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Archives of Microbiology (v.185, #2)
Dissimilation of C3-sulfonates by Alasdair M. Cook; Karin Denger; Theo H. M. Smits (pp. 83-90).
Cysteate and sulfolactate are widespread natural products in the environment, while propanesulfonate, 3-aminopropanesulfonate and propane-1,3-disulfonate are xenobiotics. While some understanding of the bacterial assimilation of cysteate sulfur has been achieved, details of the dissimilation of cysteate and sulfolactate by microbes together with information on the degradation of the xenobiotics have only recently become available. This minireview centres on bacterial catabolism of the carbon moiety in these C3-sulfonates and on the fate of the sulfonate group. Three mechanisms of desulfonation have been established. Firstly, cysteate is converted via sulfopyruvate to sulfolactate, which is desulfonated to pyruvate and sulfite; the latter is oxidized to sulfate by a sulfite dehydrogenase and excreted as sulfate in Paracoccus pantotrophus NKNCYSA. Secondly, sulfolactate can be converted to cysteate, which is cleaved in a pyridoxal 5′-phosphate-coupled reaction to pyruvate, sulfite and ammonium ions; in Silicibacter pomeroyi DSS-3, the sulfite is excreted largely as sulfite. Both desulfonation reactions seem to be widespread. The third desulfonation mechanism is oxygenolysis of, e.g. propanesulfonate(s), about which less is known.
Keywords: l-Cysteate; Desulfonation; Sulfite dehydrogenase; 3-Sulfolactate; 3-Sulfopyruvate; Sulfate exporters; Sulfite exporters
Dissimilation of C3-sulfonates by Alasdair M. Cook; Karin Denger; Theo H. M. Smits (pp. 83-90).
Cysteate and sulfolactate are widespread natural products in the environment, while propanesulfonate, 3-aminopropanesulfonate and propane-1,3-disulfonate are xenobiotics. While some understanding of the bacterial assimilation of cysteate sulfur has been achieved, details of the dissimilation of cysteate and sulfolactate by microbes together with information on the degradation of the xenobiotics have only recently become available. This minireview centres on bacterial catabolism of the carbon moiety in these C3-sulfonates and on the fate of the sulfonate group. Three mechanisms of desulfonation have been established. Firstly, cysteate is converted via sulfopyruvate to sulfolactate, which is desulfonated to pyruvate and sulfite; the latter is oxidized to sulfate by a sulfite dehydrogenase and excreted as sulfate in Paracoccus pantotrophus NKNCYSA. Secondly, sulfolactate can be converted to cysteate, which is cleaved in a pyridoxal 5′-phosphate-coupled reaction to pyruvate, sulfite and ammonium ions; in Silicibacter pomeroyi DSS-3, the sulfite is excreted largely as sulfite. Both desulfonation reactions seem to be widespread. The third desulfonation mechanism is oxygenolysis of, e.g. propanesulfonate(s), about which less is known.
Keywords: l-Cysteate; Desulfonation; Sulfite dehydrogenase; 3-Sulfolactate; 3-Sulfopyruvate; Sulfate exporters; Sulfite exporters
Genetic diversity of phenazine- and pyoluteorin-producing pseudomonads isolated from green pepper rhizosphere by Haiming Liu; Dexian Dong; Huasong Peng; Xuehong Zhang; Yuquan Xu (pp. 91-98).
The genetic diversity among indigenous phenazine-1-carboxylic acid (PCA)-producing and pyoluteorin (Plt)-producing isolates of pseudomonads screened from green pepper rhizosphere was exploited in this study. A total of 48 bacterium isolates producing one or both of these antibiotics were screened from green pepper rhizosphere in diverse regions in China. Among these isolates, 45 could produce PCA, 3 could produce both PCA and Plt, and none could produce Plt only. Based on the restriction patterns of partial 16S and 16S-23S internal transcribed spacer (ITS) PCR fragments generated by enzyme HaeIII or HinfI, these isolates fell into 19 or 17 distinct groups respectively, indicating that there was a significant diversity among them. Polygenetic analysis of the partial 16S rDNA and 16S-23S ITS sequence from the representative in each group in the context of similar sequence from previously described bacterial species indicated that most isolates were closely related to the species of Pseudomonas fluorescens, P. putida, and Stenotrophomonas maltophilia. Some of these representatives of these isolates, then, are likely to be novel strains or species in these two genera.
