Dataset Information

Effect of Trichococcus flocculiformis on a Co-culture of Syntrophomonas wolfei and Methanospirillum hungatei

ABSTRACT: The study investigated the physiological response of a butyrate-oxidizing co-culture (comprised of Syntrophomonas wolfei and Methanospirillum hungatei) to the addition of a fermentative microorganism, Trichococcus flocculiformis.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Methanospirillum Hungatei Jf-1 Syntrophomonas Wolfei Subsp. Wolfei Str. Goettingen G311 Trichococcus Sp. Es5

SUBMITTER: Sjef Boeren

LAB HEAD: Sjef Boeren

PROVIDER: PXD033054 | Pride | 2022-05-31

REPOSITORIES: Pride

ACCESS DATA

Similar Datasets

Project description:One of the key issues affecting yields of biovanillin in cell factories such as Escherichia coli is product toxicity, the mechanisms of which are poorly understood. To identify targets for engineering improved strains, we have studied mechanisms of vanillin toxicity in E. coli using a two-pronged apparoach: (i) a global proteomic analysis supported by multiple physiological experiments and mutant analyses (ii) adaptive laboratory evolution (ALE) of vanillin tolerance combined with genome sequencing. We identified 147 proteins that exhibited a significant change in abundance in response to vanillin. Upregulated proteins included enzymes capable of converting aldehydes to potentially less toxic compounds; pentose phosphate pathways enzymes, fumarase C and enzymes of the glyoxylate shunt; proteins involved in several key stress responses, in particular oxidative stress; proteins mediating uptake and processing of metal ions. 1H-NMR confirmed that E. coli detoxifies vanillin by reduction to vanillyl alcohol; the aldehyde reductases YqhD and DkgA were highly upregulated by vanillin and the purified enzymes reduced vanillinin an NADPH dependent manner. Vanillin caused accumulation of reactive oxygen species and activated an oxidative stress response through SoxRS, OxyR and MarA pathways. Slow vanillin dependent copper (II) to copper (I) reduction lead to upregulation of the copA gene and growth in the presence of vanillin was hypersensitive to inhibition by copper ions. RT-PCR and mutant growth data suggested AcrD and AaeAB as potential vanillin efflux systems. Vanillin-tolerant strains isolated by ALE had distinct non-synonymous SNPs in the citrate synthase gene gltA, in addition to strain specific mutations in cpdA, rob and marC. One strain had a large ~10 kb deletion including the marRAB region. Purifed variant GltA enzymes all showed higher activity due to a lowered Km for oxaloacetate. Our data provide new understanding and novel gene targets for engineering vanillin-tolerant strains of E. coli, including deletion of the efflux pumps eamA, acrA, and acrB, enhancing oxidative stress defences and NADPH production, improving copper homeostasis and increasing citrate synthase activity using variant enzymes.

Dataset Information

Effect of Trichococcus flocculiformis on a Co-culture of Syntrophomonas wolfei and Methanospirillum hungatei

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure