Transcriptome analysis and anaerobic C4-dicarboxylate transport in Actinobacillus succinogenes

Mi Na Rhie, Byeonghyeok Park, Hyeok Jin Ko, In-Geol Choi, Ok Bin Kim

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

A global transcriptome analysis of the natural succinate producer Actinobacillus succinogenes revealed that 353 genes were differentially expressed when grown on various carbon and energy sources, which were categorized into six functional groups. We then analyzed the expression pattern of 37 potential C4-dicarboxylate transporters in detail. A total of six transporters were considered potential fumarate transporters: three transporters, Asuc_1999 (Dcu), Asuc_0304 (DASS), and Asuc_0270-0273 (TRAP), were constitutively expressed, whereas three others, Asuc_1568 (DASS), Asuc_1482 (DASS), and Asuc_0142 (Dcu), were differentially expressed during growth on fumarate. Transport assays under anaerobic conditions with [14C]fumarate and [14C]succinate were performed to experimentally verify that A. succinogenes possesses multiple C4-dicarboxlayte transport systems with different substrate affinities. Upon uptake of 5 mmol/L fumarate, the systems had substrate specificity for fumarate, oxaloacetate, and malate, but not for succinate. Uptake was optimal at pH 7, and was dependent on both proton and sodium gradients. Asuc_1999 was suspected to be a major C4-dicarboxylate transporter because of its noticeably high and constitutive expression. An Asuc_1999 deletion (∆1999) decreased fumarate uptake significantly at approximately 5 mmol/L fumarate, which was complemented by the introduction of Asuc_1999. Asuc_1999 expressed in Escherichia coli catalyzed fumarate uptake at a level of 21.6 μmol·gDW−1·min−1. These results suggest that C4-dicarboxylate transport in A. succinogenes is mediated by multiple transporters, which transport various types and concentrations of C4-dicarboxylates.

Original languageEnglish
Article numbere00565
JournalMicrobiologyOpen
Volume7
Issue number3
DOIs
Publication statusPublished - 2018 Jun 1

Fingerprint

Actinobacillus
Fumarates
Gene Expression Profiling
Dicarboxylic Acid Transporters
Succinic Acid
Oxaloacetic Acid
Substrate Specificity
Protons
Carbon
Sodium
Escherichia coli

Keywords

  • Actinobacillus succinogenes
  • C-dicarboxylate transport
  • fumarate
  • transcriptome analysis

ASJC Scopus subject areas

  • Microbiology

Cite this

Transcriptome analysis and anaerobic C4-dicarboxylate transport in Actinobacillus succinogenes. / Rhie, Mi Na; Park, Byeonghyeok; Ko, Hyeok Jin; Choi, In-Geol; Kim, Ok Bin.

In: MicrobiologyOpen, Vol. 7, No. 3, e00565, 01.06.2018.

Research output: Contribution to journalArticle

Rhie, Mi Na ; Park, Byeonghyeok ; Ko, Hyeok Jin ; Choi, In-Geol ; Kim, Ok Bin. / Transcriptome analysis and anaerobic C4-dicarboxylate transport in Actinobacillus succinogenes. In: MicrobiologyOpen. 2018 ; Vol. 7, No. 3.
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