Keywords: Genetic diversity; 16S-23S ITS; Phenazine-1-carboxylic acid; Pyoluteorin; Pseudomonads; Green pepper
Genetic diversity of phenazine- and pyoluteorin-producing pseudomonads isolated from green pepper rhizosphere by Haiming Liu; Dexian Dong; Huasong Peng; Xuehong Zhang; Yuquan Xu (pp. 91-98).
The genetic diversity among indigenous phenazine-1-carboxylic acid (PCA)-producing and pyoluteorin (Plt)-producing isolates of pseudomonads screened from green pepper rhizosphere was exploited in this study. A total of 48 bacterium isolates producing one or both of these antibiotics were screened from green pepper rhizosphere in diverse regions in China. Among these isolates, 45 could produce PCA, 3 could produce both PCA and Plt, and none could produce Plt only. Based on the restriction patterns of partial 16S and 16S-23S internal transcribed spacer (ITS) PCR fragments generated by enzyme HaeIII or HinfI, these isolates fell into 19 or 17 distinct groups respectively, indicating that there was a significant diversity among them. Polygenetic analysis of the partial 16S rDNA and 16S-23S ITS sequence from the representative in each group in the context of similar sequence from previously described bacterial species indicated that most isolates were closely related to the species of Pseudomonas fluorescens, P. putida, and Stenotrophomonas maltophilia. Some of these representatives of these isolates, then, are likely to be novel strains or species in these two genera.
Keywords: Genetic diversity; 16S-23S ITS; Phenazine-1-carboxylic acid; Pyoluteorin; Pseudomonads; Green pepper
Immunocytochemical localization of membrane-bound ammonia monooxygenase in cells of ammonia oxidizing bacteria by Claudia Fiencke; Eberhard Bock (pp. 99-106).
The intracellular location of the membrane-bound ammonia monooxygenase (AMO) in all genera of ammonia oxidizing bacteria (Nitrosomonas, Nitrosococcus and Nitrosospira) was determined by electron microscopic immunocytochemistry. Polyclonal antibodies recognizing the two subunits, AmoA- and AmoB-proteins, were used for post-embedding labeling. Ultrathin sections revealed that the AmoB-protein was located in all genera on the cytoplasmic membrane. In cells of Nitrosomonas and Nitrosococus additional but less AmoB-labeling was found on the intracytoplasmic membrane (ICM). In contrast to the detection of AmoB-protein, the AmoA-antibodies failed to detect the AmoA-protein. Based on quantitative immunoblots the extent of ICM in Nitrosomonas eutropha was correlated with the amount of AmoA in the cells. The highest extent of ICM and amount of AmoA was found at low ammonium substrate concentrations.
Keywords: Ammonia monooxygenase; AMO; Nitrosomonas ; Nitrosococcus ; Nitrosospira ; Antibodies; Immunogold labeling
Immunocytochemical localization of membrane-bound ammonia monooxygenase in cells of ammonia oxidizing bacteria by Claudia Fiencke; Eberhard Bock (pp. 99-106).
The intracellular location of the membrane-bound ammonia monooxygenase (AMO) in all genera of ammonia oxidizing bacteria (Nitrosomonas, Nitrosococcus and Nitrosospira) was determined by electron microscopic immunocytochemistry. Polyclonal antibodies recognizing the two subunits, AmoA- and AmoB-proteins, were used for post-embedding labeling. Ultrathin sections revealed that the AmoB-protein was located in all genera on the cytoplasmic membrane. In cells of Nitrosomonas and Nitrosococus additional but less AmoB-labeling was found on the intracytoplasmic membrane (ICM). In contrast to the detection of AmoB-protein, the AmoA-antibodies failed to detect the AmoA-protein. Based on quantitative immunoblots the extent of ICM in Nitrosomonas eutropha was correlated with the amount of AmoA in the cells. The highest extent of ICM and amount of AmoA was found at low ammonium substrate concentrations.
Keywords: Ammonia monooxygenase; AMO; Nitrosomonas ; Nitrosococcus ; Nitrosospira ; Antibodies; Immunogold labeling
Molecular identification and evolution of the cyclic peptide hepatotoxins, microcystin and nodularin, synthetase genes in three orders of cyanobacteria by Anne-Dorothee Jungblut; Brett A. Neilan (pp. 107-114).
