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Structure of 60827-45-4
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The BI-3802 was designed by Boehringer Ingelheim and could be obtained free of charge through the Boehringer Ingelheim open innovation portal opnMe.com, associated with its negative control.
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| CAS No. : | 60827-45-4 |
| Formula : | C3H7ClO2 |
| M.W : | 110.54 |
| SMILES Code : | OC[C@H](O)CCl |
| MDL No. : | MFCD00210270 |
| InChI Key : | SSZWWUDQMAHNAQ-GSVOUGTGSA-N |
| Pubchem ID : | 148904 |
| GHS Pictogram: |
|
| Signal Word: | Danger |
| Hazard Statements: | H301-H312-H318 |
| Precautionary Statements: | P264-P270-P280-P301+P310-P302+P352-P305+P351+P338-P310-P322-P330-P363-P405-P501 |
| Class: | 6.1 |
| UN#: | 2689 |
| Packing Group: | Ⅲ |
| Num. heavy atoms | 6 |
| Num. arom. heavy atoms | 0 |
| Fraction Csp3 | 1.0 |
| Num. rotatable bonds | 2 |
| Num. H-bond acceptors | 2.0 |
| Num. H-bond donors | 2.0 |
| Molar Refractivity | 23.65 |
| TPSA ? Topological Polar Surface Area: Calculated from |
40.46 Ų |
| Log Po/w (iLOGP)? iLOGP: in-house physics-based method implemented from |
1.17 |
| Log Po/w (XLOGP3)? XLOGP3: Atomistic and knowledge-based method calculated by |
-0.49 |
| Log Po/w (WLOGP)? WLOGP: Atomistic method implemented from |
-0.42 |
| Log Po/w (MLOGP)? MLOGP: Topological method implemented from |
-0.18 |
| Log Po/w (SILICOS-IT)? SILICOS-IT: Hybrid fragmental/topological method calculated by |
0.16 |
| Consensus Log Po/w? Consensus Log Po/w: Average of all five predictions |
0.05 |
| Log S (ESOL):? ESOL: Topological method implemented from |
-0.08 |
| Solubility | 91.0 mg/ml ; 0.823 mol/l |
| Class? Solubility class: Log S scale |
Very soluble |
| Log S (Ali)? Ali: Topological method implemented from |
0.11 |
| Solubility | 142.0 mg/ml ; 1.28 mol/l |
| Class? Solubility class: Log S scale |
Highly soluble |
| Log S (SILICOS-IT)? SILICOS-IT: Fragmental method calculated by |
-0.14 |
| Solubility | 79.7 mg/ml ; 0.721 mol/l |
| Class? Solubility class: Log S scale |
Soluble |
| GI absorption? Gatrointestinal absorption: according to the white of the BOILED-Egg |
High |
| BBB permeant? BBB permeation: according to the yolk of the BOILED-Egg |
No |
| P-gp substrate? P-glycoprotein substrate: SVM model built on 1033 molecules (training set) |
No |
| CYP1A2 inhibitor? Cytochrome P450 1A2 inhibitor: SVM model built on 9145 molecules (training set) |
No |
| CYP2C19 inhibitor? Cytochrome P450 2C19 inhibitor: SVM model built on 9272 molecules (training set) |
No |
| CYP2C9 inhibitor? Cytochrome P450 2C9 inhibitor: SVM model built on 5940 molecules (training set) |
No |
| CYP2D6 inhibitor? Cytochrome P450 2D6 inhibitor: SVM model built on 3664 molecules (training set) |
No |
| CYP3A4 inhibitor? Cytochrome P450 3A4 inhibitor: SVM model built on 7518 molecules (training set) |
No |
| Log Kp (skin permeation)? Skin permeation: QSPR model implemented from |
-7.32 cm/s |
| Lipinski? Lipinski (Pfizer) filter: implemented from |
0.0 |
| Ghose? Ghose filter: implemented from |
None |
| Veber? Veber (GSK) filter: implemented from |
0.0 |
| Egan? Egan (Pharmacia) filter: implemented from |
0.0 |
| Muegge? Muegge (Bayer) filter: implemented from |
2.0 |
| Bioavailability Score? Abbott Bioavailability Score: Probability of F > 10% in rat |
0.55 |
| PAINS? Pan Assay Interference Structures: implemented from |
0.0 alert |
| Brenk? Structural Alert: implemented from |
1.0 alert: heavy_metal |
| Leadlikeness? Leadlikeness: implemented from |
No; 1 violation:MW<1.0 |
| Synthetic accessibility? Synthetic accessibility score: from 1 (very easy) to 10 (very difficult) |
