CN103834609A - Application of nano-liposome technology in bioconversion - Google Patents
Application of nano-liposome technology in bioconversion Download PDFInfo
- Publication number
- CN103834609A CN103834609A CN201310485034.XA CN201310485034A CN103834609A CN 103834609 A CN103834609 A CN 103834609A CN 201310485034 A CN201310485034 A CN 201310485034A CN 103834609 A CN103834609 A CN 103834609A
- Authority
- CN
- China
- Prior art keywords
- substrate
- transformation
- nanometer liposome
- microbial
- application
- 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.)
- Pending
Links
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the technical field of microorganisms and relates to an application of a nano-liposome technology in bioconversion. The application adopts lecithin as a carrier to prepare hydrophobic substrate into nano lipsome, and then carries out microbial conversion. The application is especially applicable to microbial conversion of a hydrophobic compound, or microbial conversion of the substrate which has toxicity to the microbe and inhibiting the growth of the microbe. The application has the advantages that by utilization of the characteristic of the lecithin for increasing the permeability of a cell membrane, the advantages of good biocompatibility of nano liposome and improvement on the water solubility and large special surface area can be exerted, the transportation of the substrate into cells is enhanced, and the amount of the substrate entering the microbial cells, so that the feeding concentration and the conversion rate of the substrate in a conversion system are increased. The invention provides a new method for microbial conversion of the hydrophobic compound.
Description
Technical field
The invention belongs to biological technical field, be specifically related to the application of nanometer liposome in bio-transformation.
Background technology
Prior art discloses that to utilize microbial transformation hydrophobic compound to need in actual applications the bottleneck of breaking through be that the water-insoluble of substrate causes the concentration that feeds intake low, and transformation efficiency is not high.Microbial transformation belongs to two stage fermentations, and it comprises: first need in microbial strains liquid medium within, to cultivate appropriate time (mid-term of growth or later stage), accumulation transforms needed enzyme; Then drop into and need the substrate transforming to start conversion.The liquid culture of microbial strains is carried out conventionally in water, the solubleness of substrate in the aqueous solution is directly connected to the efficiency of microbial transformation reaction, due to microbial transformation enzyme, system generally belongs to intracellular enzyme, and the substrate that is only dissolved into liquid phase could fully contact and enter cell with microorganism cells.In steroidal microbial transformation process, having a general phenomenon is the raising along with the concentration that feeds intake, and transformation efficiency can obviously decline, and this is mainly to produce restraining effect because too much insoluble substrate can and transform thalli growth.The main organic solvent of means that addresses this problem at present, tensio-active agent, aqueous two-phase system etc.
Nano lipsome body technique is applied in pharmaceutical preparation conventionally, wherein, can adopt Yelkin TTS that insoluble drug is prepared into nanometer liposome, can improve significantly the water-soluble and bioavailability of medicine.Nanometer liposome has following advantage as pharmaceutical carrier: be made up of good biocompatibility the coated water vesica of phosphatide bimolecular; Fat-soluble medicine can be dissolved in adipose membrane; Phosphatide itself is cell membrane component, therefore to organism nontoxicity.The huge specific surface area obtaining based on its small size in addition is more easily carried out reaction.Studies show that, utilizing in microbiological deterioration sterol side chain, can change Cell membrane lipids bilayer structure as the Yelkin TTS of nano-medicament carrier and increase the permeability of cytolemma, promote that hydrophobic substrate enters in born of the same parents, thereby improve sterol side chain Degradation and Transformation rate.Based on the above-mentioned advantage of nanometer liposome, the present invention intends providing the new application of the nanometer liposome of preparing based on hydrophobic substrate in bio-transformation.
Summary of the invention
The object of this invention is to provide the new application of nano lipsome body technique, be specifically related to the new application of nano lipsome body technique in bio-transformation, relate in particular to the new application of the nanometer liposome of preparing based on hydrophobic substrate in bio-transformation.
In the present invention, adopting Yelkin TTS is that hydrophobic substrate is prepared into substrate nanometer liposome by carrier, then the conversion that feeds intake.
