CN110591077A - Method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol through enzyme catalysis - Google Patents
Method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol through enzyme catalysis Download PDFInfo
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Abstract
The invention relates to a method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol by enzyme catalysis, belonging to the technical field of enzyme catalysis. And (2) catalyzing and synthesizing tyrosine oligopeptide by using protease as a catalyst, and grafting monomethoxypolyethylene glycol to the amino terminal of the oligomeric tyrosine oligopeptide to prepare the tyrosine oligopeptide-monomethoxypolyethylene glycol copolymer. The yield reaches 15-55%. The catalyst in the polypeptide synthesis step is enzyme, has the advantages of mild reaction conditions, more environment-friendly property, less side reaction and the like, and avoids the complicated protection and deprotection steps, thereby providing a novel method for preparing the amino acid polypeptide-monomethoxy polyethylene glycol.
Description
Technical Field
The invention relates to a method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol by enzyme catalysis, belonging to the technical field of enzyme catalysis.
Background
The artificial polypeptide is a polyamino compound formed by taking natural amino acid as a monomer to carry out polymerization reaction through amido bond, and the properties of the artificial polypeptide are similar to those of natural peptide, polyamino acid and protein. The material formed by the artificial polypeptide has good biocompatibility and degradability. The polypeptide or the modified oligomer thereof shows unique structural properties such as self-assembly, liquid crystal behavior and the like, has stronger plasticity, and has great application potential in the fields of biology, medicine and natural high polymer materials. The polytyrosine can be applied to the fields of drug delivery, gene therapy and the like after being modified by pegylation. In addition, each phenolic hydroxyl group of the polytyrosine molecule generates two electrons in the oxidation process, so the polytyrosine molecule is used as an electroactive marker for signal amplification of an electrochemical biosensor.
Examples of the polymerization method of amino acids include a solid phase method, an NCA method and an enzyme catalysis method. Compared with other polymerization methods, the enzyme catalysis method has the advantages of mild reaction conditions, more environment-friendly property, less side reactions and the like, and avoids fussy protection and deprotection steps.
The invention aims to synthesize tyrosine oligopeptide under the catalysis of protease as a catalyst, and then graft monomethoxypolyethylene glycol on the amino terminal of the oligomeric tyrosine oligopeptide to prepare the tyrosine oligopeptide-monomethoxypolyethylene glycol copolymer.
Disclosure of Invention
The invention aims to overcome the defects and provide a method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol through enzyme catalysis, which has mild reaction conditions and is more environment-friendly.
The technical scheme of the invention is a method for preparing tyrosine oligopeptide and grafted monomethoxy polyethylene glycol by enzyme catalysis, which comprises the following steps: (1) synthesizing tyrosine oligopeptide under the catalysis of protease; (2) preparing monomethoxy polyethylene glycol succinimide carbonate; (3) and (3) grafting the tyrosine oligopeptide with monomethoxy polyethylene glycol to prepare a tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer.
(1) Enzymatic synthesis of tyrosine oligopeptides: adding L-tyrosine methyl ester hydrochloride, dimethyl sulfoxide and protease into a phosphate buffer solution, placing a reaction mixture into a constant-temperature oscillator for reaction, centrifuging after the reaction is finished, taking a precipitate, washing with deionized water and ice ethanol twice respectively, and freeze-drying to obtain a product tyrosine oligopeptide.
The method comprises the specific steps of adding 0.5-2.0 g of L-tyrosine methyl ester hydrochloride and 2-5U of protease into a disodium hydrogen phosphate-citric acid buffer solution with the volume of 2.0-3.0 mL, the pH value of 5.5 ~ 8.0.0 and the concentration of 0.1 ~ 0.2.2M, adding dimethyl sulfoxide accounting for 2.5-25% of the buffer solution, placing the mixture into a constant temperature oscillator at 200 ~ 600rpm and at 30-70 ℃ for reaction for 5-10 hours, after the reaction is finished, centrifuging the reaction mixture at 5000 ~ 10000rpm for 3 ~ 10 minutes, washing precipitates twice with deionized water and glacial ethanol, and freeze-drying to obtain the tyrosine oligopeptide.
The protease is any one of bromelain, papain, trypsin, neutral protease and alkaline protease.
The phosphate buffer is 0.1 ~ 0.2.2M disodium hydrogen phosphate-citric acid buffer, and the pH value of the phosphate buffer is 5.5 ~ 8.0.0.
