* 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.
Reference：
[1] European Journal of Medicinal Chemistry, 2006, vol. 41, # 9, p. 1059 - 1072
17
[ 2280-44-6 ]
[ 83-46-5 ]
[ 474-58-8 ]
Yield
Reaction Conditions
Operation in experiment
82%
With trimethylsilyl trifluoromethanesulfonate In ethylbenzene at 85℃; for 15 h;
β-sitosterol124.4g0.3mol),D-glucose (18.0 g, 0.1 mol) dissolved in ethylbenzene (600 mL).At 80°C,TMSOTf (8.9 g/7.3 mL, 40 mmol)was added and underprotection carried out constant temperature reaction until the TLC detected almost complete disappearance of D-glucose (about 15h). After the reaction was concentrated , it wassubjected to silica gel column chromatography (silica gel 100-200 mesh) using dichlromethaneas eluent. TLC assay until the eluent is free of β-sitosterol, the eluent was concentrated to give β-sitosterol(83.0g, 0.2mol), continue the elution with ethyl acetateas eluent, and TLC assay until there is no title compoundin the eluent. The eluent was concentrated to give the title compound (47.3 g) in 82percent yield and HPLC purity 97.8percent.
Stage #1: With sodium hydrogencarbonate; hydrazine hydrate; acetic acid In water for 5 h; Reflux Stage #2: With sodium periodate; acetic acid In water at 20℃; for 16 h;
Quninoxaline-2-carbaldehyde was synthesized according to the previous report 22 and literature known procedure. At room temperature, glacial acetic acid (1.5 ml), o-phenylenediamine (5gm, 46mmol), hydrazine hydrate (1.5ml) and a pinch of Sodium bicarbonate were added to solution of D-glucose (8.5gm, 46mmol) in water (12.5ml) and the reaction was heated under reflux for 5hrs on sand bath the product, 2-(D-arabinotetrahydroxybutyl)-quinoxaline, which got precipitated on cooling the solution in ice, was filtered and washed with water. It was further
Reference：
[1] Bioorganic and Medicinal Chemistry Letters, 2017, vol. 27, # 10, p. 2174 - 2180
23
[ 2280-44-6 ]
[ 65-23-6 ]
Reference：
[1] Bioscience, biotechnology, and biochemistry, 2001, vol. 65, # 8, p. 1789 - 1795
24
[ 131623-13-7 ]
[ 2280-44-6 ]
[ 486-60-2 ]
Reference：
[1] Chemical and Pharmaceutical Bulletin, 1990, vol. 38, # 9, p. 2498 - 2502
[2] Chemical and Pharmaceutical Bulletin, 1990, vol. 38, # 9, p. 2498 - 2502
25
[ 2280-44-6 ]
[ 2041-14-7 ]
Reference：
[1] Phosphorus, Sulfur and Silicon and the Related Elements, 1993, vol. 76, # 1-4, p. 111 - 114
26
[ 2280-44-6 ]
[ 64-17-5 ]
[ 3198-49-0 ]
Reference：
[1] Journal of Carbohydrate Chemistry, 2008, vol. 27, # 5, p. 300 - 308
[2] Synthesis, 1991, # 7, p. 533 - 535
[3] Indian Journal of Chemistry - Section B Organic and Medicinal Chemistry, 2007, vol. 46, # 2, p. 314 - 319
[4] Patent: US2005/261484, 2005, A1, . Location in patent: Page/Page column 4
[5] Research on Chemical Intermediates, 2018, vol. 44, # 3, p. 1627 - 1645
27
[ 2280-44-6 ]
[ 1125-88-8 ]
[ 30688-66-5 ]
Yield
Reaction Conditions
Operation in experiment
53.7%
With toluene-4-sulfonic acid In N,N-dimethyl-formamide at 60℃; for 6 h;
Glucose (1.00 g, 5.55 mmol) was added to a N, N-dimethylformamide solution (11.0 mL)It was heated to 60 ° C and dissolved.To this was added benzaldehyde dimethyl acetal (1.24 mL, 8.33 mmol),Paratoluenesulfonic acid monohydrate (10 mg) was added and the mixture was stirred at 60 ° C for 6 hours.The pressure reduction operation was carried out for 10 minutes every hour. After 6 hours, the reaction solution was evaporated under reduced pressure.The obtained residue was subjected to silica gel column chromatography, and as a white crystal from a fraction eluted with methanol / ethyl acetate (1/10)4,6-O-Benzyliden-D-glucopyranose (799.5 mg, 2.98 mmol, yield 53.7percent). As glucose, those of D - (+) - glucose (manufactured by Kanto Kagaku Co., Ltd.) were used.
