Characterization of a secondary uptake system for ?-glutamate in Corynebacterium glutamicum

ABSTRACT Gram-positive soil bacterium Corynebacterium glutamicumuses the compatible solutes glycine betaine, proline, and ectoine for protection against hyperosmotic shock. Osmoregulated glycine betaine carrier BetP and proline permease PutP have been previously characterized; we have identified and characterized two additional osmoregulated secondary transporters for compatible solutes in C. glutamicum, namely, the proline/ectoine carrier, ProP, and the ectoine/glycine betaine/proline carrier, EctP. A ΔbetP ΔputP ΔproP ΔectP mutant was unable to respond to hyperosmotic stress, indicating that no additional uptake system for these compatible solutes is present. Osmoregulated ProP consists of 504 residues and preferred proline (Km , 48 μM) to ectoine (Km , 132 μM). The proP gene could not be expressed from its own promoter in C. glutamicum; however, expression was observed in Escherichia coli. ProP belongs to the major facilitator superfamily, whereas EctP, together with the betaine carrier, BetP, is a member of a newly established subfamily of the sodium/solute symporter superfamily. The constitutively expressed ectP codes for a 615-residue transporter. EctP preferred ectoine (Km , 63 μM) to betaine (Km , 333 μM) and proline (Km , 1,200 μM). Its activity was regulated by the external osmolality. The related betaine transporter, BetP, could be activated directly by altering the membrane state with local anesthetics, but this was not the case for EctP. Furthermore, the onset of osmotic activation was virtually instantaneous for BetP, whereas it took about 10 s for EctP.

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Cloning of the trkAH gene cluster and characterization of the Trk K+-uptake system of Vibrio alginolyticus

Microbiology ◽

10.1099/00221287-144-8-2281 ◽

1998 ◽

Vol 144 (8) ◽

pp. 2281-2289 ◽

Cited By ~ 30

Author(s):

T. Nakamura ◽

N. Yamamuro ◽

S. Stumpe ◽

T. Unemoto ◽

E. P. Bakker

Keyword(s):

Gene Cluster ◽

Vibrio Alginolyticus ◽

Uptake System ◽

K Uptake

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Identification and application of a different glucose uptake system that functions as an alternative to the phosphotransferase system in Corynebacterium glutamicum

Applied Microbiology and Biotechnology ◽

10.1007/s00253-011-3210-x ◽

2011 ◽

Vol 90 (4) ◽

pp. 1443-1451 ◽

Cited By ~ 51

Author(s):

Masato Ikeda ◽

Yuta Mizuno ◽

Shin-ichi Awane ◽

Masahiro Hayashi ◽

Satoshi Mitsuhashi ◽

...

Keyword(s):

Glucose Uptake ◽

Corynebacterium Glutamicum ◽

Uptake System ◽

Phosphotransferase System

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Characterization of a 24-kb plasmid pCGR2 newly isolated from Corynebacterium glutamicum

Applied Microbiology and Biotechnology ◽

10.1007/s00253-010-2701-5 ◽

2010 ◽

Vol 87 (5) ◽

pp. 1855-1866 ◽

Cited By ~ 10

Author(s):

Yoshiki Tsuchida ◽

Sakurako Kimura ◽

Nobuaki Suzuki ◽

Masayuki Inui ◽

Hideaki Yukawa

Keyword(s):

Corynebacterium Glutamicum

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Characterization of simultaneous uptake of xylose and glucose in Caldicellulosiruptor kronotskyensis for optimal hydrogen production

Biotechnology for Biofuels ◽

10.1186/s13068-021-01938-6 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Thitiwut Vongkampang ◽

Krishnan Sreenivas ◽

Jonathan Engvall ◽

Carl Grey ◽

Ed W. J. van Niel

Keyword(s):

