CN103981228B - A kind of lipase-catalyzed benzene glycolylurea is prepared the method for N-carbamyl phenylglycine - Google Patents

Abstract

The invention discloses a method for preparing N-carbamoylphenylglycine from phenylhydantoin catalyzed by lipase. The method comprises the following steps: using mesodimenhydantoin as a raw material, isopropanol as a solvent, and lipase as a catalyst. A reaction system with a pH value of 6.4-6.8 is formed in a chitosan-containing buffer solution, and the stirring reaction is carried out at 40-50°C. After the reaction is complete, the reaction solution is separated and purified to obtain N-carbamoylphenylglycine; The invention uses lipase as the hydrolytic enzyme, and the coexistence of chitosan and lipase makes the lipase have good hydrolysis activity in the pH range of 6.4-6.8, and the yield of N-carbamoylphenylglycine reaches 41.0-49.5 of the amount of raw materials %; The method of the present invention not only has little environmental pollution, high product purity, low production cost, but also has industrial application value.

Description

Method for preparing N-carbamoylphenylglycine from phenylhydantoin catalyzed by a kind of lipase

(1) Technical field

The invention relates to a method for preparing N-carbamoylphenylglycine, in particular to a method for preparing N-carbamoylphenylglycine by lipase-catalyzed hydrolysis of phenylhydantoin.

(2) Background technology

Dihydantoin is used as raw material to prepare N-carbamoylphenylglycine by enzymatically hydrolyzing its amide bond with hydantoin. The technology is relatively mature. There are not only a large number of research reports at home and abroad, but also industrial practical applications. It is the key process for the production of phenylglycine. step, N-carbamoylphenylglycine is also a key intermediate. The technical process has less impact on the environment, the product has high purity, and has development prospects. However, the main disadvantage of this technology is that hydantoinase production cost is high, so the economic benefit is relatively poor, and the actual industrial production is difficult. For this reason, reducing its production cost is an urgent problem to be solved.

(3) Contents of the invention

The purpose of the present invention is to reduce the economic cost of preparing N-carbamoylphenylglycine by enzymatically hydrolyzing the amide bond of phenylhydantoin, replacing hydantoin enzyme with lipase, and hydrolyzing the amide bond of phenylhydantoin to obtain N-carbamoylphenylglycine, fat The production cost of the enzyme is much lower than that of hydantoin, and the source is extensive, which can obviously reduce the economic cost.

The technical scheme adopted in the present invention is:

The invention provides a lipase-catalyzed method for preparing N-carbamoylphenylglycine from dihydantoin. The method comprises the following steps: using mesodimenhydantoin as a raw material, isopropanol as a solvent, and lipase as a catalyst in a Chitosan buffer constitutes a reaction system with a pH value of 6.4 to 6.8, and the stirring reaction is carried out at 40 to 50°C (preferably stirring at 200 to 300rpm). After the reaction is completed, the reaction solution is separated and purified to obtain N -carbamoylphenylglycine; the buffer solution containing chitosan is after chitosan is mixed with the acetic acid aqueous solution of volume concentration 1% into the chitosan solution of chitosan mass concentration 2.5～3.6mg/ml, adjusts pH value to 5.6 ~ 6.2, then add sodium monohydrogen phosphate and sodium dihydrogen phosphate with a final concentration of 0.025mol/L, stir evenly, and make a buffer solution containing chitosan; the amount of lipase is 1200 ～2150U/L reaction system, the initial concentration of the mesodihydantoin is 7.0～8.8g/L reaction system, the volume final concentration of the isopropanol is 49～62%, the chitosan-containing The amount of the buffer is 0.8-2.0 g/L of the reaction system based on the mass of the chitosan.

Further, preferably, the lipase is porcine pancreatic lipase (with an activity of 2500 U/g) or Candida lipase (with an activity of 5000 U/g).

Further, preferably, the degree of deacetylation of the chitosan is greater than 90% (preferably 91.5%-92.1%).