The present invention is prepared into nanometer liposome by hydrophobic substrate and carries out bio-transformation again, can utilize Yelkin TTS to increase the characteristic of permeability of cell membrane, can bring into play again nanometer liposome good biocompatibility, improve the advantage of water-soluble and huge specific surface area, promote that substrate transports in cell, its amount entering in microorganism cells is increased, concentration and transformation efficiency thereby raising transformation system substrate feeds intake.
Nano lipsome body technique disclosed by the invention is applied the microbial transformation that is particularly suitable for hydrophobic compound in bio-transformation, as sterol microorganism side chain cleavage, steroidal microorganism hydroxylation reaction etc.
In the present invention, described microbial transformation refers to the microbial transformation of hydrophobic compound, or substrate has the microbial transformation of toxicity, inhibition microorganism growth to microorganism.
The novel method that the present invention further provides nanometer liposome to be applied to bio-transformation, wherein, take the microorganism 11 α hydroxylation reactions of left-handed ethyl steroid alkene diketone as model.
In the present invention, described hydroxylation reaction is most important reaction in steroidal microbial transformation reaction, can provide important intermediate for steroid drugs chemosynthesis, in view of steroidal extremely low solubleness (lower than 0.1mM) in water, its microbial transformation reaction is the typical case of bio-transformation hydrophobic compound.
11 α hydroxy derivatives of left-handed ethyl steroid alkene diketone are the key intermediates of the potent progestogen desogestrel of preparation.Desogestrel is the main component of oral contraceptive Marvelon, because the side effect of Marvelon is little, reliable for effect, and its application is more and more extensive.It is starting raw material that the disclosed desogestrel synthesis technique of United States Patent (USP) (US2005/0234251A1) adopts left-handed ethyl steroid alkene diketone, first utilizes Aspergillus ochraceous to introduce 11 α hydroxyls through bio-transformation and carries out the synthetic desogestrel of series of chemical again.This patent is dissolved the left-handed ethyl steroid of substrate alkene diketone with organic solvent DMF and is fed intake, and while feeding intake concentration 2g/L-6g/L, hydroxylation reaction transformation efficiency is 35%-60%.
Bio-transformation model employing Metarhizium anisopliae mutant strain (Metarhizium anisopliae) the 11490(CCTCC M2011240 of system of the present invention), on left-handed ethyl steroid alkene diketone, introduce 11 α hydroxyls, described bacterial strain patent applied for, number of patent application 201110199550.7.
More specifically, nanometer liposome of the present invention is used for the method for bio-transformation, it is characterized in that, this method for transformation comprises the steps:
(1) prepare substrate nanometer liposome
Take Yelkin TTS as carrier, left-handed substrate ethyl steroid alkene diketone (DES1) is made to nanometer liposome; Optimize after prescription and preparation technology; Investigate particle diameter and structure, substrate encapsulation rate and the release conditions of nanometer liposome.
(2) microbial transformation
Microbial strains: Metarhizium anisopliae11490(culture presevation CCTCC M2011240), this bacterial classification can be introduced 11 α hydroxyls on left-handed ethyl steroid alkene diketone;
Slant culture: bacterial classification is inoculated in to potato dextrose agar (PDA) substratum, wherein adds mass percent and be 0.25% dried silkworm chrysalis meal, cultivate 5～10 days for 26～30 ℃;
Fermentation culture: above-mentioned slant strains is encircled in 40mL liquid nutrient medium (10g/L glucose, 10g/L analysis for soybean powder, 5g/L dried silkworm chrysalis meal are housed with inoculation articulating 1 under aseptic condition, pH6.5) in 250mL shaking flask, under 28 ℃ of conditions, rotary shaking table 220r/min cultivates 24 hours, obtains seed liquor;
Bio-transformation: the substrate nanometer liposome making is dropped in the above-mentioned seed liquor making, transform 60～72h under the same terms, get conversion fluid analysis.
