scholarly journals Sialic Acid Catabolism and Transport Gene Clusters Are Lineage Specific in Vibrio vulnificus

2012 ◽  
Vol 78 (9) ◽  
pp. 3407-3415 ◽  
Author(s):  
Jean-Bernard Lubin ◽  
Joseph J. Kingston ◽  
Nityananda Chowdhury ◽  
E. Fidelma Boyd
Keyword(s):  
Sialic Acid ◽  
Carbon Source ◽  
Vibrio Vulnificus ◽  
Pathogenicity Island ◽  
Gene Clusters ◽  
Acid Uptake ◽  
Acid Transport ◽  
Content Type ◽  
Natural Isolates ◽  
Chromosome Ii

ABSTRACTSialic or nonulosonic acids are nine-carbon alpha ketosugars that are present in all vertebrate mucous membranes. Among bacteria, the ability to catabolize sialic acid as a carbon source is present mainly in pathogenic and commensal species of animals. Previously, it was shown that severalVibriospecies carry homologues of the genes required for sialic acid transport and catabolism, which are genetically linked. InVibrio choleraeon chromosome I, these genes are carried on theVibriopathogenicity island-2 region, which is confined to pathogenic isolates. We found that among the three sequencedVibrio vulnificusclinical strains, these genes are present on chromosome II and are not associated with a pathogenicity island. To determine whether the sialic acid transport (SAT) and catabolism (SAC) region is universally present withinV. vulnificus, we examined 67 natural isolates whose phylogenetic relationships are known. We found that the region was present predominantly among lineage I ofV. vulnificus, which is comprised mainly of clinical isolates. We demonstrate that the isolates that contain this region can catabolize sialic acid as a sole carbon source. Two putative transporters are genetically linked to the region inV. vulnificus, the tripartite ATP-independent periplasmic (TRAP) transporter SiaPQM and a component of an ATP-binding cassette (ABC) transporter. We constructed an in-frame deletion mutation insiaM, a component of the TRAP transporter, and demonstrate that this transporter is essential for sialic acid uptake in this species. Expression analysis of the SAT and SAC genes indicates that sialic acid is an inducer of expression. Overall, our study demonstrates that the ability to catabolize and transport sialic acid is predominately lineage specific inV. vulnificusand that the TRAP transporter is essential for sialic acid uptake.

scholarly journals Uptake of N-acetylneuraminic acid by Escherichia coli K-235. Biochemical characterization of the transport system

1987 ◽  
Vol 246 (2) ◽  
pp. 287-294 ◽  
Author(s):  
L B Rodríguez-Aparicio ◽  
A Reglero ◽  
J M Luengo
Keyword(s):  
Escherichia Coli ◽  
Sialic Acid ◽  
Transport System ◽  
Biochemical Characterization ◽  
Osmotic Shock ◽  
Metabolic Inhibitors ◽  
Acid Uptake ◽  
Uptake System ◽  
Acid Transport ◽  
Acetylneuraminic Acid

Kinetic measurement of the uptake of N-acetyl[4,5,6,7,8,9-14C]neuraminic acid by Escherichia coli K-235 was carried out in vivo at 37 degrees C in 0.1 M-Tris/maleate buffer, pH 7.0. Under these conditions uptake was linear for at least 30 min and the Km calculated for sialic acid was 30 microM. The transport system was osmotic-shock-sensitive and was strongly inhibited by uncouplers of oxidative phosphorylation [2,4-dinitrophenol (100%); NaN3 (66%]) and by the metabolic inhibitors KCN (84%) and sodium arsenate (76%). The thiol-containing compounds mercaptoethanol, glutathione, cysteine, dithiothreitol and cysteine had no significant effect on the sialic acid-transport rate, whereas the thiol-modifying reagents N-ethylmaleimide, iodoacetate and p-chloromercuribenzoate almost completely blocked (greater than 94%) the uptake of this N-acetyl-sugar. N-Acetylglucosamine inhibited non-competitively the transport of N-acetylneuraminic acid, whereas other carbohydrates (hexoses, pentoses, hexitols, hexuronic acids, disaccharides, trisaccharides) and N-acetyl-sugars or amino acid derivatives (N-acetylmannosamine, N-acetylcysteine, N-acetylproline and N-acetylglutamic acid) did not have any effect. Surprisingly, L-methionine and its non-sulphur analogue L-norleucine partially blocked the transport of this sugar (50%), whereas D-methionine, D-norleucine, several L-methionine derivatives (L-methionine methyl ester, L-methionine ethyl ester, L-methionine sulphoxide) and other amino acids did not affect sialic acid uptake. The N-acetylneuraminic acid-transport system is induced by sialic acid and is strictly regulated by the carbon source used for E. coli growth, arabinose, lactose, glucose, fructose and glucosamine being the carbohydrates that cause the greatest repressions in this system. Addition of cyclic AMP to the culture broth reversed the glucose effect, indicating that the N-acetylneuraminic acid-uptake system is under catabolic regulation. Protein synthesis is not needed for sialic acid transport.