Further, it is preferred that the conversion reaction time is 52 to 60 hours. After the conversion reaction is completed, liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid-sodium acetate buffer and methanol at a volume ratio of 99:1) Phase, flow rate 0.5 ml/min, column temperature 30°C, injection volume 5 microliters, ultraviolet light detector, wavelength 254nm) to detect the reaction solution, dihydantoin was used to quantify the standard substance to determine the conversion of the product.

Further, it is preferred that the amount of lipase is 1500-1700U/L reaction system, the initial concentration of mesodihydantoin is 7.7-7.9g/L reaction system, and the volume final concentration of isopropanol is 55 -57%, the amount of the chitosan-containing buffer is 1.0-1.8g/L reaction system based on the mass of chitosan.

Further, the method for separating and purifying the reaction liquid is preferably as follows: after the reaction, the reaction liquid is vacuum-recovered with isopropanol at 50° C. under 0.1 atmospheric pressure to obtain a concentrated liquid, and petroleum ether is added to the concentrated liquid for extraction, and the mixture is allowed to stand for stratification. Finally, take the petroleum ether layer and extract it with an ethanol aqueous solution with a volume concentration of 30%, take the ethanol aqueous solution phase, recover the ethanol in a vacuum at 50°C under 0.1 atmospheric pressure and dry it to obtain N-carbamoylphenylglycine.

Furthermore, the lipase-catalyzed method for preparing N-carbamoylphenylglycine from dihydantoin of the present invention is recommended to be carried out as follows: (1) Dissolving mesodihydantoin in isopropanol to prepare mesodimenhydrin Dihydantoin solution; (2) After mixing chitosan with acetic acid aqueous solution with a volume concentration of 1% to form a chitosan solution with a chitosan mass concentration of 2.5 to 3.6 mg/ml, use 1.0M sodium hydroxide Adjust the pH value of the aqueous solution to 5.6-6.2, then add sodium monohydrogen phosphate and sodium dihydrogen phosphate with a final concentration of 0.025mol/L, stir evenly, and make a buffer solution containing chitosan, and then mix lipase with Chitosan buffer is mixed to make a lipase mixture; (3) then mix the mesodihydantoin solution prepared in step (1) with the lipase mixture prepared in step (2), and use 1.0M The concentration of sodium hydroxide aqueous solution adjusts the pH value to 6.4-6.8 to form a reaction system, and the stirring reaction is carried out at 40-50°C and 200-300rpm. After the reaction is complete, the reaction solution is separated and purified to obtain N-carbamoylbenzene Glycine; the degree of deacetylation of the chitosan is greater than 90%; the amount of the lipase is 1500-1700U/L reaction system (preferably 1600U/L), and the initial concentration of the mesodihydantoin is 7.7- 7.9g/L reaction system (preferably 7.8g/L reaction system), the volume final concentration of described isopropanol is 55～57% (preferably 56%), the consumption of described chitosan-containing buffer is expressed as chitosan The mass of sugar is 1.0-1.8g/L reaction system (preferably 1.3g/L); the lipase is pig pancreas lipase or Candida lipase.

The enzymatic preparation of N-carbamoylphenylglycine from dihydantoin has the advantages of less environmental pollution and high product purity.

Compared with the prior art, the beneficial effects of the present invention are mainly reflected in: the present invention uses lipase as a hydrolase, and uses alkaline polysaccharide-chitosan and lipase to coexist to adjust the suitable hydrolysis pH value of lipase, so that fat The enzyme has good hydrolysis activity in the medium pH range of 6.4-6.8, the yield of porcine pancreas lipase to N-carbamoyl phenylglycine reaches 49.0-49.5% of the raw material amount, and Candida lipase to N-carbamoyl phenylglycine The productive rate of phenylglycine reaches 41.0～41.5% of raw material amount; The method of the present invention not only keeps the advantages of little environmental pollution and high product purity, but also has wide source of lipase, low price and mature industrial production, which makes the production cost of the process obvious Reduced, with industrial application value. At the same time, the application scope of the lipase is also broadened.

