CN102212586B - Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase - Google Patents
Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase Download PDFInfo
- Publication number
- CN102212586B CN102212586B CN2011101104305A CN201110110430A CN102212586B CN 102212586 B CN102212586 B CN 102212586B CN 2011101104305 A CN2011101104305 A CN 2011101104305A CN 201110110430 A CN201110110430 A CN 201110110430A CN 102212586 B CN102212586 B CN 102212586B
- Authority
- CN
- China
- Prior art keywords
- lipase
- surface display
- yeast surface
- gene
- ssas
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Abstract
The invention discloses a method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast surface display lipase. The method comprises the following steps: dissolving starch of grain in water; adding dodecenly succinic anhydride ethanol solution and yeast surface display lipase after imbibition; stirring the mixture to react at 55-65 DEG C for 1-1.5 hours; and carrying out separation and purification to prepare the SSAS. The yeast surface display lipase is prepared by transforming recombinant plasmid which is subjected to linearization into Pichia pastoris GS115, inoculating the obtained transformant in a BMMY culture medium, carrying out inducing culture for 72-144 hours, centrifuging to collect a thallus, and carrying out washing, bio-imprinting and freeze-drying on the thallus. By displaying the lipase outside the cell and utilizing the lipase preparation to catalytically synthesize the SSAS, the invention improves the transformation efficiency, shortens the reaction time and lowers the production cost.
Description
Technical field
The present invention relates to technical field of bioengineering, relate in particular to the method for the lipase-catalyzed synthetic laurylene base succinic acid starch ester of a kind of yeast surface.
Background technology
Laurylene base succinic acid starch ester (Starch Sodium Alkenylsuccinate, SSAS) be that starch is through dodecenylsuccinic anhydride (Dodecenly Succinic Anhydride, DDSA) after the esterification sex change, introduced the hydrophilic and oleophilic amphiprotic group at starch molecular chain, thereby make starch molecule have hydrophilic, hydrophobic dual-use function, can be used as thickening material, emulsifying agent, sizing agent and microcapsule wall material and be applied to industry such as food, weaving, papermaking and pharmacy.SSAS is generated through esterification under weak basic condition by DDSA and starch slurry at present, aspect the preparation method, compared to shortcomings such as the equipment requirements height of dry process, reaction heterogeneity, foreign matter content height, wet processing reaction homogeneous, stability height, but the products therefrom substitution value is not high, and reaction efficiency is not high, the reaction times is long, causes generation and the product hydrolysis of side reaction, thereby influence esterification efficient, and make the side reaction product that mixes in the SSAS finished product.
Compared to the synthetic laurylene base succinic acid starch ester of chemical method, biological enzyme can obtain the reaction product of high substitution value within a short period of time.With regard to the same reaction system, biological enzyme is compared chemical method and is significantly promoted reaction efficiency and productive rate, all once also is significantly increased with stability.Simultaneously, adopt biological enzyme to avoid chemical method severe reaction conditions (strong acid, base catalysis, High Temperature High Pressure), no specificity, product complexity, drawbacks such as by product is many, separation is difficult, poor stability.Lipase be present esterification synthetic in most popular enzyme, but production cost height, numerous and diverse time-consuming its commercial applications of having limited to greatly of immobilization process.
Summary of the invention
The invention provides the method for the lipase-catalyzed synthetic laurylene base succinic acid starch ester of a kind of yeast surface display, this method can improve laurylene base succinic acid starch ester synthetic esterification efficient and productive rate, reduces synthetic cost.
The method of the lipase-catalyzed synthetic laurylene base succinic acid starch ester of a kind of yeast surface display comprises:
Cereal starch is water-soluble, add volumetric concentration behind the imbibition and be 10～20% dodecenylsuccinic anhydride ethanolic soln and yeast surface display lipase, 55～65 ℃ of following stirring reactions 1～1.5 hour, separate, purifying makes laurylene base succinic acid starch ester;
In weight part, reacting each proportioning raw materials can be as follows: 100～120 parts in water, 32～35 parts of cereal starchs, 1～1.2 part of dodecenylsuccinic anhydride ethanolic soln, 1～2 part in yeast surface display lipase.
