CN101624612B - Method for catalytic synthesis of glycoside esters compound by immobilized penicillium expansum lipase - Google Patents
Method for catalytic synthesis of glycoside esters compound by immobilized penicillium expansum lipase Download PDFInfo
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- CN101624612B CN101624612B CN2009100417023A CN200910041702A CN101624612B CN 101624612 B CN101624612 B CN 101624612B CN 2009100417023 A CN2009100417023 A CN 2009100417023A CN 200910041702 A CN200910041702 A CN 200910041702A CN 101624612 B CN101624612 B CN 101624612B
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Abstract
The invention discloses a method for catalytic synthesis of glycoside esters compound by immobilized penicillium expansum lipase, comprising glycoside and acyl donor which have the mole ratio of 1:1-20:1 are added into non-aqueous medium, and the concentration of the glycoside is 1-40mg/mL; the immobilized penicillium expansum lipase is added, and the mass ratio between the immobilized penicillium expansum lipase and the glycoside is 10:1-1:10; under the conditions that the temperature is 30-60 DEG C, the shaking speed is 100-300rpm, and the pressure is normal pressure, transesterification is carried out for 0.5-72h, and the glycoside esters compound can be obtained. The compound has the structure shown in the general formula (I). The invention utilizes the immobilized penicillium expansum lipase preparation with low price to synthesize the glycoside esters compound through the transesterification, so that the yield reaches up to 85-98%, and the cost is greatly reduced. The method has the advantages of mild reaction condition, environmental protection, high regioselectivity, simple and controllable reaction process, easily separated products and low cost.
Description
Technical field
The invention belongs to the enzyme chemical field, particularly a kind of method of utilizing immobilized penicillium expansum lipase catalysis glucosides acidylate synthesis of glycoside esters compound.
Background technology
Glucosides extensively is present in the organism, and wherein a lot of glucosides are being undertaken important biological function because of having special biological activity.Many glycoside compounds; Like Gastrodine, arbutin, glucose helicidum, homoarbutin, different homoarbutin, salicylic aldehyde-β-D-glucoside etc., caused people's attention owing to having specific pharmaceutical use (for example antitumor, antimicrobial, reducing blood-fat, anti-oxidant, enhance immunity function, calming soporific etc.).Glycoside compounds is one type of active compound; Exist a plurality of hydroxyls on the structure, can be connected, when aliphatics or aromatic acyl are gone up in the hydroxyl connection of sugar ring with the compound of other different structure; Can strengthen the fat-soluble of compound; Thereby improve its absorption in cell and intestines, improve its oral administration biaavailability, strengthen drug effect.Therefore, the synthetic and modification of this compounds is then become a main direction of studying of glucosides class medicine.Arbutin is antibiotic except having, the antibechic, multiple pharmacological effect such as hypoglycemic; Or the suppressor factor of tyrosine oxidase in the human melanoma cell; Can block the synthetic of DOPA and DOPA quinone, thereby suppress melanic generation effectively, the Pear Power effect; And skin is not had pungency, and toxic side effect is little.Yet in use, there is fat-soluble poor, problem such as bioavailability is low in arbutin.Pharmacology activity research shows that the carbon undecylenic acid of arbutin and laurate verivate all show higher antityrosinase and anti-linolic acid oxidation activity than parental generation compound.It is numerous that glucoside compound sugar ring is gone up hydroxyl, and reactive behavior is similar.When adopting general chemical process direct esterification, regioselectivity is low, is prone to produce a large amount of by products, product separation difficulty; Be the synthetic product of confirming the position acidylate, need encircle specific hydroxyl to sugar and protect and go operation such as protection, many, the complex process of reactions step; This method adopts basic catalyst simultaneously, is prone to produce alkali waste, and environment is caused severe contamination.And enzyme process synthesizes the reactions step of complicacies such as having avoided loaded down with trivial details protection and gone to protect, and reaction conditions is gentle, environmentally friendly, and products therefrom purity is high.Therefore, enzyme catalysis glucosides regioselectivity acidylate is an alternative route with bright prospects.Document (Tokiwa et al., Med.Chem.Lett.2007,17:3105-3108 were once arranged; Nakajima et al., J.Biosci.Bioeng.1997,87:105-107; Watanabe et al., Biochem.Eng.J.2009,43:261-265; Chigorimbo-Murefu et al.; J.Mol.Catal.B:Enzym., 2009,56:277-282) compound method of enzymatic arbutin ester derivative is disclosed; But its used zymin is expensive commercialization proteolytic enzyme or lypase, and catalytic efficiency (is not high.As utilize subtilisin catalysis to synthesize arbutin carbon undecylenate, and its reaction times is longer, and efficient is lower; Be used to come from the lipase-catalyzed arbutin laurate of antarctic candida and synthesizing of ferulic acid ester, its maximum conversion rate is merely 50%; Be used to come from lipase-catalyzed arbutin laurate synthetic of pseudomonas cepacia, its yield is merely 28%.Based on the structure of the lypase of different sources and the difference of function, the present invention selects for use cheap immobilized penicillium expansum lipase preparation catalysis glucosides acidylate with preparation glucosides ester cpds, and it is yield high (85%-98%) not only, and cost reduces greatly.