2.32 |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With potassium phosphate; In dichloromethane; for 3h;Heating / reflux;Product distribution / selectivity; | To 1.2 L of a methylene chloride solution of (S)-3-chloro-l,2-propanediol (200 g, 99.5% ee) was added 519 g of potassium phosphate tribasic, and then the obtained solution was refluxed, under stirring, for 3 hours. The resulting solution was cooled to 0C, and 220 g of trie thy lamine, 4 g of 4-(dimethylamino)pyridine, and 315 g of butanoic acid anhydride were dropwisely added to the solution. After additional stirring for 1 hour at a room temperature, the reaction mixture was successively washed with 2.2 L of 5% aqueous potassium carbonate solution, 2 L of IN aqueous hydrogen chloride solution, and 1 L of water. The organic layer was dried with 50 g of anhydrous sodium sulfate and filtrated. The methylene chloride was evaporated under reduced pressure. Fractional distillation (90C/19 mmHg) of the resulting residue gave 242 g of the targeted compound:- Yield: 92.7%- Chemical purity: 99.4%- Optical purity (GC) 99.5% ee. | |
| With potassium carbonate; In dichloromethane; for 25h;Heating / reflux;Product distribution / selectivity; | To 1.2 L of a methylene chloride solution of (S)-3-chloro-l,2-propanediol (200 g, 99.5% ee) was added 338 g of potassium carbonate, and then the obtained solution was refluxed, under stirring, for 25 hours. The resulting solution was cooled to 0C, and 220 g of triethylamine, 4 g of 4-(dimethylamino)pyridine, and 315 g of butanoic acid anhydride were dropwisely added to the solution. After additional stirring for 1 hour at a room temperature, the reaction mixture was successively washed with 2.2 L of 5% aqueous potassium carbonate solution, 2 L of IN aqueous hydrogen chloride solution, and 1 L of water. The organic layer was dried with 50 g of anhydrous sodium sulfate and filtrated. The methylene chloride was evaporated under reduced pressure. Fractional distillation (90C/19 mmHg) of the resulting residue gave ID g of the targeted compound:- Yield: 65.0%- Chemical purity: 97.4%- Optical purity (GC) 98.1% ee. | |
| With potassium carbonate; In dichloromethane; at 20℃; for 17.5h; | 200 g of 3-chloro-1,2-propanediol was dissolved in 2000 mL of dichloromethane, 625 g of potassium carbonate was added, and the mixture was stirred at room temperature for 17.5 hours.The precipitated insoluble materials were filtered,By washing the filtration residue with 1,000 mL of dichloromethane,(S) -oxiran-2-ylmethanolWas obtained.To the obtained solution,198 g of dihydropyran was added,After cooling to 0 C.,5.16 g of p-toluenesulfonic acid monohydrate was added at 10 C. or less,And the mixture was stirred at room temperature for 1.5 hours. 408 g of a 7.5% sodium bicarbonate aqueous solution was added,After stirring, the organic layer and the aqueous layer were separated. The obtained organic layer was concentrated,Distillation gave 247.8 g of the title compound as a colorless oil. |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 26% | With Aspergillus niger epoxide hydrolases immobilized onto modified Eupergit C; In aq. phosphate buffer; dimethyl sulfoxide; at 25℃; for 3h;pH 6.5;Enzymatic reaction;Kinetics; | General procedure: Asymmetric hydrolysis of (R/S)-SO, (R/S)-PO and (R/S)-ECH were examined in a batch type reactor (1.1 cm × 5 cm). To 1.5 mL of 100 mM phosphate buffer (pH = 7.0 for the free and EHIL; pH = 6.5 for EHIF and EHIE), 100 L of the free EH solution (1 mg mL-1) or 30 mg of each immobilized EH was loaded and the mixture kept at 25 C for 2 min. The reaction was initiated by the addition 0.4 mL of each racemic epoxide solution (0.5 M in DMSO). A hundred microliters of aliquots withdrawn at different time intervals (15, 30,60, 90, 120, 180 and 240 min) were mixed with 400 L of diethylether and analyzed by a Shodex ORpak CDC-453 HQ chiral HPLC column (4.6 mm × 150 mm) according to Yildirim et al. [23]. The enantiomers of styrene oxide and their vicinal diols were detected at 220 nm. The enantiomers of propylene oxide, epichlorohydrin and their vicinal diols were detected using a refractive index detector (Shimadzu RID-10A). The optical configurations of remaining epoxides and formed diols were identified by comparing the retention time of these compounds with their