Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Front Microbiol
2022 Apr 14;13:855736. doi: 10.3389/fmicb.2022.855736.
Show Gene links
Show Anatomy links
A Novel Efficient L-Lysine Exporter Identified by Functional Metagenomics.
Malla S
,
van der Helm E
,
Darbani B
,
Wieschalka S
,
Förster J
,
Borodina I
,
Sommer MOA
.
???displayArticle.abstract???
Lack of active export system often limits the industrial bio-based production processes accumulating the intracellular product and hence complexing the purification steps. L-lysine, an essential amino acid, is produced biologically in quantities exceeding two million tons per year; yet, L-lysine production is challenged by efficient export system at high titers during fermentation. To address this issue, new exporter candidates for efficient efflux of L-lysine are needed. Using metagenomic functional selection, we identified 58 genes encoded on 28 unique metagenomic fragments from cow gut microbiome library that improved L-lysine tolerance. These genes include a novel L-lysine transporter, belonging to a previously uncharacterized EamA superfamily, which is further in vivo characterized as L-lysine exporter using Xenopus oocyte expression system as well as Escherichia coli host. This novel exporter improved L-lysine tolerance in E. coli by 40% and enhanced yield, titer, and the specific production of L-lysine in an industrial Corynebacterium glutamicum strain by 7.8%, 9.5%, and 12%, respectively. Our approach allows the sequence-independent discovery of novel exporters and can be deployed to increase titers and productivity of toxicity-limited bioprocesses.
FIGURE 3. Functional characterization of MglE transporter activity in Xenopus oocytes, E.coli and Corynebacterium glutamicum. (A) L-lysine export assay of MglE transporter in Xenopus oocytes in Kulori buffer (i) at pH 5 and (ii) at pH 7.4. The bars represent lysine contents in the oocytes or the buffer (meansâ±âSD, 3â4 biological replicates each involving 20 oocytes). Significant changes are in comparison with the controls marked by asterisks (** pâ<â0.01; Fishers one-way ANOVA). (B) The extracellular L-lysine concentration of reference E. coli W3110 and its isogenic mutant strain DMLC along with the cell OD600nm values confirming the exporter activity of the MglE protein in (i) LB media and (ii) in M9 minimal media supplemented with yeast extract (* pâ<â0.0001, t-test). (C) Fold enhancement of specific production of L-lysine by MglE protein in the industrial C. glutamicum VL5 strain compared to the empty vector control. Error bars are s.e.m., * denotes pâ=â0.023, nâ=â4.
Archer,
The genome sequence of E. coli W (ATCC 9637): comparative genome analysis and an improved genome-scale reconstruction of E. coli.
2011, Pubmed
Archer,
The genome sequence of E. coli W (ATCC 9637): comparative genome analysis and an improved genome-scale reconstruction of E. coli.
2011,
Pubmed
Baba,
Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection.
2006,
Pubmed
Becker,
Systems and synthetic metabolic engineering for amino acid production - the heartbeat of industrial strain development.
2012,
Pubmed
Bellmann,
Expression control and specificity of the basic amino acid exporter LysE of Corynebacterium glutamicum.
2001,
Pubmed
Blattner,
The complete genome sequence of Escherichia coli K-12.
1997,
Pubmed
Borodina,
Understanding metabolite transport gives an upper hand in strain development.
2019,
Pubmed
Cock,
Biopython: freely available Python tools for computational molecular biology and bioinformatics.
2009,
Pubmed
Darbani,
Energetic evolution of cellular Transportomes.
2018,
Pubmed
Darbani,
The biosynthetic gene cluster for the cyanogenic glucoside dhurrin in Sorghum bicolor contains its co-expressed vacuolar MATE transporter.
2016,
Pubmed
,
Xenbase
Darbani,
Engineering energetically efficient transport of dicarboxylic acids in yeast Saccharomyces cerevisiae.
2019,
Pubmed
,
Xenbase
Dassler,
Identification of a major facilitator protein from Escherichia coli involved in efflux of metabolites of the cysteine pathway.
2000,
Pubmed
Eikmanns,
A family of Corynebacterium glutamicum/Escherichia coli shuttle vectors for cloning, controlled gene expression, and promoter probing.
1991,
Pubmed
Finn,
Pfam: the protein families database.
2014,
Pubmed
Forsberg,
Identification of Genes Conferring Tolerance to Lignocellulose-Derived Inhibitors by Functional Selections in Soil Metagenomes.
2016,
Pubmed
Franke,
YfiK from Escherichia coli promotes export of O-acetylserine and cysteine.
2003,
Pubmed
Gunji,
Enhancement of L-lysine production in methylotroph Methylophilus methylotrophus by introducing a mutant LysE exporter.
2006,
Pubmed
Hayashi,
Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110.
2006,
Pubmed
Hemberger,
RibM from Streptomyces davawensis is a riboflavin/roseoflavin transporter and may be useful for the optimization of riboflavin production strains.
2011,
Pubmed
Imaizumi,
Improved production of L-lysine by disruption of stationary phase-specific rmf gene in Escherichia coli.
2005,
Pubmed
Jensen,
Seven gene deletions in seven days: Fast generation of Escherichia coli strains tolerant to acetate and osmotic stress.
2015,
Pubmed
Käll,
Advantages of combined transmembrane topology and signal peptide prediction--the Phobius web server.
2007,
Pubmed
Keilhauer,
Isoleucine synthesis in Corynebacterium glutamicum: molecular analysis of the ilvB-ilvN-ilvC operon.
1993,
Pubmed
Kelle,
Reaction engineering analysis of L-lysine transport by Corynebacterium glutamicum.
1996,
Pubmed
Kikuchi,
Characterization of a second lysine decarboxylase isolated from Escherichia coli.
1997,
Pubmed
Lee,
Systems strategies for developing industrial microbial strains.
2015,
Pubmed
Lutz,
Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements.
1997,
Pubmed
Malla,
Self-resistance mechanism in Streptomyces peucetius: overexpression of drrA, drrB and drrC for doxorubicin enhancement.
2010,
Pubmed
Munck,
Limited dissemination of the wastewater treatment plant core resistome.
2015,
Pubmed
Omasits,
Protter: interactive protein feature visualization and integration with experimental proteomic data.
2014,
Pubmed
Pathania,
Distinct Paths for Basic Amino Acid Export in Escherichia coli: YbjE (LysO) Mediates Export of L-Lysine.
2015,
Pubmed
Rouanet,
The PecM protein of the phytopathogenic bacterium Erwinia chrysanthemi, membrane topology and possible involvement in the efflux of the blue pigment indigoidine.
2001,
Pubmed
Sommer,
A functional metagenomic approach for expanding the synthetic biology toolbox for biomass conversion.
2010,
Pubmed
Sommer,
Functional characterization of the antibiotic resistance reservoir in the human microflora.
2009,
Pubmed
UniProt Consortium,
UniProt: a hub for protein information.
2015,
Pubmed
van der Helm,
deFUME: Dynamic exploration of functional metagenomic sequencing data.
2015,
Pubmed
Vrljic,
The LysE superfamily: topology of the lysine exporter LysE of Corynebacterium glutamicum, a paradyme for a novel superfamily of transmembrane solute translocators.
1999,
Pubmed
Vrljic,
A new type of transporter with a new type of cellular function: L-lysine export from Corynebacterium glutamicum.
1996,
Pubmed