In the present invention, described analytical procedure comprises:
1.HPLC
Moving phase: A water, B acetonitrile
Wavelength: 215nm/254nm
Flow velocity: 1.2mL/min
Gradient elution:
0-5min A:B=70:30 keeps 5min
5-15min gradient A:B=70:30 → A:B=40:60 keeps 10min
15-25min A:B=40:60 keeps 10min
Sample dissolve with methanol, substrate DES1 and hydroxylation product D ES2 result thereof are as shown in Figure 1;
2.TLC and TLC Scanner
Get fermented liquid 2mL and add 2mL ethyl acetate soaked overnight, get 5uL point sample in GF254 silica-gel plate, get substrate and the contrast of product standard substance (1mg/mL) 1uL point sample, developping agent CH2Cl2:CH3OH(20:1 simultaneously), CAMAG Scanner-3 scanning analysis;
3. substrate nanometer liposome characterizes
Adopt film dispersion method to prepare Yelkin TTS-substrate complex suspension, high pressure homogenizer is prepared nanometer liposome, measures particle diameter and size-grade distribution with laser particle size potentiometer, adopts dialysis method to measure encapsulation rate.After PBS damping fluid dialysis purifying, transmission electron microscope observing form;
Result demonstration,
(1) prepare substrate nanometer liposome,
Adopt film dispersion method that left-handed ethyl steroid alkene diketone is made to nanometer liposome take Yelkin TTS as carrier through high pressure homogenizer, median size 131.6nm(is as shown in Figure 2 after measured), encapsulation rate is greater than 93%, and TEM (transmission electron microscope) analysis result is as shown in Figure 3;
(2) substrate absorption test
Analyze the specific absorption of mycelia to substrate in nanometer liposome and free substrate, by the free substrate of different concns or substrate nanometer liposome, to hatch 6h(hydroxylase at 28 ℃ be inducible enzyme with being suspended in mycelia in damping fluid, induction time is 6-8h), results mycelia, through washing, high pressure homogenizer smudge cells, after centrifuging, HPLC measures substrate content in mycelia, wherein, the specific absorption of free substrate in mycelia is only 13% left and right, and the specific absorption of nanometer liposome is up to 75% left and right (as shown in table 1), show that nanometer liposome has very strong promoter action to substrate to intracellular transport,
Table 1 different concns substrate and nanometer liposome thereof the specific absorption in mycelia
(3) promoter action of nanometer liposome to microbial transformation
The substrate nanometer liposome of preparation is dropped into the mycelium culture liquid of cultivating by the concentration that feeds intake of 2g/L (20mg substrate drops into 10mL bacterial culture fluid) and carry out steroidal hydroxylation reaction, and with free substrate feeding mode (DMF dissolving) and substrate+Yelkin TTS (not being prepared into liposome) feeding mode comparison, result as shown in Figure 4, through TLC scanning analysis, result shows, after being prepared into nanometer liposome, substrate feeds intake (product amount 17.44mg), product amount improves nearly 3 times than traditional DMF feeding mode (product amount 6.22mg), and substrate is almost converted into product completely.And substrate feeds intake and add equivalent Yelkin TTS (not being prepared into nanometer liposome) simultaneously, output and DMF feed intake basic identical (product amount 5.99mg); Further extend transformation time to 116h, under this feeding mode, output can reach 10.44mg.Thereby this description of test is prepared into nanometer liposome by substrate and feeds intake and can accelerate substrate and significantly improve Microbial Transformed Efficiency within a short period of time to shifting in born of the same parents, add separately Yelkin TTS and can not improve transformation efficiency within the same conversion time, but extend transformation time can suitably improve transformation efficiency (with DMF feed ratio), thereby this can increase permeability of cell membrane with bibliographical information Yelkin TTS and is conducive to substrate and is consistent to shifting raising transformation efficiency in born of the same parents.
Further relatively nanometer liposome feed intake and DMF feeding mode under the impact (Fig. 5) of concentration on the final transformation efficiency of product that feed intake, result shows that, under nanometer liposome feeding mode, the concentration that feeds intake is brought up to 6g/L, transformation efficiency still can maintain 60% left and right.And under DMF feeding mode, transformation efficiency obviously declines along with the raising of the concentration that feeds intake, product yield only 13.1% in the time of 4g/L.So nanometer liposome can not only fine raising transformation efficiency, and can effectively improve the substrate concentration that feeds intake.
Advantage of the present invention has:
Adopt nano lipsome body technique can promote hydrophobic substrate to intracellular transport, improve the concentration that feeds intake, and then improve hydrophobic compound microbial transformation reaction efficiency.Compared with the prior art: nano lipsome body technique is in more the improving for pharmaceutical preparation system, drug delivery mode of field of medicaments, the present invention is applied to the bio-transformation of the insoluble materials such as steroidal compounds, for the bio-transformation of hydrophobic compound provides novel method.
