CN105039439A - Method for biosynthesizing conjugated linoleic acid isomer in organic medium by adopting coated thalluses - Google Patents
Method for biosynthesizing conjugated linoleic acid isomer in organic medium by adopting coated thalluses Download PDFInfo
- Publication number
- CN105039439A CN105039439A CN201510423414.XA CN201510423414A CN105039439A CN 105039439 A CN105039439 A CN 105039439A CN 201510423414 A CN201510423414 A CN 201510423414A CN 105039439 A CN105039439 A CN 105039439A
- Authority
- CN
- China
- Prior art keywords
- thalline
- thalluses
- dressing
- linoleic acid
- conjugated linoleic
- 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.)
- Granted
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fats And Perfumes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for biosynthesizing conjugated linoleic acid isomer in an organic medium by adopting coated thalluses. The method comprises the following steps: (1) resuspending each gram of lactobacillus casei CGMCC 1.574 thalluses into 5mL of permeabilized treating fluid, treating for 20min at 37 DEG C, centrifuging, and collecting the permeabilized thalluses; (2) washing the permeabilized thalluses by adopting 50mM phosphate buffer with the pH being 5.8, centrifuging, collecting the thalluses, resuspending each gram of the permeabilized wet thalluses into 20mL 50 mM phosphate buffer with the pH being 5.8, adding with Tween-20 with volume being 0.5-2.0% of that of the thallus suspension liquid, treating for 2-6h at 4-40 DEG C, centrifuging, collecting the thalluses, and carrying out freeze drying on the thalluses to obtain coated thalluses; (3) adding each gram of the coated thalluses into 600mL of normal hexane, stirring uniformly, adding linoleic acid with volume being 0.1-0.3% of that of normal hexane, and carrying out reacting for 1-12 h at 4-40 DEG C; and (4) centrifuging for separating the coated thalluses, and recovering normal hexane, and thus obtaining the product c8, t11- conjugated linoleic acidisomer. According to the method, the reaction time is short, the coated thalluses can be repeatedly used for a plurality of times, the yield is obviously improved, no environmental pollution is generated, and the production cost is obviously reduced.
Description
Technical field
The invention belongs to biomedicine technical field.
Background technology
Conjugated linolic acid (conjugatedlinoleicacid, CLA) is the general designation of one group of CLA, and conjugated double bond has 4 kinds of position isomerisms (8,10; 9,11; 10,12; 11,13) and 4 kinds of rotamerisms (
cis,
cis;
cis,
trans;
trans,
cis;
trans,
trans), total nearly 16 kinds of isomer, as:
c9,
t11-CLA and
t10,
c12-CLA isomer etc.Research finds that CLA has the physiological functions such as anticancer, prevention of arterial is atherosis, enhancing body immunizing power, anti-diabetic, and increasing research shows that its physiologically active has isomer specificity, generally believes now
c9,
t11-CLA isomer has anticancer and that prevention of arterial is atherosis effect.At occurring in nature, CLA is mainly present in the fat in the milk of ruminating animal and meat, primarily of
c9,
t11-CLA is with a small amount of
t9,
t11-CLA,
t10,
cthe isomer such as 11-CLA is formed, but its content is all less than 10mg/g fat usually, cannot meet to keep healthy and Application and Development for the purpose of medical treatment.
For realizing a large amount of preparations of CLA isomer, people prepare CLA to fermentable and have carried out more research.At present, fermentable carries out all in aqueous, because added fermentation substrate-linolic acid is lipid-soluble substance, therefore linolic acid needs through emulsification before adding fermented liquid, and addition is restricted, from fermented liquid, obtain CLA product need through organic solvent extraction, and whole production technique exists the shortcoming that fermentation period is long, production cost is high, yield poorly.Patent of invention CN201110105610 proposes a kind of method of bioconverting conjugated linoleic acid by using Lactobacillus plantarum, and substrate addition is 25mg/mL, and fermentation time reaches 120h, and the output of CLA only has 1.02mg/mL, and the transformation efficiency of substrate only has 4%.
Summary of the invention
The object of the invention is for the deficiencies in the prior art, propose the method for a kind of dressing thalline biosynthesizing conjugated linoleic acid isomers in organic medium, significantly shorten generated time, improve output and transformation efficiency, reduction production cost.