Reference：
[1] Tetrahedron Letters, 2011, vol. 52, # 49, p. 6587 - 6590
[2] European Journal of Organic Chemistry, 2015, vol. 2015, # 23, p. 5075 - 5078
[3] Patent: JP2018/30828, 2018, A, . Location in patent: Paragraph 0051-0052
[4] Bioscience, Biotechnology and Biochemistry, 2001, vol. 65, # 3, p. 542 - 547
[5] Tetrahedron Asymmetry, 2008, vol. 19, # 2, p. 258 - 264
28
[ 2280-44-6 ]
[ 100-52-7 ]
[ 30688-66-5 ]
Reference：
[1] Journal of Organic Chemistry, 1995, vol. <8> 60, p. 2537 - 2548
at 150 - 280℃; under 90009.0 Torr; for 0.0333333h;Heating / reflux;
A mixture of ethanol and glucose was initially charged in a stirred vessel A. The fraction of glucose was 30% of the ethanol fraction. The mixture was conveyed by a suitable pump B at a pressure of 120 bar continuously through the reactor C. The tubular reactor C was heated to a temperature of 150 C. by the heater D. A carrier stream comprising pure ethanol from the reservoir F was heated to approx. 280 C. using the preheater G. At the mixing point H, the carrier stream was mixed with the reactant stream and fed to the reactor. The preheating temperature was sufficiently high that reaction temperature was already present at the mixing point, so that the temperature gradient in the reactor was sufficiently low to prevent carbonization at the wall. The residence time in the tubular reactor was approx. 2 min. With the aid of the control valve E, the pressure was kept at the target value mentioned. At the outlet of the system, the reacted product mixture was collected. According to HPLC, the product mixture contained 10% product
With toluene-4-sulfonic acid; In N,N-dimethyl-formamide; at 60℃; for 6h;
A suspension of D-glucose (1) (1.00 g, 5.55 mmol) in DMF(11 mL) was heated to 60 C to dissolve the solid. Benzaldehydedimethyl acetal (1.24 mL, 8.33 mmol) and p-toluenesulfonic acidmonohydrate (10 mg) were added. The resulting solution wasstirred at 60 C for 6 h, depressurizing 10 min for each hour in orderto remove MeOH. The mixture was concentrated in vacuo. Theresidue was purified by silica gel column chromatography (MeOH/AcOEt 1:10, v/v) to give glucopyranose 2 (799.5 mg, 2.98 mmol,53.7%) as a white solid.2: [a]20D 12.9 (c 0.20, MeOH); 1H-NMR (400 MHz, CD3OD):d 7.46e7.42 (m, 2H), 7.38e7.35 (m, 3H), 6.31 (d, J 4.1 Hz, 0.5H),5.79 (d, J 7.8 Hz, 0.5H), 5.60 (t, J 9.9 Hz, 0.5H), 5.52 (s, 0.5H), 5.51(s, 0.5H), 5.37 (t, J 9.2 Hz, 0.5H), 5.17e5.10 (m, 1H), 4.39 (dd,J 10.3, 4.6 Hz, 0.5H), 4.32 (dd, J 10.5, 5.0 Hz, 0.5H), 4.08e4.00(m, 0.5H), 3.80e3.63 (m, 2.5H), 2.19 (s, 1.5H), 2.11 (s, 1.5H), 2.08 (s,1.5H), 2.06 (s,1.5H), 2.05 (s,1.5H), 2.04 (s,1.5H); 13C-NMR (100 MHz,CD3OD): d 139.2, 139.1, 129.9, 129.0, 127.5, 103.0, 102.9, 99.0, 94.7,83.1, 82.4, 77.2, 74.7, 74.4, 71.8, 70.3, 69.8, 67.7, 63.5; HRMS (ESI):calcd for C13H16O6Na ([MNa]): 291.0845, found: 291.0842.
43.3%
With toluene-4-sulfonic acid; In N,N-dimethyl-formamide; at 60℃; for 5h;
Compound 3 was prepared according to the literature procedure (mp 176-178 C as reported).