Preferably, described separation, purification process are: reacting liquid pH value is adjusted to 6.5～7.0, through dehydration, alcohol washes, dry, pulverize, screening namely obtains product.
Preferably, described stir speed (S.S.) is 200～300 rev/mins.
Described yeast surface display lipase prepares by the following method:
Change the recombinant plasmid of linearization process over to pichia spp (Pichia pastoris) GS115, the gained transformant is inoculated in the BMMY substratum, inducing culture is centrifugal collection thalline after 72～144 hours, and thalline makes yeast surface display lipase through flushing, biological trace and lyophilize.
Described recombinant plasmid is by initial carrier pPIC9K and insert the lipase gene in MF α 1 signal peptide downstream among the initial carrier pPIC9K successively and the cell walls α agglutinin gene of pichia spp GS115 is formed.
Described lipase gene can be selected for use and be the sequence of AF229435 Genbank number, and pichia spp (Pichia pastoris) GS115 is commercially produced product, can buy from Invitrogen company.Genbank number of its cell walls α agglutinin gene sequence is M28164.
Carrier pPIC9K is commercially produced product (as Invitrogen company), and there is MF α 1 signal peptide sequence in it, and (Genbank number: M17301), there is the AOX1 promotor in the signal peptide sequence upstream (Genbank number: Z46233) in this carrier simultaneously.The purpose of recombinant plasmid linearization process be for born of the same parents in genome generation homologous recombination, improve expression stability.
Preferably, the used part of described biological trace is oleic acid, and it can the inducible enzyme structural modification, improves transformation efficiency.
The present invention is by importing pichia spp cell GS115 with lipase gene and cell walls α agglutinin gene, and after the pichia spp cell induction was cultivated, lipase was expressed justacrine outside born of the same parents, utilizes cell walls α lectin that this lipase is fixed on cell surface simultaneously.Utilize this yeast to represent lipase esterification is carried out catalysis, operational stability, thermotolerance and repeatability have not only been improved, reaction times also shortens greatly, dropped to 1～2 hour from original 12 hours, this yeast surface display lipase production cost is low in addition, catalyzes and synthesizes the transformation efficiency height.
Embodiment
Embodiment 1 preparation yeast surface display lipase
Method by synthetic, the lipase gene of synthetic Rhizopus oryzae (Rhizopus oryzae) (Genbank number: AF229435) and the cell walls α agglutinin gene (Genbank number is M28164) of pichia spp GS115, add connection peptides sequence GSSGGSGGSGGSGGSGS (linker) at lipase gene C end simultaneously, obtain nucleotide sequence pro-ROL-linker-α-agglutinin after the connection, add EcoR I and Not I restriction enzyme site simultaneously at the sequence two ends, wherein pro-ROL is lipase gene, and α-agglutinin is cell walls α agglutinin gene.
Be template with above-mentioned artificial synthesized sequence, utilize following primer right, carry out pcr amplification,
Upstream primer: 5 '-AAGGAAAAAAGAATTCGTTCCAGTTTCTGG-3 ';
Downstream primer: 5 '-TTTTCCTTTTGCGGCCGCTAATGAAACG-3 '
The PCR reaction system is: template DNA is 1 μ l, high-fidelity DNA polymerase 0.5 μ l, and dNTP (50mM) 0.4 μ l, each 0.5 μ l of upstream and downstream primer, 10 * PCR damping fluid, 5 μ l add water to 50 μ l.
The PCR operational conditions is: 94 ℃ 3 minutes, 35 circulations (94 ℃ 30 seconds, 60 ℃ 1 minute, 72 ℃ 30 seconds), 72 ℃ 10 minutes.