The present invention includes following steps.
(1) permeabilized treatment of thalline: collect be in logarithmic phase lactobacterium casei (
lactobacilluscasei) CGMCC1.574 thalline, be resuspended in 5mL permeabilized treatment liquid by every gram of wet thallus to fall into a trap, wet thallus is resuspended in permeabilized treatment liquid that (described permeabilized treatment liquid is the triton x-100 of volume percent 0.5-3.0%, the linolic acid of 5-50 μM, the phosphate buffered saline buffer of 20-200mM, pH5.8), resuspended thalline processes 20min through 37 DEG C, collected by centrifugation permeability thalline.
(2) thalline Cotton seeds: permeability thalline is with after the phosphate buffered saline buffer washing of 50mM, pH5.8, collected by centrifugation thalline, the phosphate buffered saline buffer being resuspended in 20mL50mM, pH5.8 by every gram of wet thallus is fallen into a trap, permeability wet thallus is resuspended in phosphate buffered saline buffer, the tween 20 adding bacteria suspension volume percent 0.5-2.0% mixes, in 4-40 DEG C of process 2-6h, collected by centrifugation thalline, obtains dressing thalline by after thalline lyophilize.
(3) biosynthesizing in organic medium: join 600mL normal hexane by every gram of freeze-dried coated thalline and fall into a trap, dressing thalline is joined in normal hexane, stirs, add the linolic acid of normal hexane volume percent 0.1-0.3%, react 1-12h at 4-40 DEG C.
(4) product collection: centrifugation dressing thalline, carries out underpressure distillation by hexane solution, and reclaim normal hexane, product is
c9,
t11-conjugated linoleic acid isomers.
In step of the present invention (1), permeabilized treatment liquid is preferably the triton x-100 of volume percent 1.0-2.0%, the linolic acid of 15-35 μM, the phosphate buffered saline buffer of 30-100mM, pH5.8.
In step of the present invention (2), the add-on of tween 20 is preferably the 0.75-1.5% of bacteria suspension volume percent.
In step of the present invention (2), treatment temp is preferably 4-25 DEG C.
In step of the present invention (2), the treatment time is preferably 3-5h.
Preferably add the linolic acid of normal hexane volume percent 0.125-0.25% in step of the present invention (3), at 15-30 DEG C, react 2-6h.
In step of the present invention (4), dressing thalline can be recycled and reused for biosynthesizing in organic medium
c9,
t11-conjugated linoleic acid isomers.
The present invention's lactobacterium casei used (
lactobacilluscasei) CGMCC1.574, be China General Microbiological culture presevation administrative center (CGMCC) preservation strain, be numbered 1.574, this bacterial strain can produce specific linoleate isomerase, linoleic acid can be become by biological isomerization
c9,
t11-CLA isomer.At present, the linoleate isomerase sterling of definite catalytic activity is not also separated to.Therefore, this isomerization reaction may be completed by multi-enzyme system co-catalysis.
By this dressing thalline biosynthetic means in organic medium, with optimal conditions, synthesized by rear containing the reaction of volume percent 0.15% linoleic hexane solution
c9,
tthe content of 11-CLA isomer is volume percent 0.1395%, and linoleic transformation efficiency is 93.0%.In step of the present invention (4), dressing thalline can be recycled and reused for biosynthesizing in the organic medium of step (3)
c9,
t11-CLA isomer, after dressing thalline reuses five times, its catalytic activity is still greater than 90%.
Carry out all in aqueous for existing fermentable synthesis CLA, it is long to there is fermentation period in whole production technique, and from fermented liquid, extract CLA product difficult, thalline can not be reused, and production cost is high, and the shortcomings such as yield poorly.The present invention proposes and in organic medium, linolic acid is catalyzed and synthesized by microbial cells
c9,
tthe novel method of 11-CLA.The method is first by triton x-100 process lactobacterium casei CGMCC1.574, while maintenance thalline integrity, improve somatic cells wall and membrane passage, can guarantee that intracellular linoleate isomerase surface energy forms complete coatings when follow-up Cotton seeds on the one hand, the speed of reaction substrate linolic acid and product C LA turnover thalline can be improved on the other hand, minimizing high concentration substrate and product are to the restraining effect of catalyzed reaction, greatly Reaction time shorten.