With camphor-10-sulfonic acid; In N,N-dimethyl-formamide; at 20℃; for 25h;
Glucose (90 g) and N, N-dimethylformamide(DMF) (600 ml) were stirred at room temperature,Benzaldehyde dimethyl acetal (90 ml) and camphorsulfonic acid (CSA) (6.18 g) were added in order,And the mixture was stirred at room temperature under reduced pressure for 25 hours.After completion of the reaction, DMF was distilled off under reduced pressure, and triethylamine (4.5 ml) and pyridine (450 ml) were added to and dissolved in the obtained concentrated residue. The solution was cooled to 0 to 5 C.,Acetic anhydride (450 ml) was added, then the temperature was returned to room temperature and stirred for 15 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, toluene was added to the residue, azeotropy was carried out, and pyridine was distilled off. Dichloromethane (600 ml) was added to the obtained concentrated residue and dissolved, and the mixture was washed with saturated aqueous sodium bicarbonate (450 ml) and saturated brine (450 ml), and dried over magnesium sulfate. After filtration, the filtrate was concentrated under reduced pressure, and the concentrated residue was crystallized from hot methanol to obtain compound (11) (90.75 g, yield 45.9%).
With toluene-4-sulfonic acid; In N,N-dimethyl-formamide; at 60℃; for 6h;Inert atmosphere;
To a stirred solution of 44 (15 g, 83.3 mmol) in DMF (150 mL), was added PhCH(OCH3)2 (19 mL, 91.6 mmol, 1.1 eq) p-TSA (0.200 g, 8.33 mmol) and the reaction mixture was stirred at 60 C for 6 h. The mixture was dried under vacuum to obtain compound 45 (10 g, 45%) as a white solid. The combined organic layer was concentrated under reduced pressure crude compound was obtained purified with combi flash eluted with 60% ethyl acetate: Hexane. NMR (400 MHz, CD3OD): d 7.45-7.36 (m, 2H), 7.28-7.21 (m, 3H), 5.46 (s, 0.7H), 5.04 (s, 0.3H), 4.49 (dd, J = 10.8, 4.4 Hz, 0.3H), 4.07 (dd, J = 10.8, 4.4 Hz, 0.7H), 3.90-3.83 (m, 0.7H), 3.77 (t, j = 9.2 Hz, 0.7H), 3.68-3.57 (m, 1.3H), 3.52 (t, j = 9.2 Hz, 0.3H), 3.40-3.30 (m, 2H), 3.17-3.12 (m, 0.3H).
With para-dodecylbenzenesulfonic acid; at 80.0℃; for 24.0h;Green chemistry;
General procedure: A typical reaction was carried out by adding 3.6 g (0.02 mol)glucose to a 100 ml glass reactor containing 0.2 mol of correspondingalcohol and 0.33 g (0.01 mol) dodecylbenzenesulfonic acid. The resultingsuspension was agitated intensively (700 rpm) at desired temperaturefor 24 h. After that the reaction mixture (clear liquid) was cooleddown to room temperature and analysed (GPC and GC/MS).
glucose [R]-3-hydroxybutyrate tetraester[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
novozyme 435; at 70℃; for 48h;
EXAMPLE 1.The synthesis of K.TX 0310 by the esterificarion of glucose with(R)-3-hvdroxvbutyric acid in the presence of CAL-B.; To a round-bottomed flask, lg glucose and 4.6g 3-hydroxybutync acid were added. The mixture was heated at 80C to obtain a homogenous solution. The temperature was lowered to 70C and 1.1 g (20% w/v of the total mixture) CaL B was added. The mixture was stirred at 70C for 48 hrs to yield glucose 3-hydroxybutyrate tri- and tetra-esters as shown in Figure 1.The material was separated by column chromatography based on its polarity. The column was packed in pure chloroform and the polarity was increased using methanol The desired product was eluted using chloroform: methanol: water (9 2: 0.3).The product was a water-soluble syrup and was obtained at a yield of 0.3g (30%). A mixture of tri- and tetra-substituted products was formed (substitution factor between 3 and 4 with 1 to 2 free hydroxyls left per monosaccharide ring). The structure of the compound was verified by LC/MS.
With alpha-L-rhamnosidase from Alternaria alternata SK37.001; In aq. acetate buffer; at 40℃; for 0.166667h;pH 5.5;Enzymatic reaction;
Two millilitres of 0.1% naringin in 0.5 M sodium acetate buffer (pH 5.5) was incubated with the crude alpha-L-rhamnosidase at 40 C for 10 min. The crude enzyme dosage was 15 U g-1 of naringin. After the reaction was stopped by placement in a boiling water bath for 10 min, the reaction solution was mixed with 1.9 ml of methanol and centrifuged at 8000g for 5 min. The supernatant was analyzed using high performance liquid chromatography (HPLC) equipped with Sepax C18 column at 35 C. The mobile phase was methanol:water 50:50 (v:v) at a flow rate of 1 ml min-1. The products were detected using an ultraviolet detector at 280 nm.