Summary of the invention
In order to solve the weak point of above-mentioned prior art, primary and foremost purpose of the present invention is to provide a kind of method of immobilized penicillium expansum lipase catalytic synthesis of glycoside esters compound.
Another object of the present invention is to provide aforesaid method synthetic glycoside esters compound.
The object of the invention is realized through following technical proposals: the method for the lipase-catalyzed synthesis of glycoside esters compound of a kind of immobilized penicillium expansum (Penicilliumexpansum); Comprise following operation steps: in non-aqueous media; The adding mol ratio is 1: 1～20: 1 acry radical donor and a glucosides; Obtain solution, the concentration of glucosides is 1～40mg/mL in this solution; Adding mass ratio with glucosides and be 10: 1～1: 10 immobilized penicillium expansum lipase, is that 30～60 ℃, hunting speed are under 100～300rpm and the condition of normal pressure in temperature, and transesterification 0.5～72 hour obtains glycoside esters compound;
Said acry radical donor is vinyl carboxylates, acid anhydrides, carboxylate methyl ester or carboxylic acid, ethyl ester;
Said glucosides is for having the compound of following general formula (II):
Wherein, R
1Be hydrogen, hydroxyl, methoxyl group, aldehyde radical, methylol, formyl radical or ethanoyl; R
2Be hydrogen, methyl or aldehyde radical; R
3Be hydrogen or methyl;
Said non-aqueous media is aprotic organic solvent or contains in the low anionic ionic liquid of nucleophilicity one or both.
Said glucosides is arbutin, Gastrodine, glucose helicidum, homoarbutin, different homoarbutin or salicylic aldehyde-β-D-glucoside.
The preparation of said immobilized penicillium expansum lipase comprises following operation steps:
(1) will contain the thick enzyme powder of penicillium expansum lipase that mass percent is 95% starch (available from the green little health biotechnology in Shenzhen ltd) is dissolved in glycocoll-sodium hydrate buffer solution; Be mixed with every milliliter of damping fluid and contain the suspension-s of the thick enzyme powder of 0.3g lypase, under 35 ℃ of temperature and rotating speed 150rpm condition, stir 1h; The centrifugal supernatant that gets under 4 ℃ of temperature and rotating speed 6000rpm condition; The weight ratio of glycocoll and sodium hydroxide is 2.9: 1 in said glycocoll-sodium hydrate buffer solution;
(2) use mass percent to be 95% alcohol immersion 24h macroporous adsorbent resin (D4020) after, be washed till the filtrating clarification with deionized water, obtain filtrating;
(3) step (2) gained filtrating is added in step (1) the gained supernatant, enzyme concentration is the thick enzyme powder of 9g/g resin; Under 35 ℃ of temperature and rotating speed 150rpm condition, stir 4h, filter with sand core funnel, with glycocoll-sodium hydrate buffer solution flushing, the suction filtration postlyophilization obtains immobilized penicillium expansum lipase; Preserve subsequent use in 4 ℃ of refrigerators immobilized penicillium expansum lipase.