scholarly journals Host-Like Carbohydrates Promote Bloodstream Survival of Vibrio vulnificusIn Vivo

2015 ◽  
Vol 83 (8) ◽  
pp. 3126-3136 ◽  
Author(s):  
Jean-Bernard Lubin ◽  
Warren G. Lewis ◽  
Nicole M. Gilbert ◽  
Cory M. Weimer ◽  
Salvador Almagro-Moreno ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Vibrio Vulnificus ◽  
Polymyxin B ◽  
The United States ◽  
Sialic Acids ◽  
Natural Resistance ◽  
Bacterial Survival ◽  
Content Type ◽  
Life Threatening

Sialic acids are found on all vertebrate cell surfaces and are part of a larger class of molecules known as nonulosonic acids. Many bacterial pathogens synthesize related nine-carbon backbone sugars; however, the role(s) of these non-sialic acid molecules in host-pathogen interactions is poorly understood.Vibrio vulnificusis the leading cause of seafood-related death in the United States due to its ability to quickly access the host bloodstream, which it can accomplish through gastrointestinal or wound infection. However, little is known about how this organism persists systemically. Here we demonstrate that sialic acid-like molecules are present on the lipopolysaccharide ofV. vulnificus, are required for full motility and biofilm formation, and also contribute to the organism's natural resistance to polymyxin B. Further experiments in a murine model of intravenousV. vulnificusinfection demonstrated that expression of nonulosonic acids had a striking benefit for bacterial survival during bloodstream infection and dissemination to other tissuesin vivo. In fact, levels of bacterial persistence in the blood corresponded to the overall levels of these molecules expressed byV. vulnificusisolates. Taken together, these results suggest that molecules similar to sialic acids evolved to facilitate the aquatic lifestyle ofV. vulnificusbut that their emergence also resulted in a gain of function with life-threatening potential in the human host.

scholarly journals Genomic and Metabolic Profiling of Nonulosonic Acids in Vibrionaceae Reveal Biochemical Phenotypes of Allelic Divergence in Vibrio vulnificus

2011 ◽  
Vol 77 (16) ◽  
pp. 5782-5793 ◽  
Author(s):  
Amanda L. Lewis ◽  
Jean-Bernard Lubin ◽  
Shilpa Argade ◽  
Natasha Naidu ◽  
Biswa Choudhury ◽  
...  
Keyword(s):  
Vibrio Vulnificus ◽  
Hot Spot ◽  
Gene Clusters ◽  
Human Pathogens ◽  
Environmental Isolates ◽  
Content Type ◽  
The Family ◽  
Domains Of Life ◽  
Biochemical Analyses ◽  
And Function

ABSTRACTNonulosonic acids (NulOs) encompass a large group of structurally diverse nine-carbon backbone α-keto sugars widely distributed among the three domains of life. Mammals express a specialized version of NulOs called sialic acids, which are displayed in prominent terminal positions of cell surface and secreted glycoconjugates. Within bacteria, the ability to synthesize NulOs has been demonstrated in a number of human pathogens and is phylogenetically widespread. Here we examine the distribution, diversity, evolution, and function of NulO biosynthesis pathways in members of the familyVibrionaceae. Among 27 species ofVibrionaceaeexamined at the genomic level, 12 species containednabgene clusters. We document examples of duplication, divergence, horizontal transfer, and recombination ofnabgene clusters in differentVibrionaceaelineages. Biochemical analyses, including mass spectrometry, confirmed that many species do, in fact, produce di-N-acetylated NulOs. A library of clinical and environmental isolates ofVibrio vulnificusserved as a model for further investigation ofnaballele genotypes and levels of NulO expression. The data show that lineage I isolates produce about 20-fold higher levels of NulOs than lineage II isolates. Moreover,nabgene alleles found in a subset ofV. vulnificusclinical isolates express 40-fold higher levels of NulOs thannaballeles associated with environmental isolates. Taken together, the data implicate the familyVibrionaceaeas a “hot spot” of NulO evolution and suggest that these molecules may have diverse roles in environmental persistence and/or animal virulence.