3-O-β-D-galactopyranosyl-(1->2)-[α-L-arabinopyranosyl-(1->3)]-β-D-glucopyranosyl quillaic acid 28-O-α-L-arabinopyranosyl-(1->3)-β-D-xylopyranosyl-(1->4)-[β-D-glucopyranosyl-(1->3)]-α-L-rhamnopyranosyl-(1->2)-[α-L-rhamnopyranosyl-(1->4)]-β-D-fucopyranosyl ester[ No CAS ]
cyanidin 3-O-[2-O-(2-O-(trans-feruloyl)-glucosyl)-6-O-(trans-p-coumaroyl)-glucoside]-5-O-[6-O-(malonyl)-glucoside][ No CAS ]
[ 2280-44-6 ]
[ 1135-24-6 ]
[ 141-82-2 ]
[ 7400-08-0 ]
[ 528-58-5 ]
Yield
Reaction Conditions
Operation in experiment
With hydrogenchloride; water; at 100℃; for 1.0h;
General procedure: The identification of anthocyanins was carried out by standard procedures and both alkaline and acid hydrolyses (Harborne 1984). Acid hydrolysis of pigments (ca. 1 mg each) was carried out with 2N HCl (1 mL) at 100 C for 1 h. Alkaline hydrolysis of pigments (ca. 1 mg each) was carried out with 2N NaOH solution (1 mL) under degassed syringe allowed to stand for 15 min. The solution was next acidified with 2N HCl (1.1 mL) and evaporated in vacuo to dryness. The data of TLC (Rf values), HPLC (Rt-min, method 1), UV-Vis (lambdamax), and FABMS spectra are shown in Section 4.4.1.-4.4.4.
peonidin 3-O-[2-O-(2-O-(trans-feruloyl)-glucosyl)-6-O-(cis-p-coumaroyl)-glucoside]-5-O-[6-O-(malonyl)-glucoside][ No CAS ]
[ 2280-44-6 ]
[ 1135-24-6 ]
[ 141-82-2 ]
[ 501-98-4 ]
[ 134-01-0 ]
Yield
Reaction Conditions
Operation in experiment
With hydrogenchloride; water; at 100℃; for 1h;
General procedure: The identification of anthocyanins was carried out by standard procedures and both alkaline and acid hydrolyses (Harborne 1984). Acid hydrolysis of pigments (ca. 1 mg each) was carried out with 2N HCl (1 mL) at 100 C for 1 h. Alkaline hydrolysis of pigments (ca. 1 mg each) was carried out with 2N NaOH solution (1 mL) under degassed syringe allowed to stand for 15 min. The solution was next acidified with 2N HCl (1.1 mL) and evaporated in vacuo to dryness. The data of TLC (Rf values), HPLC (Rt-min, method 1), UV-Vis (lambdamax), and FABMS spectra are shown in Section 4.4.1.-4.4.4.
General procedure: Isolated glycosides (5 mg, each) were dissolved in 0.2 N H2SO4 (5 mL) and heated at 95 C for 1 h. After cooling, the reaction mixture was extracted with Et2O (2 mL x 3). The aqueous layer was neutralized with NaHCO3, evaporated to dryness, and extracted with pyridine. The pyridine extract was then analyzed on silica gel TLC (EtOAc-MeOH-H2O-AcOH 13:3:3:4). Rhamnose (Rf 0.52), Apiose (Rf 0.48) and glucose (Rf 0.32) were detected by comparison with authentic samples.
With hydrogenchloride; In water; at 100.0℃;
The absolute configuration of monosaccharides was determined after acid hydrolysis of each isolated compound (1-2, 4-15, 17-18) in 1 mL of 4M HCl solution at 100 C. The sugar-containing aqueous layer, obtained after extraction with ethyl acetate, was neutralized with Amberlite IRA-400 (OH-form). Released monosaccharides were derivatizedand determined according to the modified method of Tanakaet al. (2007) (Prez et al., 2014). On the basis of the retention time of authentic standards of glucose (D-Glc 10.40 min, L-Glc 10.20 min), rhamnose (D-Rha 9.96 min, L-Rha 11.56 min) and xylose (D-Xyl 10.72 min, L-Xyl 10.78 min), the three monosaccharide isomers: D-glucose, D-xylose and L-rhamnose, were identified among isolated compounds.