Said aprotic organic solvent is THF, acetone, acetonitrile 、 diox, the trimethyl carbinol, DMSO 99.8MIN., N, dinethylformamide, pyridine, isopropyl ether, octane-iso, normal hexane, normal heptane, octane, positive nonane or hexanaphthene;
The said negatively charged ion that contains in the low anionic ionic liquid of nucleophilicity is tetrafluoroborate ion, hexafluorophosphoricacid acid ions, two (trifluoromethyl sulphonyl) imines ion or amino acid negatively charged ion; Positively charged ion is 1-alkyl-3-Methylimidazole ion, 1-alkyl-2,3-methylimidazole ion, quaternary ammonium ion, quaternary phosphonium ion, amino acid positively charged ion or N-alkyl pyridine ion.
Said non-aqueous media is two kinds in the above-mentioned aprotic organic solvent, and two kinds of aprotic organic solvent volume ratios are 1～19: 1.
Said non-aqueous media is a kind of in the above-mentioned aprotic organic solvent and above-mentionedly contains a kind of in the low anionic ionic liquid of nucleophilicity that the volumn concentration that wherein contains the low anionic ionic liquid of nucleophilicity is 5～95%.
The glycoside esters compound of method for preparing has following general formula (I):
Wherein R is the alkyloyl of 2～18 carbon atoms or the unsaturated aromaticacyl radical of 9～11 carbon atoms; R
1Be hydrogen, hydroxyl, methoxyl group, aldehyde radical, methylol, formyl radical or ethanoyl; R
2Be hydrogen, methyl or aldehyde radical; R
3Be hydrogen or methyl.
Said alkyloyl is the saturated or undersaturated alkyloyl of straight or branched; Said unsaturated aromaticacyl radical is unsaturated aromaticacyl radical of cis or trans unsaturated aromaticacyl radical.
The relative prior art of the present invention has following advantage and beneficial effect:
(1) employing is cheap, biological catalyst---self-control immobilized penicillium expansum lipase catalysis glucosides ester derivative is synthetic efficiently.The enzymatic reaction selectivity is high, and product structure is prone to control, therefore overcome traditional chemical method selectivity low, be prone to generate by product, or need protection and shortcomings such as deprotection operation and productive rate are low; The immobilized lipase zymin catalysis glucosides ester derivative that the present invention selects for use is cheap, derive from Penicilllum expansum synthetic with existing enzyme catalysis arbutin ester derivative synthetic compared with techniques, has yield height (85%～98%), the advantage that cost is low.
(2) regioselectivity of the present invention is high, and productive rate is high, product purity is high, reaction process is simple and easy to control, and product is easily separated.
(3) the present invention is that 30～60 ℃, hunting speed are under 100～300rpm, the condition of normal pressure in temperature, utilizes enzyme catalysis to prepare the glucosides ester derivative, and reaction conditions is gentle, environmental friendliness.
Embodiment
Below in conjunction with embodiment the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
Embodiment 1: arbutin 6 '-butyryl ester synthetic