With EocR I and Not I simultaneously enzyme cut PCR product and pPIC9K plasmid, and under the effect of T4 ligase enzyme, spend the night and be connected to form the pPIC9K-ROL plasmid, by the electrophoresis check and reclaim plasmid.For making goal gene and pichia spp GS115 that His 4 unit points displacement reorganization take place, carry out linearization for enzyme restriction with the pPIC9K-ROL plasmid of Sal I and handle.The about 15 μ l of goal gene that linearization for enzyme restriction is handled well join in the previously prepd pichia spp GS115 competent cell, change in the electric revolving cup ice bath 15min over to, then at 1500V, 400 Ω, the 10ms that shocks by electricity under the 25 μ F conditions, and the sorbyl alcohol of the about 1ml precooling of adding.The electricity of the above-mentioned mixing about 400 μ l of thing that change the line of production are applied on the MD flat board, and the screening positive transformant is applied to positive transformant on the G418 flat board of different concns then, the resistance screening of G418 is gone out the positive recombinant bacterial strain of multiple copied of Mut phenotype according to positive transformant.
The positive recombinant bacterial strain of multiple copied is seeded in fermentation culture 30h in the BMGY substratum, centrifugal collecting cell; Again cell is placed the BMMY substratum inducing culture 144h that contains 0.5% (volume percent) methyl alcohol, centrifugal collecting cell, after the water flushing, be seeded to 30 ℃ of cultivation 120h in the YGC substratum, 3000g collected thalline in centrifugal minute then, wash with 50mM pH7.0 phosphoric acid buffer again behind the distilled water wash, mix with 2 times of volume oleic acid then, after-80 ℃ of following pre-freezes again through the dry 24h of German Christ vacuum freeze drier, remove oleic acid with hexane wash, carry out vacuum-drying more again and remove hexane, namely obtain the yeast surface display lipase of handling through biological trace.
The lipase-catalyzed synthetic laurylene base succinic acid starch ester of embodiment 2 yeast surface display
Example 1 is got 350g cereal starch water and is mixed with 35% starch milk (starch butt), makes the starch milk water-swelling in 65 ℃ of processing 15min that absorb water down, the cooling back adds the yeast surface display lipase of the above-mentioned preparation of 1g, add 10ml 15% (volume) dodecenylsuccinic anhydride ethanolic soln then in batches, place 85-1 type magnetic stirring apparatus to stir and begin reaction, rotating speed is 200 rev/mins, temperature of reaction remains on 60 ℃, stop to stir behind the reaction 1h, regulate pH value to 7.0, through dehydration, alcohol wash, dry, pulverize, sieve and namely obtain product.
Example 2 is got 400g cereal starch water and is mixed with 40% starch milk (starch butt), makes the starch milk water-swelling in 65 ℃ of processing 15min that absorb water down, the cooling back adds the yeast surface display lipase of the above-mentioned preparation of 2g, add 15ml 15% dodecenylsuccinic anhydride ethanolic soln then in batches, place 85-1 type magnetic stirring apparatus to stir and begin reaction, rotating speed is 250 rev/mins, temperature of reaction remains on 62 ℃, stop to stir behind the reaction 1.5h, regulate pH to 6.5～7.0, through dehydration, alcohol wash, dry, pulverize, screening namely obtains product.
Embodiment 3 adopts the synthetic laurylene base succinic acid starch ester of traditional chemical method
Example 1 is got 350g cereal starch water and is mixed with 35% starch milk (starch butt), makes the starch milk water-swelling in 65 ℃ of processing 15min that absorb water down, add 10ml 15% dodecenylsuccinic anhydride ethanolic soln then in batches, place 85-1 type magnetic stirring apparatus to stir and begin reaction, rotating speed is 200 rev/mins, temperature of reaction remains on 60 ℃, reaction stops behind the 1h stirring, and regulates pH value to 7.0, through dehydration, alcohol wash, dry, pulverize, sieve and namely obtain product.
