Mechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids

Christopher J. Vavricka, Takanobu Yoshida, Yuki Kuriya, Shunsuke Takahashi, Teppei Ogawa, Fumie Ono, Kazuko Agari, Hiromasa Kiyota, Jianyong Li, Jun Ishii, Kenji Tsuge, Hiromichi Minami, Michihiro Araki, Tomohisa Hasunuma, Akihiko Kondo

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Previous studies have utilized monoamine oxidase (MAO) and L-3,4-dihydroxyphenylalanine decarboxylase (DDC) for microbe-based production of tetrahydropapaveroline (THP), a benzylisoquinoline alkaloid (BIA) precursor to opioid analgesics. In the current study, a phylogenetically distinct Bombyx mori 3,4-dihydroxyphenylacetaldehyde synthase (DHPAAS) is identified to bypass MAO and DDC for direct production of 3,4-dihydroxyphenylacetaldehyde (DHPAA) from L-3,4-dihydroxyphenylalanine (L-DOPA). Structure-based enzyme engineering of DHPAAS results in bifunctional switching between aldehyde synthase and decarboxylase activities. Output of dopamine and DHPAA products is fine-tuned by engineered DHPAAS variants with Phe79Tyr, Tyr80Phe and Asn192His catalytic substitutions. Balance of dopamine and DHPAA products enables improved THP biosynthesis via a symmetrical pathway in Escherichia coli. Rationally engineered insect DHPAAS produces (R,S)-THP in a single enzyme system directly from L-DOPA both in vitro and in vivo, at higher yields than that of the wild-type enzyme. However, DHPAAS-mediated downstream BIA production requires further improvement.

Original languageEnglish
Article number2015
Pages (from-to)2015
JournalNature communications
Volume10
Issue number1
DOIs
Publication statusPublished - May 1 2019

Fingerprint

Benzylisoquinolines
dopa
Tetrahydropapaveroline
alkaloids
insects
Levodopa
Alkaloids
Carboxy-Lyases
Insects
Tuning
tuning
enzymes
dopamine
oxidase
Monoamine Oxidase
Dopamine
Enzymes
biosynthesis
Bombyx
bypasses

ASJC Scopus subject areas

  • Chemistry(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physics and Astronomy(all)

Cite this

Mechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids. / Vavricka, Christopher J.; Yoshida, Takanobu; Kuriya, Yuki; Takahashi, Shunsuke; Ogawa, Teppei; Ono, Fumie; Agari, Kazuko; Kiyota, Hiromasa; Li, Jianyong; Ishii, Jun; Tsuge, Kenji; Minami, Hiromichi; Araki, Michihiro; Hasunuma, Tomohisa; Kondo, Akihiko.

In: Nature communications, Vol. 10, No. 1, 2015, 01.05.2019, p. 2015.

Research output: Contribution to journalArticle

Vavricka, CJ, Yoshida, T, Kuriya, Y, Takahashi, S, Ogawa, T, Ono, F, Agari, K, Kiyota, H, Li, J, Ishii, J, Tsuge, K, Minami, H, Araki, M, Hasunuma, T & Kondo, A 2019, 'Mechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids', Nature communications, vol. 10, no. 1, 2015, pp. 2015. https://doi.org/10.1038/s41467-019-09610-2
Vavricka, Christopher J. ; Yoshida, Takanobu ; Kuriya, Yuki ; Takahashi, Shunsuke ; Ogawa, Teppei ; Ono, Fumie ; Agari, Kazuko ; Kiyota, Hiromasa ; Li, Jianyong ; Ishii, Jun ; Tsuge, Kenji ; Minami, Hiromichi ; Araki, Michihiro ; Hasunuma, Tomohisa ; Kondo, Akihiko. / Mechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids. In: Nature communications. 2019 ; Vol. 10, No. 1. pp. 2015.
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