scholarly journals The Fungal Pathogen Candida albicans Autoinduces Hyphal Morphogenesis by Raising Extracellular pH

mBio ◽  
2011 ◽  
Vol 2 (3) ◽  
Author(s):  
Slavena Vylkova ◽  
Aaron J. Carman ◽  
Heather A. Danhof ◽  
John R. Collette ◽  
Huaijin Zhou ◽  
...  
Keyword(s):  
Amino Acids ◽  
Candida Albicans ◽  
Amino Acid ◽  
Carbon Source ◽  
Fungal Pathogen ◽  
Host Cells ◽  
Acid Uptake ◽  
Content Type ◽  
Virulence Trait ◽  
Ph Adaptation

ABSTRACTpH homeostasis is critical for all organisms; in the fungal pathogenCandida albicans, pH adaptation is critical for virulence in distinct host niches. We demonstrate that beyond adaptation,C. albicansactively neutralizes the environment from either acidic or alkaline pHs. Under acidic conditions, this species can raise the pH from 4 to >7 in less than 12 h, resulting in autoinduction of the yeast-hyphal transition, a critical virulence trait. Extracellular alkalinization has been reported to occur in several fungal species, but under the specific conditions that we describe, the phenomenon is more rapid than previously observed. Alkalinization is linked to carbon deprivation, as it occurs in glucose-poor media and requires exogenous amino acids. These conditions are similar to those predicted to exist inside phagocytic cells, and we find a strong correlation between the use of amino acids as a cellular carbon source and the degree of alkalinization. Genetic and genomic approaches indicate an emphasis on amino acid uptake and catabolism in alkalinizing cells. Mutations in four genes,STP2, a transcription factor regulating amino acid permeases,ACH1(acetyl-coenzyme A [acetyl-CoA] hydrolase),DUR1,2(urea amidolyase), andATO5, a putative ammonia transporter, abolish or delay neutralization. The pH changes are the result of the extrusion of ammonia, as observed in other fungi. We propose that nutrient-deprivedC. albicanscells catabolize amino acids as a carbon source, excreting the amino nitrogen as ammonia to raise environmental pH and stimulate morphogenesis, thus directly contributing to pathogenesis.IMPORTANCECandida albicansis the most important fungal pathogen of humans, causing disease at multiple body sites. The ability to switch between multiple morphologies, including a rounded yeast cell and an elongated hyphal cell, is a key virulence trait in this species, as this reversible switch is thought to promote dissemination and tissue invasion in the host. We report here thatC. albicanscan actively alter the pH of its environment and induce its switch to the hyphal form. The change in pH is caused by the release of ammonia from the cells produced during the breakdown of amino acids. This phenomenon is unprecedented in a human pathogen and may substantially impact host physiology by linking morphogenesis, pH adaptation, carbon metabolism, and interactions with host cells, all of which are critical for the ability ofC. albicansto cause disease.

scholarly journals Lactobacillus casei Ferments theN-Acetylglucosamine Moiety of Fucosyl-α-1,3-N-Acetylglucosamine and Excretes l-Fucose

2012 ◽  
Vol 78 (13) ◽  
pp. 4613-4619 ◽  
Author(s):  
Jesús Rodríguez-Díaz ◽  
Antonio Rubio-del-Campo ◽  
María J. Yebra
Keyword(s):  
Carbon Source ◽  
Lactobacillus Casei ◽  
Genomic Sequence ◽  
Protein A ◽  
Physiological Role ◽  
Gene Clusters ◽  
Transcriptional Analysis ◽  
Phosphotransferase System ◽  
Content Type

