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

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

doi:10.1007/s10529-011-0575-y

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

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

^*Corresponding author for this work

Department of Biology

Research output: Contribution to journal › Journal article › peer-review

16 Citations (Scopus)

Abstract

Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11) converts hyoscyamine to scopolamine in the last step of scopolamine biosynthetic pathway. The gene encoding H6H in Anisodus acutangulus was cloned and expressed in Escherichia coli and the recombinant proteins fused with His-tag or GST-tag at its N-terminal were purified and then confirmed by Western bolt analysis. The biofunctional assay revealed that the His-AaH6H and GST-AaH6H converted hyoscyamine (40 mg/l) to scopolamine at 32 and 31 mg/l, respectively. This is the first report on AaH6H expression, purification and functional characterization facilitates further genetic improvement of scopolamine yield in A. acutangulus.

Original language	English
Pages (from-to)	1361-1365
Number of pages	5
Journal	Biotechnology Letters
Volume	33
Issue number	7
Early online date	5 Mar 2011
DOIs	https://doi.org/10.1007/s10529-011-0575-y
Publication status	Published - Jul 2011

User-Defined Keywords

Anisodus acutangulus
Hyoscyamine
Hyoscyamine 6β-hydroxylase
Scopolamine

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10.1007/s10529-011-0575-y

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

abstract = "Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11) converts hyoscyamine to scopolamine in the last step of scopolamine biosynthetic pathway. The gene encoding H6H in Anisodus acutangulus was cloned and expressed in Escherichia coli and the recombinant proteins fused with His-tag or GST-tag at its N-terminal were purified and then confirmed by Western bolt analysis. The biofunctional assay revealed that the His-AaH6H and GST-AaH6H converted hyoscyamine (40 mg/l) to scopolamine at 32 and 31 mg/l, respectively. This is the first report on AaH6H expression, purification and functional characterization facilitates further genetic improvement of scopolamine yield in A. acutangulus.",

keywords = "Anisodus acutangulus, Hyoscyamine, Hyoscyamine 6β-hydroxylase, Scopolamine",

author = "Guoyin Kai and Yuanyuan Liu and Xiaoyun Wang and Sheng Yang and Xueqing Fu and Xiuqin Luo and Pan Liao",

note = "Funding Information: This work was supported by National Natural Science Fund (30900110), National Transgenic Organism New Variety Culture Key Project (2009ZX08012-002B), Project from Ministry of Science and Technology of China (NC2010AE0075, NC2010AE0372), Shanghai Science and Technology Committee Project (10JC1412000, 09QH1401900, 06QA14038, 08391911800, 073158202, 075405117, 065458022, 05ZR14093), Kunshan Municipal Science & Technology Project (KS0914), Zhejiang Provincial Natural Science Fund (Y2080621), Program from Shanghai Municipal Education Commission (06DZ015, 09ZZ138), Fujian Science and Technology Committee Key Special Project (2008NZ0001-4), Leading Academic Discipline Project of Shanghai Municipal Education Commission (J50401) and Project from Shanghai Normal University. Publisher copyright: {\textcopyright} 2011 Elsevier B.V., All rights reserved. ",

year = "2011",

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doi = "10.1007/s10529-011-0575-y",

language = "English",

volume = "33",

pages = "1361--1365",

journal = "Biotechnology Letters",

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TY - JOUR

T1 - Functional identification of hyoscyamine 6β-hydroxylase from Anisodus acutangulus and overproduction of scopolamine in genetically-engineered Escherichia coli

AU - Kai, Guoyin

AU - Liu, Yuanyuan

AU - Wang, Xiaoyun

AU - Yang, Sheng

AU - Fu, Xueqing

AU - Luo, Xiuqin

AU - Liao, Pan

N1 - Funding Information: This work was supported by National Natural Science Fund (30900110), National Transgenic Organism New Variety Culture Key Project (2009ZX08012-002B), Project from Ministry of Science and Technology of China (NC2010AE0075, NC2010AE0372), Shanghai Science and Technology Committee Project (10JC1412000, 09QH1401900, 06QA14038, 08391911800, 073158202, 075405117, 065458022, 05ZR14093), Kunshan Municipal Science & Technology Project (KS0914), Zhejiang Provincial Natural Science Fund (Y2080621), Program from Shanghai Municipal Education Commission (06DZ015, 09ZZ138), Fujian Science and Technology Committee Key Special Project (2008NZ0001-4), Leading Academic Discipline Project of Shanghai Municipal Education Commission (J50401) and Project from Shanghai Normal University. Publisher copyright: © 2011 Elsevier B.V., All rights reserved.

PY - 2011/7

Y1 - 2011/7

N2 - Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11) converts hyoscyamine to scopolamine in the last step of scopolamine biosynthetic pathway. The gene encoding H6H in Anisodus acutangulus was cloned and expressed in Escherichia coli and the recombinant proteins fused with His-tag or GST-tag at its N-terminal were purified and then confirmed by Western bolt analysis. The biofunctional assay revealed that the His-AaH6H and GST-AaH6H converted hyoscyamine (40 mg/l) to scopolamine at 32 and 31 mg/l, respectively. This is the first report on AaH6H expression, purification and functional characterization facilitates further genetic improvement of scopolamine yield in A. acutangulus.

AB - Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11) converts hyoscyamine to scopolamine in the last step of scopolamine biosynthetic pathway. The gene encoding H6H in Anisodus acutangulus was cloned and expressed in Escherichia coli and the recombinant proteins fused with His-tag or GST-tag at its N-terminal were purified and then confirmed by Western bolt analysis. The biofunctional assay revealed that the His-AaH6H and GST-AaH6H converted hyoscyamine (40 mg/l) to scopolamine at 32 and 31 mg/l, respectively. This is the first report on AaH6H expression, purification and functional characterization facilitates further genetic improvement of scopolamine yield in A. acutangulus.

KW - Anisodus acutangulus

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JO - Biotechnology Letters

JF - Biotechnology Letters

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ER -

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

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