CN105112461A - Method for biosynthesizing conjugate gamma-linolenic acid isomer from coated thalli in organic medium - Google Patents
Method for biosynthesizing conjugate gamma-linolenic acid isomer from coated thalli in organic medium Download PDFInfo
- Publication number
- CN105112461A CN105112461A CN201510424009.XA CN201510424009A CN105112461A CN 105112461 A CN105112461 A CN 105112461A CN 201510424009 A CN201510424009 A CN 201510424009A CN 105112461 A CN105112461 A CN 105112461A
- Authority
- CN
- China
- Prior art keywords
- thalline
- gamma
- linolenic acid
- thalli
- dressing
- 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
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Fats And Perfumes (AREA)
Abstract
A method for biosynthesizing conjugate a gamma-linolenic acid isomer from coated thalli in an organic medium comprises the following steps: 1, re-suspending Lactobacillus casei CGMCC 1.574 thalli in a permeabilization treatment liquid according to a ratio of 1g/5mL, treating at 37DEG C for 20min, centrifuging, and collecting permeabilized thalli; 2, washing by using a phosphate buffer solution with the concentration of 50mM and the pH value of 5.8, centrifuging, collecting thalli, re-suspending the permeabilized wet thalli in the phosphate buffer solution with the concentration of 50mM and the pH value of 5.8 according to a ratio of 1g/20mL, adding tween-20 accounting for 0.5-2.0% of the volume of the obtained thallus suspension, uniformly mixing, treating at 4-40DEG C for 2-6h, centrifuging, collecting thalli, and freeze-drying to obtain coated thalli; 3, adding the coated thalli to n-hexane according to a ratio of 1g/600mL, uniformly stirring, adding gamma-linolenic acid accounting for 0.1-0.3% of the volume of n-hexane, and reacting at 4-40DEG C for 1-12h; and 4, centrifuging, separating coated thalli, and recovering n-hexane to obtain the product c6,c9,t11-conjugate gamma-linolenic acid isomer. The method has the advantages of short reaction time, repeated use of the coated thalli, great improvement of the output, no environmental pollution, and great reduction of the production cost.
Description
Technical field
The invention belongs to biomedicine technical field.
Background technology
Conjugation gamma-linolenic acid is the conjugated isomers of gamma-linolenic acid, is the general designation of one group of CLnA, has multiple position isomerism and geometrical isomer, as:
c6,
c9,
t11-conjugation gamma-linolenic acid and
c6,
t10,
c12-conjugation gamma-linolenic acid isomer etc.Research shows that conjugation gamma-linolenic acid has the physiological functions such as anticancer, prevention of arterial is atherosis, fat-reducing, and its physiologically active has isomer specificity, as:
c6,
c9,
t11-conjugation gamma-linolenic acid isomer has anticancer effect.At occurring in nature, conjugation gamma-linolenic acid is mainly present in the milk of ruminating animal and the fat of meat, primarily of
c6,
c9,
tthe isomer such as 11-conjugation gamma-linolenic acid is formed, but its content is usually very low, cannot meet to keep healthy and Application and Development for the purpose of medical treatment.
Also do not have the report of conjugation gamma-linolenic acid preparation method at present, people have just carried out some researchs to fermentable synthesis of conjugate alpha-linolenic acid.At present, fermentable carries out all in aqueous, because fermentation substrate-gamma-linolenic acid is lipid-soluble substance, therefore gamma-linolenic acid needs through emulsification before adding fermented liquid, and addition is restricted, and conjugation gamma-linolenic acid product will be obtained from fermented liquid need through organic solvent extraction, there is the shortcoming that fermentation period is long, production cost is high, yield poorly in whole production technique.
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 conjugation gamma-linolenic acid isomer 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 gamma-linolenic 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 permeability 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, stir, add the gamma-linolenic acid of normal hexane volume percent 0.1-0.3%, at 4-40 DEG C, react 1-12h.
(4) product collection: centrifugation dressing thalline, carries out underpressure distillation by hexane solution, and reclaim normal hexane, product is
c6,
c9,
t11-conjugation gamma-linolenic acid isomer.
In step of the present invention (1), permeabilized treatment liquid is preferably the triton x-100 of volume percent 1.0-2.0%, the gamma-linolenic 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 gamma-linolenic 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
c6,
c9,
t11-conjugation gamma-linolenic acid isomer.