ABSTRACTWe have previously characterized fromLactobacillus caseiBL23 three α-l-fucosidases, AlfA, AlfB, and AlfC, which hydrolyzein vitronatural fucosyl-oligosaccharides. In this work, we have shown thatL. caseiis able to grow in the presence of fucosyl-α-1,3-N-acetylglucosamine (Fuc-α-1,3-GlcNAc) as a carbon source. Interestingly,L. caseiexcretes thel-fucose moiety during growth on Fuc-α-1,3-GlcNAc, indicating that only theN-acetylglucosamine moiety is being metabolized. Analysis of the genomic sequence ofL. caseiBL23 shows that downstream fromalfB, which encodes the α-l-fucosidase AlfB, a gene,alfR, that encodes a transcriptional regulator is present. Divergently fromalfB, three genes,alfEFG, that encode proteins with homology to the enzyme IIAB (EIIAB), EIIC, and EIID components of a mannose-class phosphoenolpyruvate:sugar phosphotransferase system (PTS) are present. Inactivation of eitheralfBoralfFabolishes the growth ofL. caseion Fuc-α-1,3-GlcNAc. This proves that AlfB is involved in Fuc-α-1,3-GlcNAc metabolism and that the transporter encoded byalfEFGparticipates in the uptake of this disaccharide. A mutation in the PTS general component enzyme I does not eliminate the utilization of Fuc-α-1,3-GlcNAc, suggesting that the transport via the PTS encoded byalfEFGis not coupled to phosphorylation of the disaccharide. Transcriptional analysis withalfRandccpAmutants shows that the two gene clustersalfBRandalfEFGare regulated by substrate-specific induction mediated by the inactivation of the transcriptional repressor AlfR and by carbon catabolite repression mediated by the catabolite control protein A (CcpA). This work reports for the first time the characterization of the physiological role of an α-l-fucosidase in lactic acid bacteria and the utilization of Fuc-α-1,3-GlcNAc as a carbon source for bacteria.

scholarly journals The Acquisition of the scr Gene Cluster Encoding Sucrose Metabolization Enzymes Enables Strains of Vibrio parahaemolyticus and Vibrio vulnificus to Utilize Sucrose as Carbon Source

2021 ◽  
Vol 12 ◽  
Author(s):  
Jens Andre Hammerl ◽  
Cornelia Göllner ◽  
Claudia Jäckel ◽  
Fatima Swidan ◽  
Helena Gutmann ◽  
...  
Keyword(s):  
Carbon Source ◽  
Vibrio Parahaemolyticus ◽  
Genomic Island ◽  
Vibrio Vulnificus ◽  
Shuttle Vector ◽  
E Coli ◽  
Gene Coding ◽  
Chromosome Ii ◽  
Pcr Targeting ◽  
Sucrose Utilization

Most strains of Vibrio parahaemolyticus are unable to utilize sucrose as carbon source, though few exceptions exist. We investigated a sucrose-positive V. parahaemolyticus strain by whole-genome sequencing (WGS) and confirmed the presences of a genomic island containing sucrose utilization genes. A 4.7 kb DNA cluster consisting of three genes: scrA encoding a sucrose uptake protein, scrK encoding a fructokinase, and scrB coding for a sucrose-6-phosphate hydrolase, was PCR amplified and inserted into the Vibrio/Escherichia coli shuttle vector pVv3. Two recombinant plasmids, only differing in the orientation of the insert with respect to the pVv3-lacZα-fragment, conferred the E. coli K12 transformants the ability to utilize sucrose. The introduction of the two plasmids into sucrose-negative V. parahaemolyticus and V. vulnificus strains also results in a change of the sucrose utilization phenotype from negative to positive. By performing a multiplex PCR targeting scrA, scrK, and scrB, 43 scr-positive V. parahaemolyticus isolates from our collection of retail strains were detected and confirmed to be able to use sucrose as carbon source. Strains unable to utilize the disaccharide were negative by PCR for the scr genes. For in-depth characterization, 17 sucrose-positive V. parahaemolyticus were subjected to WGS. A genomic island with a nucleotide identity of >95% containing scrA, scrB, scrK and three additional coding sequences (CDS) were identified in all strains. The additional genes were predicted as a gene coding for a transcriptional regulator (scrR), a porin encoding gene and a CDS of unknown function. Sequence comparison indicated that the genomic island was located in the same region of the chromosome II in all analyzed V. parahaemolyticus strains. Structural comparison of the genomes with sequences of the sucrose utilizing species V. alginolyticus revealed the same genomic island, which indicates a possible distribution of this genetic structure by horizontal gene transfer. The comparison of all genome sequences based on SNP differences reveals that the presence of sucrose utilizing genes is found in genetically diverse V. parahaemolyticus strains and is not restricted to a subset of closely related strains.