optically active standard forms. The enantiomeric excess (ee) values of formed vicinal diol and remaining epoxide were calculated from the equations: eeepoxide=([S-R]epoxide)/([S+R]epoxide) and eediol=([R-S]diol)/([R+S]diol) The enantiomeric ratio values (E) of free and immobilized EHs were calculated from the equation proposed by Chen et al. [27]. E=(Vmax(R)/Km(R))/(Vmax(S)/Km(S)) where Vmax(R) and Km(R) values are maximum velocity and Michealis-Menten constant of free and immobilized EH preparations toward (R)-enantiomer of epoxide and Vmax(S) and Km(S) are corresponding values toward (S)-enantiomer of epoxide. |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| EXAMPLE 3 This Example shows the conversion of (S)-3-amino-1,2-dihydroxypropane isopropylidene acetal 11 to (S)-1-chloro-2,3-dihydroxypropane 3. | ||
| EXAMPLE 2 Sodium periodate (9.31 g) was added slowly to a cooled and stirred solution of 6,6'-dichloro-6,6'-dideoxy-sucrose (5.0 g) in water (250 ml). After stirring at room temperature for 2 hours, the solution was diluted with ethanol (750 ml) and the resulting suspension was stirred at room temperature for a further 2 hours. The suspension was then filtered and the filtrate was concentrated to a syrup (25 ml) which was diluted with water (100 ml). A solution of sodium borohydride (2.04 g) in water (50 ml) was added slowly with cooling to the solution of oxidised product. After the addition, the reaction mixture was stirred at room temperature for 20 hours, and then a few drops of acetic acid were added. Concentration in vacuo at 40 gave a semi-solid mass which was then dissolved in ethyl acetate (AR):ethanol (9:1) and eluted through a column of silica gel. Evaporation of the elude yielded a semi-solid mass (5.1 g). T.l.c. showed that this was a mixture. The components were identified by g.l.c. to be alpha-chlorohydrin, dihydroxyacetone (dimer) and hydroxyacetaldehyde. Chromatography and distillation of the product mixture gave S-3-chloro-1,2-propanediol identical with that obtained in Example 1. | ||
| EXAMPLE 3 This Example shows the conversion of (S)-3-amino-1,2-dihydroxypropane isopropylidene acetal 11 to (S)-1-chloro-2,3-dihydroxypropane 3 . |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With potassium tert-butylate; acetic acid; In tetrahydrofuran; hexane; water; ethyl acetate; | To a solution of 1056 g (3.64 mol) of 2-Methylpropyl (4-bromo-3-fluorophenyl)carbamate in 6.65 L of THF cooled to -15 C. in a 22 L round bottom flask was added a solution of 428 g (4.55 mol, 1.25 equiv.) of lithium t-amylate over 10 min via an addition funnel, maintaining -15 to -12 C. In a separate 5 L flask, a solution of 438 g (4.37 mol, 1.20 equiv.) of (S)-3-chloro-1,2-propanediol in 1.75 L of THF was cooled to -25 C. and treated with a 20% t-BuOK solution in THF (2645 mL, 4.29 mol, 1.18 equiv) over 25 min, resulting in a thick but stirrable slurry. This was allowed to warm to 10 over 75 min and then poured into the 22 L flask containing the carbamate solution. The resulting slurry was allowed to warm from -7 to 7 over 1.5 h, monitoring reaction progress by HPLC. Upon completion (~2.5% each of remaining 2-Methylpropyl (4-bromo-3-fluorophenyl)carbamate and the over addition product) a quench solution composed of 1.05 AcOH and 3.5 L of water was added. The layers were separated. The aqueous layer was back-extracted with 1 L of THF, and the combined organic layers were washed with brine. The volatile were removed, giving white solids wet with acetic acid. This material was slurried in 1.6 L of EtOAc. Hexane (4L) was added over 1 h. The resulting slurry was cooled to 2 over 1 h and filtered, giving 916 g (87%) of (5R)-3-(4-bromo-3-fluorophenyl)-5-(hydroxymethyl)-1,3-oxazolidin-2-one as coarse white crystals: TLC Rf=0.008 (50% EtOAc/hexane); HPLC rt=2.55 min; mp 114-121 ; [alpha]D=+52.2 c. =1, MeOH); 1H NMR (DMSO) delta7.49 (m, 2H), 7.15 (d,1H, J=8.5 Hz), 5.30 (br s, 1H), 4.54 (m,1H), 3.89 (t, 1H, J=8.6 Hz), 3.67 (m, 1H), 3.50 (dd, 1H, J=3.0, 11.8 Hz), 3.39 (dd, 1H, 3.8, 11.8 Hz); 13C NMR (DMSO) delta158.1 (s, JCF=241 hz), 154.2 (S), 139.7 (S, JCF=10 hz), 133.3 (D), 114.9 (D), 105.9 (d, JCF=29 Hz), 100.9 (s, JCF=21 Hz), 73.3(d), 61.5(t), 45.9 (t); Anal. calc'd for C10H9BrFNO3: C, 41.40; H, 3.13; N, 4.83: Br, 27.55; found: C, 41.11; H, 3.06; N, 4.83: Br, 26.97. |
