcription of citP was even higher in the presence of citrate (3.8-fold compared with 2.0-fold). The effect of citrate has been overlooked by other researchers because they determined the effect of citrate using M17 medium, which already contains 0.80 ± 0.07 mM citrate. The plasmid copy number increased in cells grown under amino acid limitation (1.6-fold) and/or at low pH (1.4-fold). No significant differences in citP mRNA processing were found. Citrate utilization rates increased from approximately 1 to 65 ?mol min-1 gDW-1 from lowest to highest citP expression. Acetoin formation increased during growth in an acidic environment due to induction of the acetoin pathway. Quantification of the relative contributions allowed us to construct a model for regulation of citrate utilization in L. lactis biovar diacetylactis. This knowledge will help to select conditions to improve flavour formation from citrate.

SUBMITTER: van Mastrigt O

PROVIDER: S-EPMC5812246 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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Citrate, low pH and amino acid limitation induce citrate utilization in Lactococcus lactis biovar diacetylactis.

van Mastrigt Oscar O Mager Emma E EE Jamin Casper C Abee Tjakko T Smid Eddy J EJ

Microbial biotechnology 20171207 2

In Lactococcus lactis subsp. lactis biovar diacetylactis, citrate transport is facilitated by the plasmid-encoded citrate permease (CitP). In this work, we analysed regulation of citrate utilization by pH, nutrient limitation and the presence of citrate at four different levels: (i) plasmid copy number, (ii) citP transcription, (iii) citP mRNA processing and (iv) citrate utilization capacity. Citrate was supplied as cosubstrate together with lactose. The citP gene is known to be induced in cells ...[more]

PMID: 29215194

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Project description:BACKGROUND:Important industrial traits have been linked to plasmids in Lactococcus lactis. RESULTS:The dairy isolate L. lactis subsp. lactis biovar diacetylactis FM03P was sequenced revealing the biggest plasmidome of all completely sequenced and published L. lactis strains up till now. The 12 plasmids that were identified are: pLd1 (8277 bp), pLd2 (15,218 bp), pLd3 (4242 bp), pLd4 (12,005 bp), pLd5 (7521 bp), pLd6 (3363 bp), pLd7 (30,274 bp), pLd8 (47,015 bp), pLd9 (15,313 bp), pLd10 (39,563 bp), pLd11 (9833 bp) and pLd12 (3321 bp). Structural analysis of the repB promoters and the RepB proteins showed that eleven of the plasmids replicate via the theta-type mechanism, while only plasmid pLd3 replicates via a rolling-circle replication mechanism. Plasmids pLd2, pLd7 and pLd10 contain a highly similar operon involved in mobilisation of the plasmids. Examination of the twelve plasmids of L. lactis FM03P showed that 10 of the plasmids carry putative genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as lactose utilisation (lacR-lacABCDFEGX), citrate uptake (citQRP), peptide degradation (pepO and pepE) and oligopeptide uptake (oppDFBCA), uptake of magnesium and manganese (2 mntH, corA), exopolysaccharides production (eps operon), bacteriophage resistance (1 hsdM, 1 hsdR and 7 different hsdS genes of a type I restriction-modification system, an operon of three genes encoding a putative type II restriction-modification system and an abortive infection gene) and stress resistance (2 uspA, cspC and cadCA). Acquisition of these plasmids most likely facilitated the adaptation of the recipient strain to the dairy environment. Some plasmids were already lost during a single propagation step signifying their instability in the absence of a selective pressure. CONCLUSIONS:Lactococcus lactis FM03P carries 12 plasmids important for its adaptation to the dairy environment. Some of the plasmids were easily lost demonstrating that propagation outside the dairy environment should be minimised when studying dairy isolates of L. lactis.

Dataset Information

Citrate, low pH and amino acid limitation induce citrate utilization in Lactococcus lactis biovar diacetylactis.

Publications

Citrate, low pH and amino acid limitation induce citrate utilization in Lactococcus lactis biovar diacetylactis.

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