Sleep research article
Engineering a heme-dependent tryptophan hydroxylase pathway in <i>E. coli</i> for enhanced melatonin production.
Authors: Zhang L , Yin G , Pan S , Zheng X , Zhou S , Du G , Li J , Chen J , Xu R , Kang Z
One-line summary
A sleep science research article on Engineering a heme-dependent tryptophan hydroxylase pathway in <i>E. coli</i> for enhanced melatonin production..
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Original abstract
Melatonin is a high-value bioactive indoleamine broadly applied in the pharmaceutical, food and nutraceutical industries, yet its microbial production remains constrained by pathway complexity and low enzymatic efficiency. Here, a streamlined melatonin biosynthetic pathway was established in <i>Escherichia coli</i> by introducing the heme-dependent tryptophan hydroxylase Luz15. We further enhanced the intracellular heme availability and increased melatonin production by 111.45%. Fusion-tag engineering improved the solubility of all heterologous enzymes and boosted production by 50.64%. Structure-guided rational design of Luz15 subsequently yielded a beneficial D299S/W376H mutant, enhancing hydroxylation activity and contributing a 33.81% increase. To divert metabolic flux toward melatonin, a high-producing chassis was constructed via gene-editing and systematic sRNA library screening. The final strain produced 753.78 mg/L melatonin in a 5-L fed-batch bioreactor from glucose, a 38.78-fold improvement over the initial. This study establishes an efficient and scalable microbial platform for the sustainable biosynthesis of melatonin and other hydroxylated tryptophan-derived compounds.
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