[ 1011475-45-8 ]
[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In water; isopropyl alcohol; at 60℃; for 72h; | Example 14 To a solution of 4-[5-(5,6-diethyl-pyridin-3-yl)-[1,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenol (100 mg, 0.296 mmol) in isopropanol (10 mL) and 3 N aq. NaOH (3 mL), (S)-3-chloro-1,2-propanediol (98 mg, 0.89 mmol) is added. The mixture is stirred at 60 C. for 24 h before another portion of (S)-3-chloro-1,2-propanediol (98 mg, 0.89 mmol) is added. Stirring is continued at 60 C. for 2 days. The mixture is diluted with EA and washed with sat. aq. NaHCO3 solution. The org. extract is dried over MgSO4, filtered and evaporated. The crude product is purified by chromatography on prep. TLC plates with EA-heptane to give (S)-3-{4-[5-(5,6-diethyl-pyridin-3-yl)-[1,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenoxy}-propane-1,2-diol (14 mg) as a red oil; LC-MS: tR=0.93 min, [M+1]+=412.16. | |
| With sodium hydroxide; In water; isopropyl alcohol; at 60℃; for 72h; | To a solution of 4-[5-(5,6-diethyl-pyridin-3-yl)-[1 ,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenol (100 mg, 0.296 mmol) in isopropanol (10 ml.) and 3 N aq. NaOH (3 ml_), (S)-3-chloro-1 ,2- propanediol (98 mg, 0.89 mmol) is added. The mixture is stirred at 600C for 24 h before another portion of (S)-3-chloro-1 ,2-propanediol (98 mg, 0.89 mmol) is added. Stirring is continued at 600C for 2 days. The mixture is diluted with EA and washed with sat. aq. NaHCOs solution. The org. extract is dried over MgSO4, filtered and evaporated. The crude product is purified by chromatography on prep. TLC plates with EA-heptane to give (S)-3- {4-[5-(5,6-diethyl-pyridin-3-yl)-[1 ,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenoxy}-propane-1 ,2- diol (14 mg) as a red oil; LC-MS: tR = 0.93 min, [M+1]+ = 412.16. |
[ 23010-10-8 ]
[ 60827-45-4 ]

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 66.8% | In Na4e(CN)610H2; water; ethyl acetate; toluene; | 6.9 Controlled Potential Electrocatalytic ADH of allyl Chloride A divided glass H-cell as described in Example-6.1 which is been equipped with a platinum-on-titanium metal anode (5 cm2 and a nickel foil cathode is charged in the cathode compartment with 90 ML of saturated aqueous NaH2O4olution and in the anode compartment with Na4e(CN)610H2 (4.84 g, 10.0 mmol), K2O313.8 g, 100 mmol) and t-BuOH (45 ML)/water (55 ML), the chiral ligand, PHAL-DHQD (292 mg. 0.375 mmol), OsO40.196 M in toluene, 0.128 ML, 0.025 mmol) and allyl chloride (3.83 g, 50 mmol). The mixture is electrolyzed at 0.4 V (vs ref. Ag/AgCl) at 15 C until about 9090 coulombs (19 hrs, ca. 94% conversion) of electricity is used. After stirring without electricity for one hour, the anode reaction mixture is transferred to a flask (250 ML) and saturated with NaCl. The reaction mixture is then quenched with 5 g of Na2O3 and extracted with 3 x 50 ML of ethyl acetate. The combined organic solution is washed with 10 ML of 5% H3O4queous solution saturated with NaCl, 15 ML of saturated NaCl solution and dried over 5 g of anhydrous Na2O4 After filtration and removal of solvent under vacuum, (S)-3-chloro-1,2-propanediol is obtained as a pale yellow oil (3.48 g, 66.8% yield based on 94% conversion). The ee of the diol is 54.9% by HPLC analysis of the bis-acetate derivative of the diol on a Chiracell OD column). A sample of this product is analyzed, subsequently, to provide the following physical characteristics: [alpha] [25/D] = +4.43 (c = 2.1, MeOH); literature: EP 0286059 for R-isomer, [alpha] [25/D] = -6.9 (c = 2.0, H2); 1 NMR: (CDCl3 delta 3.6-4.2O); (m, 7H); IR (neat) 3357, 2946, 1432, 1303, 1102, 747 cm-1 |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With Serratia-derived glycerol dehydrogenase; NADH; at 30℃; for 2h;pH 6.5;Aqueous phophate buffer; Enzymatic reaction; | (Example 9) Method for synthesizing (S)-3-chloro-1,2-propanediol A Serratia-derived glycerol dehydrogenase solution was obtained from a culture broth of Serratia marcescens IFO12648 strains by a purification operation comprising ammonium sulfate fractionation and ion-exchange chromatography. 