Engineered synthesis of rosmarinic acid in Escherichia coli resulting production of a new intermediate, caffeoyl-phenyllactate

Jiang, Jingjie; Bi, Huiping; Zhuang, Yibin; Liu, Shaowei; Liu, Tao; Ma, Yanhe

doi:10.1007/s10529-015-1945-7

Cited by 32 publications

(25 citation statements)

References 9 publications

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“…RA levels in some in vitro culture systems are higher than in the parent plant. Engineered production of RA and its analogues have also been achieved in an Escherichia coli culture system 22–24 .…”

Section: Introductionmentioning

confidence: 99%

A tyrosine aminotransferase involved in rosmarinic acid biosynthesis in Prunella vulgaris L

Wang

Bai

et al. 2017

Sci Rep

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Rosmarinic acid (RA) and its derivants are medicinal compounds that comprise the active components of several therapeutics. We isolated and characterised a tyrosine aminotransferase of Prunella vulgaris (PvTAT). Deduced PvTAT was markedly homologous to other known/putative plant TATs. Cytoplasmic localisation of PvTAT was observed in tobacco protoplasts. Recombinantly expressed and purified PvTAT had substrates preference for L-tyrosine and phenylpyruvate, with apparent K m of 0.40 and 0.48 mM, and favoured the conversion of tyrosine to 4-hydroxyphenylpyruvate. In vivo activity was confirmed by functional restoration of the Escherichia coli tyrosine auxotrophic mutant DL39. Agrobacterium rhizogenes-mediated antisense/sense expression of PvTAT in hairy roots was used to evaluate the contribution of PvTAT to RA synthesis. PvTAT were reduced by 46–95% and RA were decreased by 36–91% with low catalytic activity in antisense transgenic hairy root lines; furthermore, PvTAT were increased 0.77–2.6-fold with increased 1.3–1.8-fold RA and strong catalytic activity in sense transgenic hairy root lines compared with wild-type counterparts. The comprehensive physiological and catalytic evidence fills in the gap in RA-producing plants which didn’t provide evidence for TAT expression and catalytic activities in vitro and in vivo. That also highlights RA biosynthesis pathway in P. vulgaris and provides useful information to engineer natural products.

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Section: Introductionmentioning

confidence: 99%

A tyrosine aminotransferase involved in rosmarinic acid biosynthesis in Prunella vulgaris L

Wang

Bai

et al. 2017

Sci Rep

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“…Such substrate flexibility has been explored for RA synthase from Coleus blumei and allowed biosynthesis of 13 RA analogues in E. coli [10]. Heterelogous pathways for the synthesis of the two acceptors, 4-hydroxyphenyllactate and 3,4-dihydroxyphenyllactate, from an inexpensive renewable carbon source has already been demonstrated in E. coli [8, 9] and could be implemented in yeast for sustainable and economical biosynthesis. We also demonstrated that expression of tyrosine ammonia-lyase (FjTAL) in addition to 4CL5 and LaAT1 results in the production of p -coumaroyl 4′-hydroxyphenyllactate when the FjTAL-4CL5-LaAT1 strain is fed with only 4-hydroxyphenyllactate (Additional file 1: Figure S1A).…”