With arbutin (109mg; 0.4mmol), vinyl butyrate (3mmol), 20mL anhydrous tetrahydro furan add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 1000mg immobilized penicillium expansum lipase; In 35 ℃, the constant temperature oscillator of 300rpm, vibrate under the normal pressure, utilize thin layer chromatography (TLC) monitoring reaction.Behind the reaction 1h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying get white powder product 116mg, and yield is 85%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.04(C4″)，18.52(C3″)，36.02(C2″)，63.99(C6′)，70.64(C4′)，73.79(C2′)，74.22(C5′)，76.93(C3′)，102.05(C1′)，116.01(C3+C5)，118.24(C2+C6)，150.70(C1)，152.91(C4)，173.17(C1″)。
1H?NMR：δppm?0.88(t，3H，H4″)，1.54(q，2H，H3″)，2.27(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)，3.51(t，1H，H5′)，4.06(dd，1H，H6′)，4.33(d，1H，H6′)，4.68(d，1H，H1′)，5.14(d，1H，OH4′)，5.24(d，1H，OH3′)，5.30(d，1H，OH2′)，6.65(d，2H，H3+H5)，6.84(d，2H，H2+H6)，9.01(s，1H，OH4)。
Embodiment 2: arbutin 6 '-butyryl ester synthetic
With arbutin (218mg; 0.8mmol), butyryl oxide (5.6mmol), 20mL normal hexane/THF (1/4; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 900mg immobilized penicillium expansum lipase, in 40 ℃, the constant temperature oscillator of 200rpm, vibrate under the normal pressure, utilize thin layer chromatography (TLC) monitoring reaction.Behind the reaction 10h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying get white powder product 230mg, and yield is 84%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.04(C4″)，18.52(C3″)，36.02(C2″)，63.99(C6′)，70.64(C4′)，73.79(C2′)，74.22(C5′)，76.93(C3′)，102.05(C1′)，116.01(C3+C5)，118.24(C2+C6)，150.70(C1)，152.91(C4)，173.17(C1″)。
1H?NMR：δppm?0.88(t，3H，H4″)，1.54(q，2H，H3″)，2.27(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)，3.51(t，1H，H5′)，4.06(dd，1H，H6′)，4.33(d，1H，H6′)，4.68(d，1H，H1′)，5.14(d，1H，OH4′)，5.24(d，1H，OH3′)，5.30(d，1H，OH2′)，6.65(d，2H，H3+H5)，6.84(d，2H，H2+H6)，9.01(s，1H，OH4)。
Embodiment 3: arbutin 6 '-capronate synthetic
With arbutin (20mg; 0.07mmol), vinyl caproate (0.5mmol), 20mL isopropyl ether/THF (1/19; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 10mg immobilized penicillium expansum lipase, in 35 ℃, the constant temperature oscillator of 150rpm, vibrate under the normal pressure, with TLC monitoring reaction.Behind the reaction 24h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.25) purifying get white powder product 22.4mg, and yield is 86%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.37(C6″)，22.34(C5″)，24.69(C3″)，31.24(C4″)，34.08(C2″)，63.98(C6′)，70.62(C4′)，73.78(C2′)，74.22(C5′)，76.93(C3′)，102.07(C1′)，116.01(C3+C5)，118.23(C2+C6)，150.71(C1)，152.92(C4)，173.32(C1″)。
1H?NMR：δppm?0.85(t，3H，H6″)，1.25(s，4H，H4″+H5″)，1.52(q，2H，H3″)，2.28(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)，3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.33(d，1H，H6′)，4.67(d，1H，H1′)，5.14(d，1H，OH4′)，5.23(d，1H，OH3′)，5.31(d，1H，OH2′)，6.65(d，2H，H3+H5)，6.84(d，2H，H2+H6)，9.01(s，1H，OH4)。
Embodiment 4: arbutin 6 '-capronate synthetic
With arbutin (109mg; 0.4mmol), NSC 8882 (3.6mmol), 20mL isopropyl ether/acetonitrile (1/5; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 800mg immobilized penicillium expansum lipase, in 45 ℃, the constant temperature oscillator of 250rpm, vibrate under the normal pressure, with TLC monitoring reaction.Behind the reaction 4.5h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.25) purifying get white powder product 127mg, and yield is 85%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.37(C6″)，22.34(C5″)，24.69(C3″)，31.24(C4″)，34.08(C2″)，63.98(C6′)，70.62(C4′)，73.78(C2′)，74.22(C5′)，76.93(C3′)，102.07(C1′)，116.01(C3+C5)，118.23(C2+C6)，150.71(C1)，152.92(C4)，173.32(C1″)。