The present invention is when permeabilized treatment thalline, add the linolic acid of 5-50 μM, linolic acid can combine with thalline Linoleic acid Isomerases catalyze active centre, when thalline Cotton seeds, molecular imprinting can be formed at the catalytic active center of enzyme, make thalline catalytic active center conformation of enzyme after lyophilize constant, higher catalytic activity can be kept in organic medium.Meanwhile, the present invention is combined in thalline Cotton seeds process and selects suitable phosphate buffered saline buffer, tween 20 concentration and dressing temperature, time, makes gained dressing thalline still have higher catalytic capability in organic medium.After dressing, in thalline, the thermostability of enzyme improves, and every batch of reaction times only needs 4h, time a couple of days in traditional aqueous needed for Batch fermentation.
Gained dressing thalline of the present invention has higher catalytic activity in normal hexane, only needs centrifugation dressing thalline, hexane solution is carried out underpressure distillation, reclaims normal hexane, can obtain product
c9,
t11-CLA.Centrifugal gained dressing thalline can repeatedly be recycled and reused for biosynthesizing in organic medium
c9,
t11-CLA isomer, significantly shorten the production time, improves output, reduces production cost.In addition, normal hexane is the organic solvent that food oils industry is commonly used, and this guarantees gained of the present invention
c9,
tthe safety in utilization of 11-CLA.
The present invention compared with prior art tool has the following advantages.
The present invention directly carries out Cotton seeds to thalline, is used for catalyzed reaction as immobilized enzyme, decreases the cost of enzyme purification on the one hand, avoids the loss of enzyme activity when extracting; Can multi-enzyme system be wrapped in together on the other hand, make whole reaction process smooth; Moreover dressing thalli granule is greater than dressing enzyme, be easy to separate from reaction soln, and filtration resistance is less, therefore, dressing thalline is suitable for the column reactor of successive reaction more.
The present invention carries out catalyzed reaction by dressing thalline in organic medium, avoids the problem that substrate linoleic acid solubleness in traditional aqueous reaction system is low and product C LA extraction is difficult.In dressing thalline, the thermostability of enzyme improves, and every batch of reaction times only needs 4h, time a couple of days of fermenting required in traditional aqueous.Dressing thalline is repeatedly reusable, significantly improves product production, and non-environmental-pollution, significantly can reduce production cost; And the thalline in traditional aqueous system fermented liquid can only use once, production cost is high, and Wastewater treating costly, easy contaminate environment.
Embodiment
The present invention is further described by following examples.
Embodiment 1.
By activation lactobacterium casei (
lactobacilluscasei) CGMCC1.574 strain inoculation is in MRS substratum, 20h is cultivated at 30 DEG C, the centrifugal 10min of 5000g collects thalline, 10 grams of wet thallus are resuspended in 50mL containing in the permeabilized treatment liquid of the triton x-100 of volume percent 1.5%, the linolic acid of 25 μMs and 50mM phosphate buffered saline buffer (pH5.8), 37 DEG C of centrifugal 10min of water bath processing 20min, 5000g obtain permeability thalline.
Permeability thalline is with after 50mL50mM phosphate buffered saline buffer (pH5.8) washing, the centrifugal 10min of 6000g collects thalline, wet thallus is resuspended in 200mL50mM phosphate buffered saline buffer (pH5.8), add 2mL tween 20 and mix, in 4 DEG C of stir process 4h, the centrifugal 10min of 6000g collects thalline, and thalline carries out lyophilize after-80 DEG C of pre-freezes, obtains dressing thalline.
1 gram of freeze-dried coated thalline is joined in 600mL normal hexane, stirs, add 0.9mL linolic acid, stirring reaction 4h at 25 DEG C.6000g centrifugation dressing thalline, carries out underpressure distillation by hexane solution at 40 DEG C, reclaims normal hexane, obtains 0.9mL product, wherein
c9,
tthe content of 11-CLA is 93.0%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c9,
tthe content of 11-CLA is 90.5%.
Embodiment 2.