(4) Specific implementation methods

The present invention is further described below in conjunction with specific embodiment, but protection scope of the present invention is not limited thereto:

Chitosan of the present invention was purchased from Zhejiang Jinke Biochemical Co., Ltd., porcine pancreatic lipase was purchased from Shanghai Yixin Biological Co., Ltd., isopropanol was purchased from Hangzhou Shuanglin Chemical Reagent Factory, and mesodihydantoin was purchased from Macheng, Hubei Tengfei Biotechnology Co., Ltd. Candida lipase was purchased from Beijing Kaitai New Century Biology Co., Ltd.

Example 1:

(1) Take 99ml of distilled water and add 1.0ml of chemically pure acetic acid to make 1.0% volume concentration of acetic acid aqueous solution. Add 0.3 g of chitosan with a deacetylation degree of 92.1% (purchased from Zhejiang Jinke Biochemical Co., Ltd.) to the aqueous acetic acid solution, stir until the chitosan is completely dissolved, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of dihydrogen phosphate Sodium, stir and dissolve evenly, adjust the pH value to 5.9 with 1.0mol/L sodium hydroxide aqueous solution to make a chitosan-containing buffer solution. Add 0.15 g of porcine pancreatic lipase (purchased from Shanghai Yixin Biological Co., Ltd.) with an enzyme activity of 2500 U/g into the buffer solution containing chitosan, stir evenly, and then stir at 90 rpm for 15 minutes to make fat Enzyme Mix. (2) Add 1.8 g of mesodimenhydantoin (purchased from Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 130 ml of chemically pure isopropanol (purchased from Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to produce dihydantoin solution. (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.6 with 1.0mol/L sodium hydroxide aqueous solution to form a reaction system with a total volume of 233ml, and place in a water bath Keep at 45°C, mechanically stir at 300 rpm, and stop the reaction for 56 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , UV light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified by the standard substance, that is, 0.89 g of N-carbamoylphenylglycine was generated, and the reaction yield was 49.4%.

Reaction solution 232ml isopropanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure. After the recovery was completed, 100ml petroleum ether was added to extract for 15 minutes. The aqueous ethanol phase was removed, and the ethanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure and dried to obtain 0.72 g of N-carbamoylphenylglycine with a yield of 80.9%.

Example 2:

(1) Take 99ml of distilled water and add 1.0ml of chemically pure acetic acid to make 1.0% volume concentration of acetic acid aqueous solution. Add 0.36 g of chitosan with a deacetylation degree of 91.7% (purchased from Zhejiang Jinke Biochemical Co., Ltd.) to the aqueous acetic acid solution, stir until the chitosan is completely dissolved, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of dihydrogen phosphate Sodium, stir and dissolve evenly, adjust the pH value to 5.6 with 1.0mol/L sodium hydroxide aqueous solution to make a chitosan-containing buffer solution. Add 0.12 g of porcine pancreatic lipase with an enzyme activity of 2500 units/g (purchased from Shanghai Yixin Biological Co., Ltd.) into a buffer solution containing chitosan, stir evenly, and then stir at 100 rpm for 15 minutes to make fat Enzyme Mix. (2) Mix 1.4 g of mesodimenhydantoin (Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 100ml of chemically pure isopropanol (purchased from Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to make a dihydantoin solution . (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.4 with 1.0mol/L aqueous sodium hydroxide solution to form a reaction system with a total volume of 203ml, and place in a water bath Keep at 40°C, mechanically stir at 200 rpm, and stop the reaction for 52 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , ultraviolet light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified by the standard substance, that is, 0.69 g of N-carbamoylphenylglycine was generated, and the reaction yield was 49.3%.

Reaction solution 202ml isopropanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure. After the recovery was completed, 100ml petroleum ether was added to extract for 15 minutes. The aqueous ethanol phase was taken out, and the ethanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure and dried to obtain 0.55 g of N-carbamoylphenylglycine with a yield of 79.7%.

