CN105603009A - Microorganism converting method - Google Patents
Microorganism converting method Download PDFInfo
- Publication number
- CN105603009A CN105603009A CN201610086443.6A CN201610086443A CN105603009A CN 105603009 A CN105603009 A CN 105603009A CN 201610086443 A CN201610086443 A CN 201610086443A CN 105603009 A CN105603009 A CN 105603009A
- Authority
- CN
- China
- Prior art keywords
- thalline
- phe
- proteus
- phenyllactic acid
- concentration
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/42—Hydroxy-carboxylic acids
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for producing R-(+)-2-hydroxyl-3-phenylpropionic acid by converting L-phenylalanine with Proteus microorganisms. The method is simple in process and has significant industrial application value.
Description
Technical field
The present invention adopts proteus to transform L-Phe and produces D-phenyllactic acid, belongs to industrial microorganism field.
Background technology
D-phenyllactic acid, formal name used at school R-(+)-PLA, English by name: R-(+)-2-hydroxy-3-phenylpropanoicacid、D-(+)-3-Phenyllacticacid。
Phenyllactic acid is a kind of New Biological Preservatives that discovered in recent years can be produced by the secretion of part lactic acid bacteria.It has D-phenyllactic acid and two kinds of mapping allosome bodies of L-phenyllactic acid, can effectively suppress the multiple food spoilage that causesBacterium and toxigenic fungi. The bacteriostasis that research shows D-phenyllactic acid is a little more than L-phenyllactic acid.
As a kind of New Biological Preservatives, the using value that phenyllactic acid tool in food industry is important. CurrentMain by the production of large intestine genetic engineering bacterium expression lactic dehydrogenase enzymatic conversion phenylpyruvic acid, as Chinese patentCN201410818165.X, CN201410393768.X and CN201210338378.3. Also have by all kinds ofLactic acid bacteria transforms phenylpyruvic acid or phenylalanine and produces, as Chinese patent CN201510089823.0,CN201210005544.8 etc. Also there is the method that adopts bacillus to transform phenylpyruvic acid product phenyllactic acid(EfficientConversionofPhenylpyruvicAcidtoPhenyllacticAcidbyUsingWholeCellsofBacilluscoagulansSDM，2011，PLoSOne.6(4):e19030)。
Based on the significant application value of D-phenyllactic acid, this patent has proposed the microbial conversion of employing ProteusL-Phe is produced the scheme of D-phenyllactic acid. L-Phe is current main by Production by Microorganism Fermentation,Abundant raw material is easy to get.
Proteus (Proteus) is to be extensively distributed in occurring in nature, as soil, water, rubbish, corrupt organicMicroorganism in thing and human or animal's enteron aisle. Existing 5 kinds: proteus vulgaris (Proteusvulgaris),Proteus mirabilis (Proteusmirabilis), the glutinous proteus (Proteusmyxofaciens) of product, Pan Shi distortionBacillus (Proteuspenneri) and Hao Shi proteus (Proteushauseri). Proteus has powerful oxygenChange deamination ability (SherrisMedicalMicrobiology, 2004,4thed.), L-Phe can be becomeShape bacillus oxidative deamination becomes phenylpyruvic acid. Pertinent literature thinks that proteus microorganism belonging to genus can only be by L-phenylpropyl alcohol ammoniaAcid oxidase becomes phenylpyruvic acid, and cannot further be reduced into phenyllactic acid (α-ketoacidsarenovelsiderophoresinthegeneraproteus,providencia,andmorganellaandareproducedbyAminoaciddeaminases, 1993, Journalofbacteriology, 175 (9), 2727-2733), this canCondition is selected improper relevant with it.
The present invention can change into optically pure D-phenyllactic acid by L-Phe by proteus microorganism belonging to genus.Although phenylpyruvic acid is more direct substrate, price is higher, and therefore L-Phe is best substrate.Proteus is a kind of facultative anaerobe, can under anaerobic condition or aerobic condition, transform and produce D-phenyllactic acid,There is high conversion and highly purified feature.