Accompanying drawing explanation
Fig. 1: substrate (DES1) and product (DES2) HPLC collection of illustrative plates.
Fig. 2: left-handed ethyl steroid alkene diketone nanometer liposome size distribution.
Fig. 3: left-handed ethyl steroid alkene diketone nanometer liposome TEM (transmission electron microscope) analysis.
Fig. 4: nanometer liposome feeds intake, substrate+Yelkin TTS (not being prepared into liposome) feeds intake and DMF dissolves the product amount comparison feeding intake after substrate.
Fig. 5: nanometer liposome feed intake and DMF feeding mode under the impact of concentration on the final transformation efficiency of product that feed intake.
Embodiment
The 11 α hydroxylation reactions of embodiment 1:Metarhizium anisopliae11490 to left-handed ethyl steroid alkene diketone nanometer liposome, the concentration that feeds intake 2g/L.
(1) prepare substrate nanometer liposome
Take Yelkin TTS as carrier, left-handed substrate ethyl steroid alkene diketone is made to nanometer liposome, median size 131.6nm.
(2) microbial transformation
Microbial strains: Metarhizium anisopliae11490, culture presevation CCTCC M2011240, this bacterial classification can be introduced 11 α hydroxyls on left-handed ethyl steroid alkene diketone.
(3) substratum
Slant culture: potato dextrose agar (PDA) substratum, wherein adds mass percent and is 0.25% dried silkworm chrysalis meal;
Fermention medium: 10g/L glucose, 10g/L analysis for soybean powder, 5g/L dried silkworm chrysalis meal, pH6.5;
(4) microorganism culturing
Bacterial classification, under 28 ℃ of conditions, is cultivated 24 hours under rotary shaking table 220r/min, 40mL/250mL shaking flask liquid amount condition, obtains seed liquor;
(5) bio-transformation
The left-handed ethyl steroid alkene diketone nanometer liposome making is dropped in the above-mentioned seed liquor making of 10mL by the concentration that feeds intake of 2g/L, under the same terms, transform 66h, get conversion fluid analysis;
Result demonstration, 11 α hydroxy derivatives output of left-handed ethyl steroid alkene diketone are 17.44mg, transformation efficiency 87.2%.
The 11 α hydroxylation reactions of embodiment 2:Metarhizium anisopliae11490 to left-handed ethyl steroid alkene diketone nanometer liposome, the concentration that feeds intake 4g/L.
(1) prepare substrate nanometer liposome
Take Yelkin TTS as carrier, left-handed substrate ethyl steroid alkene diketone is made to nanometer liposome, median size 131.6nm.
(2) microbial transformation
Microbial strains: Metarhizium anisopliae11490, culture presevation CCTCC M2011240, this bacterial classification can be introduced 11 α hydroxyls on left-handed ethyl steroid alkene diketone.
(3) substratum
Slant culture: potato dextrose agar (PDA) substratum, wherein adds mass percent and is 0.25% dried silkworm chrysalis meal;
Fermention medium: 10g/L glucose, 10g/L analysis for soybean powder, 5g/L dried silkworm chrysalis meal, pH6.5;
(4) microorganism culturing
Bacterial classification, under 28 ℃ of conditions, is cultivated 24 hours under rotary shaking table 220r/min, 40mL/250mL shaking flask liquid amount condition, obtains seed liquor;
(5) bio-transformation
The left-handed ethyl steroid alkene diketone nanometer liposome making is dropped in the above-mentioned seed liquor making of 10mL by the concentration that feeds intake of 4g/L, under the same terms, transform 70h, get conversion fluid analysis;
Result demonstration, 11 α hydroxy derivatives output of left-handed ethyl steroid alkene diketone are 27.24mg, transformation efficiency 68.1%.
The 11 α hydroxylation reactions of embodiment 3:Metarhizium anisopliae11490 to left-handed ethyl steroid alkene diketone nanometer liposome, the concentration that feeds intake 6g/L.
(1) prepare substrate nanometer liposome
Take Yelkin TTS as carrier, left-handed substrate ethyl steroid alkene diketone is made to nanometer liposome, median size 131.6nm.
