 Chemical Structure.png)
Physicochemical Properties
| Exact Mass | 463.16 |
| Elemental Analysis | C, 54.30; H, 7.59; Cl, 15.26; N, 9.05; S, 13.80 |
| CAS # | 675825-78-2 |
| Related CAS # | 675825-78-2 (HCl); 189950-11-6 |
| PubChem CID | 16752681 |
| Appearance | Typically exists as solid at room temperature |
| Hydrogen Bond Donor Count | 4 |
| Hydrogen Bond Acceptor Count | 5 |
| Rotatable Bond Count | 10 |
| Heavy Atom Count | 28 |
| Complexity | 434 |
| Defined Atom Stereocenter Count | 4 |
| InChi Key | HZDNFBVWGKDFIP-GCMGGMMESA-N |
| InChi Code | InChI=1S/C21H34ClN3S2.ClH/c1-24-18-6-7-21(24)20(15-25(11-13-27)10-8-23-9-12-26)19(14-18)16-2-4-17(22)5-3-16;/h2-5,18-21,23,26-27H,6-15H2,1H3;1H/t18-,19+,20-,21+;/m0./s1 Create Date: 2007-10-29 |
| Chemical Name | 2-[2-[[(1R,2R,3S,5S)-3-(4-chlorophenyl)-8-methyl-8-azabicyclo[3.2.1]octan-2-yl]methyl-(2-sulfanylethyl)amino]ethylamino]ethanethiol;hydrochloride |
| Synonyms | Trodat 1; DLK3EG08UH; UNII-DLK3EG08UH; 675825-78-2; Ethanethiol, 2-((2-((((1R,2R,3S,5S)-3-(4-chlorophenyl)-8-methyl-8-azabicyclo(3.2.1)oct-2-yl)methyl)(2-mercaptoethyl)amino)ethyl)amino)-, hydrochloride (1:1) |
| 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) |
Biological Activity
| Targets | Chelating agent |
| ln Vitro | Propanethiol (PT) is a hazardous pollutant that poses risks to both the environment and human well-being. Pseudomonas putida S-1 has been identified as a microorganism capable of utilizing PT as its sole carbon source. However, the metabolic pathway responsible for PT degradation in P. putida S-1 has remained poorly understood, impeding its optimization and practical application. In this study, we investigated the catabolic network involved in PT desulfurization with P. putida S-1 and identified key gene modules crucial to this process. Notably, propanethiol oxidoreductase (PTO) catalyzes the initial degradation of PT, a pivotal step for P. putida S-1's survival on PT. PTO facilitates the oxidation of PT, resulting H2S, H2O2, and propionaldehyde (PA). Catalase-peroxidase catalyzes the conversion of H2O2 to oxygen and water, while PA undergoes gradual conversion to Succinyl-CoA, which is subsequently utilized in the tricarboxylic acid cycle. H2S is digested in a comprehensive desulfurization network where sulfide-quinone oxidoreductase (SQOR) predominantly converts it to sulfane sulfur. The transcriptome analysis suggests that sulfur can be finally converted to sulfite or sulfate and exported out of the cell. [1] |
| ln Vivo | The Propanethiol (PT) degradation capacity of P. putida S-1 was enhanced by increasing the transcription level of PTO and SQOR genes in vivo.IMPORTANCEThis work investigated the PT catabolism pathway in Pseudomonas putida S-1, a microorganism capable of utilizing PT as the sole carbon source. Critical genes that control the initiation of PT degradation were identified and characterized, such as pto and sqor. By increasing the transcription level of pto and sqor genes in vivo, we have successfully enhanced the PT degradation efficiency and growth rate of P. putida S-1. This work does not only reveal a unique PT degradation pathway but also highlights the potential of enhancing the microbial desulfurization process in the bioremediation of thiol-contaminated environment. [1] |
| Enzyme Assay |
Strains and plasmids [1] Strain Pseudomonas putida S-1 was originally isolated from active sludge and grew at 30°C and 180 rpm for Propanethiol (PT) degradation. Strain E. coli DH5α was used as host for plasmid construction. Strain E. coli BL21 (DE3) was used as host for protein expression. The auxotroph strain E. coli WM3064 was used for genome editing, and it was cultivated in LB with a supplementation of 2,6-diaminopimelic acid (57 mg/L). Enzyme activity assays [1] Steady-state kinetics of PTO and SQOR was analyzed according to the classic Michaelis-Menten equation. The protein concentration of PTO and SQOR was diluted to 100 µM for catalytic reaction. The concentration of FAD and CoQ1 was also set to 100 µM. The catalytic reactions were conducted at certain substrate concentrations, and the initial rate was calculated for generation of MM plot. The data collected was fitted by using the software Graph Pad Prism version 8. |
| References |
[1]. The critical roles of propanethiol oxidoreductase and sulfide-quinone oxidoreductase in the propanethiol catabolism pathway in Pseudomonas putida S-1. Appl Environ Microbiol . 2024 Feb 21;90(2):e0195923. |
| Additional Infomation |