Summary of the invention
The present invention, by cultivating Proteus microbial cells, transforms L-Phe and produces D-phenyllactic acid,Technical scheme is as follows:
1, bacterial strain
The present invention's bacterial strain used have purchased from the ProteusmirabilisATCC29906 of U.S. ATCC strain library,ProteusmyxofaciensATCC19692、ProteushauseriATCC700826、ProteuspenneriATCC33519、ProteusmirabilisATCC25933、ProteusmirabilisATCC33946、ProteusvulgarisATCC19181, ProteusvulgarisATCC27972 and micro-purchased from Chinese industrialProteusvulgarisCICC10401, the ProteusmirabilisCICC of biological inoculum preservation administrative center22928。
2, thalline is cultivated
Liquid state fermentation culture medium consists of: carbon source 0-50g/L, and nitrogenous source 0-50g/L, diammonium hydrogen phosphate 0-2g/L,Potassium dihydrogen phosphate 0-1g/L, magnesium sulfate 0-0.5g/L, ferrous sulfate 0-0.5g/L, common salt 0-5g/L, canRegulate pH to 2-8, also pH nature; Inoculum concentration is 5-20%, and fermentation temperature is 20-40 DEG C, fermentation weekPhase is 24-72 hour. Seed and fermentation all adopt this culture medium.
For the carbon source of cultivating bacterial classification can be glucose, fructose, maltose, wood sugar, sucrose, galactolipin,Glycerine. Cultivate with nitrogenous source can be ammonium sulfate, ammonium chloride, ammonium nitrate, peptone, yeast extract, urea,The organic nitrogen sources such as corn steep liquor. Carbon and nitrogen sources can be one or more combination.
Liquid process of cultivating thalline can adopt anaerobism or aerobic mode.
3, transform and produce D-phenyllactic acid
The scheme that conversion process adopts has 2 kinds:
(1) the liquid incubation of cell, adds L-Phe substrate in above-mentioned cultivating system;L-Phe addition is 0-20g/L.
(2) centrifugal liquid state fermentation culture removal zymotic fluid is obtained to thalline, thalline is put into and contained L-benzeneIn the reactant aqueous solution system of alanine substrate, carry out; Conversion process temperature 20-40 DEG C, pH2-8, transformation time 1-24 hour. In conversion fluid, L-Phe initial concentration is 0.1-20g/L.
4, the detection analysis of sample
Conversion fluid adopts PerkinElmerSeries200 high performance liquid chromatograph to detect and analyzes, and chromatographic condition is:Mobile phase is methyl alcohol-0.1% formic acid water (40:60), adopt the MegresC18 of Chinese nation chromatographic column (4.6 × 250mm,5 μ m), flow velocity 0.6ml/min, 30 DEG C of column temperatures, sample size 20 μ l, detector wavelength 200nm.
Adopt the DAC-HB50 preparative chromatography post preparation of Hanbon Sci. & Tech. Co., Ltd. to transform sample, preparationChromatographic condition is: mobile phase 50% methyl alcohol, column temperature nature, flow velocity 3ml/min, sample size 5ml. Sample reachesTo chromatographically pure 99.9%, sample introduction separates the product obtaining vacuum rotating evaporate to dryness at 50 DEG C repeatedly. Take preparationThe sample 0.5g obtaining, is dissolved in deionized water and is settled to 50ml, adopts Japan to like to delay AP-300 entirely certainlyMoving polarimeter is measured degree of giving out light. Further adopt VarianGemini2000 (VnmrS600MHz,600/54/ASP) the nuclear magnetic data of analytic sample. Sample further adopts the analysis of UPLC-QTOF-MS method to divideSon amount, instrument is the WatersMALDISYNAPTQTOFMS liquid chromatogram series connection quadrupole rod flight timeMass spectrograph.
Detailed description of the invention
Embodiment 1
The analysis of converted product is measured.
Preparation culture medium is as follows: yeast extract 5g/L, peptone 10g/L, sodium chloride 5g/L. 500ml triangleIn bottle, liquid amount is 100ml, totally 50 bottles, and 120 DEG C, sterilizing in 20 minutes. Get ProteusmirabilisATCC29906 glycerine pipe seed liquor 1mL inoculation, 30 DEG C of fermentation temperatures, shaking speed 200rpm. TrainingAfter supporting 24, by centrifugal zymotic fluid (rotating speed 10000rpm, time 10min), remove supernatant and obtain thalline,Twice of sterile water wash of the thalline of centrifugal acquisition. Getting 20g wet thallus, to put into L-Phe concentration be 10In the 1000ml solution of g/L, mix. Vibrate (100rpm) after 24 hours in 37 DEG C of shaking tables, 100 DEG CHeating water bath is killed thalline for 3 minutes. Centrifugal (rotating speed 10000rpm, time 10min) removes thalline again,Arrive the supernatant after transforming. Liquid phase analysis D-phenyllactic acid concentration is 4.1g/L, and residue L-Phe concentration is4.5g/L. Adopt preparative chromatography to obtain 3.4g sterling.
