Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

Kai, Guoyin; Liu, Yuanyuan; Wang, Xiaoyun; Yang, Sheng; Fu, Xueqing; Luo, Xiuqin; Liao, Pan

doi:10.1007/s10529-011-0575-y

Cited by 17 publications

(8 citation statements)

References 13 publications

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“…Additionally, specific hydroxylase and epoxidase activity: 2.60 ± 0.19 nKat mg −1 and 0.24 ± 0.02 nKat mg −1 for these two compounds were observed, respectively. The H6H from Anisodus acutangulus was cloned and expressed in E. coli fused with either a His- or GST-tag at the N -terminus [109]. A bifunctional assay revealed that both recombinant enzymes converted up to 80% of fed hyoscyamine to scopolamine, however, reaction kinetics were not analysed.…”

Section: Biotechnological Approaches Of Scopolamine Production Andmentioning

confidence: 99%

Tropane Alkaloids: Chemistry, Pharmacology, Biosynthesis and Production

2019

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Tropane alkaloids (TA) are valuable secondary plant metabolites which are mostly found in high concentrations in the Solanaceae and Erythroxylaceae families. The TAs, which are characterized by their unique bicyclic tropane ring system, can be divided into three major groups: hyoscyamine and scopolamine, cocaine and calystegines. Although all TAs have the same basic structure, they differ immensely in their biological, chemical and pharmacological properties. Scopolamine, also known as hyoscine, has the largest legitimate market as a pharmacological agent due to its treatment of nausea, vomiting, motion sickness, as well as smooth muscle spasms while cocaine is the 2nd most frequently consumed illicit drug globally. This review provides a comprehensive overview of TAs, highlighting their structural diversity, use in pharmaceutical therapy from both historical and modern perspectives, natural biosynthesis in planta and emerging production possibilities using tissue culture and microbial biosynthesis of these compounds.

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Section: Biotechnological Approaches Of Scopolamine Production Andmentioning

confidence: 99%

Tropane Alkaloids: Chemistry, Pharmacology, Biosynthesis and Production

2019

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“…However, our previous results found that AaH6H did not have the activity to convert scopolamine into anisodine. 26 In fact anisodine and scopolamine have similar chemical structures, but anisodine possesses a hydroxyl-group on the a-carbon atom of the tropic acid moiety. 27 The biosynthetic pathway of anisodine is not yet clear, but it can be deduced that it may be synthesized from scopolamine by an unknown hydroxylase in view of the chemical structure.…”

Section: Discussionmentioning

confidence: 99%

Enhancing the production of tropane alkaloids in transgenic Anisodus acutangulus hairy root cultures by over-expressing tropinone reductase I and hyoscyamine-6β-hydroxylase

et al. 2012

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Tropane alkaloids (TA) including hyoscyamine, anisodamine, scopolamine and anisodine, are used medicinally as anticholinergic agents with increasing market demand, so it is very important to improve TA production by metabolic engineering strategy. Here, we report the simultaneous introduction of genes encoding the branch-controlling enzyme tropinone reductase I (TRI, EU424321) and the downstream rate-limiting enzyme hyoscyamine-6β-hydroxylase (H6H, EF187826) involved in TA biosynthesis into Anisodus acutangulus hairy roots by Agrobacterium-mediated gene transfer technology. Transgenic hairy root lines expressing both TRI and H6H (TH lines) produced significantly higher (P < 0.05) levels of TA compared with the control and single gene transformed lines (T or H lines). The best double gene transformed line (TH53) produced 4.293 mg g(-1) TA, which was about 4.49-fold higher than that of the control lines (0.96 mg g(-1)). As far as it is known, this is the first report on simultaneous introduction of TRI and H6H genes into TA-producing plant by biotechnological approaches. Besides, the content of anisodine was also greatly improved in A. acutangulus by over-expression of AaTRI and AaH6H genes. The average content of anisodine in TH lines was 0.984 mg g(-1) dw, about 18.57-fold of BC lines (0.053 mg g(-1) dw). This is the first time that this phenomenon has been found in TA-producing plants.

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“…Hyoscyamine 6 β -hydroxylase (H6H) catalyzes hyoscyamine to produce anisodamine through 6 β -hydroxylation, and subsequently oxidizes anisodamine to generate scopolamine ( Hashimoto and Yamada, 1986 ) The H6H genes have been identified from H. niger ( Hashimoto and Yamada, 1986 ), A. belladonna ( Li et al, 2012 ), A. acutangulus ( Kai et al, 2011a ), and S. luridus ( Lan et al, 2018 ). Of these H6H enzymes, the H. niger one (HnH6H) showed the highest efficiency in converting hyoscyamine to scopolamine, and HnH6H is the best choice in engineering scopolamine production.…”

Section: Engineering Medicinal Tropane Alkaloid Production In Hairy R...mentioning

confidence: 99%

Biotechnological Approaches on Engineering Medicinal Tropane Alkaloid Production in Plants

Gong

Lan³

et al. 2022

Front. Plant Sci.

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Hyoscyamine and scopolamine, belonging to medicinal tropane alkaloids (MTAs), are potent anticholinergic drugs. Their industrial production relies on medicinal plants, but the levels of the two alkaloids are very low in planta. Engineering the MTA’s production is an everlasting hot topic for pharmaceutical industry. With understanding the MTA’s biosynthesis, biotechnological approaches are established to produce hyoscyamine and scopolamine in an efficient manner. Great advances have been obtained in engineering MTA’s production in planta. In this review, we summarize the advances on the biosynthesis of MTAs and engineering the MTA’s production in hairy root cultures, as well in plants. The problems and perspectives on engineering the MTA’s production are also discussed.

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Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

Cited by 17 publications

References 13 publications

Tropane Alkaloids: Chemistry, Pharmacology, Biosynthesis and Production

Tropane Alkaloids: Chemistry, Pharmacology, Biosynthesis and Production

Enhancing the production of tropane alkaloids in transgenic Anisodus acutangulus hairy root cultures by over-expressing tropinone reductase I and hyoscyamine-6β-hydroxylase

Biotechnological Approaches on Engineering Medicinal Tropane Alkaloid Production in Plants

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