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, can by polyunsaturated fatty acid
c9,
c12 double-bond isomerisms become
c9,
t11 conjugated double bonds, by biological isomerization can by gamma-linolenic acid (
c6,
c9,
c12-18:3) change into the novel substance having charateristic avsorption band at 268nm place, and 268nm place absorption peak is the characteristic peak of conjugated triene, shows that gamma-linolenic acid can change into by this bacterium
c6,
c9,
t11-conjugation gamma-linolenic acid isomer.At present, from this bacterium, be not also separated to the isomerase sterling of definite catalytic activity.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, containing volume percent 0.15% gamma-linolenic acid hexane solution reaction after synthesized by
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid isomer is volume percent 0.1388%, and the transformation efficiency of gamma-linolenic acid is 92.5%.In step of the present invention (4), dressing thalline can be recycled and reused for biosynthesizing in the organic medium of step (3)
c6,
c9,
t11-conjugation gamma-linolenic acid isomer, after dressing thalline reuses five times, its catalytic activity is still greater than 90%.
Carry out all in aqueous for existing fermentable synthesis of conjugate gamma-linolenic acid, it is long to there is fermentation period in whole production technique, and from fermented liquid, extract conjugation gamma-linolenic acid product difficulty, thalline can not be reused, production cost is high, and the shortcomings such as yield poorly.The present invention proposes novel method gamma-linolenic acid being catalyzed and synthesized conjugation gamma-linolenic acid by microbial cells in organic medium.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 isomerase surface energy forms complete coatings when follow-up Cotton seeds on the one hand, the speed of reaction substrate gamma-linolenic acid and product conjugation gamma-linolenic acid 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 gamma-linolenic acid of 5-50 μM, gamma-linolenic 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
c6,
c9,
t11-conjugation gamma-linolenic acid.Centrifugal gained dressing thalline can repeatedly be recycled and reused for biosynthesizing in organic medium
c6,
c9,
t11-conjugation gamma-linolenic acid isomer, significantly shorten the production time, improves output, reduce 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
c6,
c9,
tthe safety in utilization of 11-conjugation gamma-linolenic acid.
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 gamma-linolenic acid solubleness in traditional aqueous reaction system is low and the extraction of product conjugation gamma-linolenic acid 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 gamma-linolenic 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 gamma-linolenic 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
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 92.5%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid 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 gamma-linolenic 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 gamma-linolenic 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
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 79.2%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 68.9%.
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 gamma-linolenic 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 gamma-linolenic 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
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 88.3%.The dressing thalline collected is recycled and reused for above-mentioned building-up reactions, when using continuously five times in product
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 80.1%.
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 gamma-linolenic 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 gamma-linolenic acid is joined in 300mL normal hexane and mixes, by pump by the hexane solution injecting reactor containing gamma-linolenic acid, 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
c6,
c9,
tthe content of 11-conjugation gamma-linolenic acid is 90.9%.By being cascaded by several pillar bio-reactor, continuous seepage can be realized.
Claims (6)
1. the method for dressing thalline biosynthesizing conjugation gamma-linolenic acid isomer 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 permeability 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 gamma-linolenic 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
c6,
c9,
t11-conjugation gamma-linolenic acid isomer;
Permeabilized treatment liquid described in step (1) is the triton x-100 of volume percent 0.5-3.0%, the gamma-linolenic 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 conjugation gamma-linolenic acid isomer 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 gamma-linolenic 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 conjugation gamma-linolenic acid isomer 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 conjugation gamma-linolenic acid isomer 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 conjugation gamma-linolenic acid isomer 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 conjugation gamma-linolenic acid isomer in organic medium, it is characterized in that the gamma-linolenic acid adding normal hexane volume percent 0.125-0.25% in described step (3), at 15-30 DEG C, react 2-6h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510424009.XA CN105112461B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugation acid and gamma-linolenic isomers in organic media |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510424009.XA CN105112461B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugation acid and gamma-linolenic isomers in organic media |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105112461A true CN105112461A (en) | 2015-12-02 |