The cyanobacterial hepatotoxins, microcystin and nodularin, are produced by a wide range of cyanobacteria. Microcystin production has been reported in the four cyanobacterial orders: Oscillatoriales, Chroococcales, Stigonematales, and Nostocales. The production of nodularin is a distinct characteristic of the Nostocales genus Nodularia. A single rapid method is needed to reliably detect cyanobacteria that are potentially capable of producing these hepatotoxins. To this end, a PCR was designed to detect all potential microcystin and nodularin-producing cyanobacteria from laboratory cultures as well as in harmful algal blooms. The aminotransferase (AMT) domain, which is located on the modules mcyE and ndaF of the microcystin and nodularin synthetase enzyme complexes, respectively, was chosen as the target sequence because of its essential function in the synthesis of all microcystins as well as nodularins. Using the described PCR, it was possible to amplify a 472 bp PCR product from the AMT domains of all tested hepatotoxic species and bloom samples. Sequence data provided further insight into the evolution of the microcystin and nodularin synthetases through bioinformatic analyses of the AMT in microcystin and nodularin synthetases, with congruence between the evolution of 16S rRNA and the AMT domain.
Keywords: Cyanobacteria; Microcystin synthetase; Nodularin synthetase; Cyclic peptide hepatotoxin; Aminotransferase
Molecular identification and evolution of the cyclic peptide hepatotoxins, microcystin and nodularin, synthetase genes in three orders of cyanobacteria by Anne-Dorothee Jungblut; Brett A. Neilan (pp. 107-114).
The cyanobacterial hepatotoxins, microcystin and nodularin, are produced by a wide range of cyanobacteria. Microcystin production has been reported in the four cyanobacterial orders: Oscillatoriales, Chroococcales, Stigonematales, and Nostocales. The production of nodularin is a distinct characteristic of the Nostocales genus Nodularia. A single rapid method is needed to reliably detect cyanobacteria that are potentially capable of producing these hepatotoxins. To this end, a PCR was designed to detect all potential microcystin and nodularin-producing cyanobacteria from laboratory cultures as well as in harmful algal blooms. The aminotransferase (AMT) domain, which is located on the modules mcyE and ndaF of the microcystin and nodularin synthetase enzyme complexes, respectively, was chosen as the target sequence because of its essential function in the synthesis of all microcystins as well as nodularins. Using the described PCR, it was possible to amplify a 472 bp PCR product from the AMT domains of all tested hepatotoxic species and bloom samples. Sequence data provided further insight into the evolution of the microcystin and nodularin synthetases through bioinformatic analyses of the AMT in microcystin and nodularin synthetases, with congruence between the evolution of 16S rRNA and the AMT domain.
Keywords: Cyanobacteria; Microcystin synthetase; Nodularin synthetase; Cyclic peptide hepatotoxin; Aminotransferase
Global gene regulation in Yersinia enterocolitica: effect of FliA on the expression levels of flagellar and plasmid-encoded virulence genes by Shelley M. Horne; Birgit M. Prüß (pp. 115-126).
This study describes the involvement of the sigma factor of the flagellar system, FliA, in global gene regulation of Yersinia enterocolitica. In addition to exhibiting a positive effect upon the expression levels of eight class III flagellar operons, FliA also exhibited a negative effect upon the expression levels of four virulence operons that are located on the pYV virulence plasmid. These are yadA, virC, yopQ, and the insertion element ISYen1. While the positive effect on class III flagellar operons by FliA is most likely direct, the negative effect on the virulence operons appears to require the known transcriptional activator of these genes, VirF. This was determined using microarray analysis, quantitative PCR and a search for putative binding sites for FliA. In addition to the FliA regulation of flagellar and plasmid-encoded virulence genes, we studied temperature regulation of these genes. While wild-type cells exhibited increased expression levels of flagellar genes and decreased expression levels of plasmid-encoded virulence genes at 25°C (as compared to 37°C), temperature dependence of gene expression was much reduced in the fliA mutants. We conclude that FliA contributes to the inverse temperature regulation of flagellar and plasmid-encoded virulence genes. We present a network of transcriptional regulation around FlhD/FlhC and FliA.
Keywords: FliA; FlhD/FlhC; Flagellar genes; Plasmid-encoded virulence genes; Microarray analysis; Global regulation; Transcriptional network
Global gene regulation in Yersinia enterocolitica: effect of FliA on the expression levels of flagellar and plasmid-encoded virulence genes by Shelley M. Horne; Birgit M. Prüß (pp. 115-126).