Embodiment 4 measures substitution value and phosphorus content
Take by weighing sample 1.5g in the beaker of 100ml, add the ethanolic soln acidified sample of 20ml2.5mol/L hydrochloric acid with the wetting back of 1.5ml ethanol.Stir the ethanol that adds 40ml 90% after 30 minutes, continue to move into suction filtration in the sand core funnel behind the stirring 10min, till having chlorion, (do not use AgNO with 90% ethanol drip washing
3The solution check), then the acid SSAS in the sand core funnel is moved in the beaker of 500ml, add the standard sodium hydroxide solution of 7.5ml 0.25mol/L, add 150ml distilled water.Add 10 1% phenolphthalein indicator behind the boiling water bath 20min, after stirring while hot sulphuric acid soln titration to the solution changes color with 0.05mol/L be terminal point, record consumes the volume number of sulfuric acid.Make blank with the sample that does not add the acid anhydrides preparation simultaneously, the calculation formula of substitution value is as follows:
M
0＝7.5×M
1-V
0×M
2
M
x＝[(7.5×M
1-V
0×M
2)-M
0]/(2×W)
DS＝162M
x/(1000-265×M
x)
In the formula: M
0The mmole number of-sodium hydroxide that every gram blank sample consumes; M
xThe mmole number of the sodium hydroxide that consumes behind the every gram sample in the blank back of-deduction; M
1The volumetric molar concentration of-sodium hydroxide solution; M
2The volumetric molar concentration of-sulphuric acid soln; V
0The volume of the sulphuric acid soln that-blank sample consumes (ml); V
xThe volume of the sulphuric acid soln that-sample consumes (ml); The quality of W-sample (g); 162: the molecular weight of each glucosyl residue; 265: the molecular weight of dodecenylsuccinic anhydride.
Substitution reaction efficient=actual substitution value/theoretical substitution value * 100%
According to aforesaid method the laurylene base succinic acid starch ester sample that makes is detected, among the embodiment 2, substitution value and the reaction efficiency of example 1 product are respectively 0.029 and 86.6%, and substitution value and the reaction efficiency of example 2 products are respectively 0.031 and 82.9%.And substitution value and the reaction efficiency of embodiment 3 synthetic laurylene base succinic acid starch esters only are 0.022 and 75%.
Claims (1)
1. the method for the lipase-catalyzed synthetic laurylene base succinic acid starch ester of yeast surface display comprises:
The lipase gene of synthetic Rhizopus oryzae (Rhizopus oryzae) and the cell walls α agglutinin gene of pichia spp GS115, add the gene fragment of coding connection peptides GSSGGSGGSGGSGGSGS simultaneously at lipase gene C end, connection obtains nucleotide sequence pro-ROL-linker-α-agglutinin; Pro-ROL is lipase gene, and Genbank number is AF229435, and α-agglutinin is cell walls α agglutinin gene, and Genbank number is M28164, and linker is the gene fragment of coding connection peptides GSSGGSGGSGGSGGSGS;
Be template with above-mentioned artificial synthesized sequence, utilize following primer right, carry out pcr amplification,
Upstream primer: 5 '-AAGGAAAAAAGAATTCGTTCCAGTTTCTGG-3 ';
Downstream primer: 5 '-TTTTCCTTTTGCGGCCGCTAATGAAACG-3 '
With EocR I and Not I simultaneously enzyme cut PCR product and pPIC9K plasmid, and under the effect of T4 ligase enzyme, spend the night and be connected to form the pPIC9K-ROL plasmid, by the electrophoresis check and reclaim plasmid;
Carry out linearization for enzyme restriction with the pPIC9K-ROL plasmid of Sal I and handle, then 15 μ l linearization plasmids are joined in the pichia spp GS115 competent cell, change in the electric revolving cup, ice bath 15min, then at 1500V, 400 Ω, 10ms and the sorbyl alcohol of adding 1ml precooling shock by electricity under the 25 μ F conditions;
The electricity thing 400 μ l that change the line of production are applied on the MD flat board, and the screening positive transformant is applied to positive transformant on the G418 flat board of different concns then, the resistance screening of G418 is gone out the positive recombinant bacterial strain of multiple copied of Mut phenotype according to positive transformant;