scholarly journals Sialic Acid Transport Contributes to Pneumococcal Colonization

2010 ◽  
Vol 79 (3) ◽  
pp. 1262-1269 ◽  
Author(s):  
Carolyn Marion ◽  
Amanda M. Burnaugh ◽  
Shireen A. Woodiga ◽  
Samantha J. King
Keyword(s):  
Sialic Acid ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Upper Respiratory Tract ◽  
Acid Transport ◽  
Upper Respiratory ◽  
Pneumococcal Colonization ◽  
Free Carbohydrates ◽  
Airway Colonization

ABSTRACTStreptococcus pneumoniaeis a major cause of pneumonia and meningitis. Airway colonization is a necessary precursor to disease, but little is known about how the bacteria establish and maintain colonization. Carbohydrates are required as a carbon source for pneumococcal growth and, therefore, for colonization. Free carbohydrates are not readily available in the naso-oropharynx; however, N- and O-linked glycans are common in the airway. Sialic acid is the most common terminal modification on N- and O-linked glycans and is likely encountered frequently byS. pneumoniaein the airway. Here we demonstrate that sialic acid supports pneumococcal growth when provided as a sole carbon source. Growth on sialic acid requires import into the bacterium. Three genetic regions have been proposed to encode pneumococcal sialic acid transporters: one sodium solute symporter and two ATP binding cassette (ABC) transporters. Data demonstrate that one of these,satABC, is required for transport of sialic acid. AsatABCmutant displayed significantly reduced growth on both sialic acid and the human glycoprotein alpha-1. The importance ofsatABCfor growth on human glycoprotein suggests that sialic acid transport may be importantin vivo. Indeed, thesatABCmutant was significantly reduced in colonization of the murine upper respiratory tract. This work demonstrates thatS. pneumoniaeis able to use sialic acid as a sole carbon source and that utilization of sialic acid is likely important during pneumococcal colonization.

scholarly journals Comparative Analyses of the Lipooligosaccharides from Nontypeable Haemophilus influenzae and Haemophilus haemolyticus Show Differences in Sialic Acid and Phosphorylcholine Modifications

2016 ◽  
Vol 84 (3) ◽  
pp. 765-774 ◽  
Author(s):  
Deborah M. B. Post ◽  
Margaret R. Ketterer ◽  
Jeremy E. Coffin ◽  
Lorri M. Reinders ◽  
Robert S. Munson ◽  
...  
Keyword(s):  
Sialic Acid ◽  
Haemophilus Influenzae ◽  
Mass Spectrometric ◽  
Lower Airway ◽  
Titration Calorimetry ◽  
Acid Uptake ◽  
Nontypeable Haemophilus Influenzae ◽  
Content Type ◽  
Comparative Analyses ◽  
Tract Infections

Haemophilus haemolyticusand nontypeableHaemophilus influenzae(NTHi) are closely related upper airway commensal bacteria that are difficult to distinguish phenotypically. NTHi causes upper and lower airway tract infections in individuals with compromised airways, whileH. haemolyticusrarely causes such infections. The lipooligosaccharide (LOS) is an outer membrane component of both species and plays a role in NTHi pathogenesis. In this study, comparative analyses of the LOS structures and corresponding biosynthesis genes were performed. Mass spectrometric and immunochemical analyses showed that NTHi LOS contained terminal sialic acid more frequently and to a higher extent thanH. haemolyticusLOS did. Genomic analyses of 10 strains demonstrated thatH. haemolyticuslacked the sialyltransferase geneslic3Aandlic3B(9/10) andsiaA(10/10), but all strains contained the sialic acid uptake genessiaPandsiaT(10/10). However, isothermal titration calorimetry analyses of SiaP from twoH. haemolyticusstrains showed a 3.4- to 7.3-fold lower affinity for sialic acid compared to that of NTHi SiaP. Additionally, mass spectrometric and immunochemical analyses showed that the LOS fromH. haemolyticuscontained phosphorylcholine (ChoP) less frequently than the LOS from NTHi strains. These differences observed in the levels of sialic acid and ChoP incorporation in the LOS structures fromH. haemolyticusand NTHi may explain some of the differences in their propensities to cause disease.

scholarly journals Differential transcription of expanded gene families in central carbon metabolism of Streptomyces coelicolor A3(2)