1 ml of a 0.1M phosphate buffer (PH 6.5) containing 10 mul of the thus-obtained glycerol dehydrogenase solution, 1.5 mg of 1-chloro-3-hydroxyacetone, and 10 mg of NADH was stirred at 30C for 2 hours.. After the reaction was completed, the reaction solution was saturated with ammonium sulfate, and extracted with an equivalent volume of ethyl acetate.. The amounts of the substrate and product in the extract were measured by gas chromatography (GC) analysis to determine the conversion rate (%).. Also, the optical purity (%ee) of the product was measured by high-performance liquid chromatography (HPLC) analysis after the primary hydroxyl group of the product was tosylated with tosyl chloride.. As a result, (S)-3-chloro-1,2-propanediol was produced with a conversion rate of 100% and an optical purity of 100%ee.. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With D-glucose; NAD; In water;pH 7.0;Microbiological reaction; | (Example 12) Method for synthesizing (S)-3-chloro-1,2-propanediol Recombinant Escherichia coli HB101 (PNTSGG1) Accession No. FERM P-18449 were inoculated into 50 ml of sterilized 2*YT medium (16 g of tryptone, 10 g of yeast extract, 5 g of sodium chloride, and 1L of water, PH 7.0 before sterillization) in a 500 ml Sakaguchi flask, and cultured with shaking at 37C for 18 hours.. Then, 10 mg of 1-chloro-3-hydroxyacetone, 0.6 mg of NAD+, and 25 mg of glucose were added to 1 ml of the resultant culture broth, and the mixture was then stirred at 30C for 2 hours.. After the reaction was completed, the conversion rate and optical purity of the reaction product (S)-3-chloro-1,2-propanediol were analyzed by the same method as in Example 9.. As a result, the conversion rate was 97.1%, and the optical purity was 100%ee. | |
| With Cellulomonas-derived glycerol dehydrogenase; NADH;Enzymatic reaction; | (Example 10) Method for synthesizing (S)-3-chloro-1,2-propanediol 1 ml of a 0.1M phosphate buffer (PH 6.5) containing 0.05 mg of Cellulomonas-derived glycerol dehydrogenase (produced by SIGMA), 1.5 mg of 1-chloro-3-hydroxyacetone, and 10 mg of NADH was stirred at 30C for 2 hours.. After the reaction was completed, the conversion rate and optical purity of the reaction product (S)-3-chloro-1,2-propanediol were analyzed by the same method as in Example 9.. As a result, the conversion rate was 100%, and the optical purity was 99.7%ee. |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With potassium dihydrogenphosphate; D-glucose; In water; at 30℃; for 24h;pH 5.5;Aqueous phophate buffer; Microbiological reaction; | (Example 1) A liquid medium (pH 7.0) containing 4% of glucose, 0.3% of yeast extract, 1.3% of KH2PO4, 0.7% of (NH4)2HPO4, 0.01% of NaCl, 0.08% of MgSO4·7H2O, 0.006% of ZnSO4·7H2O, 0.009% of FeSO4·7H2O, 0.0005% of CuSO4·5H2O, and 0.001% of MnSO4·4-5H2O was prepared, and 5 ml of the liquid medium was injected into each of large test tubes and then sterilized with steam at 120C for 20 minutes. Then, one platinum loop of the microorganisms of each of the genera shown in Table 1 and 2 was inoculated into the liquid medium, and cultured with shaking at 30C for 2 to 3 days. Cells were collected from each of the culture broths by centrifugation, washed with water and then suspended in 1 ml of a 0.1M phosphate buffer (pH 5.5). Then, 0.5 ml of the cell suspension was mixed with 0.5 ml of a 0.1M KH2PO4 aqueous solution containing 5 mg of 1-chloro-3-hydroxyacetone and 20 mg of glucose, and the resultant mixture was placed in a test tube with a stopper and shaken at 30C for 24 hours. After completion of reaction, a saturated amount of ammonium sulfate was added to the reaction solution, and extraction was performed with an equivalent volume of ethyl acetate. | |