Example 3:

(1) Take 99ml of distilled water and add 1.0ml of chemically pure acetic acid to make 1.0% volume concentration of acetic acid aqueous solution. Add 0.26 g of chitosan with a deacetylation degree of 91.5% (Zhejiang Jinke Biochemical Co., Ltd.) to the aqueous acetic acid solution, stir until the chitosan is completely dissolved, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of sodium dihydrogen phosphate, Stir to dissolve evenly, and adjust the pH value to 6.2 with 1.0 mol/L aqueous sodium hydroxide solution to prepare a chitosan-containing buffer solution. Add 0.13 g of porcine pancreatic lipase (Shanghai Yixin Biological Co., Ltd.) with an enzyme activity of 2500 units/g to the glycan buffer, stir evenly, and then stir at 90 rpm for 20 minutes to prepare a lipase mixture. (2) Mix 2.0 g of mesodimenhydantoin (Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 160 ml of chemically pure isopropanol (Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to prepare a dihydantoin solution. (3) Mix the dihydantoin solution and the chitosan mixture prepared in step (1) evenly, adjust the pH value to 6.8 with a 1.0mol/L aqueous sodium hydroxide solution to form a reaction system with a total volume of 263ml, and place in a water bath Keep 50°C in the middle, stir mechanically at 250 rpm, and stop the reaction for 60 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , ultraviolet light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified due to the standard substance, that is, 0.98 g of N-carbamoylphenylglycine was generated, and the yield was 49.0%.

Reaction solution 262ml isopropanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure. After the recovery was completed, 100ml petroleum ether was added to extract for 15 minutes. The aqueous ethanol phase was taken out, and the ethanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure and dried to obtain 0.79 g of N-carbamoylphenylglycine with a yield of 80.6%.

Example 4:

(1) Take 99ml of distilled water and add 1.0ml of chemically pure acetic acid to make 1.0% volume concentration of acetic acid aqueous solution. Add 0.3 g of chitosan with a deacetylation degree of 92.1% (Zhejiang Jinke Biochemical Co., Ltd.) to the aqueous acetic acid solution, stir until the chitosan is completely dissolved, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of sodium dihydrogen phosphate, Stir and dissolve evenly, adjust the pH value to 5.9 with 1.0 mol/L sodium hydroxide aqueous solution to prepare a buffer solution containing chitosan. Mix 0.10 g of Candida lipase (Beijing Kaitai New Century Biological Co., Ltd.) with an enzyme activity of 5000 units/g into chitosan buffer, stir well, and then stir at 90 rpm for 20 minutes to make fat Enzyme Mix. (2) Mix 2.0 g of mesodimenhydantoin (Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 130 ml of chemically pure isopropanol (Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to prepare a dihydantoin solution. (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.8 with 1.0mol/L aqueous sodium hydroxide solution to form a reaction system with a total volume of 233ml, and place in a water bath Keep at 45°C, mechanically stir at 300 rpm, and stop the reaction for 60 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , ultraviolet light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified due to the standard substance, that is, 0.82 g of N-carbamoylphenylglycine was generated, and the yield was 41.0%.

Reaction solution 232ml isopropanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure. After the recovery was completed, 100ml petroleum ether was added to extract for 15 minutes. The aqueous ethanol phase was removed, and the ethanol was recovered under vacuum at 50°C under 0.1 atmospheric pressure and dried to obtain 0.66 g of N-carbamoylphenylglycine with a yield of 80.5%.

Comparative example 1

Take 100ml of distilled water, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of sodium dihydrogen phosphate, stir and dissolve evenly, adjust the pH value of the chitosan solution to 5.9 with 1.0 mol/L aqueous sodium hydroxide solution to make a buffer . Add 0.15 g of porcine pancreatic lipase with an enzyme activity of 2500 U/g (purchased from Shanghai Yixin Biological Co., Ltd.) into the buffer, stir evenly, and then stir at 90 rpm for 15 minutes to prepare a lipase mixture. (2) Add 1.8 g of mesodimenhydantoin (purchased from Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 130 ml of chemically pure isopropanol (purchased from Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to produce dihydantoin solution. (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.6 with 1.0mol/L sodium hydroxide aqueous solution to form a reaction system with a total volume of 232ml, and place in a water bath Keep at 45°C, mechanically stir at 300 rpm, and stop the reaction for 56 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , ultraviolet light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified by the standard substance, that is, 0.42 g of N-carbamoylphenylglycine was generated, and the reaction yield was 23.3%.