Polarimeter is analyzed its optical activityNuclear magnetic data is: 1HNMR (CDCl3600MHz):δ7.32(2H,dJ7.6Hz),7.28(1H,d,J7.6Hz),7.25(2H,d,J7.6Hz)，4.48(1H,dd,J4.3,7.3Hz)，4.24(2H,OHs,broad)，3.22(1H,dd,J4.3,14.0Hz)，2.97(1H,dd,J7.3,14.0Hz)。13CNMR(CDCl3,600MHz):δ177.56，135.92，129.51，129.51，128.56,128.63,127.14,71.11,40.12. The mass spectrometric data of converted product is: (ESI,negativemode)m/z:165.0[M-1]。
According to above data, determine that its molecule and optical texture and D-phenyllactic acid are in full accord.
Embodiment 2
Aerobic cultivation and the comparison of detesting cultivation. Preparation culture medium: glucose 30g/L, peptone 20g/L,Diammonium hydrogen phosphate 1g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.3g/L, ferrous sulfate 0.3g/L; InitialIt is 100ml that pH and sweat pH are liquid amount in nature 500ml triangular flask, totally 2 bottles, and 120 DEG C,Sterilizing in 20 minutes.
Get 1 bottle of ProteusmirabilisCICC22928 glycerine pipe seed liquor 1mL inoculation, cultivate in anaerobismIn case, cultivate 72 hours twice of sterile water wash of the thalline of centrifugal acquisition for 35 DEG C. Getting 0.1g wet thallus putsEntering L-Phe concentration is in the 4ml solution of 2g/L, mixes, and in anaerobic culture box, 35 DEG C leave standstillTransform after 24 hours, 100 DEG C of heating water baths are killed thalline for 3 minutes. Again centrifugal (rotating speed 10000rpm, timeBetween 10min) remove thalline, obtain transform after supernatant. Liquid phase analysis D-phenyllactic acid concentration is 1.4g/L,Residue L-Phe concentration is 0.4g/L.
Get 1 bottle of ProteusmirabilisCICC22928 glycerine pipe seed liquor 1mL inoculation, in shaking table(200rpm) in, cultivate 24 hours for 35 DEG C. By centrifugal zymotic fluid (rotating speed 10000rpm, time 10min), goExcept supernatant obtains thalline, twice of sterile water wash of the thalline of centrifugal acquisition. Get 0.1g wet thallus and put into L-Concentration of phenylalanine is in the 4ml solution of 2g/L, mixes. In 35 DEG C of shaking table vibration (100rpm) 12After hour, 100 DEG C of heating water baths are killed thalline for 3 minutes. Centrifugal (rotating speed 10000rpm, time 10min) againRemove thalline, obtain the supernatant after transforming. Liquid phase analysis D-phenyllactic acid concentration is 1.2g/L, residue L-benzeneAlanine concentration is 0.4g/L.
Embodiment 3
Culture medium consists of: glucose 1g/L, urea 1g/L, diammonium hydrogen phosphate 1g/L, potassium dihydrogen phosphate0.1g/L, magnesium sulfate 0.1g/L, ferrous sulfate 0.1g/L. In 250ml triangular flask, liquid amount is 50ml,120 DEG C, sterilizing in 20 minutes. Getting ProteusvulgarisATCC19181 glycerine pipe seed liquor 0.5mL connectsKind, 35 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, by centrifugal zymotic fluid (rotating speed10000rpm, time 10min), remove supernatant and obtain thalline, the thalline sterile water wash of centrifugal acquisitionTwice. Get 0.05g wet thallus and put into the 2ml solution that L-Phe concentration is 0.1g/L, and regulate pHTo 6, mix. Static conversion 24 hours in anaerobic culture box, filtering with microporous membrane conversion fluid, liquidIn analysis of hplc conversion fluid, D-phenyllactic acid concentration is that 0.06g/L and L-Phe residual quantity are 0.01g/L.