| With D-glucose; NAD; In water; at 30℃; for 2h;Microbiological reaction; | 5(Example 13) Method for synthesizing (S)-3-chloro1,2-propanediol[0091] Recombinant Escherichia coli HB101 (pNTCRG) Accession No. FERM BP-6898 were inoculated into 50 mlof sterilized 23YT medium (16 g of tryptone, 10 g of yeast extract, 5 g of sodium chloride, and 1L of water, pH 7.0before sterillization) in a 500 ml Sakaguchi flask, and cultured with shaking at 37C for 18 hours. Then, 10 mg of1-chloro-3-hydroxyacetone, 70 mg of NAD+, and 25 mg of glucose were added to 1 ml of the resultant culture broth,and the mixture was then stirred at 30C for 2 hours. After the reaction was completed, the conversion rate and opticalpurity of the reaction product (S)-3-chloro-1,2-propanediol were analyzed by the same method as in Example 9. As aresult, the conversion rate was 42.8%, and the optical purity was 45.8%ee. |

| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 95.4%Chromat. | Example 1 Preparation of (S)-3-chloro-1,2-propanediol To a mixture of N',N'-bis(3,5-di-tertbutylsalicylidene) ethylenediaminatocobalt(II) (119 mg, 0.216 mmol) and tetrahydrofuran (THF) (2.0 ml) was added (+) camphorsulfonic acid (60.3 mg, 0.260 mmol) and the reaction system was stirred for 1 hour while being filled with air.Thereto, namely to cobalt (III) complex in THF solution was added (S)-epichlorohydrin (1.00 g, 10.8 mmol, optical purity 99% e.e.) and water (234 mul, 13.0 mmol) in order, and the mixture was stirred for 20 hours at room temperature.After the reaction, quantitative analysis and optical purity measurement were conducted by gas chromatography on the reaction solution.As a result the subject compound, (S)-3-chloro-1,2-propanediol. (1.14 g, 95.4%) was produced with optical purity at 99% e.e. |
[ 875012-84-3 ]
[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; sodium iodide; In water; isopropyl alcohol; at 90℃; for 8h; | c) To a solution of (1aS,5aR)-1,1,2-trimethyl-1,1a,5,5a-tetrahydro-3-thia-cyclopropa[a]pentalene-4-carboxylic acid 4-hydroxy-3,5-dimethyl-benzyIamide (8.9 mg, 0.025 mmol) in isopropanol (1 ml_) is added 2 N aq. NaOH (100 jaL), Nal (1 mg, 0.007 mmol) and (S)-3-chloro-propane-1,2-diol (11.1 mg, 0.1 mmol). The reaction mixture is heated to 90C and shaken for 8 h before it is cooled to it Formic acid (0.2 ml) is added and the reaction mixture is separated by prep. HPLC to give (1 aS,5aR)-1,1,2-trimethyl-1,1 a,5,5a-tetrahydro-3-thia-cyclopropa[a]pentalene-4-carboxylic acid 4-((S)-2,3-dihydroxy-propoxy)-3,5-dimethyl-benzylamide (3.7 mg) as a colourless lyophilisate. LC-MS: tR = 0.94 min, [M+1]+ = 430.27. | |
| With sodium hydroxide;sodium iodide; In isopropyl alcohol; at 90℃; for 8h; | To a solution of (1aS,5aR)-1,1,2-trimethyl-1,1a,5,5a-tetrahydro-3-thia-cyclopropa[a]pentalene-4-carboxylic acid 4-hydroxy-3,5-dimethyl-benzylamide (8.9mg, 0.025 mmol) in isopropanol (1 ml) is added 2 N aq. NaOH (100 pi), Nal (1mg, 0.007 mmol) and (S)-3-chloro-propane-1,2-diol (11.1 mg, 0.1 mmol). Thereaction mixture is heated to 90C and shaken for 8 h before it is cooled to rt.Formic acid (0.2 mL) is added and the reaction mixture is separated by prep.HPLC to give (1aS,5aR)-1,1,2-trimethyl-1,1a,5,5a-tetrahydro-3-thia-cyclopropa[a]pentalene-4-carboxylic acid 4-((S)-2,3-dihydroxy-propoxy)-3,5-dimethyl-benzylamide (3.7 mg) as a colourless lyophilisate. LC-MS: tR = 0.94 min,[M+1]+ = 430.27. |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| In N,N-dimethyl-formamide; | a (2R)-3-Azido-1,2-bis(triethylsilyloxy)propane (2S)-3-Chloro-1,2-propanediol (3.30 g) was dissolved in 15 ml of DMF. Sodium azide (2.33 g) was added to the solution. The mixture was stirred at 100 C. for 20 hr. DMF (15 ml) was added to the reaction solution. Imidazole (2.03 g) and 5.67 ml of triethylsilyl chloride were added thereto under cooling in