Natural and synthetic variants of the tricarboxylic acid cycle in cyanobacteria: Introduction of the GABA Shunt into Synechococcus sp. PCC 7002

Shuyi Zhang, Xiao Qian, Shannon Chang, G Charles Dismukes, Donald A. Bryant

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

For nearly half a century, it was believed that cyanobacteria had an incomplete tricarboxylic acid (TCA) cycle, because 2-oxoglutarate dehydrogenase (2-OGDH) was missing. Recently, a bypass route via succinic semialdehyde (SSA), which utilizes 2-oxoglutarate decarboxylase (OgdA) and succinic semialdehyde dehydrogenase (SsaD) to convert 2-oxoglutarate (2-OG) into succinate, was identified, thus completing the TCA cycle in most cyanobacteria. In addition to the recently characterized glyoxylate shunt that occurs in a few of cyanobacteria, the existence of a third variant of the TCA cycle connecting these metabolites, the γ-aminobutyric acid (GABA) shunt, was considered to be ambiguous because the GABA aminotransferase is missing in many cyanobacteria. In this study we isolated and biochemically characterized the enzymes of the GABA shunt. We show that N-acetylornithine aminotransferase (ArgD) can function as a GABA aminotransferase and that, together with glutamate decarboxylase (GadA), it can complete a functional GABA shunt. To prove the connectivity between the OgdA/SsaD bypass and the GABA shunt, the gadA gene from Synechocystis sp. PCC 6803 was heterologously expressed in Synechococcus sp. PCC 7002, which naturally lacks this enzyme. Metabolite profiling of seven Synechococcus sp. PCC 7002 mutant strains related to these two routes to succinate were investigated and proved the functional connectivity. Metabolite profiling also indicated that, compared to the OgdA/SsaD shunt, the GABA shunt was less efficient in converting 2-OG to SSA in Synechococcus sp. PCC 7002. The metabolic profiling study of these two TCA cycle variants provides new insights into carbon metabolism as well as evolution of the TCA cycle in cyanobacteria.

Original languageEnglish
Article number1972
JournalFrontiers in Microbiology
Volume7
Issue numberDEC
DOIs
Publication statusPublished - 2016

Fingerprint

2-oxoglutarate decarboxylase
Synechococcus
Citric Acid Cycle
Cyanobacteria
Succinate-Semialdehyde Dehydrogenase
gamma-Aminobutyric Acid
4-Aminobutyrate Transaminase
Succinic Acid
Ketoglutarate Dehydrogenase Complex
Aminobutyrates
Synechocystis
Glutamate Decarboxylase
Enzymes
Carbon
Genes

Keywords

  • 2-oxoglutaric acid
  • Cyanobacteria
  • GABA shunt
  • Photosynthesis
  • Succinic acid semialdehyde
  • Synechococcus sp. PCC 7002
  • Synechocystis sp. PCC 6803
  • TCA cycle

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Natural and synthetic variants of the tricarboxylic acid cycle in cyanobacteria : Introduction of the GABA Shunt into Synechococcus sp. PCC 7002. / Zhang, Shuyi; Qian, Xiao; Chang, Shannon; Dismukes, G Charles; Bryant, Donald A.

In: Frontiers in Microbiology, Vol. 7, No. DEC, 1972, 2016.

Research output: Contribution to journalArticle

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