Comparative example 2:

(1) Take 100ml of distilled water, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of sodium dihydrogen phosphate, stir and dissolve evenly, and adjust the pH value to 5.9 with 1.0 mol/L aqueous sodium hydroxide solution to make a buffer solution. Mix 0.10 g of Candida lipase (Beijing Kaitai New Century Biological Co., Ltd.) with an enzyme activity of 5000 units/g into the buffer solution, stir evenly, and then stir at 90 rpm for 20 minutes to make a lipase mixture . (2) Mix 2.0 g of mesodimenhydantoin (Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 130 ml of chemically pure isopropanol (Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to prepare a dihydantoin solution. (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.8 with 1.0mol/L sodium hydroxide aqueous solution to form a reaction system with a total volume of 232ml, and place in a water bath Keep at 45°C, mechanically stir at 300 rpm, and stop the reaction for 60 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , ultraviolet light detector, wavelength 254nm) to detect the reaction solution, benzohydantoin was quantified due to the standard substance, and 0.39 g of N-carbamoylphenylglycine was generated, with a yield of 19.5%.

Comparative example 3:

(1) Take 99ml of distilled water and add 1.0ml of chemically pure acetic acid to make 1.0% volume concentration of acetic acid aqueous solution. Add 0.3 g of chitosan with a deacetylation degree of 92.1% (purchased from Zhejiang Jinke Biochemical Co., Ltd.) to the aqueous acetic acid solution, stir until the chitosan is completely dissolved, add 0.36 g of sodium monohydrogen phosphate and 0.30 g of dihydrogen phosphate Sodium, stir and dissolve evenly, adjust the pH value of the chitosan solution to 5.9 with an aqueous sodium hydroxide solution of 1.0 mol/L concentration, and make a buffer solution containing chitosan. Add 0.15 g of Penicillium lipase (Beijing Kaitai New Century Biological Co., Ltd.) with an enzyme activity of 3000 U/g into the chitosan buffer, stir well, and then stir at 90 rpm for 15 minutes to make a lipase mixture . (2) Add 1.8 g of mesodimenhydantoin (purchased from Hubei Macheng Tengfei Biotechnology Co., Ltd.) into 130 ml of chemically pure isopropanol (purchased from Hangzhou Shuanglin Chemical Reagent Factory), and dissolve evenly to produce dihydantoin solution. (3) Mix the dihydantoin solution and the lipase mixture prepared in step (1) evenly, adjust the pH value to 6.6 with 1.0mol/L sodium hydroxide aqueous solution to form a reaction system with a total volume of 233ml, and place in a water bath Keep at 45°C, mechanically stir at 300 rpm, and stop the reaction for 56 hours. Use liquid chromatography (chromatographic conditions: C18 column, pH5.2 acetic acid: sodium acetate buffer and methanol at a volume ratio of 99:1 as the mobile phase, flow rate 0.5 ml/min, column temperature 30 ° C, injection volume 5 microliters , UV light detector, wavelength 254nm) to detect the reaction solution, benzohydrin was quantified by the standard substance, that is, 0.19 g of N-carbamoylphenylglycine was generated, and the reaction yield was 10.6%.