ice. The mixture was stirred for 10 min. Imidazole (2.23 g) and 5.95 ml of triethylsilyl chloride were added thereto. The mixture was stirred for 10 min. Brine (120 ml) was then added to the reaction solution, and the mixture was extracted with 120 ml of diethyl ether three times. The combined organic layers were dried over anhydrous magnesium sulfate, and were filtered. The solvent was removed by distillation under the reduced pressure. Thus, 11.21 g of (2R)-3-azido-1,2-bis(triethylsilyloxy)propane was prepared. NMR (CDCl3) delta: 0.57-0.67 (18H, m), 0.91-1.01 (12H, m), 3.19-3.43 (2H, m), 3.49-3.61 (2H, m), 3.80-3.87 (1H, m) |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 99% | In water; | EXAMPLE 5 Preparation of (S)-3-Chloro-1,2-propanediol with (S,S)-Co(salen): To a 3-necked, 250-mL, round-bottomed flask fitted with a vacuum adapter, an air inlet, and a temperature probe was added (S,S)-Co(salen) (1.51 g, 0.0023 mol, 0.23 mol %), propylene glycol monomethyl ether (20 mL), and p-nitrobenzoic acid (0.835 g, 0.0049 mol, 0.49 mol %). A slight vacuum was applied to pull air into the reaction medium, and the reaction was allowed to stir for 1 h. The air inlet was removed, and epichlorohydrin (92.52 g, 78.40 mL, 1.00 mol) was charged after the reaction vessel was purged with nitrogen. The reaction was cooled to 0 C., and water (8.1 g, 8.1 mL, 0.45 mol, 45 mol %) was slowly added, maintaining the reaction temperature below 5 C. The reaction was allowed to stir at room temperature for 16 h. When determined to be complete, the reaction was partitioned between toluene (80 mL) and water (80 mL). The aqueous layer was collected and washed with toluene (80 mL) to remove unreacted epichlorohydrin (isopropyl alcohol (20%) was added to facilitate the phase separation). After concentration of aqueous layer, the residue was vacuum distilled to give (S)-3-chloro-1,2-propanediol (37.4 g, 76% yield, >99% purity, 97% ee; b.p. 115-120 C. a 10 mm Hg). |
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| 70% | With hydrogenchloride; N,N-dimethylamino-pyridine; potassium carbonate; triethylamine; In water; | Comparative Example 3 To a suspension of 37.5 g of potassium carbonate (0.27 mol) and 250 ml of 1,2-dichloroethane, was added dropwise at 24-28 C. 20 g of (S)-3-chloro-1,2-propanediol (0.18 mol) (optical purity: 98.9% ee). After finishing the addition, the solution was stirred for 26 hours and cooled. To the reaction solution were added dropwise under stirring at 5-10 C. 20.1 g of triethylamine (0.2 mol) and 0.4 g of N,N-dimethylaminopyridine (0.0033 mol) and 34.5 g of p-toluenesulfonyl chloride (0.18 ml) in order. After finishing the addition the mixture was stirred for 3 hours and the resulting salt was dissolved by adding 150 ml of 3% hydrochloric acid. The organic layer was washed with 150 ml of 1% hydrochloric acid and 150 ml of water. The excess solvent was removed under vacuo. The chemical purity and optical purity at that time were 93.7%, 96.6% ee respectively. The residue was recrystallized from isopropyl alcohol/hexane=1/1 (V/V) to give 28.9 g of (S)-glycidyl tosylate (yield 70%). Chemical purity: 98.3%, optical purity: 97.5% ee |
[ 917873-20-2 ]
[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In water; isopropyl alcohol; at 65℃; for 24h; | Example 2; A solution of 3-(4-hydroxy-3,5-dimethyl-phenyl)-1 -(4-phenyl-5-trifluoromethyl- thiophen-2-yl)-propan-1 -one (8 mg, 20 mumol) in 2-propanol (0.7 mL) and 2 N aq. NaOH (0.27 mL) is treated with (S)-3-chloro-propane-1 ,2-diol (9 mg, 80 mumol) and the mixture is stirred at 65 C for 24 h. The mixture is cooled to rt, diluted with formic EPO <DP n="22"/>acid (0.25 ml_) and separated by prep. HPLC (Waters Xterra MS18 19x50mm 5mum, 10 to 95% acetonitrile in water containing 0.5 % formic acid) to give 3-[4-((S)-2,3- dihydroxy-propoxy)-3,5-dimethyl-phenyl]-1 -(4-phenyl-5-trifluoro-methyl-thiophen-2- yl)-propan-1 -one (3 mg) as a colourless lyophilisate; LC-MS: tR = 1.06 min, [M+1]+ = 479.21. |
