<i>In Vivo</i>Bioconversion of Tetrahydroisoquinoline by Recombinant Coclaurine<i>N</i>-Methyltransferase

Morishige, Takashi; Choi, Kum-Boo; Sato, Fumihiko

doi:10.1271/bbb.68.939

Cited by 15 publications

(4 citation statements)

References 8 publications

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“…, 2005), exhibit a monophyletic origin (Figure 5). In general, these NMTs accept a wide range of BIA substrates with diverse backbone structures, suggesting a potential utility in metabolic engineering and synthetic biology applications (Morishige et al. , 2004; Hawkins and Smolke, 2008; Minami et al.…”

Section: Discussionmentioning

confidence: 99%

Targeted metabolite and transcript profiling for elucidating enzyme function: isolation of novel N‐methyltransferases from three benzylisoquinoline alkaloid‐producing species

et al. 2009

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SUMMARYAn integrated approach using targeted metabolite profiles and modest EST libraries each containing approximately 3500 unigenes was developed in order to discover and functionally characterize novel genes involved in plant-specialized metabolism. EST databases have been established for benzylisoquinoline alkaloid-producing cell cultures of Eschscholzia californica, Papaver bracteatum and Thalictrum flavum, and are a rich repository of alkaloid biosynthetic genes. ESI-FTICR-MS and ESI-MS/MS analyses facilitated unambiguous identification and relative quantification of the alkaloids in each system. Manual integration of known and candidate biosynthetic genes in each EST library with benzylisoquinoline alkaloid biosynthetic networks assembled from empirical metabolite profiles allowed identification and functional characterization of four N-methyltransferases (NMTs). One cDNA from T. flavum encoded pavine N-methyltransferase (TfPavNMT), which showed a unique preference for (AE)-pavine and represents the first isolated enzyme involved in the pavine alkaloid branch pathway. Correlation of the occurrence of specific alkaloids, the complement of ESTs encoding known benzylisoquinoline alkaloid biosynthetic genes and the differential substrate range of characterized NMTs demonstrated the feasibility of bilaterally predicting enzyme function and species-dependent specialized metabolite profiles.

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Section: Discussionmentioning

confidence: 99%

Targeted metabolite and transcript profiling for elucidating enzyme function: isolation of novel N‐methyltransferases from three benzylisoquinoline alkaloid‐producing species

et al. 2009

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“…The improvement of utilization efficiency of dopamine will be needed for in vivo large-scale production. An advantage of this system is that (R,S)-reticuline was produced without the addition of a methyl group donor, S-adenosyl-L-methionine (SAM), because the regeneration of SAM in microbial cells is known to maintain in vivo methylation activity during bioconversion (24).…”

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confidence: 99%

Microbial production of plant benzylisoquinoline alkaloids

Minami

Kim

Ikezawa

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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312

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Benzylisoquinoline alkaloids, such as the analgesic compounds morphine and codeine, and the antibacterial agents berberine, palmatine, and magnoflorine, are synthesized from tyrosine in the Papaveraceae, Berberidaceae, Ranunculaceae, Magnoliaceae, and many other plant families. It is difficult to produce alkaloids on a large scale under the strict control of secondary metabolism in plants, and they are too complex for cost-effective chemical synthesis. By using a system that combines microbial and plant enzymes to produce desired benzylisoquinoline alkaloids, we synthesized (S)-reticuline, the key intermediate in benzylisoquinoline alkaloid biosynthesis, from dopamine by crude enzymes from transgenic Escherichia coli. The final yield of (S)-reticuline was 55 mg/liter within 1 h. Furthermore, we synthesized an aporphine alkaloid, magnoflorine, or a protoberberine alkaloid, scoulerine, from dopamine via reticuline by using different combination cultures of transgenic E. coli and Saccharomyces cerevisiae cells. The final yields of magnoflorine and scoulerine were 7.2 and 8.3 mg/liter culture medium. These results indicate that microbial systems that incorporate plant genes cannot only enable the mass production of scarce benzylisoquinoline alkaloids but may also open up pathways for the production of novel benzylisoquinoline alkaloids.(S)-reticuline ͉ magnoflorine ͉ scoulerine

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“…An advantage of fermentative (R,S)-reticuline is that no additional SAM (a methyldonor for methyltransferases) is required, because microbial cells regenerate SAM naturally to maintain in vivo methylation activity during bioconversion. 24 …”

Section: Metabolic Engineering Of Bias In Microbesmentioning

confidence: 99%

Fermentative Production of Plant Benzylisoquinoline Alkaloids in Microbes

Minami

2013

Bioscience, Biotechnology, and Biochemistry

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Higher plants produce diverse chemicals, including alkaloids, terpenoids, and phenolic compounds (phenylpropanoids and flavonoids) as secondary metabolites. These chemicals are widely used for human health and nutrition. Alkaloids, for example, are valued in medicine due to their high biological activities, but most of these metabolites accumulate at low levels in plant cells, resulting in poor extraction yields. Increasingly, attention is devoted to the production of plant metabolites by reconstructing plant biosynthetic pathways in microorganisms. This technology has been aided by advances in synthetic biology and metabolic engineering. Here, the review a fermentation platform for low-cost production of numerous alkaloids using bioengineered Escherichia coli and/or Saccharomyces cerevisiae.

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In VivoBioconversion of Tetrahydroisoquinoline by Recombinant CoclaurineN-Methyltransferase

Cited by 15 publications

References 8 publications

Targeted metabolite and transcript profiling for elucidating enzyme function: isolation of novel N‐methyltransferases from three benzylisoquinoline alkaloid‐producing species

Targeted metabolite and transcript profiling for elucidating enzyme function: isolation of novel N‐methyltransferases from three benzylisoquinoline alkaloid‐producing species

Microbial production of plant benzylisoquinoline alkaloids

Fermentative Production of Plant Benzylisoquinoline Alkaloids in Microbes

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