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 language | English |
|---|---|
| Article number | 2015 |
| Pages (from-to) | 2015 |
| Journal | Nature communications |
| Volume | 10 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - May 1 2019 |
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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 journal › Article
}
TY - JOUR
T1 - Mechanism-based tuning of insect 3,4-dihydroxyphenylacetaldehyde synthase for synthetic bioproduction of benzylisoquinoline alkaloids
AU - Vavricka, Christopher J.
AU - Yoshida, Takanobu
AU - Kuriya, Yuki
AU - Takahashi, Shunsuke
AU - Ogawa, Teppei
AU - Ono, Fumie
AU - Agari, Kazuko
AU - Kiyota, Hiromasa
AU - Li, Jianyong
AU - Ishii, Jun
AU - Tsuge, Kenji
AU - Minami, Hiromichi
AU - Araki, Michihiro
AU - Hasunuma, Tomohisa
AU - Kondo, Akihiko
PY - 2019/5/1
Y1 - 2019/5/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85065169335&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85065169335&partnerID=8YFLogxK
U2 - 10.1038/s41467-019-09610-2
DO - 10.1038/s41467-019-09610-2
M3 - Article
C2 - 31043610
AN - SCOPUS:85065169335
VL - 10
SP - 2015
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 2015
ER -