By activation lactobacterium casei (
lactobacilluscasei) CGMCC1.574 strain inoculation is in MRS substratum, 18h is cultivated at 30 DEG C, the centrifugal 5min of 4000g collects thalline, 10 grams of wet thallus are resuspended in 50mL containing in the permeabilized treatment liquid of the triton x-100 of volume percent 3.0%, the linolic acid of 5 μMs and 200mM phosphate buffered saline buffer (pH5.8), 37 DEG C of centrifugal 10min of water bath processing 20min, 4000g obtain permeability thalline.
Permeability thalline is with after 50mL50mM phosphate buffered saline buffer (pH5.8) washing, the centrifugal 10min of 5000g collects thalline, wet thallus is resuspended in 200mL50mM phosphate buffered saline buffer (pH5.8), add 4mL tween 20 and mix, in 4 DEG C of stir process 2h, the centrifugal 10min of 5000g collects thalline, and thalline carries out lyophilize after-80 DEG C of pre-freezes, obtains dressing thalline.
1 gram of freeze-dried coated thalline is joined in 600mL normal hexane, stirs, add 1.8mL linolic acid, stirring reaction 12h at 40 DEG C.6000g centrifugation dressing thalline, carries out underpressure distillation by hexane solution at 40 DEG C, reclaims normal hexane, obtains 1.8mL product, wherein
c9,
tthe content of 11-CLA is 81.2%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c9,
tthe content of 11-CLA is 71.3%.
Embodiment 3.
By activation lactobacterium casei (
lactobacilluscasei) CGMCC1.574 strain inoculation is in MRS substratum, 18h is cultivated at 30 DEG C, the centrifugal 5min of 4000g collects thalline, 10 grams of wet thallus are resuspended in 50mL containing in the permeabilized treatment liquid of the triton x-100 of volume percent 0.5%, the linolic acid of 50 μMs and 20mM phosphate buffered saline buffer (pH5.8), 37 DEG C of centrifugal 10min of water bath processing 20min, 4000g obtain permeability thalline.
Permeability thalline is with after 50mL50mM phosphate buffered saline buffer (pH5.8) washing, the centrifugal 10min of 5000g collects thalline, wet thallus is resuspended in 200mL50mM phosphate buffered saline buffer (pH5.8), add 1mL tween 20 and mix, in 4 DEG C of stir process 6h, the centrifugal 10min of 5000g collects thalline, and thalline carries out lyophilize after-80 DEG C of pre-freezes, obtains dressing thalline.
1 gram of freeze-dried coated thalline is joined in 600mL normal hexane, stirs, add 0.6mL linolic acid, stirring reaction 6h at 4 DEG C.6000g centrifugation dressing thalline, carries out underpressure distillation by hexane solution at 40 DEG C, reclaims normal hexane, obtains 0.6mL product, wherein
c9,
tthe content of 11-CLA is 90.9%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c9,
tthe content of 11-CLA is 86.3%.
Embodiment 4.
By activation lactobacterium casei (
lactobacilluscasei) CGMCC1.574 strain inoculation is in MRS substratum, 20h is cultivated at 30 DEG C, the centrifugal 10min of 5000g collects thalline, 10 grams of wet thallus are resuspended in 50mL containing in the permeabilized treatment liquid of the triton x-100 of volume percent 1.5%, the linolic acid of 25 μMs and 50mM phosphate buffered saline buffer (pH5.8), 37 DEG C of centrifugal 10min of water bath processing 20min, 5000g obtain permeability thalline.
Permeability thalline is with after 50mL50mM phosphate buffered saline buffer (pH5.8) washing, the centrifugal 10min of 6000g collects thalline, wet thallus is resuspended in 200mL50mM phosphate buffered saline buffer (pH5.8), add 2mL tween 20 and mix, in 4 DEG C of stir process 4h, the centrifugal 10min of 6000g collects thalline, and thalline carries out lyophilize after-80 DEG C of pre-freezes, obtains dressing thalline.
