Benzyl alcohol is a naturally occurring aromatic alcohol and has been widely used in the cosmetics and flavor/fragrance industries. The whole‐cell biotransformation for synthesis of benzyl alcohol directly from bio‐based L‐phenylalanine (L‐Phe) was herein explored using an artificial enzyme cascade in Escherichia coli. Benzaldehyde was first produced from L‐Phe via four heterologous enzymatic steps that comprises L‐amino acid deaminase (LAAD), hydroxymandelate synthase (HmaS), (S)‐mandelate dehydrogenase (SMDH) and benzoylformate decarboxylase (BFD). The subsequent reduction of benzaldehyde to benzyl alcohol was achieved by a broad substrate specificity phenylacetaldehyde reductase (PAR) from Solanum lycopersicum. We found the designed enzyme cascade could efficiently convert L‐Phe into benzyl alcohol with conversion above 99%. In addition, we also examined L‐tyrosine (L‐Tyr) and m‐fluoro‐phenylalanine (m‐f‐Phe) as substrates, the cascade biotransformation could also efficiently produce p‐hydroxybenzyl alcohol and m‐fluoro‐benzyl alcohol. In summary, the developed biocatalytic pathway has great potential to produce various high‐valued fine chemicals.

Heavy metals, that is Cu(II), are harmful to the environment. There is an increasing demand to develop inexpensive detection methods for heavy metals. Here, we developed a yeast biosensor with reduced‐noise and improved signal output for potential on‐site copper ion detection. The copper‐sensing circuit was achieved by employing a secondary genetic layer to control the galactose‐inducible (GAL) system in Saccharomyces cerevisiae. The reciprocal control of the Gal4 activator and Gal80 repressor under copper‐responsive promoters resulted in a low‐noise and sensitive yeast biosensor for copper ion detection. Furthermore, we developed a betaxanthin‐based colorimetric assay, as well as 2‐phenylethanol and styrene‐based olfactory outputs for the copper ion detection. Notably, our engineered yeast sensor confers a narrow range switch‐like behaviour, which can give a ‘yes/no’ response when coupled with a betaxanthin‐based visual phenotype. Taken together, we envision that the design principle established here might be applicable to develop other sensing systems for various chemical detections.

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Promoters that play an essential role in the gene regulation are of particular interest to the synthetic biology communities. Recent advances in high-throughput DNA sequencing have greatly increased the breadth of new genetic parts. The development of promoters with broad host properties could enable rapid phenotyping of genetic constructs in different hosts. In this study, we discovered that the GAL1/10 bidirectional promoter from Saccharomyces cerevisiae could be used for shuttle expression in Escherichia coli . Further investigation revealed that the GAL1/10 bidirectional promoter is subjected to catabolite repression in E. coli . We next constructed a set of Golden-Gate assembly vectors for shuttle expression between S. cerevisiae and E. coli . The utility of shuttle vectors was demonstrated for rapid phenotyping of a multigene pathway for cinnamyl alcohol production. Taken together, our work opens a new avenue for the future development of broad host expression systems between prokaryotic and eukaryotic hosts.

Energy crisis and global climate change have driven an increased effort toward biofuel synthesis from renewable feedstocks. Herein, purple nonsulfur photosynthetic bacterium (PNSB) of Rhodobacter capsulatus was explored as a platform for high-titer production of a terpene-based advanced biofuel–bisabolene. A multilevel engineering strategy such as promoter screening, improving the NADPH availability, strengthening the precursor supply, suppressing the side pathways, and introducing a heterologous mevalonate pathway, was used to improve the bisabolene titer in R. capsulatus. The above strategies enabled a 35-fold higher titer of bisabolene than that of the starting strain, reaching 1089.7 mg/L from glucose in a shake flask. The engineered strain produced 9.8 g/L bisabolene with a yield of >0.196 g/g-glucose under the two-phase fed-batch fermentation, which corresponds to >78% of theoretical maximum. Taken together, our work represents one of the pioneering studies to demonstrate PNSB as a promising platform for terpene-based advanced biofuel production.