The ImpX transporters of the drug/metabolite transporter superfamily were first proposed to transport riboflavin (RF; vitamin B2) based on findings of a cis-regulatory RNA element responding to flavin mononucleotide (an FMN riboswitch). Bdellovibrio exovorous JSS has a homolog belonging to this superfamily. It has 10 TMSs and shows 30% identity to the previously characterized ImpX transporter from Fusobacterium nucleatum. However, the ImpX homolog is not regulated by an FMN-riboswitch. In order to test the putative function of the ImpX homolog from B. exovorous (BexImpX), we cloned and heterologously expressed its gene. We used functional complementation, growth inhibition experiments, direct uptake experiments and inhibition studies, suggesting a high degree of specificity for RF uptake. The EC50 for growth with RF was estimated to be in the range 0.5–1 µM, estimated from the half-maximal RF concentration supporting the growth of a RF auxotrophic Escherichia coli strain, but the Khalf for RF uptake was 20 µM. Transport experiments suggested that the energy source is the proton motive force but that NaCl stimulates uptake. Thus, members of the ImpX family members are capable of RF uptake, not only in RF prototrophic species such as F.  nucleatum, but also in the B2 auxotrophic species, B. exovorous.

1.
Cao
H
,
He
S
,
Wang
H
,
Hou
S
,
Lu
L
,
Yang
X
.
Bdellovibrios, potential biocontrol bacteria against pathogenic Aeromonas hydrophila
.
Vet Microbiol
.
2012
Jan
;
154
(
3-4
):
413
8
.
[PubMed]
0378-1135
2.
Davidov
Y
,
Friedjung
A
,
Jurkevitch
E
.
Structure analysis of a soil community of predatory bacteria using culture-dependent and culture-independent methods reveals a hitherto undetected diversity of Bdellovibrio-and-like organisms
.
Environ Microbiol
.
2006
a
Sep
;
8
(
9
):
1667
73
.
[PubMed]
1462-2912
3.
Davidov
Y
,
Huchon
D
,
Koval
SF
,
Jurkevitch
E
.
A new alpha-proteobacterial clade of Bdellovibrio-like predators: implications for the mitochondrial endosymbiotic theory
.
Environ Microbiol
.
2006
b
Dec
;
8
(
12
):
2179
88
.
[PubMed]
1462-2912
4.
Deka
RK
,
Brautigam
CA
,
Biddy
BA
,
Liu
WZ
,
Norgard
MV
.
Evidence for an ABC-type riboflavin transporter system in pathogenic spirochetes
.
MBio
.
2013
Feb
;
4
(
1
):
e00615
12
.
[PubMed]
2150-7511
5.
Edgar
RC
.
MUSCLE: multiple sequence alignment with high accuracy and high throughput
.
Nucleic Acids Res
.
2004
Mar
;
32
(
5
):
1792
7
.
[PubMed]
0305-1048
6.
Fassbinder
F
,
Kist
M
,
Bereswill
S
.
Structural and functional analysis of the riboflavin synthesis genes encoding GTP cyclohydrolase II (ribA), DHBP synthase (ribBA), riboflavin synthase (ribC), and riboflavin deaminase/reductase (ribD) from Helicobacter pylori strain P1
.
FEMS Microbiol Lett
.
2000
Oct
;
191
(
2
):
191
7
.
[PubMed]
0378-1097
7.
Fischer
M
,
Bacher
A
.
Biosynthesis of Riboflavin
.
Ecosal Plus
.
2010
Sep
;
4
(
1
):
4
.
[PubMed]
2324-6200
8.
García Angulo
VA
,
Bonomi
HR
,
Posadas
DM
,
Serer
MI
,
Torres
AG
,
Zorreguieta
Á
, et al.
.
Identification and characterization of RibN, a novel family of riboflavin transporters from Rhizobium leguminosarum and other proteobacteria
.
J Bacteriol
.
2013
Oct
;
195
(
20
):
4611
9
.
[PubMed]
0021-9193
9.
Garcia Costas
AM
,
Poudel
S
,
Miller
AF
,
Schut
GJ
,
Ledbetter
RN
,
Fixen
KR
, et al.
.
Defining Electron Bifurcation in the Electron-Transferring Flavoprotein Family
.
J Bacteriol
.
2017
Oct
;
199
(
21
):
199
.