1 gram of freeze-dried coated thalline is loaded in pillar bio-reactor, 0.9mL linolic acid is joined in 300mL normal hexane and mixes, will containing linoleic hexane solution injecting reactor by pump, flow is 3mL/min, collects effluent liquid, effluent liquid is repeated injecting reactor 2 times, collect effluent liquid, carry out underpressure distillation at 40 DEG C, reclaim normal hexane, obtain 0.9mL product, wherein
c9,
tthe content of 11-CLA is 91.5%.By being cascaded by several pillar bio-reactor, continuous seepage can be realized.
Claims (6)
1. the method for dressing thalline biosynthesizing conjugated linoleic acid isomers in organic medium, is characterized in that comprising the following steps:
(1) the lactobacterium casei CGMCC1.574 thalline being in logarithmic phase is collected, be resuspended in 5mL permeabilized treatment liquid by every gram of wet thallus to fall into a trap, be resuspended in by wet thallus in permeabilized treatment liquid, resuspended thalline processes 20min through 37 DEG C, collected by centrifugation permeability thalline;
(2) permeability thalline is with after the phosphate buffered saline buffer washing of 50mM, pH5.8, collected by centrifugation thalline, the phosphate buffered saline buffer being resuspended in 20mL50mM, pH5.8 by every gram of wet thallus is fallen into a trap, permeability wet thallus is resuspended in phosphate buffered saline buffer, the tween 20 adding bacteria suspension volume percent 0.5-2.0% mixes, in 4-40 DEG C of process 2-6h, collected by centrifugation thalline, obtains dressing thalline by after thalline lyophilize;
(3) join 600mL normal hexane by every gram of freeze-dried coated thalline to fall into a trap, dressing thalline is joined in normal hexane, stirs, add the linolic acid of normal hexane volume percent 0.1-0.3%, at 4-40 DEG C, react 1-12h;
(4) centrifugation dressing thalline, carries out underpressure distillation by hexane solution, and reclaim normal hexane, product is
c9,
t11-conjugated linoleic acid isomers;
Permeabilized treatment liquid described in step (1) is the triton x-100 of volume percent 0.5-3.0%, the linolic acid of 5-50 μM, the phosphate buffered saline buffer of 20-200mM, pH5.8.
2. the method for dressing thalline according to claim 1 biosynthesizing conjugated linoleic acid isomers in organic medium, it is characterized in that in described step (1), permeabilized treatment liquid is the triton x-100 of volume percent 1.0-2.0%, the linolic acid of 15-35 μM, the phosphate buffered saline buffer of 30-100mM, pH5.8.
3. the method for dressing thalline according to claim 1 biosynthesizing conjugated linoleic acid isomers in organic medium, is characterized in that the add-on of tween 20 in described step (2) is the 0.75-1.5% of bacteria suspension volume percent.
4. the method for dressing thalline according to claim 1 biosynthesizing conjugated linoleic acid isomers in organic medium, is characterized in that in described step (2), treatment temp is 4-25 DEG C.
5. the method for dressing thalline according to claim 1 biosynthesizing conjugated linoleic acid isomers in organic medium, is characterized in that in described step (2), the treatment time is 3-5h.
6. the method for dressing thalline according to claim 1 biosynthesizing conjugated linoleic acid isomers in organic medium, is characterized in that the linolic acid adding normal hexane volume percent 0.125-0.25% in described step (3), at 15-30 DEG C, reacts 2-6h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510423414.XA CN105039439B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugated linoleic acid isomers in organic media |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510423414.XA CN105039439B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugated linoleic acid isomers in organic media |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105039439A true CN105039439A (en) | 2015-11-11 |
| CN105039439B CN105039439B (en) | 2018-06-08 |
Family
ID=54446408
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510423414.XA Active CN105039439B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugated linoleic acid isomers in organic media |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105039439B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928101A (en) * | 2006-09-07 | 2007-03-14 | 河北工业大学 | Method for preparing conjugated linoleic acid |
