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231223s2010 xx |||||o 00| ||eng c |
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|a 10.1016/j.plaphy.2010.09.003
|2 doi
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|a pubmed25n0675.xml
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|a (DE-627)NLM202421872
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|a (NLM)20951598
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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| 100 |
1 |
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|a Pramod, K K
|e verfasserin
|4 aut
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|a Biochemical and structural characterization of recombinant hyoscyamine 6β-hydroxylase from Datura metel L
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|c 2010
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 02.05.2011
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|a Date Revised 30.09.2020
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Copyright © 2010 Elsevier Masson SAS. All rights reserved.
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|a Hyoscyamine 6β-hydroxylase (H6H; EC 1.14.11.11), an important enzyme in the biosynthesis of tropane alkaloids, catalyzes the hydroxylation of hyoscyamine to give 6β-hydroxyhyoscyamine and its epoxidation in the biosynthetic pathway leading to scopolamine. Datura metel produces scopolamine as the predominant tropane alkaloid. The cDNA encoding H6H from D. metel (DmH6H) was cloned, heterologously expressed and biochemically characterized. The purified recombinant His-tagged H6H from D. metel (DmrH6H) was capable of converting hyoscyamine to scopolamine. The functionally expressed DmrH6H was confirmed by HPLC and ESI-MS verification of the products, 6β-hydroxyhyoscyamine and its derivative, scopolamine; the DmrH6H epoxidase activity was low compared to the hydroxylase activity. The K(m) values for both the substrates, hyoscyamine and 2-oxoglutarate, were 50μM each. The CD (circular dichroism) spectrum of the DmrH6H indicated a preponderance of α-helicity in the secondary structure. From the fluorescence studies, Stern-Volmer constants for hyoscyamine and 2-oxoglutarate were found to be 0.14M(-1) and 0.56M(-1), respectively. These data suggested that the binding of the substrates, hyoscyamine and 2-oxoglutarate, to the enzyme induced significant conformational changes
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|a Journal Article
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|a Research Support, Non-U.S. Gov't
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|a DNA, Complementary
|2 NLM
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| 650 |
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|a Ketoglutaric Acids
|2 NLM
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| 650 |
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|a Plant Proteins
|2 NLM
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|a Recombinant Proteins
|2 NLM
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|a Atropine
|2 NLM
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|a 7C0697DR9I
|2 NLM
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| 650 |
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|a Scopolamine
|2 NLM
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|a DL48G20X8X
|2 NLM
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| 650 |
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|a Mixed Function Oxygenases
|2 NLM
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|a EC 1.-
|2 NLM
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|a Oxidoreductases
|2 NLM
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|a EC 1.-
|2 NLM
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| 650 |
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|a hyoscyamine (6S)-dioxygenase
|2 NLM
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| 650 |
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|a EC 1.14.11.11
|2 NLM
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1 |
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|a Singh, Satpal
|e verfasserin
|4 aut
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|a Jayabaskaran, C
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t Plant physiology and biochemistry : PPB
|d 1991
|g 48(2010), 12 vom: 15. Dez., Seite 966-70
|w (DE-627)NLM098178261
|x 1873-2690
|7 nnns
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| 773 |
1 |
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|g volume:48
|g year:2010
|g number:12
|g day:15
|g month:12
|g pages:966-70
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|u http://dx.doi.org/10.1016/j.plaphy.2010.09.003
|3 Volltext
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|d 48
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