[PubMed]
0021-9193
10.
García-Angulo
VA
.
Overlapping riboflavin supply pathways in bacteria
.
Crit Rev Microbiol
.
2017
Mar
;
43
(
2
):
196
209
.
[PubMed]
1040-841X
11.
Gordon
RF
,
Stein
MA
,
Diedrich
DL
.
Heat shock-induced axenic growth of Bdellovibrio bacteriovorus
.
J Bacteriol
.
1993
Apr
;
175
(
7
):
2157
61
.
[PubMed]
0021-9193
12.
Gutiérrez-Preciado
A
,
Torres
AG
,
Merino
E
,
Bonomi
HR
,
Goldbaum
FA
,
García-Angulo
VA
.
Extensive Identification of Bacterial Riboflavin Transporters and Their Distribution across Bacterial Species
.
PLoS One
.
2015
May
;
10
(
5
):
e0126124
.
[PubMed]
1932-6203
13.
Heidari Tajabadi
F
,
Medrano-Soto
A
,
Ahmadzadeh
M
,
Salehi Jouzani
G
,
Saier
MH
Jr
.
Comparative Analyses of Transport Proteins Encoded within the Genomes of Bdellovibrio bacteriovorus HD100 and Bdellovibrio exovorus JSS
.
J Mol Microbiol Biotechnol
.
2017
;
27
(
6
):
332
49
.
[PubMed]
1464-1801
14.
Jankowitsch
F
,
Schwarz
J
,
Rückert
C
,
Gust
B
,
Szczepanowski
R
,
Blom
J
, et al.
.
Genome sequence of the bacterium Streptomyces davawensis JCM 4913 and heterologous production of the unique antibiotic roseoflavin
.
J Bacteriol
.
2012
Dec
;
194
(
24
):
6818
27
.
[PubMed]
0021-9193
15.
Jurkevitch
E
.
Isolation and classification of bdellovibrio and like organisms.
Curr Protoc Microbiol
2006
;Chapter 7:Unit 7B 1.
16.
Karpowich
NK
,
Song
J
,
Wang
DN
.
An Aromatic Cap Seals the Substrate Binding Site in an ECF-Type S Subunit for Riboflavin
.
J Mol Biol
.
2016
Jul
;
428
(
15
):
3118
30
.
[PubMed]
0022-2836
17.
Koval
SF
,
Hynes
SH
,
Flannagan
RS
,
Pasternak
Z
,
Davidov
Y
,
Jurkevitch
E
.
Bdellovibrio exovorus sp. nov., a novel predator of Caulobacter crescentus
.
Int J Syst Evol Microbiol
.
2013
Jan
;
63
(
Pt 1
):
146
51
.
[PubMed]
1466-5026
18.
Langer
S
,
Hashimoto
M
,
Hobl
B
,
Mathes
T
,
Mack
M
.
Flavoproteins are potential targets for the antibiotic roseoflavin in Escherichia coli
.
J Bacteriol
.
2013
Sep
;
195
(
18
):
4037
45
.
[PubMed]
0021-9193
19.
Lefort
V
,
Longueville
JE
,
Gascuel
O
.
SMS: Smart Model Selection in PhyML
.
Mol Biol Evol
.
2017
Sep
;
34
(
9
):
2422
4
.
[PubMed]
0737-4038
20.
Liu
LK
,
Becker
DF
,
Tanner
JJ
.
Structure, function, and mechanism of proline utilization A (PutA)
.
Arch Biochem Biophys
.
2017
Oct
;
632
:
142
57
.
[PubMed]
0003-9861
21.
Livshits
VA
,
Zakataeva
NP
,
Aleshin
VV
,
Vitushkina
MV
.
Identification and characterization of the new gene rhtA involved in threonine and homoserine efflux in Escherichia coli
.
Res Microbiol
.
2003
Mar
;
154
(
2
):
123
35
.
[PubMed]
0923-2508
22.
Magnúsdóttir
S
,
Ravcheev
D
,
de Crécy-Lagard
V
,
Thiele
I
.
Systematic genome assessment of B-vitamin biosynthesis suggests co-operation among gut microbes
.
Front Genet
.
2015
Apr
;
6
:
148
.
[PubMed]
1664-8021
23.
McAnulty
MJ
,
Wood
TK
.
YeeO from Escherichia coli exports flavins
.