General procedure: Each compound (1 mg) was treated with 2 M HCl (10 ml) under conditions of reflux at 80 C this being maintained for 4 h. Each reaction mixture was concentrated to dryness, partitioned between CHCl3 and H2O and then the H2O layer was concentrated to dryness to yield a mixture of sugars. Each mixture was dissolved in anhydrous pyridine (1 ml) and reacted with l-cysteine methyl ester hydrochloride (1 mg) in an oven at 60 C for 2 h. After the reaction mixture was evaporated under a stream of N2, trimethylsilyl imidazole (0.2 ml) was added, and the mixture warmed to 60 C for another 1 h. After drying the solution, the products were partitioned between cyclohexane and H2O. The same reactions were applied to standard sugars. Then the cyclohexane layers were analyzed by gas chromatography on a column, AT-SE-30 (0.5 mum × 0.32 mm × 30 m); column temperature, 250 C; detector temperature, 250 C; injection temperature, 230 C; carrier gas, N2. The derivatives of d-glucose, l-glucose, l-rhamnose, d-apiose, l-arabinose and d-xylose gave peaks at tR 25.67, 26.81, 16.33, 12.69, 13.09 and 12.94 min, respectively.
With pyrophosphatase, inorganic from yeast; UTP-glucose-1-phosphate galactokinase from Streptococcus pneumoniae TIGR4; UTP-glucose-1-phosphate uridylyltransferase from Streptococcus pneumoniae TIGR4; adenosine 5'-triphosphate sodium salt; magnesium chloride; In aq. buffer; at 42℃; for 24.0h;pH 8;Enzymatic reaction;
General procedure: We chose 14 monosaccharides which could be used by GalK in previous report36 as substrates for one-pot three-enzyme system to synthesize various sugar nucleotides. The reactions were carried out in the system containing 50 mM Tris-HCl (pH 8.0), 10 mM MgCl2, various monosaccharides, 10 mM ATP, 10 mM UTP, 0.5 mg/mL SpGalK, 0.5 mg/mL SpGalU, and 0.1 U PPase. The reactions were incubated at 42 C for 24 h. The reactions were terminated by heating at 100 C for 5 min, followed by centrifugated at 13,400 rpm at room temperature for 30 min to remove protein precipitation. The products were determined by TLC, CE and MS. For CE analysis, 5 muL of each sample was diluted into 50 muL and subjected to CE analysis as described above.
luteolin-7,4'-di-O-α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside[ No CAS ]
[ 2280-44-6 ]
[ 73-34-7 ]
[ 491-70-3 ]
Yield
Reaction Conditions
Operation in experiment
With trifluoroacetic acid; In acetone; at 100℃; for 2h;
Compound 1 (2 mg) was dissolved in TFA (5%) in Me2CO (5 mL), heated to 100 C (2 h), and concentrated in vacuo. The residue was dissolved in MeOH and analyzed by HPLC. The hydrolysate contained luteolin (conditions 2, tR 3.52 min), glucose, and rhamnose (conditions 3, tR 14.12 and 17.35 min) respectively.
