Natural product; Diterpenoids
The primary targets of (Z)-Abienol are not well-defined in the context of pharmacological activity. As a natural product, it has been studied for its antifungal activity, specifically its ability to inhibit the hyphal growth of P. nicotianae. The compound is also a biosynthetic precursor of important flavors and aromas in tobacco. Its mechanism of action as an antifungal agent and its specific molecular targets are not extensively characterized in the available literature.

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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In vitro, (Z)-Abienol inhibits the hyphal growth of the plant pathogen Phytophthora nicotianae. The compound's antifungal activity is demonstrated in assays where P. nicotianae is cultured in the presence of varying concentrations of (Z)-Abienol, and the inhibition of hyphal growth is measured. The compound is also used as a reference standard for labdane diterpenoids in analytical chemistry. However, detailed IC50 or EC50 values are not reported in the available literature.

In vivo data for (Z)-Abienol are not extensively reported. As a natural product primarily studied for its biosynthetic role and antifungal activity, the compound has not been extensively evaluated in animal models. It is not a pharmacological agent in the traditional sense. Its in vivo relevance is primarily in the context of plant biology and tobacco breeding.

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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In vitro enzyme assays for (Z)-Abienol involve studying the enzymes responsible for its biosynthesis. Two key enzymes have been identified: NtCPS2, a class-II terpene synthase that synthesizes 8-hydroxy-copalyl diphosphate, and NtABS, a kaurene synthase-like (KSL) protein that uses 8-hydroxy-copalyl diphosphate to produce Z-abienol. Enzyme assays involve incubating these recombinant enzymes with their substrates and measuring the production of Z-abienol by GC-MS or other analytical methods.

Cellular assays for (Z)-Abienol are performed in the context of plant biology. For example, RT-PCR results show that NtCPS2 and NtABS genes are preferentially expressed in trichomes, the glandular structures on tobacco leaves. Microscopy of NtCPS2 promoter-GUS fusion transgenics demonstrated high specificity of expression to trichome glandular cells. Ectopic expression of both genes in transgenic Nicotiana sylvestris conferred Z-abienol formation.

In vivo animal studies with (Z)-Abienol are not typically performed, as the compound is not a pharmacological agent. Its primary applications are in plant biology, tobacco breeding, and metabolic engineering. The compound is used to study the biosynthesis of labdanoid diterpenes and to develop tobacco cultivars with improved flavor and aroma characteristics.

(Z)-Abienol is a lipophilic diterpenoid with a molecular weight that is not specified in the available literature. It is typically dissolved in DMSO for in vitro studies. The compound is stable when stored as a powder at -20°C. However, detailed pharmacokinetic parameters such as absorption, distribution, metabolism, excretion, half-life, and bioavailability are not applicable for this compound, as it is not a drug.

(Z)-Abienol is a natural product that is not considered toxic in the context of its use as a flavor precursor or research compound. However, comprehensive toxicology data for this compound are not extensively reported. The compound is classified as a research-use-only chemical and is not intended for human consumption. Specific toxicological data, including acute toxicity, genotoxicity, and target organ effects, are not reported in the available literature.

[1]. Characterization of two genes for the biosynthesis of the labdane diterpene Z-abienol in tobacco (Nicotiana tabacum) glandular trichomes. Plant J. 2012 Oct;72(1):1-17.

Cis-Abidenol is a latanane diterpenoid compound with a latanane skeleton containing double bonds at C-12 and C-14 (Z-configuration at C-12) and a hydroxyl group at C-8. It is a metabolite. It is a latanane diterpenoid compound and also a tertiary alcohol. Cis-Abidenol has been reported to exist in Oriental spruce (Picea orientalis), European spruce (Picea abies), and other organisms with relevant data.

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(Z)-Abienol is a research-grade natural product not approved for clinical use. Its primary applications are in plant biology, tobacco breeding, and metabolic engineering. It is used as a biosynthetic precursor for labdanoid diterpenes and as a reference standard for analytical chemistry. The compound is also used to study the structure-function relationships of terpene synthases. Its antifungal activity against P. nicotianae makes it of interest for agricultural research.

C20H34O

290.49

290.261

C, 82.69; H, 11.80; O, 5.51

17990-16-8

643723

White to off-white solid powder

0.91g/cm3

369.9ºC at 760 mmHg

161.9ºC

5.59E-07mmHg at 25°C

1.487

5.502

1

1

3

21

433

4

C/C(=C/C[C@@H]1[C@]2(CCCC([C@@H]2CC[C@@]1(C)O)(C)C)C)/C=C

ZAZVCYBIABTSJR-SZAPHMHZSA-N

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

(1R,2R,4aS,8aS)-2,5,5,8a-tetramethyl-1-((Z)-3-methylpenta-2,4-dien-1-yl)decahydronaphthalen-2-ol

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

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

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

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)