1H?NMR：δppm?0.85(t，3H，H6″)，1.25(s，4H，H4″+H5″)，1.52(q，2H，H3″)，2.28(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)，3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.33(d，1H，H6′)，4.67(d，1H，H1′)，5.14(d，1H，OH4′)，5.23(d，1H，OH3′)，5.31(d，1H，OH2′)，6.65(d，2H，H3+H5)，6.84(d，2H，H2+H6)，9.01(s，1H，OH4)。
Embodiment 5: arbutin 6 '-octanoate synthetic
With arbutin (109mg; 0.4mmol), sad vinyl acetate (0.4mmol), 20mL [C4MIm] PF6/ THF (1/9; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 600mg immobilized penicillium expansum lipase, in 40 ℃, the constant temperature oscillator of 250rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 8h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.27) purifying get white powder product 145mg, and yield is 91%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.51(C8″)，22.61(C7″)，25.01(C3″)，28.93(C4″)，29.01(C5″)，31.68(C6″)，34.12(C2″)，63.98(C6′)，70.61(C4′)，73.79(C2′)，74.22(C5′)，76.93(C3′)，102.08(C1′)，116.01(C3+C5)，118.22(C2+C6)，150.71(C1)，152.93(C4)，173.32(C1″)。
1H?NMR：δppm?0.85(t，3H，H8″)，1.25(s，8H，H4″+H5″+H6″+H7″)，1.52(t，2H，H3″)，2.28(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.31(d，1H，H6′)，4.67(d，1H，H1′)，5.14(d，1H，OH4′)，5.22(d，1H，OH3′)，5.30(d，1H，OH2′)，6.66(d，2H，H3+H5)，6.83(d，2H，H2+H6)，9.00(s，1H，OH4)。
Embodiment 6: arbutin 6 '-octanoate synthetic
With arbutin (109mg; 0.4mmol), methyl caprylate (4mmol), 20mL [C6MIm] PF6/ THF (1/9; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 600mg immobilized penicillium expansum lipase, in 45 ℃, the constant temperature oscillator of 250rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 4h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.27) purifying get white powder product 145mg, and yield is 91%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.51(C8″)，22.61(C7″)，25.01(C3″)，28.93(C4″)，29.01(C5″)，31.68(C6″)，34.12(C2″)，63.98(C6′)，70.61(C4′)，73.79(C2′)，74.22(C5′)，76.93(C3′)，102.08(C1′)，116.01(C3+C5)，118.22(C2+C6)，150.71(C1)，152.93(C4)，173.32(C1″)。
1H?NMR：δppm?0.85(t，3H，H8″)，1.25(s，8H，H4″+H5″+H6″+H7″)，1.52(t，2H，H3″)，2.28(t，2H，H2″)，3.29-3.11(m，3H，H2′+H3′+H4′)3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.31(d，1H，H6′)，4.67(d，1H，H1′)，5.14(d，1H，OH4′)，5.22(d，1H，OH3′)，5.30(d，1H，OH2′)，6.66(d，2H，H3+H5)，6.83(d，2H，H2+H6)，9.00(s，1H，OH4)。
Embodiment 7: arbutin 6 '-decylate synthetic
With arbutin (800mg; 2.9mmol), capric acid vinyl acetate (14mmol), 20mL diox add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 520mg immobilized penicillium expansum lipase, in 60 ℃, the constant temperature oscillator of 250rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 24h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.27) purifying get white powder product 1087mg, and yield is 88%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1HNMR (400MHz) confirms:
13C?NMR：δppm?14.54(C10″)，22.67(C9″)，25.01(C3″)，29.06(C4″)，29.23(C5″)，29.28(C7″)，29.43(C6″)，31.85(C8″)，34.12(C2″)，63.99(C6′)，70.62(C4′)，73.79(C2′)，74.23(C5′)，76.94(C3′)，102.11(C1′)，116.00(C3+C5)，118.23(C2+C6)，150.72(C1)，152.94(C4)，173.31(C1″)。
1H?NMR：δppm?0.85(t，3H，H10″)，1.23(s，12H，H4″+H5″+H6″+H7″+H8″+H9″)，1.51(t，2H，H3″)，2.28(t，2H，H2″)，3.29-3.08(m，3H，H2′+H3′+H4′)，3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.31(d，1H，H6′)，4.66(d，1H，H1′)，5.14(d，1H，OH4′)，5.22(d，1H，OH3′)，5.30(d，1H，OH2′)，6.65(d，2H，H3+H5)，6.83(d，2H，H2+H6)，9.00(s，1H，OH4)。
Embodiment 8: arbutin 6 '-laurate synthetic
With arbutin (109mg, 0.4mmol), vinyl laurate (0.8mmol), 20mL [C