The positive recombinant bacterial strain of multiple copied is seeded in fermentation culture 30h in the BMGY substratum, centrifugal collecting cell; Again cell is placed the BMMY substratum inducing culture 144h that contains volume percent 0.5% methyl alcohol, centrifugal collecting cell, after the water flushing, be seeded to 30 ℃ of cultivation 120h in the YGC substratum, 3000g collected thalline in centrifugal minute then, wash with 50mM pH7.0 phosphoric acid buffer again behind the distilled water wash, mix with 2 times of volume oleic acid then, after-80 ℃ of following pre-freezes again through the dry 24h of German Christ vacuum freeze drier, remove oleic acid with hexane wash, carry out vacuum-drying more again and remove hexane, namely obtain the yeast surface display lipase of handling through biological trace;
Getting 350g cereal starch water is mixed with 35% starch milk, makes the starch milk water-swelling in 65 ℃ of processing 15min that absorb water down, the cooling back adds the yeast surface display lipase of the above-mentioned preparation of 1g, add the 10ml volumetric concentration then in batches and be 15% dodecenylsuccinic anhydride ethanolic soln, place 85-1 type magnetic stirring apparatus to stir and begin reaction, rotating speed is 200 rev/mins, temperature of reaction remains on 60 ℃, stop to stir behind the reaction 1h, regulate pH value to 7.0, through dehydration, alcohol wash, dry, pulverize, sieve and namely obtain product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101104305A CN102212586B (en) | 2011-04-28 | 2011-04-28 | Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101104305A CN102212586B (en) | 2011-04-28 | 2011-04-28 | Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102212586A CN102212586A (en) | 2011-10-12 |
| CN102212586B true CN102212586B (en) | 2013-10-09 |
Family
ID=44744118
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101104305A Expired - Fee Related CN102212586B (en) | 2011-04-28 | 2011-04-28 | Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102212586B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492751B (en) * | 2011-11-29 | 2013-10-16 | 山东轻工业学院 | Method of synthesizing starch succinate under catalysis of lipase |
| CN103333262A (en) * | 2013-06-28 | 2013-10-02 | 江南大学 | Preparation method of modified starch nanocrystal |
| CN104046660A (en) * | 2014-07-08 | 2014-09-17 | 长沙理工大学 | Synthesis method of ethyl acetate |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101285078A (en) * | 2008-06-10 | 2008-10-15 | 华南理工大学 | Process for synthesizing ethyl caproate by yeast display lipase synthesis |
| CN101792721A (en) * | 2009-09-30 | 2010-08-04 | 华南理工大学 | Pichia pastoris engineering bacteria of surface display candida antarctica lipase B (CALB) and method for catalyzing and synthesizing short-chain aromatic ester |
| CN101974572A (en) * | 2010-07-29 | 2011-02-16 | 天津科技大学 | Method for preparing starch aliphatic ester by enzyme catalysis |
-
2011
- 2011-04-28 CN CN2011101104305A patent/CN102212586B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101285078A (en) * | 2008-06-10 | 2008-10-15 | 华南理工大学 | Process for synthesizing ethyl caproate by yeast display lipase synthesis |
| CN101792721A (en) * | 2009-09-30 | 2010-08-04 | 华南理工大学 | Pichia pastoris engineering bacteria of surface display candida antarctica lipase B (CALB) and method for catalyzing and synthesizing short-chain aromatic ester |
| CN101974572A (en) * | 2010-07-29 | 2011-02-16 | 天津科技大学 | Method for preparing starch aliphatic ester by enzyme catalysis |
Non-Patent Citations (10)
| Title |
|---|
| Display of Candida antarctica lipase B on Pichia pastoris and its application to flavor ester synthesis;Guo-Dong Su et al;《Appl Microbiol Biotechnol》;20091224;第86卷;第1494页右栏第10行至第1498页左栏第7行及图1、3 * |