(2) microbial transformation
Microbial strains: Metarhizium anisopliae11490, culture presevation CCTCC M2011240, this bacterial classification can be introduced 11 α hydroxyls on left-handed ethyl steroid alkene diketone.
(3) substratum
Slant culture: potato dextrose agar (PDA) substratum, wherein adds mass percent and is 0.25% dried silkworm chrysalis meal;
Fermention medium: 10g/L glucose, 10g/L analysis for soybean powder, 5g/L dried silkworm chrysalis meal, pH6.5;
(4) microorganism culturing
Bacterial classification, under 28 ℃ of conditions, is cultivated 24 hours under rotary shaking table 220r/min, 40mL/250mL shaking flask liquid amount condition, obtains seed liquor;
(5) bio-transformation
The left-handed ethyl steroid alkene diketone nanometer liposome making is dropped in the above-mentioned seed liquor making of 10mL by the concentration that feeds intake of 6g/L, under the same terms, transform 90h, get conversion fluid analysis;
Result demonstration, 11 α hydroxy derivatives output of left-handed ethyl steroid alkene diketone are 35.04mg, transformation efficiency 58.4%.
Claims (4)
1. the application of nanometer liposome in bio-transformation, it is that hydrophobic substrate is prepared into substrate nanometer liposome by carrier that described nanometer liposome adopts Yelkin TTS.
2. by application claimed in claim 1, it is characterized in that, described bio-transformation is the microbial transformation of hydrophobic compound, or substrate has the microbial transformation of toxicity, inhibition microorganism growth to microorganism.
3. by application claimed in claim 1, it is characterized in that, described microbial transformation is sterol microorganism side chain cleavage or steroidal microorganism hydroxylation reaction.
4. nanometer liposome, for a method for bio-transformation, is characterized in that, this method for transformation comprises the steps:
(1) prepare substrate nanometer liposome
Take Yelkin TTS as carrier, substrate is made to nanometer liposome; Optimize after prescription and preparation technology; Investigate particle diameter and structure, substrate encapsulation rate and the release conditions of nanometer liposome;
(2) microbial transformation
Microbial strains: Metarhizium anisopliae11490, culture presevation CCTCC M2011240, this bacterial classification is introduced 11 α hydroxyls on left-handed ethyl steroid alkene diketone;
(3) slant culture: bacterial classification is inoculated in to potato dextrose agar (PDA) substratum, wherein adds mass percent and be 0.25% dried silkworm chrysalis meal, cultivate 5～10 days for 26～30 ℃;
(4) fermentation culture: above-mentioned slant strains is connected to liquid nutrient medium 10g/L glucose, 10g/L analysis for soybean powder, 5g/L dried silkworm chrysalis meal are housed, and in the shaking flask of pH6.5, under 28 ℃ of conditions, rotary shaking table 220r/min cultivates 48 hours, obtains seed liquor;
(5) bio-transformation: the substrate nanometer liposome making is dropped in the above-mentioned seed liquor making, transform 60～90h under the same terms, get conversion fluid analysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310485034.XA CN103834609A (en) | 2012-11-26 | 2013-10-16 | Application of nano-liposome technology in bioconversion |
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210485147.5 | 2012-11-26 | ||
| CN201210485147 | 2012-11-26 | ||
| CN201310485034.XA CN103834609A (en) | 2012-11-26 | 2013-10-16 | Application of nano-liposome technology in bioconversion |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103834609A true CN103834609A (en) | 2014-06-04 |
Family
ID=50798506
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310485034.XA Pending CN103834609A (en) | 2012-11-26 | 2013-10-16 | Application of nano-liposome technology in bioconversion |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103834609A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111593086A (en) * | 2020-05-27 | 2020-08-28 | 湖北葛店人福药业有限责任公司 | Method for reducing impurities in hydroxylation process of ethyl diketone 11a by using mixed solvent |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876582A (en) * | 2011-07-15 | 2013-01-16 | 复旦大学 | Metarhizium anisopliae and application of Metarhizium anisopliae to hydroxylation reaction of steroids |