VOSC degradation micrograms are often highly substrate specific. Previously reported studies of thiol biodegradation mainly focused on the degradation of methanethiol; a detailed characterization of the comprehensive catabolism pathway, however, has not been conducted. This study provides insights into the fine-tuned propanethiol catabolic mechanism network in Pseudomonas putida S-1 and identified critical gene modules accounting for the initiation of PT desulfurization. Based on the experimental observations made, the PT catabolism in P. putida S-1 was initiated by PTO (S1GL003403), and the reaction products H2O2, H2S, and propionaldehyde were then catalyzed by SQOR (S1GL000007 and S1GL003435) and the genes downstream. In the BRENDA Enzyme Databse (https://www.brenda-enzymes.org/index.php), oxidases with thiol as the substrates are categorized into two groups, thiol oxidase (EC 1.8.3.2) and methanethiol oxidase (MTO) (EC 1.8.3.4). Thiol oxidase converts thiols to the corresponding thioether, while MTO is restricted to one substrate, methanethiol. For example, the first MTO enzyme reported was isolated from the bacterium Hyphomicrobium sp. VS, and the Km value of this protein is around 0.3 µM. The MTO from human beings catalyzes methanethiol to sulfide in a similar mechanism and has an apparent Km of 1.8 nM, and this low Km value could be contributing to the prevention of methanethiol toxicity in the human body. Although these MTO enzymes catalyze methanethiol to sulfide with relatively high efficiency, their substrate spectrum is narrow. Propanethiol, for example, could not be catalyzed by them. The substrate specificity of PTO exhibits a high degree of selectivity. It was observed that the bacterium P. putida S-1, from which PTO was isolated, was unable to thrive solely on MT as a carbon source. If PTO were capable of degrading MT, one would expect the bacterium to utilize the resulting products as a nutritional source. The limited range of substrates for MTO or PTO implies that oxidases acting on thiols as substrates are likely to display pronounced substrate specificity. The discovery of PTO addresses a significant gap in the existing enzyme repertoire, suggesting its potential application in the bioremediation of PT pollution.
[1] After the initial desulfurization step, SQOR genes S1GL000007 and S1GL003435 conduct the conversion of H2S to sulfane sulfur, GSSH, or thiosulfate (Fig. S8). Whole-genome sequencing has also annotated three genes potentially encoded for cysteine synthase that can catalyze sulfide to cysteine, but all of them were downregulated under PT induction (Table S5). Considering cysteine was reported to form reactive oxygen species during oxidization that can damage DNA and lead to mutations or cell death, the generation of cysteine might not be the key step for sulfur circulation in P. putida S-1, although cysteine could serve as the carbon source for the bacteria. GSSH, the product of SQOR, could be oxidized by PDO to sulfite, which could be oxidized to sulfate naturally or by SDH, and some of these genes (e.g., S1GL000006) were upregulated in P. putida S-1 with PT induction (Table S5). Sulfate could be transported out of P. putida S-1 through a sulfite exporter (TAUE) or reduced to sulfide by sulfite reductase (SRD). As PT exhibits limited solubility in water, it could form disulfide in the environment and enter the cell. Unlike tauE, the only srd gene was significantly downregulated in P. putida S-1 (Table S5). Sulfite can be converted to thiosulfate by a thiosulfate sulfurtransferase, and thiosulfate can be further transferred to tetrathionate. However, the SfnB family sulfur acquisition oxidoreductase (SFNB) that catalyzes this reaction was largely downregulated, suggesting tetrathionate is not a favorable metabolite during PT catabolism. Propionaldehyde generated by PTO could be converted to propionate by aldehyde dehydrogenase. The propionate is converted to propionyl-coenzyme A (CoA) by propionate CoA-transferase. Finally, propionyl-CoA is catalyzed to succinyl-CoA by propionyl-CoA:succinyl-CoA transferase and enters the tricarboxylic acid cycle (Fig. S8). [1] In conclusion, the critical gene modules S1GL003403 (pto) and S1GL003435 (sqor) were successfully found to be responsible for the initial catabolism process of the PT catabolism pathway in Pseudomonas putida S-1. When increasing the transcription level of pto and sqor genes, PT degradation capacity of P. putida S-1 can be significantly enhanced. These findings put new insight of and suggest more strategy to enhance the microbial desulfurization process.[1] This work investigated the PT catabolism pathway in Pseudomonas putida S-1, a microorganism capable of utilizing PT as the sole carbon source. Critical genes that control the initiation of PT degradation were identified and characterized, such as pto and sqor. By increasing the transcription level of pto and sqor genes in vivo, we have successfully enhanced the PT degradation efficiency and growth rate of P. putida S-1. This work does not only reveal a unique PT degradation pathway but also highlights the potential of enhancing the microbial desulfurization process in the bioremediation of thiol-contaminated environment. |
Solubility Data
| Solubility (In Vitro) | Typically soluble in DMSO (e.g. 10 mM) |
| 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 (e.g. IP/IV/IM/SC) 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). 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) 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). 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 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. (Please use freshly prepared in vivo formulations for optimal results.) |