Section: Resultsmentioning

confidence: 99%

Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast

et al. 2016

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BackgroundBAHD acyltransferases, named after the first four biochemically characterized enzymes of the group, are plant-specific enzymes that catalyze the transfer of coenzyme A-activated donors onto various acceptor molecules. They are responsible for the synthesis in plants of a myriad of secondary metabolites, some of which are beneficial for humans either as therapeutics or as specialty chemicals such as flavors and fragrances. The production of pharmaceutical, nutraceutical and commodity chemicals using engineered microbes is an alternative, green route to energy-intensive chemical syntheses that consume petroleum-based precursors. However, identification of appropriate enzymes and validation of their functional expression in heterologous hosts is a prerequisite for the design and implementation of metabolic pathways in microbes for the synthesis of such target chemicals.ResultsFor the synthesis of valuable metabolites in the yeast Saccharomyces cerevisiae, we selected BAHD acyltransferases based on their preferred donor and acceptor substrates. In particular, BAHDs that use hydroxycinnamoyl-CoAs and/or benzoyl-CoA as donors were targeted because a large number of molecules beneficial to humans belong to this family of hydroxycinnamate and benzoate conjugates. The selected BAHD coding sequences were synthesized and cloned individually on a vector containing the Arabidopsis gene At4CL5, which encodes a promiscuous 4-coumarate:CoA ligase active on hydroxycinnamates and benzoates. The various S. cerevisiae strains obtained for co-expression of At4CL5 with the different BAHDs effectively produced a wide array of valuable hydroxycinnamate and benzoate conjugates upon addition of adequate combinations of donors and acceptor molecules. In particular, we report here for the first time the production in yeast of rosmarinic acid and its derivatives, quinate hydroxycinnamate esters such as chlorogenic acid, and glycerol hydroxycinnamate esters. Similarly, we achieved for the first time the microbial production of polyamine hydroxycinnamate amides; monolignol, malate and fatty alcohol hydroxycinnamate esters; tropane alkaloids; and benzoate/caffeate alcohol esters. In some instances, the additional expression of Flavobacterium johnsoniae tyrosine ammonia-lyase (FjTAL) allowed the synthesis of p-coumarate conjugates and eliminated the need to supplement the culture media with 4-hydroxycinnamate.ConclusionWe demonstrate in this study the effectiveness of expressing members of the plant BAHD acyltransferase family in yeast for the synthesis of numerous valuable hydroxycinnamate and benzoate conjugates.Electronic supplementary materialThe online version of this article (doi:10.1186/s12934-016-0593-5) contains supplementary material, which is available to authorized users.

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“…As for phenolic acids, recent progress in metabolic engineering shows that several L-tyrosine-derived phenolic acids, including caffeic acid, gallic acid, RA and salvianolic acid can be biosynthesised in microbial systems [86][87][88]. Jiang et al [89] successfully biosynthesised RA by feeding caffeic acid thus constructing an artificial pathway for 3,4-dihydroxyphenyllactic acid. Furthermore, unnatural compounds such as caffeoylphenyllactate were biosynthesised in engineered E. coli, too.…”

Section: Synthetic Biology-based Approaches For Producing Danshen Biomentioning

confidence: 99%

Biosynthesis of bioactive ingredients of Salvia miltiorrhiza and advanced biotechnologies for their production

Geng²,

Zhao

2018

Biotechnology & Biotechnological Equipment

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This review deals with the progress in the biosynthesis and regulation of tanshinones and salvianolic acids as well as the prospects and challenges for producing such compounds through biotechnology techniques. Tanshinones (lipophilic diterpenoids) and salvianolic acids (hydrophilic phenolic acids) are valuable natural products from danshen (Salvia miltiorrhiza Bunge) with remarkable clinical efficacy to treat cardiovascular diseases, with potential application in the treatment of cancer and possibly other disorders. The significant bioactivities of S. miltiorrhiza inspired scientists to explore its biosynthetic mechanism that promotes the production of these compounds. Computational and comparative genomics and transcriptomics have been used to analyse a number of pathway genes, transcription factors and microRNAs that are associated with the biosynthesis and regulation of tanshinones and salvianolic acids. At the same time, plant cell and tissue culture, transgenic plants, microbial biotransformation and metabolic engineering have been used to synthesise all these compounds.

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Engineered synthesis of rosmarinic acid in Escherichia coli resulting production of a new intermediate, caffeoyl-phenyllactate

Abstract: This work not only leads to high yield production of rosmarinic acid and analogues, but also sheds new light on the construction of the pathway of rosmarinic acid in E. coli.

Cited by 32 publications

References 9 publications

A tyrosine aminotransferase involved in rosmarinic acid biosynthesis in Prunella vulgaris L

A tyrosine aminotransferase involved in rosmarinic acid biosynthesis in Prunella vulgaris L

Exploiting members of the BAHD acyltransferase family to synthesize multiple hydroxycinnamate and benzoate conjugates in yeast

Biosynthesis of bioactive ingredients of Salvia miltiorrhiza and advanced biotechnologies for their production

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