-
2013
- 2013-10-16 CN CN201310485034.XA patent/CN103834609A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102876582A (en) * | 2011-07-15 | 2013-01-16 | 复旦大学 | Metarhizium anisopliae and application of Metarhizium anisopliae to hydroxylation reaction of steroids |
Non-Patent Citations (4)
| Title |
|---|
| NURY LLANES ET AL.: "Conversion of liposomal 4-androsten-3,17-dione by A. simplex immobilized cells in calcium pectate", 《JOURNAL OF STEROID BIOCHEMISTRY & MOLECULAR BIOLOGY》, 31 December 2002 (2002-12-31) * |
| 史济平 等: "金龟子绿僵菌对甾族化合物的11α-羟化", 《上海医科大学学报》, vol. 24, no. 4, 30 July 1997 (1997-07-30) * |
| 叶丽 等: "雄甾-4-烯-3, 17-二酮11α羟化突变株Metarhizium anisopliae M28-203 的代谢调控", 《复旦学报（医学版）》, vol. 38, no. 1, 31 January 2011 (2011-01-31) * |
| 龚连生 等: "阿霉素纳米脂质体制备工艺研究", 《中国现代医学杂志》, vol. 19, no. 24, 31 December 2009 (2009-12-31) * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111593086A (en) * | 2020-05-27 | 2020-08-28 | 湖北葛店人福药业有限责任公司 | Method for reducing impurities in hydroxylation process of ethyl diketone 11a by using mixed solvent |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Fang et al. | Effect of initial pH on production of ganoderic acid and polysaccharide by submerged fermentation of Ganoderma lucidum | |
| CN109439701B (en) | Method for preparing ergothioneine by biosynthesis and fermentation medium | |
| CN104342390B (en) | A kind of Sinorhizobium meliloti strain and combinations thereof and application | |
| CN101020919A (en) | Biological sterol transforming process with cyclodextrin and its application | |
| CA2028838A1 (en) | Production of beta-1, 3-glucan in algae | |
| CN101831481A (en) | New method for preparing Iturin A and homolugues thereof | |
| CN101525651A (en) | Method for preparing androstenedione by biodegradation of phytosterol in two-liquid-phase system | |
| CN107164295A (en) | A kind of selenium-enriched microbe its preparation method and application | |
| CN110790820B (en) | Lipopeptide produced by bacillus strain FJAT-52631 and preparation method thereof | |
| CN112322687B (en) | Microbial inoculum for preparing diosgenin and application thereof | |
| CN102127511B (en) | Penicillium decumbens, culture method thereof, and application thereof in transforming steroid medicines | |
| CN101191141A (en) | Fermentation culture medium for biologically synthesizing 7alpha, 15alpha-dihydroxyandrostenolone | |
| CN108866128B (en) | Method for improving biological value of kasugamycin | |
| CN102703558B (en) | Method for hydroxylating dehydroisoandrosterone by using colletotrichumlini | |
| CN103555806B (en) | Method for synthesizing 7alpha-hydroxy-androstenone by efficient utilization of colletotrichum lini | |
| CN108841889A (en) | The main part of Songgangmeisu --- the method for griseofulvin is produced using microbial fermentation | |
| CN103834609A (en) | Application of nano-liposome technology in bioconversion | |
| CN103849663A (en) | Method of preparing fumigaclavine C by symbiotic aspergillus fumigatus of sea crab and culture medium of fumigaclavine C | |
| CN103834712B (en) | The optimization method of desogestrel intermediate nanometer liposome bio-transformation | |
| CN102965300B (en) | Micromonospora strain, its preparation method and application | |
| Manosroi et al. | A novel one-step biotransformation of cortexolone-21-acetate to hydrocortisone acetate using Cunninghamella blakesleeana ATCC 8688a | |
| Chen et al. | Biotransformation of saponins to astragaloside IV from Radix Astragali by immobilized Aspergillus niger | |
| Ramakrishna et al. | Evaluation of solid, slurry, and submerged fermentations for the production of cyclodextrin glycosyl transferase by Bacillus cereus | |
| CN104371950B (en) | Oxygen resistant clostridium and application of oxygen resistant clostridium in aerobic synthesis of O-desmethylangolensin | |
| CN105483028B (en) | It is a kind of to synthesize 11 α, the method for 15 α-diOH- 16ALPHA,17ALPHA-epoxyprogesterones using gibberella |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140604 |