| Enhanced reactivity of Rhizopus oryzae lipase displayed on yeast cell surfaces in organic solvents potential as a whole-cell biocatalyst in organic solvents;Seizaburo Shiraga et al;《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》;20050831;第71卷(第8期);4335-4338页 * |
| Guo-Dong Su et al.Display of Candida antarctica lipase B on Pichia pastoris and its application to flavor ester synthesis.《Appl Microbiol Biotechnol》.2009,第86卷第1494页右栏第10行至第1498页左栏第7行及图1、3. |
| Seizaburo Shiraga et al.Enhanced reactivity of Rhizopus oryzae lipase displayed on yeast cell surfaces in organic solvents potential as a whole-cell biocatalyst in organic solvents.《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》.2005,第71卷(第8期),4335-4338页. |
| 全细胞脂肪酶生物催化剂构建及其在生物转化中的应用;阮晖 等;《中国食品科学技术学会第七届年会论文摘要集》;20101104;第127-128页第1002篇摘要部分 * |
| 十二烯基琥珀酸淀粉酯的制备;陈煦 等;《无锡轻工大学学报》;20000930;第19卷(第5期);495-497页 * |
| 张伟 等.酵母表面展示技术在蛋白质工程中的应用.《生物技术通报》.2009,(第8期),63-66页. |
| 酵母表面展示技术在蛋白质工程中的应用;张伟 等;《生物技术通报》;20090826(第8期);63-66页 * |
| 阮晖 等.全细胞脂肪酶生物催化剂构建及其在生物转化中的应用.《中国食品科学技术学会第七届年会论文摘要集》.2010,第127-128页第1002篇摘要部分. |
| 陈煦 等.十二烯基琥珀酸淀粉酯的制备.《无锡轻工大学学报》.2000,第19卷(第5期),495-497页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102212586A (en) | 2011-10-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102212586B (en) | Method for catalytically synthesizing starch sodium alkenylsuccinate (SSAS) with yeast display lipase | |
| CN102212601B (en) | Method for performing catalytic synthesis of vitamin A palmitate by using yeast display lipase | |
| CN102212583B (en) | Method for catalyzing and synthesizing octenyl succinic anhydride modified starch ester through yeast show lipase | |
| CN102212584B (en) | Method for catalyzing and synthesizing starch acetate through yeast show lipase | |
| CN102212585B (en) | Method for catalytic synthesis of starch phosphate by utilizing yeast display lipase | |
| CN102212587B (en) | Method for catalytically synthesizing starch xanthate with yeast display lipase | |
| CN102212572B (en) | Method for catalytically synthesizing L-ascorbyl oleate with yeast display lipase | |
| CN102212588B (en) | Method for performing catalytic synthesis of starch phosphate monoester by using yeast display lipase | |
| CN102296101B (en) | Method for preparing micro capsulate embedded antihypertensive peptide | |
| CN102212580B (en) | Method for catalytic synthesis of glucose laurate by utilizing yeast display lipase | |
| CN102212570B (en) | Method for catalytically synthesizing vitamin C conjugated linoleate with yeast display lipase | |
| CN102212575B (en) | Method for synthesizing L-ascorbyl palmitate by catalysis of yeast display lipase | |
| CN102212593B (en) | Method for performing catalytic synthesis of maltose stearate by using yeast display lipase | |
| CN102212591B (en) | Method for catalyzing and synthesizing sucrose stearate through yeast show lipase | |
| CN102212578B (en) | Method for synthesizing fructose palmitate by catalysis of yeast display lipase | |
| CN102212579B (en) | Method for catalyzing and synthesizing glucose myristate through yeast show lipase | |
| CN102212590B (en) | Method for catalytically synthesizing sucrose palmitate with yeast display lipase | |
| CN102321598B (en) | Method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves | |
| CN102212576B (en) | Method for synthesizing vitamin E linoleate by catalysis of yeast display lipase | |
| CN102212577B (en) | Method for catalytic synthesis of vitamin E succinate by utilizing yeast display lipase | |
| CN102226209B (en) | Method for catalyzing and synthesizing vitamin E oleate by using lipase displayed on yeast | |
| CN102212602B (en) | Method for synthesizing vitamin A lactate by catalysis of yeast display lipase | |
| CN102212592B (en) | Method for catalyzing and synthesizing maltose laurate through yeast show lipase | |
| CN102212573B (en) | Method for catalytically synthesizing L-ascorbyl docosahexaenoate with yeast display lipase | |
| CN102212574B (en) | Method for catalytic synthesis of L-ascorbic acid linoleate by utilizing yeast display lipase |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131009 Termination date: 20160428 |