This study describes the involvement of the sigma factor of the flagellar system, FliA, in global gene regulation of Yersinia enterocolitica. In addition to exhibiting a positive effect upon the expression levels of eight class III flagellar operons, FliA also exhibited a negative effect upon the expression levels of four virulence operons that are located on the pYV virulence plasmid. These are yadA, virC, yopQ, and the insertion element ISYen1. While the positive effect on class III flagellar operons by FliA is most likely direct, the negative effect on the virulence operons appears to require the known transcriptional activator of these genes, VirF. This was determined using microarray analysis, quantitative PCR and a search for putative binding sites for FliA. In addition to the FliA regulation of flagellar and plasmid-encoded virulence genes, we studied temperature regulation of these genes. While wild-type cells exhibited increased expression levels of flagellar genes and decreased expression levels of plasmid-encoded virulence genes at 25°C (as compared to 37°C), temperature dependence of gene expression was much reduced in the fliA mutants. We conclude that FliA contributes to the inverse temperature regulation of flagellar and plasmid-encoded virulence genes. We present a network of transcriptional regulation around FlhD/FlhC and FliA.
Keywords: FliA; FlhD/FlhC; Flagellar genes; Plasmid-encoded virulence genes; Microarray analysis; Global regulation; Transcriptional network
Differential regulation of Streptococcus mutans gtfBCD genes in response to copper ions by Pei-Min Chen; Jen-Yang Chen; Jean-San Chia (pp. 127-135).
To persist in the oral cavity, bacteria must be able to tolerate environmental fluctuation, particularly in pH, nutrients, and essential elements. Glucosyltransferases B, C, and D of Streptococcus mutans synthesize glucans, and play essential roles in the sucrose-dependent adhesion of the organism to tooth surfaces. Transcriptions of gtfB, gtfC, and gtfD could be differentially regulated through independent promoters. To test the hypothesis that environmental factors frequently encountered in the dental plaque might serve as effector molecules involved in regulation, transcripts of individual gtfs were identified by reverse transcriptase-polymerase chain reaction assay and confirmed by Northern blot analysis using anti-sense RNA probes. When S. mutans was grown in different medium at low pH, differential regulation of the gtfs was observed. More specifically, the transcription and translational expression of gtfD but not gtfB and gtfC was specifically induced by copper ion (Cu2+). The up-regulation was independent of the Cu2+-transport operon copYAZ. These findings support the involvement of Cu2+ as an effector molecule in the regulation of S. mutans gtfD. Nutrient change dominates influence of pH but not the effect of Cu2+.
Keywords: Streptococcus mutans ; Plaque; Glucosyltransferases; pH; Gene regulation; Cu2+
Differential regulation of Streptococcus mutans gtfBCD genes in response to copper ions by Pei-Min Chen; Jen-Yang Chen; Jean-San Chia (pp. 127-135).
To persist in the oral cavity, bacteria must be able to tolerate environmental fluctuation, particularly in pH, nutrients, and essential elements. Glucosyltransferases B, C, and D of Streptococcus mutans synthesize glucans, and play essential roles in the sucrose-dependent adhesion of the organism to tooth surfaces. Transcriptions of gtfB, gtfC, and gtfD could be differentially regulated through independent promoters. To test the hypothesis that environmental factors frequently encountered in the dental plaque might serve as effector molecules involved in regulation, transcripts of individual gtfs were identified by reverse transcriptase-polymerase chain reaction assay and confirmed by Northern blot analysis using anti-sense RNA probes. When S. mutans was grown in different medium at low pH, differential regulation of the gtfs was observed. More specifically, the transcription and translational expression of gtfD but not gtfB and gtfC was specifically induced by copper ion (Cu2+). The up-regulation was independent of the Cu2+-transport operon copYAZ. These findings support the involvement of Cu2+ as an effector molecule in the regulation of S. mutans gtfD. Nutrient change dominates influence of pH but not the effect of Cu2+.
Keywords: Streptococcus mutans ; Plaque; Glucosyltransferases; pH; Gene regulation; Cu2+
Regulation of citB expression in Bacillus subtilis: integration of multiple metabolic signals in the citrate pool and by the general nitrogen regulatory system by Hans-Matti Blencke; Irene Reif; Fabian M. Commichau; Christian Detsch; Ingrid Wacker; Holger Ludwig; Jörg Stülke (pp. 136-146).