The addition amount of the dimethyl sulfoxide is 2.5-25% of that of the phosphate buffer solution according to the volume concentration.
(2) Preparation of monomethoxypolyethylene glycol succinimide carbonate: adding monomethoxy polyethylene glycol, N' -disuccinimidyl carbonate and 4- (dimethylamino) pyridine into anhydrous tetrahydrofuran, and stirring for reaction; and (3) distilling the obtained reactant in vacuum, pouring the distilled reactant into precooled anhydrous ether, carrying out reduced pressure suction filtration, and collecting the precipitate to obtain the monomethoxy polyethylene glycol succinimide carbonate.
Adding 1-2 g of monomethoxy polyethylene glycol, 5-8 g of N, N' -disuccinimidyl carbonate and 3-6 g of 4- (dimethylamino) pyridine into 25mL of anhydrous tetrahydrofuran, reacting for 2 ~ 6 hours at the stirring speed of 200 ~ 400rpm and the temperature of 30 ~ 60 ℃, after the reaction is finished, pouring the reaction mixture into precooled anhydrous ether at the temperature of 25 ~ 40 ℃, removing the solvent through vacuum distillation, carrying out vacuum filtration at the pressure of 0 ~ 10kPa, collecting precipitates, and obtaining the precipitates which are the methoxypolyethylene glycol succinimidyl carbonate.
The monomethoxypolyethylene glycol has an average molecular weight of 200, 400 or 1000 Da.
(3) Preparation of tyrosine oligopeptide-monomethoxypolyethylene glycol copolymer: adding tyrosine oligopeptide, methoxy polyethylene glycol succinimide carbonate and triethylamine into absolute ethyl alcohol, mixing and reacting under the protection of nitrogen; standing overnight after the reaction is finished, performing vacuum filtration, performing vacuum drying, and then dissolving in acetone; standing again overnight, and carrying out vacuum filtration to obtain the grafted product tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer.
Adding 2-5 g of tyrosine oligopeptide, 2.4-10 g of monooxypolyethylene glycol succinimide carbonate and 12-60 g of triethylamine into 15mL of anhydrous methanol, reacting the reaction mixture at 30 ~ 50 ℃ for 10-24 h under the protection of nitrogen, standing the reaction mixture at 0 ℃ overnight after the reaction is finished, carrying out suction filtration under reduced pressure of 0 ~ 10kPa, carrying out vacuum drying on the filtrate at 30 ~ 70 ℃ for 6 ~ 10 h, adding acetone preheated to 30 ~ 40 ℃ until the acetone is completely dissolved, standing the obtained solution at 0 ℃ overnight, carrying out suction filtration under reduced pressure of 0 ~ 10kPa, and obtaining a precipitate which is the tyrosine oligopeptide-monomethoxypolyethylene glycol copolymer.
The weight ratio of the tyrosine oligopeptide, the monomethoxy polyethylene glycol succinimide carbonate and the triethylamine is 1:1.2: 6-1: 2: 12.
The 1HNMR detection is carried out on the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer by the following method: the 1HNMR spectra of the copolymers were determined at room temperature using an AVANCE III HD-400 MHz NMR spectrometer with CF3COOD as solvent and Tetramethylsilane (TMS) as internal standard.
The invention has the beneficial effects that: the catalyst in the polypeptide synthesis step is enzyme, has the advantages of mild reaction conditions, more environment-friendly property, less side reaction and the like, and avoids the complicated protection and deprotection steps, thereby providing a novel method for preparing the amino acid polypeptide-monomethoxy polyethylene glycol.
Drawings
FIG. 1 is a scheme for detecting a tyrosine oligopeptide-monomethoxypolyethylene glycol copolymer by HNMR in example 11.
Detailed Description
In the following examples, the protease was obtained from Waokay Biotechnology Ltd, the buffer was obtained from Nanchang rain and dew laboratory facilities Ltd, the other reagents were obtained from Chemicals group Ltd or Huaian petrochemical plant of Jiangsu province, and the 4- (dimethylamino) pyridine was obtained from Aradin Biotechnology Ltd.