| CN101565367A (en) * | 2009-05-27 | 2009-10-28 | 浙江大学 | Preparation method of conjugated linoleic acid |
| CN102206687A (en) * | 2011-04-27 | 2011-10-05 | 浙江大学 | Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum |
| CN104404092A (en) * | 2014-11-04 | 2015-03-11 | 南昌大学 | Conjugated linoleic acid isomer biological enrichment method |
-
2015
- 2015-07-20 CN CN201510423414.XA patent/CN105039439B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1928101A (en) * | 2006-09-07 | 2007-03-14 | 河北工业大学 | Method for preparing conjugated linoleic acid |
| CN101565367A (en) * | 2009-05-27 | 2009-10-28 | 浙江大学 | Preparation method of conjugated linoleic acid |
| CN102206687A (en) * | 2011-04-27 | 2011-10-05 | 浙江大学 | Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum |
| CN104404092A (en) * | 2014-11-04 | 2015-03-11 | 南昌大学 | Conjugated linoleic acid isomer biological enrichment method |
Non-Patent Citations (3)
| Title |
|---|
| SHIGENOBU KISHINO 等: "Linoleic Acid Isomerase in Lactobacillus plantarum AKU1009a Proved to Be a Multi-Component Enzyme System Requiring Oxidoreduction Cofactors", 《BIOSCIENCE, BIOTECHNOLOGY, AND BIOCHEMISTRY》 * |
| 刘晓华 等: "透性化细胞制备方法的研究进展", 《食品工业科技》 * |
| 郑钰: "保加利亚乳杆菌亚油酸异构酶合成共轭亚油酸研究", 《中国优秀硕士学位论文全文数据库（电子期刊）》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105039439B (en) | 2018-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101838672B (en) | Method for producing gamma-amino butyric acid by using immobilized lactobacillus plantarum | |
| CN101899484B (en) | Preparation method of genipin | |
| CN107418995B (en) | A kind of ellagic acid and preparation method thereof of granatanine liquid state fermentation preparation | |
| CN108611380A (en) | A kind of method of selectivity synthesis 2,5- dihydroxymethyl furans | |
| CN103484504A (en) | Method for preparing ethyl alcohol through fermentation by taking cassava as raw material | |
| CN105039440A (en) | Method for biosynthesizing conjugated Alpha-linolenic acid isomer in organic medium by coated bacteria | |
| CN108358690A (en) | A kind of preparation method of selenium-enriched plant source synergy biology complex capsule fertilizer | |
| CN105039444A (en) | Method for biosynthesizing conjugated linoleic acid isomer in organic medium | |
| CN105039439A (en) | Method for biosynthesizing conjugated linoleic acid isomer in organic medium by adopting coated thalluses | |
| CN101205523A (en) | Saccharomyces cerevisiae and its application in preparation of beta-hydroxyphenyl propionic acid ethyl | |
| CN105039443A (en) | Method for biosynthesizing conjugated gamma-linolenic acid isomer in organic medium | |
| CN105112462A (en) | Non-aqueous enzymological synthesis method of conjugated linoleic acid isomer | |
| CN105087691A (en) | Method for catalytic synthesis of conjugated linoleic acid isomer by virtue of surfactant-coated bacterium | |
| CN105112461A (en) | Method for biosynthesizing conjugate gamma-linolenic acid isomer from coated thalli in organic medium | |
| CN105039441A (en) | Nonaqueous enzymologic synthetic method of conjugated alpha-linolenic acid isomer | |
| CN105087692A (en) | Method for catalytic synthesis of conjugated alpha-linolenic acid isomer by virtue of surfactant-coated bacterium | |
| CN105039442A (en) | Method for biosynthesis of conjugated alpha-linolenic acid isomer in organic medium | |
| CN105087693A (en) | Method for catalytic synthesis of conjugated gamma-linolenic acid isomer by virtue of surfactant-coated bacterium | |
| CN105039438A (en) | Non-aqueous enzymological synthesis method of conjugated Nu-linolenic acid isomer | |
| CN104789489A (en) | Arginine deiminase high-yielding Bacillus cereus and application thereof | |
| CN102719496B (en) | Preparation method of (S)-(+)-ethyl mandelate by microbial transformed ethyl benzoylformate | |
| CN101755911B (en) | Production of yogurt rich in conjugated linoleic acid by using immobilization technology and method thereof | |
| CN101348808B (en) | Double enzyme coupling preparation of ornithine hydrochloride | |
| CN109628543B (en) | Method for preparing betulinic acid by using recombinant saccharomyces cerevisiae and inonotus obliquus mixed bacteria system | |
| CN109182408B (en) | Method for preparing natural perfume gamma-decalactone by adopting fermentation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |