Physiological Response of <i>Corynebacterium glutamicum</i> to Indole

• Main Authors: Tatjana Walter, Kareen H. Veldmann, Susanne Götker, Tobias Busche, Christian Rückert, Arman Beyraghdar Kashkooli, Jannik Paulus, Katarina Cankar, Volker F. Wendisch
• Format: Article
• Language: English
• Published: MDPI AG 2020-12-01
• Series: Microorganisms
• Subjects: <i>Corynebacterium glutamicum</i>; amino acids; indole; adaptive laboratory evolution; iron homeostasis; oxidative stress
• Online Access: https://www.mdpi.com/2076-2607/8/12/1945

The aromatic heterocyclic compound indole is widely spread in nature. Due to its floral odor indole finds application in dairy, flavor, and fragrance products. Indole is an inter- and intracellular signaling molecule influencing cell division, sporulation, or virulence in some bacteria that synthesize it from tryptophan by tryptophanase. <i>Corynebacterium glutamicum</i> that is used for the industrial production of amino acids including tryptophan lacks tryptophanase. To test if indole is metabolized by <i>C. glutamicum</i> or has a regulatory role, the physiological response to indole by this bacterium was studied. As shown by RNAseq analysis, indole, which inhibited growth at low concentrations, increased expression of genes involved in the metabolism of iron, copper, and aromatic compounds. In part, this may be due to iron reduction as indole was shown to reduce Fe³⁺ to Fe²⁺ in the culture medium. Mutants with improved tolerance to indole were selected by adaptive laboratory evolution. Among the mutations identified by genome sequencing, mutations in three transcriptional regulator genes were demonstrated to be causal for increased indole tolerance. These code for the regulator of iron homeostasis DtxR, the regulator of oxidative stress response RosR, and the hitherto uncharacterized Cg3388. Gel mobility shift analysis revealed that Cg3388 binds to the intergenic region between its own gene and the <i>iolT2-rhcM2D2</i> operon encoding inositol uptake system IolT2, maleylacetate reductase, and catechol 1,2-dioxygenase. Increased RNA levels of <i>rhcM2</i> in a <i>cg3388</i> deletion strain indicated that Cg3388 acts as repressor. Indole, hydroquinone, and 1,2,4-trihydroxybenzene may function as inducers of the <i>iolT2-rhcM2D2</i> operon in vivo as they interfered with DNA binding of Cg3388 at physiological concentrations in vitro. Cg3388 was named IhtR.