High titer production of gastrodin enabled by systematic refactoring of yeast genome and an antisense-transcriptional regulation toolkit.

Gastrodin, a phenolic glycoside, is a prominent component of Gastrodia elata, which is renowned for its sedative, hypnotic, anticonvulsant, and neuroprotective activities. Engineering heterologous production of plant natural products in microbial host represents a safe, cost-effective, and scalable alternative to plant extraction. Here, we present the construction of an engineered Yarrowia lipolytica yeast that achieves a high-titer production of gastrodin. We systematically refactored the yeast genome by enhancing the flux of the shikimate pathway and optimizing the glucosyl transfer system. We introduced more than five dozen of genetic modifications onto the yeast genome, including enzyme screening, alleviation of rate-limiting steps, promoter selection, genomic integration site optimization, downregulation of competing pathways, and elimination of gastrodin degradation. Meanwhile, we developed a Copper-induced Antisense-Transcriptional Regulation (CATR) tool. The developed CATR toolkit achieved dynamic repression and activation of violacein synthesis through the addition of copper in Y. lipolytica. This strategy was further used to dynamically regulate the pyruvate kinase node to effectively redirect glycolytic flux towards the shikimate pathway while maintaining cell growth at proper rate. Taken together, these efforts resulted in 9477.1 mg/L of gastrodin in shaking flaks and 13.4 g/L of gastrodin with a yield of 0.149 g/g glucose in a 5-L bioreactor, highlighting the potential for large-scale and sustainable production of gastrodin from microbial fermentation.

Aberrant connectivity of the lateralized readiness system in non-syndromic congenital mirror movements.

OBJECTIVES: Non-syndromic CMM has a complex phenotype. Abnormal corpus callosum and corticospinal tract processes are suggested mechanisms of the mirror movements. To further explore behavioural and neural phenotype(s) the present study tests the hypothesis that the response readiness network comprising supplementary motor area (SMA) and connections with motor cortex (M1) functions abnormally in CMM. METHODS: Twelve participants with (non-syndromic) CMM and a control group (n = 28) were tested on a probabilistic Go-NoGo task while electroencephalography (EEG) was recorded to assess possible group differences in lateralized readiness of voluntary hand movements together with measures of SMA-M1 functional connectivity. RESULTS: The CMM group demonstrated delayed lateralized readiness and stronger functional connectivity between left-brain SMA-M1 regions. Connectivity strength was correlated with measures of behavioural performance but not with extent of mirroring. CONCLUSIONS: Abnormalities in brain processes upstream of movement output likely reflect neurocompensation as a result of lifelong experience with mirroring in CMM. SIGNIFICANCE: These findings extend the known neural abnormalities in CMM to include brain networks upstream from those involved in motor output and raise the question of whether neurocompensatory plasticity might be involved.