6Py] NTf
2/ acetone (19/1, v/v) add in the 50mL tool plug triangular flask behind the mixing, in wherein adding the 100mg immobilized penicillium expansum lipase, in 60 ℃, the constant temperature oscillator of 250rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 72h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/3, Rf=0.27) purifying get white powder product 160mg, and yield is 88%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?14.51(C
8″)，22.61(C
7″)，25.01(C
3″)，28.93(C
4″)，29.01(C
5″)，31.68(C
6″)，34.12(C
2″)，63.98(C
6′)，70.61(C
4′)，73.79(C
2′)，74.22(C
5′)，76.93(C
3′)，102.09(C
1′)，116.01(C
3+C
5)，118.22(C
2+C
6)，150.71(C
1)，152.93(C
4)，173.32(C
1″)。
1H?NMR：δppm?0.85(t，3H，H
8″)，1.25(s，8H，H
4″+H
5″+H
6″+H
7″)，1.52(t，2H，H
3″)，2.28(t，2H，H
2″)，3.29-3.11(m，3H，H
2′+H3′+H
4′)3.50(t，1H，H
5′)，4.06(dd，1H，H
6′)，4.31(d，1H，H
6′)，4.67(d，1H，H
1′)，5.14(d，1H，OH
4′)，5.22(d，1H，OH
3′)，5.30(d，1H，OH
2′)，6.66(d，2H，H
3+H
5)，6.83(d，2H，H
2+H
6)，9.00(s，1H，OH
4)。
Embodiment 9: arbutin 6 '-cetylate synthetic
With arbutin (109mg; 0.4mmol), palmitinic acid vinyl acetate (4mmol), 20mL diox/octane-iso (1/1; V/v) add in the 50mL tool plug triangular flask behind the mixing; In wherein adding the 200mg immobilized penicillium expansum lipase, in 50 ℃, the constant temperature oscillator of 200rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 10h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/3, Rf=0.28) purifying get white powder product 174mg, and yield is 85%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is through 1
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?13.97(C16″)，22.13(C15″)，24.44(C3″)，28.51(C4″)，28.75(C5″)，28.93(C13″)，29.07(C6″+C7″+C8″+C9″+C10″+C11″+C12″)，31.33(C14″)，33.53(C2″)，63.43(C6′)，70.02(C4′)，73.19(C2′)，73.63(C5′)，76.34(C3′)，101.51(C1′)，115.41(C3+C5)，117.62(C2+C6)，150.13(C1)，152.36(C4)，172.72(C1″)。
1H?NMR：δppm?0.85(t，3H，H16″)1.23(s，24H，H4″+H5″+H6″+H7″+H8″+H9″+H10″+H11″+H12″+H13″+H14″+H15″)，1.50(t，2H，H3″)，2.28(t，2H，H2″)，3.23-3.14(m，3H，H2′+H3′+H4′)，3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.29(d，1H，H6′)，4.66(d，1H，H1′)，5.17(s，1H，OH4′)，5.25(d，1H，OH3′)，5.32(s，1H，OH2′)，6.65(d，2H，H3+H5)，6.83(d，2H，H2+H6)，9.01(s，1H，OH4)。
Embodiment 10: arbutin 6 '-methacrylic ester synthetic
With arbutin (109mg, 0.4mmol), methacrylic vinyl acetate (6mmol), 20mL [C
4MIm] BF
4/ acetonitrile (1/19, v/v) add 50mL tool plug triangular flask mixing after, in wherein adding the 600mg immobilized penicillium expansum lipase, in 55 ℃, the constant temperature oscillator of 150rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 72h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.25) purifying get white powder product 115mg, and yield is 85%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?17.88(C4″)，64.05(C6′)，70.17(C4′)，73.18(C2′)，73.61(C5′)，76.38(C3′)，101.35(C1′)，115.40(C3+C5)，117.47(C2+C6)，125.67(C3″)，135.84(C2″)，150.10(C1)，152.24(C4)，166.33(C1″)。
1H?NMR：δppm?1.89(s，3H，H4″)，3.32-3.15(m，3H，H2′+H3′+H4′)，3.59(t，1H，H5′)，4.06(dd，1H，H6′)，4.44(d，1H，H6′)，4.69(d，1H，H1′)，5.14(s，1H，OH4′)，5.26(d，1H，OH3′)，5.29(d，1H，OH2′)，5.69(d，1H，H3″)，6.05(s，1H，H3″)，6.63(d，2H，H3+H5)，6.84(d，2H，H2+H6)，8.98(s，1H，OH4)。
Embodiment 11: arbutin 6 '-carbon undecylenate synthetic