Claims (7)

1. lipase-catalyzed benzene glycolylurea is prepared a method for N-carbamyl phenylglycine, and its feature existsIn described method be: taking meso benzene glycolylurea as raw material, taking isopropyl alcohol as solvent, taking lipase as catalysisAgent forms pH value and is 6.4～6.8 reaction system, at 40～50 DEG C of bars in the buffer solution of chitosan-containingUnder part, carry out stirring reaction, after reacting completely, by reactant liquor separation and purification, obtain N-carbamyl benzene sweetPropylhomoserin; The buffer solution of described chitosan-containing is that the acetic acid aqueous solution of shitosan and volumetric concentration 1% is mixed intoAfter the chitosan solution of chitosan mass concentration 2.5～3.6mg/ml, regulate pH value to 5.6～6.2, thenAdd again final concentration to be disodium-hydrogen and the sodium dihydrogen phosphate of 0.025mol/L, stir, obtainThe buffer solution of chitosan-containing; The consumption of described lipase is 1200～2150U/L reaction system, disappears in describedThe initial concentration that revolves benzene glycolylurea is 7.0～8.8g/L reaction system, and the volume final concentration of described isopropyl alcohol is49～62%, the consumption of the buffer solution of described chitosan-containing is in the quality of shitosan, and the consumption of shitosan is0.8～2.0g/L reaction system.

2. the side that lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycine as claimed in claim 1Method, is characterized in that described lipase is porcine pancreatic lipase or lipase from candida sp.

3. the side that lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycine as claimed in claim 1Method, is characterized in that the deacetylation of described shitosan is greater than 90%.

4. the side that lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycine as claimed in claim 1Method, is characterized in that the described catalytic reaction time is 52～60 hours.

5. the side that lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycine as claimed in claim 1Method, the consumption that it is characterized in that described lipase is 1500～1700U/L reaction system, described meso benzeneThe initial concentration of glycolylurea is 7.7～7.9g/L reaction system, and the volume final concentration of described isopropyl alcohol is 55-57%,The consumption of the buffer solution of described chitosan-containing is in the quality of shitosan, and the consumption of shitosan is 1.0～1.8g/LReaction system.

6. the side that lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycine as claimed in claim 1Method, is characterized in that described method carries out as follows: meso benzene glycolylurea is dissolved in isopropyl alcohol by (1)In, make meso benzene glycolylurea solution; (2) shitosan is mixed with the acetic acid aqueous solution of volumetric concentration 1%After the chitosan solution of the mass concentration 2.5～3.6mg/ml of one-tenth shitosan, use the NaOH of 1.0M concentrationThe aqueous solution regulates pH value to 5.6～6.2, and then adds final concentration to be phosphoric acid one hydrogen of 0.025mol/LSodium and sodium dihydrogen phosphate, stir, and makes the buffer solution of chitosan-containing, then by lipase and poly-containing shellThe buffer solution of sugar is mixed and made into lipase mixed liquor; (3) the meso benzene sea of then being prepared by step (1)The lipase mixed liquor of preparing with step (2) because of solution mixes the rear NaOH by 1.0M concentrationThe aqueous solution regulates pH value to 6.4～6.8, forms reaction system, 40～50 DEG C, 200～300rpm conditionUnder carry out stirring reaction, after reacting completely, by reactant liquor separation and purification, obtain N-carbamyl benzene sweet ammoniaAcid; The deacetylation of described shitosan is greater than 90%; The consumption of described lipase is 1500～1700U/LReaction system, the initial concentration of described meso benzene glycolylurea is 7.7～7.9g/L reaction system, described isopropylThe volume final concentration of alcohol is 55～57%, and the consumption of the buffer solution of described chitosan-containing is with the quality of shitosanMeter, the consumption of shitosan is 1.0～1.8g/L reaction system; Described lipase is porcine pancreatic lipase or false silkYeast-lipase.

7. as described in claim 1 or 6, lipase-catalyzed benzene glycolylurea is prepared N-carbamyl phenylglycineMethod, it is characterized in that the method for described reactant liquor separation and purification is: reaction finish after, by reactant liquorUnder 0.1 atmospheric pressure, 50 DEG C of vacuum reclaim isopropyl alcohol, obtain concentrate, in concentrate, add benzinumExtract, after stratification, get petroleum ether layer and extract with the ethanol water of volumetric concentration 30%,Get ethanol water phase, under 0.1 atmospheric pressure, 50 DEG C of vacuum reclaim ethanol dry, obtain N-carbamylBase phenylglycine.