Example 1
(1) Protease catalyzed synthesis of tyrosine oligopeptides: to 2.0 mL of 0.1M disodium hydrogen phosphate-citric acid buffer pH5.8 was added 0.5 g of L-tyrosine methyl ester hydrochloride, dimethyl sulfoxide and 2U of protease. The volume fraction of dimethyl sulfoxide was 2.5% phosphate buffer. The reaction mixture was placed in a constant temperature shaker (600 rpm) and reacted at 30 ℃ for 10 hours. After the reaction is finished, centrifuging the reaction mixture for 3 minutes at 10000rpm, washing the precipitate twice by deionized water and glacial ethanol respectively, and freeze-drying to obtain the product tyrosine oligopeptide.
(2) The preparation of monomethoxy polyethylene glycol succinimide carbonate is carried out by adding 2g monomethoxy polyethylene glycol, 5g N, N' -disuccinimide carbonate and 3 g 4- (dimethylamino) pyridine into 25mL anhydrous tetrahydrofuran, reacting the reaction mixture at 30 ℃ for 6 hours under the condition of stirring (200 rpm), after the reaction is finished, distilling the reaction mixture at 40 ℃ in vacuum to remove solvent, pouring into precooled anhydrous ether, carrying out vacuum filtration under 0 ~ 10kPa, collecting precipitate, and obtaining the precipitate as monomethoxy polyethylene glycol succinimide carbonate.
(3) The preparation method of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer comprises the steps of adding 2g of tyrosine oligopeptide, methoxy polyethylene glycol succinimide carbonate, triethylamine, the weight ratio of the tyrosine oligopeptide, the methoxy polyethylene glycol succinimide carbonate and the triethylamine into 15mL of anhydrous methanol, reacting the reaction mixture at 30 ℃ for 10 hours under the protection of nitrogen, standing the reaction mixture at 0 ℃ overnight after the reaction is finished, carrying out vacuum filtration under 0 ~ 10kPa, drying the filtrate at 30 ℃ for 6 hours in vacuum, adding acetone preheated to 40 ℃ until the acetone is completely dissolved, standing the obtained solution at 0 ℃ overnight, carrying out vacuum filtration under 0 ~ 10kPa, and obtaining the precipitate which is the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer with the yield of 15%.
By passing1Detecting the substitution degree of tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer by HNMR method, specifically as shown in figure 1, wherein a is-CH in polyethylene glycol monomethyl ether chain segment3Chemical shift peak of hydrogen; f is a hydrogen chemical shift peak of a terminal methyl (-OCH3) of the tyrosine oligopeptide. The substitution degree of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer obtained from the peak area ratio is 1.1.
Example 2
(1) Protease catalyzed synthesis of tyrosine oligopeptides: to 2.5 mL of 0.15M disodium hydrogen phosphate-citric acid buffer pH7.0 was added 1 g of L-tyrosine methyl ester hydrochloride, dimethyl sulfoxide and 3.5U of papain. The volume fraction of dimethyl sulfoxide was 15% phosphate buffer. The reaction mixture was placed in a constant temperature shaker (600 rpm) and reacted at 30 ℃ for 8 hours. After the reaction is finished, centrifuging the reaction mixture for 3 minutes at 10000rpm, washing the precipitate twice by deionized water and glacial ethanol respectively, and freeze-drying to obtain the product tyrosine oligopeptide.
(2) The preparation of monomethoxy polyethylene glycol succinimide carbonate is carried out by adding 1.5g monomethoxy polyethylene glycol, 6g N, N' -disuccinimidyl carbonate, 6g 4- (dimethylamino) pyridine into 25mL anhydrous tetrahydrofuran, reacting the reaction mixture at 30 ℃ for 6 hours under stirring (200 rpm), after the reaction, distilling the reaction mixture at 40 ℃ in vacuum to remove solvent, pouring into anhydrous precooled ether, filtering under reduced pressure of 0 ~ 10kPa, collecting precipitate, and obtaining the precipitate as monomethoxy polyethylene glycol succinimide carbonate.
(3) The preparation method of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer comprises the steps of adding 3 g of tyrosine oligopeptide, methoxy polyethylene glycol succinimide carbonate, triethylamine, the weight ratio of the tyrosine oligopeptide, the methoxy polyethylene glycol succinimide carbonate and the triethylamine into 15ml of anhydrous methanol, reacting the reaction mixture at 40 ℃ for 16 h under the protection of nitrogen, standing the reaction mixture at 0 ℃ overnight after the reaction is finished, carrying out vacuum filtration under 0 ~ 10kPa, drying the filtrate at 30 ℃ for 6 h in vacuum, adding acetone preheated to 40 ℃ until the acetone is completely dissolved, standing the obtained solution at 0 ℃ overnight, carrying out vacuum filtration under 0 ~ 10kPa, and obtaining the precipitate which is the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer with the yield of 30%.