Bioengineered
.
2014
;
5
(
6
):
386
92
.
[PubMed]
2165-5979
24.
McNeely
D
,
Chanyi
RM
,
Dooley
JS
,
Moore
JE
,
Koval
SF
.
Biocontrol of Burkholderia cepacia complex bacteria and bacterial phytopathogens by Bdellovibrio bacteriovorus
.
Can J Microbiol
.
2017
Apr
;
63
(
4
):
350
8
.
[PubMed]
0008-4166
25.
Mironov
AS
,
Gusarov
I
,
Rafikov
R
,
Lopez
LE
,
Shatalin
K
,
Kreneva
RA
, et al.
.
Sensing small molecules by nascent RNA: a mechanism to control transcription in bacteria
.
Cell
.
2002
Nov
;
111
(
5
):
747
56
.
[PubMed]
0092-8674
26.
Munro
AW
,
Noble
MA
.
Fluorescence analysis of flavoproteins
.
Methods Mol Biol
.
1999
;
131
:
25
48
.
[PubMed]
1064-3745
27.
Ott
E
,
Stolz
J
,
Lehmann
M
,
Mack
M
.
The RFN riboswitch of Bacillus subtilis is a target for the antibiotic roseoflavin produced by Streptomyces davawensis
.
RNA Biol
.
2009
Jul-Aug
;
6
(
3
):
276
80
.
[PubMed]
1547-6286
28.
Pedrolli
D
,
Langer
S
,
Hobl
B
,
Schwarz
J
,
Hashimoto
M
,
Mack
M
.
The ribB FMN riboswitch from Escherichia coli operates at the transcriptional and translational level and regulates riboflavin biosynthesis
.
FEBS J
.
2015
Aug
;
282
(
16
):
3230
42
.
[PubMed]
1742-464X
29.
Plegaria
JS
,
Sutter
M
,
Ferlez
B
,
Aussignargues
C
,
Niklas
J
,
Poluektov
OG
, et al.
.
Structural and Functional Characterization of a Short-Chain Flavodoxin Associated with a Noncanonical 1,2-Propanediol Utilization Bacterial Microcompartment
.
Biochemistry
.
2017
Oct
;
56
(
42
):
5679
90
.
[PubMed]
0006-2960
30.
Rodionov
DA
,
Hebbeln
P
,
Eudes
A
,
ter Beek
J
,
Rodionova
IA
,
Erkens
GB
, et al.
.
A novel class of modular transporters for vitamins in prokaryotes
.
J Bacteriol
.
2009
Jan
;
191
(
1
):
42
51
.
[PubMed]
0021-9193
31.
Rodionova
IA
,
Li
X
,
Plymale
AE
,
Motamedchaboki
K
,
Konopka
AE
,
Romine
MF
, et al.
.
Genomic distribution of B-vitamin auxotrophy and uptake transporters in environmental bacteria from the Chloroflexi phylum
.
Environ Microbiol Rep
.
2015
Apr
;
7
(
2
):
204
10
.
[PubMed]
1758-2229
32.
Stietz
MS
,
Lopez
C
,
Osifo
O
,
Tolmasky
ME
,
Cardona
ST
.
Evaluation of the electron transfer flavoprotein as an antibacterial target in Burkholderia cenocepacia
.
Can J Microbiol
.
2017
Oct
;
63
(
10
):
857
63
.
[PubMed]
0008-4166
33.
Sun
EI
,
Rodionov
DA
.
Computational analysis of riboswitch-based regulation
.
Biochim Biophys Acta
.
2014
Oct
;
1839
(
10
):
900
7
.
[PubMed]
0006-3002
34.
Vitreschak
AG
,
Rodionov
DA
,
Mironov
AA
,
Gelfand
MS
.
Regulation of riboflavin biosynthesis and transport genes in bacteria by transcriptional and translational attenuation
.
Nucleic Acids Res
.
2002
Jul
;
30
(
14
):
3141
51
.
[PubMed]
0305-1048
35.
Yamamoto
K
,
Nonaka
G
,
Ozawa
T
,
Takumi
K
,
Ishihama
A
.
Induction of the Escherichia coli yijE gene expression by cystine
.
Biosci Biotechnol Biochem
.
2015
;
79
(
2
):
218
22
.
[PubMed]
0916-8451
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