| CN105112461B CN105112461B (en) | 2018-04-17 |
Family
ID=54660569
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510424009.XA Expired - Fee Related CN105112461B (en) | 2015-07-20 | 2015-07-20 | It is coated the method for thalline biosynthesis conjugation acid and gamma-linolenic isomers in organic media |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105112461B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1629302A (en) * | 2004-07-28 | 2005-06-22 | 南昌大学 | Process for preparing conjugated fatty acid |
| CN104404092A (en) * | 2014-11-04 | 2015-03-11 | 南昌大学 | Conjugated linoleic acid isomer biological enrichment method |
| CN104480150A (en) * | 2014-11-04 | 2015-04-01 | 南昌大学 | Biological enrichment method of conjugated linolenic acid isomer |
-
2015
- 2015-07-20 CN CN201510424009.XA patent/CN105112461B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1629302A (en) * | 2004-07-28 | 2005-06-22 | 南昌大学 | Process for preparing conjugated fatty acid |
| CN104404092A (en) * | 2014-11-04 | 2015-03-11 | 南昌大学 | Conjugated linoleic acid isomer biological enrichment method |
| CN104480150A (en) * | 2014-11-04 | 2015-04-01 | 南昌大学 | Biological enrichment method of conjugated linolenic acid isomer |
Non-Patent Citations (5)
| Title |
|---|
| JUN OGAWA等: "Production of conjugated fatty acids by lactic acid bacteria", 《JOURNAL OF BIOSCIENCE AND BIOENGINEERING》 * |
| S.KISHINO等: "Microbial production of conjugated γ-linolenic acid from γ-linolenic acid by Lactobacillus plantarum AKU 1009a", 《JOURNAL OF APPLIED MICROBIOLOGY》 * |
| 刘晓华等: "透性化细胞制备方法的研究进展", 《食品工业科技》 * |
| 宋宝东等: "谷氨酸二烷酯核糖醇的合成及应用", 《石油化工》 * |
| 高红丽等: "共轭亚麻酸的制备、表征和生物活性研究进展", 《化学通报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105112461B (en) | 2018-04-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101838672B (en) | Method for producing gamma-amino butyric acid by using immobilized lactobacillus plantarum | |
| CN105349587B (en) | The method of EPA and DHA content in a kind of raising glyceride type fish oil | |
| CN107418995B (en) | A kind of ellagic acid and preparation method thereof of granatanine liquid state fermentation preparation | |
| CN102206687B (en) | Method for bioconverting conjugated linoleic acid by using Lactobacillus plantarum | |
| CN101538595B (en) | Method for producing gamma-aminobutyric acid by separated fermentation of enterococcus faecium | |
| CN101205523B (en) | Saccharomyces cerevisiae and its application in preparation of beta-hydroxyphenyl propionic acid ethyl | |
| CN101205548B (en) | Use of saccharomyces cerevisiae in preparation of (S)-(-)-3-chlorine-1-phenylpropanol | |
| CN105039440A (en) | Method for biosynthesizing conjugated Alpha-linolenic acid isomer in organic medium by coated bacteria | |
| CN105039443A (en) | Method for biosynthesizing conjugated gamma-linolenic acid isomer in organic medium | |
| CN104762360A (en) | High-content nicotinamide synthesis induced by new-feature nitrile hydratase | |
| CN105039444A (en) | Method for biosynthesizing conjugated linoleic acid isomer in organic medium | |
| CN105112461A (en) | Method for biosynthesizing conjugate gamma-linolenic acid isomer from coated thalli in organic medium | |
| CN101265453B (en) | Beer yeast strain containing CMP kinase and CDP kinase and application | |
| CN105039438A (en) | Non-aqueous enzymological synthesis method of conjugated Nu-linolenic acid isomer | |
| CN105087693A (en) | Method for catalytic synthesis of conjugated gamma-linolenic acid isomer by virtue of surfactant-coated bacterium | |
| CN105039442A (en) | Method for biosynthesis of conjugated alpha-linolenic acid isomer in organic medium | |
| CN105039441A (en) | Nonaqueous enzymologic synthetic method of conjugated alpha-linolenic acid isomer | |
| CN105112462A (en) | Non-aqueous enzymological synthesis method of conjugated linoleic acid isomer | |
| CN105039439A (en) | Method for biosynthesizing conjugated linoleic acid isomer in organic medium by adopting coated thalluses | |
| CN105087692A (en) | Method for catalytic synthesis of conjugated alpha-linolenic acid isomer by virtue of surfactant-coated bacterium | |
| CN105087691A (en) | Method for catalytic synthesis of conjugated linoleic acid isomer by virtue of surfactant-coated bacterium | |
| CN101215590A (en) | Method for removing monosaccharide component from oligomerization galactose by utilizing immobilized yeast | |
| CN102199636A (en) | Efficient preparation process of gamma-amino-n-butyric acid | |
| CN104789489A (en) | Arginine deiminase high-yielding Bacillus cereus and application thereof | |
| CN102191293A (en) | Method for inversing microbe to prepare ethyl (S)-3-hydroxy-3-(2-thienyl)-propanoate |
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 | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180417 Termination date: 20210720 |