By passing1The substitution degree of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer detected by HNMR means is 1.01.
Example 3
(1) Protease catalyzed synthesis of tyrosine oligopeptides: to 2.5 mL of 0.2M, pH 8.0.0 disodium hydrogen phosphate-citric acid buffer was added 1 g of L-tyrosine methyl ester hydrochloride, dimethyl sulfoxide and 3.5U of alkaline protease. The volume fraction of dimethyl sulfoxide was 25% phosphate buffer. The reaction mixture was placed in a constant temperature shaker (600 rpm) and reacted at 70 ℃ for 10 hours. After the reaction is finished, centrifuging the reaction mixture for 3 minutes at 10000rpm, washing the precipitate twice by deionized water and glacial ethanol respectively, and freeze-drying to obtain the product tyrosine oligopeptide.
(2) The preparation of monomethoxy polyethylene glycol succinimide carbonate is carried out by adding 2g monomethoxy polyethylene glycol, 8g N, N' -disuccinimide carbonate and 5g 4- (dimethylamino) pyridine into 25mL anhydrous tetrahydrofuran, reacting the reaction mixture at 30 ℃ for 6 hours under the condition of stirring (200 rpm), after the reaction is finished, distilling the reaction mixture at 40 ℃ in vacuum to remove solvent, pouring into precooled anhydrous ether, carrying out vacuum filtration under 0 ~ 10kPa, collecting precipitate, and obtaining the precipitate as monomethoxy polyethylene glycol succinimide carbonate.
(3) The preparation method of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer comprises the steps of adding 5g of tyrosine oligopeptide, oxypolyethylene glycol succinimide carbonate, triethylamine, tyrosine oligopeptide, methoxypolyethylene glycol succinimide carbonate and triethylamine into 15mL of anhydrous methanol, wherein the weight ratio of the tyrosine oligopeptide to the methoxypolyethylene glycol succinimide carbonate to the triethylamine is 1: 2:12, reacting the reaction mixture at 50 ℃ for 24h under the protection of nitrogen, standing the reaction mixture at 0 ℃ overnight after the reaction is finished, carrying out vacuum filtration at 0 ~ 10kPa, drying the filtrate at 30 ℃ for 6 h in vacuum, adding acetone preheated to 40 ℃ until the acetone is completely dissolved, standing the obtained solution at 0 ℃ overnight, carrying out vacuum filtration at 0 ~ 10kPa, and obtaining the precipitate which is the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer with the yield of 55%.
By passing1The substitution degree of the tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer detected by HNMR means is 1.06.
Claims (8)
1. The method for grafting tyrosine oligopeptide by using monomethoxypolyethylene glycol is characterized by comprising the following steps of: adding tyrosine oligopeptide, monomethoxy polyethylene glycol succinimide carbonate and triethylamine into absolute ethyl alcohol, mixing and reacting under the protection of nitrogen; standing overnight after the reaction is finished, performing vacuum filtration, performing vacuum drying, and then dissolving in acetone; standing again overnight, and carrying out vacuum filtration to obtain the grafted product tyrosine oligopeptide-monomethoxy polyethylene glycol copolymer.
2. The method for grafting tyrosine oligopeptide through monomethoxy polyethylene glycol as claimed in claim 1, which is characterized in that the method comprises the following specific steps of adding 2-5 g of tyrosine oligopeptide, 2.4-10 g of monooxygroup polyethylene glycol succinimide carbonate and 12-60 g of triethylamine into 15mL of anhydrous methanol, reacting the reaction mixture at 30 ~ 50 ℃ for 10-24 h under the protection of nitrogen, standing the reaction mixture at 0 ℃ overnight after the reaction is finished, carrying out vacuum filtration at 0 ~ 10kPa, drying the filtrate at 30 ~ 70 ℃ for 6 ~ 10 h, adding acetone preheated to 30 ~ 40 ℃ until all the acetone is dissolved, standing the obtained solution at 0 ℃ overnight, carrying out vacuum filtration at 0 ~ 10kPa, and obtaining the precipitate as the tyrosine-monomethoxy polyethylene glycol copolymer.
