| Size | Price | |
|---|---|---|
| 5mg | ||
| 10mg | ||
| Other Sizes |
| Targets |
Natural product; Diterpenoids
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|---|---|
| ln Vitro |
Leaves of tobacco (Nicotiana tabacum) are covered with glandular trichomes that produce sucrose esters and diterpenoids in varying quantities, depending on cultivar type. The bicyclic diterpene Z-abienol is the major labdanoid present in some oriental tobacco cultivars, where it constitutes a precursor of important flavours and aromas. We describe here the identification and characterization of two genes governing the biosynthesis of Z-abienol in N. tabacum. As for other angiosperm labdanoid diterpenes, the biosynthesis of Z-abienol proceeds in two steps. NtCPS2 encodes a class-II terpene synthase that synthesizes 8-hydroxy-copalyl diphosphate, and NtABS encodes a kaurene synthase-like (KSL) protein that uses 8-hydroxy-copalyl diphosphate to produce Z-abienol. Phylogenetic analysis indicates that NtABS belongs to a distinct clade of KSL proteins that comprises the recently identified tomato (Solanum habrochaites) santalene and bergamotene synthase. RT-PCR results show that both genes are preferentially expressed in trichomes. Moreover, microscopy of NtCPS2 promoter-GUS fusion transgenics demonstrated a high specificity of expression to trichome glandular cells. Ectopic expression of both genes, but not of either one alone, driven by a trichome-specific promoter in transgenic Nicotiana sylvestris conferred Z-abienol formation to this species, which does not normally produce it. Furthermore, sequence analysis of over 100 tobacco cultivars revealed polymorphisms in NtCPS2 that lead to a prematurely truncated protein in cultivars lacking Z-abienol, thus establishing NtCPS2 as a major gene controlling Z-abienol biosynthesis in tobacco. These results offer new perspectives for tobacco breeding and the metabolic engineering of labdanoid diterpenes, as well as for structure-function relationship studies of terpene synthases.[1]
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| Enzyme Assay |
To identify the genes involved in the biosynthesis of Z-abienol in tobacco, and in the absence of a complete tobacco genomic sequence, we hypothesized that this biosynthesis should proceed in two sequential steps, each catalyzed by a distinct enzyme. According to this hypothesis and the current prevailing view on the biosynthesis of Z-abienol (Carman and Duffield, 1993; Guo and Wagner, 1995), the first enzyme, with a CPS-like activity, should yield 8-α-hydroxy-copalyl diphosphate (8-OH-CPP) of normal configuration (3), which would then be converted to Z-abienol by KS-like activity (see Figure 1). It is unlikely that normal copalyl diphosphate without an 8-hydroxyl group is the intermediate because the molecular weight of Z-abienol is 290, and not 288, indicating that the hydroxyl group is the result of the neutralization of a carbocation by water capture during the first cyclization, rather than a subsequent oxidation by cytochrome P450 or another oxidase. In addition, a CPS-like enzyme from Cistus creticus was shown to catalyse the synthesis of 8-OH-CPP (Falara et al., 2010), and a bifunctional Z-abienol synthase from balsam fir (Abies balsamea) was recently characterized (Zerbe et al., 2012), lending further support to this pathway hypothesis. However, to the best of our knowledge, no KSL enzyme able to use 8-OH-CPP has been reported so far, and the identification of such enzymes would expand the repertoire of substrates of terpene synthases, and open the door for the identification of other related enzymes producing labdanoids with an 8-hydroxyl group, like sclareol or labdenediol. Here we describe the cloning and characterization of two genes from tobacco, NtCPS2 and NtABS, necessary and sufficient for Z-abienol biosynthesis from GGPP in glandular trichomes. In addition, we show that the absence of Z-abienol in certain tobacco cultivars can be accounted for by sequence polymorphisms in the NtCPS2 gene.[1]
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| References | |
| Additional Infomation |
Cis-abienol is a labdane diterpenoid in which the labdane skeleton has double bonds at C-12 and C-14 (the former with Z-stereochemistry) and carries a hydroxy group at position C-8. It has a role as a metabolite. It is a labdane diterpenoid and a tertiary alcohol.
cis-Abienol has been reported in Picea orientalis, Picea abies, and other organisms with data available. |
| Molecular Formula |
C20H34O
|
|---|---|
| Molecular Weight |
290.49
|
| Exact Mass |
290.261
|
| Elemental Analysis |
C, 82.69; H, 11.80; O, 5.51
|
| CAS # |
17990-16-8
|
| PubChem CID |
643723
|
| Appearance |
White to off-white solid powder
|
| Density |
0.91g/cm3
|
| Boiling Point |
369.9ºC at 760 mmHg
|
| Flash Point |
161.9ºC
|
| Vapour Pressure |
5.59E-07mmHg at 25°C
|
| Index of Refraction |
1.487
|
| LogP |
5.502
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
1
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
21
|
| Complexity |
433
|
| Defined Atom Stereocenter Count |
4
|
| SMILES |
C/C(=C/C[C@@H]1[C@]2(CCCC([C@@H]2CC[C@@]1(C)O)(C)C)C)/C=C
|
| InChi Key |
ZAZVCYBIABTSJR-SZAPHMHZSA-N
|
| InChi Code |
InChI=1S/C20H34O/c1-7-15(2)9-10-17-19(5)13-8-12-18(3,4)16(19)11-14-20(17,6)21/h7,9,16-17,21H,1,8,10-14H2,2-6H3/b15-9-/t16-,17+,19-,20+/m0/s1
|
| Chemical Name |
(1R,2R,4aS,8aS)-2,5,5,8a-tetramethyl-1-((Z)-3-methylpenta-2,4-dien-1-yl)decahydronaphthalen-2-ol
|
| Synonyms |
Labda-12,14-dien-8-ol; (Z)- (Z)-Abienol; (+)-cis-Abienol; cis-Abienol; (Z)-Abienol; 17990-16-8; (12Z)-Abienol; Abienol, cis-; (Z)-Labda-12,14-dien-8-ol; 9H-(Labda-12,14-dien-8-ol); (12Z)-Abienol
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.4425 mL | 17.2123 mL | 34.4246 mL | |
| 5 mM | 0.6885 mL | 3.4425 mL | 6.8849 mL | |
| 10 mM | 0.3442 mL | 1.7212 mL | 3.4425 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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