Glucose Uptake ◽

Growth Patterns ◽

Sugar Uptake ◽

Uptake System ◽

Sugar Mixtures ◽

Xylose Uptake ◽

Growth Profiles ◽

High Concentrations ◽

Xylose Transporter

Abstract Background Caldicellulosiruptor kronotskyensis has gained interest for its ability to grow on various lignocellulosic biomass. The aim of this study was to investigate the growth profiles of C. kronotskyensis in the presence of mixtures of glucose–xylose. Recently, we characterized a diauxic-like pattern for C. saccharolyticus on lignocellulosic sugar mixtures. In this study, we aimed to investigate further whether C. kronotskyensis has adapted to uptake glucose in the disaccharide form (cellobiose) rather than the monosaccharide (glucose). Results Interestingly, growth of C. kronotskyensis on glucose and xylose mixtures did not display diauxic-like growth patterns. Closer investigation revealed that, in contrast to C. saccharolyticus, C. kronotskyensis does not possess a second uptake system for glucose. Both C. saccharolyticus and C. kronotskyensis share the characteristics of preferring xylose over glucose. Growth on xylose was twice as fast (μmax = 0.57 h−1) as on glucose (μmax = 0.28 h−1). A study of the sugar uptake was made with different glucose–xylose ratios to find a kinetic relationship between the two sugars for transport into the cell. High concentrations of glucose inhibited xylose uptake and vice versa. The inhibition constants were estimated to be KI,glu = 0.01 cmol L−1 and KI,xyl = 0.001 cmol L−1, hence glucose uptake was more severely inhibited by xylose uptake. Bioinformatics analysis could not exclude that C. kronotskyensis possesses more than one transporter for glucose. As a next step it was investigated whether glucose uptake by C. kronotskyensis improved in the form of cellobiose. Indeed, cellobiose is taken up faster than glucose; nevertheless, the growth rate on each sugar remained similar. Conclusions C. kronotskyensis possesses a xylose transporter that might take up glucose at an inferior rate even in the absence of xylose. Alternatively, glucose can be taken up in the form of cellobiose, but growth performance is still inferior to growth on xylose. Therefore, we propose that the catabolism of C. kronotskyensis has adapted more strongly to pentose rather than hexose, thereby having obtained a specific survival edge in thermophilic lignocellulosic degradation communities.

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Crystal Structure and Biochemical Characterization of Tetrahydrodipicolinate N-Succinyltransferase from Corynebacterium glutamicum

Journal of Agricultural and Food Chemistry ◽

10.1021/acs.jafc.5b04785 ◽

2015 ◽

Vol 63 (49) ◽

pp. 10641-10646 ◽

Cited By ~ 1

Author(s):

Hye-Young Sagong ◽

Kyung-Jin Kim

Keyword(s):

Crystal Structure ◽

Corynebacterium Glutamicum ◽

Biochemical Characterization

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Characterization of the monoamine uptake system in catecholamine storage vesicles isolated from a pheochromocytoma taken from a child

Biochemical Pharmacology ◽

10.1016/0006-2952(84)90662-2 ◽

1984 ◽

Vol 33 (14) ◽

pp. 2245-2252 ◽

Cited By ~ 4

Author(s):

M.P. Roisin ◽

M.F. Isambert ◽

J.P. Henry ◽

M. Guillot ◽

G. Lenoir

Keyword(s):

Uptake System ◽

Monoamine Uptake ◽

Storage Vesicles ◽

Catecholamine Storage

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Characterization of a Sinorhizobium melilotiATP-Binding Cassette Histidine Transporter Also Involved in Betaine and Proline Uptake

Journal of Bacteriology ◽

10.1128/jb.182.13.3717-3725.2000 ◽

2000 ◽

Vol 182 (13) ◽

pp. 3717-3725 ◽

Cited By ~ 31

Author(s):

Eric Boncompagni ◽

Laurence Dupont ◽

Tam Mignot ◽

Magne Østeräs ◽

Annie Lambert ◽

...

Keyword(s):

Glycine Betaine ◽

Sinorhizobium Meliloti ◽

Binding Protein ◽

Compatible Solutes ◽

Site Directed Mutagenesis ◽

Uptake System ◽

Periplasmic Binding Protein ◽

Gene Encoding ◽

Inhibition Of Growth

ABSTRACT The symbiotic soil bacterium Sinorhizobium melilotiuses the compatible solutes glycine betaine and proline betaine for both protection against osmotic stress and, at low osmolarities, as an energy source. A PCR strategy based on conserved domains in components of the glycine betaine uptake systems from Escherichia coli(ProU) and Bacillus subtilis (OpuA and OpuC) allowed us to identify a highly homologous ATP-binding cassette (ABC) binding protein-dependent transporter in S. meliloti. This system was encoded by three genes (hutXWV) of an operon which also contained a fourth gene (hutH2) encoding a putative histidase, which is an enzyme involved in the first step of histidine catabolism. Site-directed mutagenesis of the gene encoding the periplasmic binding protein (hutX) and of the gene encoding the cytoplasmic ATPase (hutV) was done to study the substrate specificity of this transporter and its contribution in betaine uptake. These mutants showed a 50% reduction in high-affinity uptake of histidine, proline, and proline betaine and about a 30% reduction in low-affinity glycine betaine transport. When histidine was used as a nitrogen source, a 30% inhibition of growth was observed inhut mutants (hutX and hutH2). Expression analysis of the hut operon determined using ahutX-lacZ fusion revealed induction by histidine, but not by salt stress, suggesting this uptake system has a catabolic role rather than being involved in osmoprotection. To our knowledge, Hut is the first characterized histidine ABC transporter also involved in proline and betaine uptake.

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