2020 ◽  
Vol 2 (6) ◽  
Author(s):  
Jana K. Schniete ◽  
Richard Reumerman ◽  
Leena Kerr ◽  
Nicholas P. Tucker ◽  
Iain S. Hunter ◽  
...  
Keyword(s):  
Carbon Source ◽  
Streptomyces Coelicolor ◽  
Carbon Sources ◽  
Gene Families ◽  
Gene Clusters ◽  
Central Carbon Metabolism ◽  
Rna Seq ◽  
Content Type ◽  
Link Type ◽  
Enzymatic Function

Background. Streptomycete bacteria are prolific producers of specialized metabolites, many of which have clinically relevant bioactivity. A striking feature of their genomes is the expansion of gene families that encode the same enzymatic function. Genes that undergo expansion events, either by horizontal gene transfer or duplication, can have a range of fates: genes can be lost, or they can undergo neo-functionalization or sub-functionalization. To test whether expanded gene families in Streptomyces exhibit differential expression, an RNA-Seq approach was used to examine cultures of wild-type Streptomyces coelicolor grown with either glucose or tween as the sole carbon source. Results. RNA-Seq analysis showed that two-thirds of genes within expanded gene families show transcriptional differences when strains were grown on tween compared to glucose. In addition, expression of specialized metabolite gene clusters (actinorhodin, isorenieratane, coelichelin and a cryptic NRPS) was also influenced by carbon source. Conclusions. Expression of genes encoding the same enzymatic function had transcriptional differences when grown on different carbon sources. This transcriptional divergence enables partitioning to function under different physiological conditions. These approaches can inform metabolic engineering of industrial Streptomyces strains and may help develop cultivation conditions to activate the so-called silent biosynthetic gene clusters.

scholarly journals Genes of theN-Methylglutamate Pathway Are Essential for Growth of Methylobacterium extorquens DM4 with Monomethylamine

2014 ◽  
Vol 80 (11) ◽  
pp. 3541-3550 ◽  
Author(s):  
Christelle Gruffaz ◽  
Emilie E. L. Muller ◽  
Yousra Louhichi-Jelail ◽  
Yella R. Nelli ◽  
Gilles Guichard ◽  
...  
Keyword(s):  
Carbon Source ◽  
Nitrogen Source ◽  
Gene Clusters ◽  
Sole Source ◽  
Nadh:Ubiquinone Oxidoreductase ◽  
Methylotrophic Bacteria ◽  
Methylobacterium Extorquens ◽  
Content Type ◽  
Carbon And Nitrogen ◽  
In The Wild

ABSTRACTMonomethylamine (MMA, CH3NH2) can be used as a carbon and nitrogen source by many methylotrophic bacteria.Methylobacterium extorquensDM4 lacks the MMA dehydrogenase encoded bymaugenes, which inM. extorquensAM1 is essential for growth on MMA. Identification and characterization of minitransposon mutants with an MMA-dependent phenotype showed that strain DM4 grows with MMA as the sole source of carbon, energy, and nitrogen by theN-methylglutamate (NMG) pathway. Independent mutations were found in a chromosomal region containing the genesgmaS,mgsABC, andmgdABCDfor the three enzymes of the pathway, γ-glutamylmethylamide (GMA) synthetase, NMG synthase, and NMG dehydrogenase, respectively. Reverse transcription-PCR confirmed the operonic structure of the two divergent gene clustersmgsABC-gmaSandmgdABCDand their induction during growth with MMA. The genesmgdABCDandmgsABCwere found to be essential for utilization of MMA as a carbon and nitrogen source. The genegmaSwas essential for MMA utilization as a carbon source, but residual growth of mutant DM4gmaSgrowing with succinate and MMA as a nitrogen source was observed. Plasmid copies ofgmaSand thegmaShomolog METDI4690, which encodes a protein 39% identical to GMA synthetase, fully restored the ability of mutants DM4gmaSand DM4gmaSΔmetdi4690 to use MMA as a carbon and nitrogen source. Similarly, chemically synthesized GMA, the product of GMA synthetase, could be used as a nitrogen source for growth in the wild-type strain, as well as in DM4gmaSand DM4gmaSΔmetdi4690 mutants. The NADH:ubiquinone oxidoreductase respiratory complex component NuoG was also found to be essential for growth with MMA as a carbon source.

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