β-D-glucopyranose-1,2,3,4,6-pentakis[3,4-bis(phenylmethoxy)benzoate][ No CAS ]
α-D-glucopyranose-1,2,3,4,6-pentakis[3,4-bis(phenylmethoxy)benzoate][ No CAS ]
Yield
Reaction Conditions
Operation in experiment
With dmap; dicyclohexyl-carbodiimide; In dichloromethane; at 40℃;
General procedure: A suspension of D-glucose (0.03 g,0.16 mmol), 4-(benzyloxy)-3-methoxybenzoic acid (0.34 g,1.30 mmol), dicyclohexylcarbodiimide (DCC) (0.28 g, 1.32 mmol) andN,N-(di-methylamino) pyridine (DMAP) (0.16 g, 1.36 mmol) in freshlydistilled dichloromethane (DCM) was refluxed for 16?18 hrs (monitoredby TLC). After cooling reaction mixture at room temperature,urea by-product was removed by gravity filtration and the filtratewas evaporated. Compound 11 was purified by column chromatographyeluting with a mixture of dichloromethane, hexane, andethyl acetate. The product was collected, evaporated and precipitatedusing toluene and cyclohexane
With sodium dihydrogenphosphate; citric acid; In water; at 37℃; for 24h;pH 7.0;Microbiological reaction;Kinetics;
Transfructosylation reactions were carried out using uninduced,inulin-induced and sucrose-induced cells in a reaction volumeof 5 ml. The reaction mixture contained 100 mg lyophilizedcells and 10% (w/v) sucrose in 5 ml phosphate citrate buffer (pH7.0). The mixture was incubated at 37 C for 24 h at 220 rpm in areciprocating shaker (SI-300R Jeo Tech). At the end of the reaction,cells were separated out by centrifugation and the vials heated in aboiling water bath. The clear supernatant was analysed for residualsucrose, glucose and synthesized fructo-oligosaccharides
In water; acetone; for 24h;pH 7.0;Microbiological reaction;
General procedure: All of the filamentous fungi were pre-incubated in potato medium to get enough fungi cells. The glucosylation medium was sodium phosphate buffer: 13.62 g NaH2PO4, 2.36 g NaOH, and 20 g D-glucose in1000 mL water. 1.5 g R. oryzae AS 3.2380 fungus cells were preincubatedin the glucosylation medium (200 mL), 30 C, 130 rpm for12 h. Then, estrogen substrates (10 mg) dissolved in acetone were injected into the medium with the continued incubation for 24 h [19].When the fungus cells were filtered, the incubation culture was subjected to a MCI (Middle chromatogram isolated gel) column, which was eluted by 30% ethanol aqueous, 50% ethanol aqueous, and 95%ethanol, successively. The estrogen 3beta-glucosides could be obtained in the 50% ethanol aqueous elution with the purity > 95%.
In water; acetone; for 24h;pH 7;Microbiological reaction;
General procedure: All of the filamentous fungi were pre-incubated in potato medium to get enough fungi cells. The glucosylation medium was sodium phosphate buffer: 13.62 g NaH2PO4, 2.36 g NaOH, and 20 g D-glucose in1000 mL water. 1.5 g R. oryzae AS 3.2380 fungus cells were preincubatedin the glucosylation medium (200 mL), 30 C, 130 rpm for12 h. Then, estrogen substrates (10 mg) dissolved in acetone were injected into the medium with the continued incubation for 24 h [19].When the fungus cells were filtered, the incubation culture was subjected to a MCI (Middle chromatogram isolated gel) column, which was eluted by 30% ethanol aqueous, 50% ethanol aqueous, and 95%ethanol, successively. The estrogen 3beta-glucosides could be obtained in the 50% ethanol aqueous elution with the purity > 95%.
In water; acetone; for 24h;pH 7.0;Microbiological reaction;
General procedure: All of the filamentous fungi were pre-incubated in potato medium to get enough fungi cells. The glucosylation medium was sodium phosphate buffer: 13.62 g NaH2PO4, 2.36 g NaOH, and 20 g D-glucose in1000 mL water. 1.5 g R. oryzae AS 3.2380 fungus cells were preincubatedin the glucosylation medium (200 mL), 30 °C, 130 rpm for12 h. Then, estrogen substrates (10 mg) dissolved in acetone were injected into the medium with the continued incubation for 24 h [19].When the fungus cells were filtered, the incubation culture was subjected to a MCI (Middle chromatogram isolated gel) column, which was eluted by 30percent ethanol aqueous, 50percent ethanol aqueous, and 95percentethanol, successively. The estrogen 3beta-glucosides could be obtained in the 50percent ethanol aqueous elution with the purity > 95percent.
In water; acetone; for 24.0h;pH 7.0;Microbiological reaction;
General procedure: All of the filamentous fungi were pre-incubated in potato medium to get enough fungi cells. The glucosylation medium was sodium phosphate buffer: 13.62 g NaH2PO4, 2.36 g NaOH, and 20 g D-glucose in1000 mL water. 1.5 g R. oryzae AS 3.2380 fungus cells were preincubatedin the glucosylation medium (200 mL), 30 C, 130 rpm for12 h. Then, estrogen substrates (10 mg) dissolved in acetone were injected into the medium with the continued incubation for 24 h [19].When the fungus cells were filtered, the incubation culture was subjected to a MCI (Middle chromatogram isolated gel) column, which was eluted by 30% ethanol aqueous, 50% ethanol aqueous, and 95%ethanol, successively. The estrogen 3beta-glucosides could be obtained in the 50% ethanol aqueous elution with the purity > 95%.
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