With arbutin (109mg; 0.4mmol), after carbon undecylenic acid vinyl acetate (6mmol), 20mL THF add 50mL tool plug triangular flask mixing; In wherein adding the 600mg immobilized penicillium expansum lipase, in 45 ℃, the constant temperature oscillator of 200rpm, vibrate under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 0.5h, (sherwood oil (PE)/ETHYLE ACETATE (EA)=4/1, Rf=0.25) purifying get white powder product 172mg, and yield is 98%, and purity is greater than 99% for filtration, vacuum concentrated filtrate, column chromatography.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
1H?NMR：δppm?1.23(s，8H，H4′+H5″+H6″+H7″)，1.32-1.30(m，2H，H8″)，1.51(s，2H，H3″)，1.99(q，2H，H9″)，2.27(t，2H，H2″)，3.27-3.12(m，3H，H2′+H3′+H4′)，3.50(t，1H，H5′)，4.06(dd，1H，H6′)，4.31(d，1H，H6′)，4.66(d，1H，H1′)，5.00-4.90(m，2H，H11″)，5.13(s，1H，OH4′)，5.21(d，1H，OH3′)，5.29(s，1H，OH2′)，5.82-5.72(m，1H，H10″)，6.65(d，2H，H3+H5)，6.83(d，2H，H2+H6)，8.99(s，1H，OH4)。
13C?NMR：δppm?24.38(C3″)，28.25(C8″)，28.41(C6″+C7″)，28.60(C5″)，28.65(C4″)，33.12(C9″)，33.52(C2″)，63.38(C6′)，70.05(C4′)，73.19(C2′)，73.65(C5′)，76.36(C3′)，101.57(C1′)，114.51(C11″)，115.40(C3+C5)，117.66(C2+C6)，138.77(C10″)，150.12(C1)，152.34(C4)，172.66(C1″)。
Embodiment 12: arbutin 6 '-benzoic ether synthetic
With arbutin (109mg; 0.4mmol), vinyl benzoate (8mmol), 20mL octane/THF (19/1; V/v) behind the adding 50mL tool plug triangular flask mixing; In wherein adding the 700mg immobilized penicillium expansum lipase, place vibration in 60 ℃, the constant temperature oscillator of 200rpm under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 72h, filtration, vacuum concentrated filtrate, column chromatography (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying, product 130mg, yield is 86%, white powder, purity is greater than 99%.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms:
13C?NMR：δppm?165.51(C1″)，152.25(C4)，150.05(C1)，133.34(C5″)，129.68(C2″)，129.14(C3′+C7″)，128.68(C4″+C6″)，117.56(C2+C6)，115.40(C3+C5)，101.36(C1′)，76.45(C3′)，73.65(C5′)，73.24(C2′)，70.25(C4′)，64.32(C6′)。
1H?NMR：δppm?9.02(s，1H，OH4)，7.98(d，2H，H3″+H7″)，7.70-7.66(m，1H，H5″)，7.55(t，2H，H4″+H6″)，6.88(d，2H，H2+H6)，6.60(d，2H，H3+H5)，5.37-5.36(m，2H，OH2′+OH3′)，5.18(t，1H，OH4′)，4.76(t，1H，H1′)，4.63(d，1H，H6′)，4.31-4.27(m，1H，H6′)，3.54(t，1H，H5′)，3.38-3.28(m，3H，H2′+H3′+H4′)。
Embodiment 13: Gastrodine 6 '-capronate synthetic
With Gastrodine (115mg; 0.4mmol), vinyl caproate (4mmol), 20mL hexanaphthene/THF (1/4; V/v) behind the adding 50mL tool plug triangular flask mixing; In wherein adding the 500mg immobilized penicillium expansum lipase, place vibration in 50 ℃, the constant temperature oscillator of 200rpm under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 6h, filtration, vacuum concentrated filtrate, column chromatography (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying, product 131mg, yield is 85%, white powder, purity is greater than 99%.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms.
Embodiment 14: glucose helicidum 6 '-octanoate synthetic
With glucose helicidum (115mg; 0.4mmol), sad vinyl acetate (6mmol), 20mL octane-iso/trimethyl carbinol (1/5; V/v) behind the adding 50mL tool plug triangular flask mixing; In wherein adding the 400mg immobilized penicillium expansum lipase, place vibration in 55 ℃, the constant temperature oscillator of 200rpm under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 4h, filtration, vacuum concentrated filtrate, column chromatography (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying, product 145mg, yield is 88%, white powder, purity is greater than 99%.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms.