The tricarboxylic acid (TCA) cycle is one of the major routes of carbon catabolism in Bacillus subtilis. The syntheses of the enzymes performing the initial reactions of the cycle, citrate synthase, and aconitase, are synergistically repressed by rapidly metabolizable carbon sources and glutamine. This regulation involves the general transcription factor CcpA and the specific repressor CcpC. In this study, we analyzed the expression and intracellular localization of CcpC. The synthesis of citrate, the effector of CcpC, requires acetyl-CoA. This metabolite is located at a branching point in metabolism. It can be converted to acetate in overflow metabolism or to citrate. Manipulations of the fate of acetyl-CoA revealed that efficient citrate synthesis is required for the expression of the citB gene encoding aconitase and that control of the two pathways utilizing acetyl-CoA converges in the control of citrate synthesis for the induction of the TCA cycle. The citrate pool seems also to be controlled by arginine catabolism. The presence of arginine results in a severe CcpC-dependent repression of citB. In addition to regulators involved in sensing the carbon status of the cell, the pleiotropic nitrogen-related transcription factor, TnrA, activates citB transcription in the absence of glutamine.
Keywords: Bacillus subtilis ; Aconitase; Catabolite repression; Nitrogen regulation; CcpA; TnrA
Regulation of citB expression in Bacillus subtilis: integration of multiple metabolic signals in the citrate pool and by the general nitrogen regulatory system by Hans-Matti Blencke; Irene Reif; Fabian M. Commichau; Christian Detsch; Ingrid Wacker; Holger Ludwig; Jörg Stülke (pp. 136-146).
The tricarboxylic acid (TCA) cycle is one of the major routes of carbon catabolism in Bacillus subtilis. The syntheses of the enzymes performing the initial reactions of the cycle, citrate synthase, and aconitase, are synergistically repressed by rapidly metabolizable carbon sources and glutamine. This regulation involves the general transcription factor CcpA and the specific repressor CcpC. In this study, we analyzed the expression and intracellular localization of CcpC. The synthesis of citrate, the effector of CcpC, requires acetyl-CoA. This metabolite is located at a branching point in metabolism. It can be converted to acetate in overflow metabolism or to citrate. Manipulations of the fate of acetyl-CoA revealed that efficient citrate synthesis is required for the expression of the citB gene encoding aconitase and that control of the two pathways utilizing acetyl-CoA converges in the control of citrate synthesis for the induction of the TCA cycle. The citrate pool seems also to be controlled by arginine catabolism. The presence of arginine results in a severe CcpC-dependent repression of citB. In addition to regulators involved in sensing the carbon status of the cell, the pleiotropic nitrogen-related transcription factor, TnrA, activates citB transcription in the absence of glutamine.
Keywords: Bacillus subtilis ; Aconitase; Catabolite repression; Nitrogen regulation; CcpA; TnrA
Molecular characterisation of ABC transporter type FtsE and FtsX proteins of Mycobacterium tuberculosis by Mushtaq Ahmad Mir; Haryadi S. Rajeswari; Usha Veeraraghavan; Parthasarathi Ajitkumar (pp. 147-158).
Elicitation of drug resistance and various survival strategies inside host macrophages have been the hallmarks of Mycobacterium tuberculosis as a successful pathogen. ATP Binding Cassette (ABC) transporter type proteins are known to be involved in the efflux of drugs in bacterial and mammalian systems. FtsE, an ABC transporter type protein, in association with the integral membrane protein FtsX, is involved in the assembly of potassium ion transport proteins and probably of cell division proteins as well, both of which being relevant to tubercle bacillus. In this study, we cloned ftsE gene of M. tuberculosis, overexpressed and purified. The recombinant MtFtsE-6xHis protein and the native MtFtsE protein were found localized on the membrane of E. coli and M. tuberculosis cells, respectively. MtFtsE-6xHis protein showed ATP binding in vitro, for which the K42 residue in the Walker A motif was found essential. While MtFtsE-6xHis protein could partially complement growth defect of E. coli ftsE temperature-sensitive strain MFT1181, co-expression of MtFtsE and MtFtsX efficiently complemented the growth defect, indicating that the MtFtsE and MtFtsX proteins might be performing an associated function. MtFtsE and MtFtsX-6xHis proteins were found to exist as a complex on the membrane of E. coli cells co-expressing the two proteins.