3. The preparation method of the tyrosine oligopeptide through enzyme catalysis synthesis is characterized in that: adding L-tyrosine methyl ester hydrochloride, dimethyl sulfoxide and protease into a phosphate buffer solution, placing a reaction mixture into a constant-temperature oscillator for reaction, centrifuging after the reaction is finished, taking a precipitate, washing with deionized water and ice ethanol twice respectively, and freeze-drying to obtain a product tyrosine oligopeptide.
4. The method for preparing tyrosine oligopeptide by enzymatic synthesis according to claim 3, wherein the method comprises the steps of adding 0.5-2.0 g of L-tyrosine methyl ester hydrochloride and 2-5U of protease into a disodium hydrogen phosphate-citric acid buffer solution with the volume of 2.0-3.0 mL, the pH value of 5.5 ~ 8.0.0 and the concentration of 0.1 ~ 0.2.2M, adding dimethyl sulfoxide accounting for 2.5-25% of the volume of the buffer solution, placing the mixture in a constant temperature oscillator at 200 ~ 600rpm and 30-70 ℃ for reaction for 5-10 hours, centrifuging the reaction mixture at 5000 ~ 10000rpm for 3 ~ 10 minutes after the reaction is finished, washing the precipitate twice with deionized water and glacial ethanol, and freeze-drying to obtain the tyrosine oligopeptide.
5. The method for preparing tyrosine oligopeptide by enzymatic synthesis according to claim 3, wherein: the protease is bromelain, papain, trypsin, neutral protease or alkaline protease.
6. The preparation method of the monomethoxy polyethylene glycol succinimide carbonate is characterized by comprising the following steps: adding monomethoxy polyethylene glycol, N' -disuccinimidyl carbonate and 4- (dimethylamino) pyridine into anhydrous tetrahydrofuran, and stirring for reaction; and (3) distilling the obtained reactant in vacuum, pouring the distilled reactant into precooled anhydrous ether, carrying out reduced pressure suction filtration, and collecting the precipitate to obtain the monomethoxy polyethylene glycol succinimide carbonate.
7. The preparation method of monomethoxy polyethylene glycol succinimide carbonate as claimed in claim 6, which comprises adding 1-2 g monomethoxy polyethylene glycol, 5-8 g N, N' -disuccinimide carbonate, 3-6 g 4- (dimethylamino) pyridine into 25mL anhydrous tetrahydrofuran, reacting at 30 ~ 60 ℃ for 2 ~ 6 hours under 200 ~ 400rpm stirring speed, after the reaction is over, distilling the reaction mixture at 25 ~ 40 ℃ under vacuum to remove the solvent, pouring into precooled anhydrous ether, filtering under reduced pressure at 0 ~ 10kPa to collect the precipitate, and obtaining the precipitate as methoxy polyethylene glycol succinimide carbonate.
8. The process for preparing methoxypolyethylene glycol succinimide carbonate according to claim 6, wherein: the methoxypolyethylene glycol has an average molecular weight of 200, 400 or 1000 Da.
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| CN112251484A (en) * | 2020-10-26 | 2021-01-22 | 江南大学 | Method for preparing glycine oligopeptide-polyethylene glycol copolymer under catalysis of lipase |
Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5281698A (en) * | 1991-07-23 | 1994-01-25 | Cetus Oncology Corporation | Preparation of an activated polymer ester for protein conjugation |
| US5324844A (en) * | 1989-04-19 | 1994-06-28 | Enzon, Inc. | Active carbonates of polyalkylene oxides for modification of polypeptides |
| CN1673230A (en) * | 2004-03-24 | 2005-09-28 | 中国科学院过程工程研究所 | Prepn of recombinant composite interferon-polyglycol conjugate and conjugate product |
| CN102827365A (en) * | 2006-11-07 | 2012-12-19 | 中国药科大学 | Amino acid connected with polyethylene glycol and preparation method and application for amino acid |
| CN104109235A (en) * | 2014-05-30 | 2014-10-22 | 厦门赛诺邦格生物科技有限公司 | Mono-functionalized polyethylene glycol with nitrogen atom branched center and its preparation method and biologically-relevant matter |
| CN104144705A (en) * | 2011-11-03 | 2014-11-12 | 拜耳知识产权有限责任公司 | Tyrosine based linkers for the releasable connection of peptides |