[ 917873-23-5 ]
[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In water; isopropyl alcohol; at 70℃; for 24h; | Example 5; A solution of 3-(4-hydroxy-3,5-dimethyl-phenyl)-1 -(5-methyl-4-phenyl-thiophen-2-yl)- propan-1 -one (60 mg, 0.172 mmol) in 2-propanol (1 ml_) and 3 N aq. NaOH (0.3 ml_) is treated with (S)-3-chloro-propane-1 ,2-diol (95 mg, 0.86 mmol) and the mixture is stirred at 70C for 24 h. The solvent is evaporated and the residue is purified on prep. TLC plates with EA to give 3-[4-((S)-2,3-dihydroxy-propoxy)-3,5-dimethyl- phenyl]-1 -(5-methyl-4-phenyl-thiophen-2-yl)-propan-1 -one (44 mg) as an oil; LC-MS: tR = 1.00 min, [M+I]+ = 425.23; 1H NMR (CDCI3): delta 7.60 (s, 1 H), 7.46-7.32 (m, 5H), 6.88 (s, 2H), 4.1 1 -4.06 (m, 1 H), 3.86-3.80 (m, 4H), 3.19-3.1 1 (m, 2H), 2.98-2.91 (m, 2H), 2.72 (d, J = 5.3 Hz, 1 H), 2.53 (s, 3H), 2.25 (s, 6H), 2.1 1 -2.05 (m, 1 H). |
[ 917873-30-4 ]
[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In water; isopropyl alcohol; at 70℃; for 15h; | Example 10; A solution of 3-(4-hydroxy-3,5-dimethyl-phenyl)-1 -(5-ethyl-4-phenyl-thiophen-2-yl)- propan-1 -one (52 mg, 0.143 mmol) in 2-propanol (1 mL) and 3 N aq. NaOH (0.3 mL) is treated with (S)-3-chloro-propane-1 ,2-diol (79 mg, 0.71 mmol) and the mixture is stirred at 70C for 15 h. The reaction mixture is diluted with water and extracted with DCM. The solvent of the organic extract is evaporated and the crude product is purified on prep. TLC plates with heptane:EA 1 :3 to give 3-[4-((S)-2,3-dihydroxy- EPO <DP n="28"/>propoxy)-3,5-dimethyl-phenyl]-1 -(5-ethyl-4-phenyl-thiophen-2-yl)-propan-1 -one (14 mg) as a colourless resin; LC-MS: tR = 1.03 min, [M+1 ]+ = 439.25; 1H NMR (CDCI3): delta 7.58 (s, 1 H), 7.45-7.30 (m, 5H), 6.88 (s, 2H), 4.1 1 -4.04 (m, 1 H), 3.87-3.80 (m, 4H), 3.67-3.60 (m, 2H), 3.60-3.56 (m, 1 H), 3.48-3.40 (m, 1 H), 3.20-3.10 (m, 2H), 2.98- 2.87 (m, 4H), 2.25 (s, 6H), 1.30 (t, J = 7.6 Hz, 3H). |

[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In ethanol; water; at 60℃; for 14h; | Example 20; EPO <DP n="36"/>A solution of 1 -(5-ethyl-4-p-tolyl-thiophen-2-yl)-3-(4-hydroxy-3,5-dimethyl-phenyl)- propan-1 -one (18 mg, 48 mumol) in ethanol (1.5 ml_) and 3 N aq. NaOH (0.3 ml_) is treated with (S)-3-chloro-propane-1 ,2-diol (30 mul_) and the mixture is stirred at 60C for 14 h. The solvent is removed in vacuo and the residue is separated on prep. TLC plates with heptane:EA 2:1 to give 3-[4-((S)-2,3-dihydroxy-propoxy)-3,5-dimethyl- phenyl]-1 -(5-ethyl-4-o-tolyl-thiophen-2-yl)-propan-1 -one (4 mg) as a colourless resin; LC-MS: tR = 1.04 min, [M+I ]+ = 453.28; 1H NMR (CDCI3): delta 7.45 (s, 1 H), 7.29-7.08 (m, 4H), 6.87 (s, 2H), 4.1 1 -4.04 (m, 1 H), 3.87-3.80 (m, 4H), 3.16-3.10 (m, 2H), 2.97- 2.90 (m, 2H), 2.64 (q, J = 7.6 Hz, 2H), 2.24 (s, 3H), 2.17 (s, 6H), 1.20 (t, J = 7.6 Hz, 3H). |

[ 60827-45-4 ]
| Yield | Reaction Conditions | Operation in experiment |
|---|---|---|
| With sodium hydroxide; In ethanol; water; at 60℃; for 140h; | Example 22; A solution of 1 -(5-ethyl-4-p-tolyl-thiophen-2-yl)-3-(4-hydroxy-3,5-dimethyl-phenyl)- propan-1 -one (47 mg, 0.125 mmol) in ethanol (2 mL) and 3 N aq. NaOH (0.5 mL) is treated with (S)-3-chloro-propane-1 ,2-diol (90 muL) and the mixture is stirred at 60C for 20 h. Another portion of (S)-3-chloro-propane-1 ,2-diol (60 muL) is added and stirring is continued at 60C for 5 days. The reaction mixture is diluted with water and extracted with DCM. The solvent of the organic extract is evaporated and the crude product is purified on prep. TLC plates with heptane:EA 2:1 to give 3-[4-((S)- 2,3-dihydroxy-propoxy)-3,5-dimethyl-phenyl]-1 -(5-ethyl-4-p-tolyl-thiophen-2-yl)- propan-1 -one (6 mg) as a colourless resin; LC-MS: tR = 1.05 min, [M+1 ]+ = 453.25; 1H NMR (CDCI3): delta 7.56 (s, 1 H), 7.30-7.21 (m, 4H), 6.88 (s, 2H), 4.1 1 -4.05 (m, 1 H), 3.87-3.80 (m, 4H), 3.18-3.1 1 (m, 2H), 2.98-2.85 (m, 4H), 2.39 (s, 3H), 2.25 (s, 6H), 1.29 (t, J = 7.6 Hz, 3H). |

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