Embodiment 15: glucose helicidum 6 '-laurate synthetic
With glucose helicidum (115mg; 0.4mmol), hot LAURIC ACID 99 MIN vinyl acetate (4mmol), the positive nonane/acetone (1/5 of 20mL; V/v) behind the adding 50mL tool plug triangular flask mixing; In wherein adding the 520mg immobilized penicillium expansum lipase, place vibration in 30 ℃, the constant temperature oscillator of 250rpm under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 4h, filtration, vacuum concentrated filtrate, column chromatography (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying, product 142mg, yield is 86%, white powder, purity is greater than 99%.Structure is passed through
13C NMR (100.5MHz) and
1H NMR (400MHz) confirms.
Embodiment 16: salicylic aldehyde-β-D-glucoside 6 '-crotonate synthetic
With salicylic aldehyde-β-D-glucoside (115mg; 0.4mmol), Ba Dousuan vinyl acetate (4mmol), 20mL normal heptane/acetonitrile (1/5; V/v) behind the adding 50mL tool plug triangular flask mixing; In wherein adding the 800mg immobilized penicillium expansum lipase, place vibration in 55 ℃, the constant temperature oscillator of 100rpm under the normal pressure, utilize TLC monitoring reaction.Behind the reaction 72h, filtration, vacuum concentrated filtrate, column chromatography (sherwood oil (PE)/ETHYLE ACETATE (EA)=1/4, Rf=0.25) purifying, product 145mg, yield is 88%, white powder, purity is greater than 99%.Structure is passed through
13C NMR (100.5MHz) and
1HNMR (400MHz) confirms.
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (1)
1. the method for the lipase-catalyzed synthesis of glycoside esters compound of an immobilized penicillium expansum (Penicillium expansum); It is characterized in that comprising following operation steps: in non-aqueous media; The adding mol ratio is 1: 1～20: 1 acry radical donor and a glucosides; Obtain solution, the concentration of glucosides is 1～40mg/mL in this solution; Adding mass ratio with glucosides and be 10: 1～1: 10 immobilized penicillium expansum lipase, is that 30～60 ℃, hunting speed are under 100～300rpm and the condition of normal pressure in temperature, and transesterification 0.5～72 hour obtains glycoside esters compound;
Said acry radical donor is vinyl carboxylates or butyryl oxide;
Said glucosides is arbutin, Gastrodine, glucose helicidum, homoarbutin, different homoarbutin or salicylic aldehyde-β-D-glucoside;
Said non-aqueous media is two kinds in the aprotic organic solvent, or in the aprotic organic solvent a kind of with contain a kind of in the low anionic ionic liquid of nucleophilicity, the volumn concentration that wherein contains the low anionic ionic liquid of nucleophilicity is 5～95%;
Said aprotic organic solvent is THF, acetone, acetonitrile 、 diox, the trimethyl carbinol, isopropyl ether, octane-iso, normal hexane, normal heptane, octane, positive nonane or hexanaphthene;
The said negatively charged ion that contains in the low anionic ionic liquid of nucleophilicity is tetrafluoroborate ion, hexafluorophosphoricacid acid ions or two (trifluoromethyl sulphonyl) imines ion; Positively charged ion is a 1-alkyl-2,3-methylimidazole ion;
The preparation of said immobilized penicillium expansum lipase comprises following operation steps:
(1) will contain mass percent is that the thick enzyme powder of penicillium expansum lipase of 95% starch is dissolved in glycocoll-sodium hydrate buffer solution; Be mixed with every milliliter of damping fluid and contain the suspension-s of the thick enzyme powder of 0.3g lypase, under 35 ℃ of temperature and rotating speed 150rpm condition, stir 1h; The centrifugal supernatant that gets under 4 ℃ of temperature and rotating speed 6000rpm condition; The weight ratio of glycocoll and sodium hydroxide is 2.9: 1 in said glycocoll-sodium hydrate buffer solution;
(2) use mass percent to be 95% alcohol immersion macroporous adsorbent resin D4020 after, be washed till the filtrating clarification with deionized water, obtain filtrating;
(3) step (2) gained filtrating is added in step (1) the gained supernatant, enzyme concentration is the thick enzyme powder of 9g/g resin; Under 35 ℃ of temperature and rotating speed 150rpm condition, stir 4h, filter, with glycocoll-sodium hydrate buffer solution flushing, the suction filtration postlyophilization obtains immobilized penicillium expansum lipase.
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