Keywords: FtsE; FtsX; ABC transporter type protein; Walker A motif; ATP binding; Mycobacterium tuberculosis ; cell division
Molecular characterisation of ABC transporter type FtsE and FtsX proteins of Mycobacterium tuberculosis by Mushtaq Ahmad Mir; Haryadi S. Rajeswari; Usha Veeraraghavan; Parthasarathi Ajitkumar (pp. 147-158).
Elicitation of drug resistance and various survival strategies inside host macrophages have been the hallmarks of Mycobacterium tuberculosis as a successful pathogen. ATP Binding Cassette (ABC) transporter type proteins are known to be involved in the efflux of drugs in bacterial and mammalian systems. FtsE, an ABC transporter type protein, in association with the integral membrane protein FtsX, is involved in the assembly of potassium ion transport proteins and probably of cell division proteins as well, both of which being relevant to tubercle bacillus. In this study, we cloned ftsE gene of M. tuberculosis, overexpressed and purified. The recombinant MtFtsE-6xHis protein and the native MtFtsE protein were found localized on the membrane of E. coli and M. tuberculosis cells, respectively. MtFtsE-6xHis protein showed ATP binding in vitro, for which the K42 residue in the Walker A motif was found essential. While MtFtsE-6xHis protein could partially complement growth defect of E. coli ftsE temperature-sensitive strain MFT1181, co-expression of MtFtsE and MtFtsX efficiently complemented the growth defect, indicating that the MtFtsE and MtFtsX proteins might be performing an associated function. MtFtsE and MtFtsX-6xHis proteins were found to exist as a complex on the membrane of E. coli cells co-expressing the two proteins.
Keywords: FtsE; FtsX; ABC transporter type protein; Walker A motif; ATP binding; Mycobacterium tuberculosis ; cell division
Gene cloning, expression and functional characterization of an acyl carrier protein AcpV from Vibrio anguillarum by Qin Liu; Yue Ma; Lingyun Zhou; Yuanxing Zhang (pp. 159-163).
Acyl carrier protein (ACP) is a small acidic protein that acts as an essential cofactor in many biosynthetic pathways depending on acyl transfer reactions. In this work, a Vibrio anguillarum ACP encoding gene, acpV, was first cloned from the chromosome of a virulent V. anguillarum strain MVM425. acpV was over-expressed in Escherichia coli and the resultant protein AcpV was purified. The purified AcpV was incubated with purified phosphopantetheinyl transferase (PPtase) in the presence of CoA to assay the 4′-phosphopantetheinylation of AcpV in vitro; and on the other hand, the acpV gene was co-expressed with PPtase-encoding gene in E. coli to examine the 4′-phosphopantetheinylation of AcpV in vivo. Our results suggested that acpV encoded a functional ACP of V. anguillarum, which can be 4′-phosphopantetheinylated well by AcpS-type PPtase (E. coli AcpS) both in vitro and in vivo, but cannot serve as a good substrate for Sfp-type PPtase (V. anguillarum AngD).
Keywords: Vibrio anguillarrum ; Acyl carrier protein; Phosphopantetheinyl transferase
Gene cloning, expression and functional characterization of an acyl carrier protein AcpV from Vibrio anguillarum by Qin Liu; Yue Ma; Lingyun Zhou; Yuanxing Zhang (pp. 159-163).
Acyl carrier protein (ACP) is a small acidic protein that acts as an essential cofactor in many biosynthetic pathways depending on acyl transfer reactions. In this work, a Vibrio anguillarum ACP encoding gene, acpV, was first cloned from the chromosome of a virulent V. anguillarum strain MVM425. acpV was over-expressed in Escherichia coli and the resultant protein AcpV was purified. The purified AcpV was incubated with purified phosphopantetheinyl transferase (PPtase) in the presence of CoA to assay the 4′-phosphopantetheinylation of AcpV in vitro; and on the other hand, the acpV gene was co-expressed with PPtase-encoding gene in E. coli to examine the 4′-phosphopantetheinylation of AcpV in vivo. Our results suggested that acpV encoded a functional ACP of V. anguillarum, which can be 4′-phosphopantetheinylated well by AcpS-type PPtase (E. coli AcpS) both in vitro and in vivo, but cannot serve as a good substrate for Sfp-type PPtase (V. anguillarum AngD).
Keywords: Vibrio anguillarrum ; Acyl carrier protein; Phosphopantetheinyl transferase