| CN104693433A (en) * | 2015-02-04 | 2015-06-10 | 国家纳米科学中心 | PEGylation modified amifostine as well as preparation method and use of PEGylation modified amifostine |
| CN104774891A (en) * | 2015-01-15 | 2015-07-15 | 南京工业大学 | Technology for efficiently synthesizing benzyloxycarbonyl aspartame through using enzyme method |
| CN107857749A (en) * | 2017-10-26 | 2018-03-30 | 湖南华腾制药有限公司 | A kind of poly glycol monomethyl ether statin compound and preparation method thereof |
| CN108503857A (en) * | 2018-03-16 | 2018-09-07 | 东华大学 | A kind of double cross connection mussel for tissue adhesive bonds albumen biomim betatic and preparation method thereof |
| CN108796020A (en) * | 2018-07-08 | 2018-11-13 | 宋冠冠 | A kind of plant source polypeptide with tyrosinase inhibitory activity and containing the cosmetics of the polypeptide |
| CN109207542A (en) * | 2018-09-27 | 2019-01-15 | 武汉轻工大学 | A kind of method of modifying of soya-bean polypeptides |
| CN109234343A (en) * | 2018-09-30 | 2019-01-18 | 中国海洋大学 | A kind of methionine oligopeptide and its preparation method and application |
| CN109535247A (en) * | 2018-12-24 | 2019-03-29 | 中国药科大学 | Polyethyleneglycol modified phycocyanin and preparation method thereof and pharmacy application |
-
2019
- 2019-10-17 CN CN201910985766.2A patent/CN110591077B/en active Active
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5324844A (en) * | 1989-04-19 | 1994-06-28 | Enzon, Inc. | Active carbonates of polyalkylene oxides for modification of polypeptides |
| US5281698A (en) * | 1991-07-23 | 1994-01-25 | Cetus Oncology Corporation | Preparation of an activated polymer ester for protein conjugation |
| CN1673230A (en) * | 2004-03-24 | 2005-09-28 | 中国科学院过程工程研究所 | Prepn of recombinant composite interferon-polyglycol conjugate and conjugate product |
| CN102827365A (en) * | 2006-11-07 | 2012-12-19 | 中国药科大学 | Amino acid connected with polyethylene glycol and preparation method and application for amino acid |
| CN104144705A (en) * | 2011-11-03 | 2014-11-12 | 拜耳知识产权有限责任公司 | Tyrosine based linkers for the releasable connection of peptides |
| CN104109235A (en) * | 2014-05-30 | 2014-10-22 | 厦门赛诺邦格生物科技有限公司 | Mono-functionalized polyethylene glycol with nitrogen atom branched center and its preparation method and biologically-relevant matter |
| CN104774891A (en) * | 2015-01-15 | 2015-07-15 | 南京工业大学 | Technology for efficiently synthesizing benzyloxycarbonyl aspartame through using enzyme method |
| CN104693433A (en) * | 2015-02-04 | 2015-06-10 | 国家纳米科学中心 | PEGylation modified amifostine as well as preparation method and use of PEGylation modified amifostine |
| CN107857749A (en) * | 2017-10-26 | 2018-03-30 | 湖南华腾制药有限公司 | A kind of poly glycol monomethyl ether statin compound and preparation method thereof |
| CN108503857A (en) * | 2018-03-16 | 2018-09-07 | 东华大学 | A kind of double cross connection mussel for tissue adhesive bonds albumen biomim betatic and preparation method thereof |
| CN108796020A (en) * | 2018-07-08 | 2018-11-13 | 宋冠冠 | A kind of plant source polypeptide with tyrosinase inhibitory activity and containing the cosmetics of the polypeptide |
| CN109207542A (en) * | 2018-09-27 | 2019-01-15 | 武汉轻工大学 | A kind of method of modifying of soya-bean polypeptides |
| CN109234343A (en) * | 2018-09-30 | 2019-01-18 | 中国海洋大学 | A kind of methionine oligopeptide and its preparation method and application |
| CN109535247A (en) * | 2018-12-24 | 2019-03-29 | 中国药科大学 | Polyethyleneglycol modified phycocyanin and preparation method thereof and pharmacy application |
Non-Patent Citations (1)
| Title |
|---|
| 刘严华等: "L-苯丙氨酸聚肽的酶催化合成", 《化学通报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112251484A (en) * | 2020-10-26 | 2021-01-22 | 江南大学 | Method for preparing glycine oligopeptide-polyethylene glycol copolymer under catalysis of lipase |
| CN112251484B (en) * | 2020-10-26 | 2022-05-24 | 江南大学 | Method for preparing glycine oligopeptide-polyethylene glycol copolymer under catalysis of lipase |
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