Embodiment 4
Culture medium consists of: fructose 50g/L, ammonium sulfate 5g/L, diammonium hydrogen phosphate 1g/L, biphosphatePotassium 1g/L, ferrous sulfate 0.5g/L. In 250ml triangular flask, liquid amount is 50ml, totally 10 bottles, and 120 DEG C,Sterilizing in 20 minutes. Get ProteushauseriATCC700826 glycerine pipe seed liquor 0.5mL inoculation, fermentation20 DEG C of temperature, shaking speed 200rpm. Cultivate after 72, by centrifugal zymotic fluid (rotating speed 10000rpm, timeBetween 10min), remove supernatant and obtain thalline, twice of sterile water wash for the thalline of centrifugal acquisition. Get 1gWet thallus is put into the 2ml solution that L-Phe concentration is 20g/L, mixes. Shake in 20 DEG C of shaking tablesSwing (100rpm) after 24 hours, filtering with microporous membrane conversion fluid, D-phenyllactic acid in liquid-phase chromatographic analysis conversion fluidConcentration is that 8.5g/L and L-Phe residual quantity are 7.8g/L.
Embodiment 5
Culture medium consists of: wood sugar 50g/L, ammonium sulfate 1g/L, diammonium hydrogen phosphate 1g/L, biphosphatePotassium 1g/L, magnesium sulfate 0.2g/L. In 250ml triangular flask, liquid amount is 50ml, 120 DEG C, within 20 minutes, goes outBacterium. Get ProteusvulgarisCICC10401 glycerine pipe seed liquor 0.5mL inoculation, 40 DEG C of fermentation temperatures,Shaking speed 200rpm. After cultivating 48, by centrifugal zymotic fluid (rotating speed 10000rpm, time 10min), goExcept supernatant obtains thalline, twice of sterile water wash of the thalline of centrifugal acquisition. Get 0.1g wet thallus and put into L-Concentration of phenylalanine is in the 2ml solution of 2g/L, mixes. In 40 DEG C of shaking table vibration (100rpm) 24After hour, filtering with microporous membrane conversion fluid, in liquid-phase chromatographic analysis conversion fluid, D-phenyllactic acid concentration is 0.7g/LAnd L-Phe residual quantity is 1.1g/L.
Embodiment 6
Culture medium consists of: glycerine 15g/L, ammonium nitrate 1g/L, diammonium hydrogen phosphate 1g/L, biphosphatePotassium 0.1g/L, magnesium sulfate 0.1g/L, ferrous sulfate 0.1g/L. In 250ml triangular flask, liquid amount is 50ml,120 DEG C, sterilizing in 20 minutes. Get ProteusmyxofaciensATCC19692 glycerine pipe seed liquor 0.5mLInoculation, 30 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, by centrifugal zymotic fluid (rotating speed10000rpm, time 10min), remove supernatant and obtain thalline, the thalline sterile water wash of centrifugal acquisitionTwice. Get 0.1g wet thallus and put into the 2ml solution that L-Phe concentration is 2g/L, mix.Static conversion 24 hours in 20 DEG C of anaerobic culture boxes, filtering with microporous membrane conversion fluid, liquid-phase chromatographic analysisIn conversion fluid, D-phenyllactic acid concentration is that 0.6g/L and L-Phe residual quantity are 1.2g/L.
Embodiment 7
Culture medium consists of: maltose 15g/L, ammonium chloride 1g/L, diammonium hydrogen phosphate 1g/L, di(2-ethylhexyl)phosphateHydrogen potassium 0.1g/L, magnesium sulfate 0.1g/L, ferrous sulfate 0.1g/L, regulates pH to 5. In 250ml triangular flaskLiquid amount is 50ml, 120 DEG C, and sterilizing in 20 minutes. Get ProteusvulgarisATCC27972 glycerine pipeSeed liquor 0.5mL inoculation, 35 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, by zymotic fluidCentrifugal (rotating speed 10000rpm, time 10min), removes supernatant and obtains thalline, and the thalline of centrifugal acquisition is usedTwice of sterile water wash. Get 0.1g wet thallus and put into the 2ml solution that L-Phe concentration is 3g/L,Mix. Static conversion 24 hours in 40 DEG C of anaerobic culture boxes, filtering with microporous membrane conversion fluid, liquidIn analysis of hplc conversion fluid, D-phenyllactic acid concentration is that 1.7g/L and L-Phe residual quantity are 1.0g/L.
Embodiment 8
Culture medium consists of: galactolipin 1g/L, urea 1g/L, diammonium hydrogen phosphate 1g/L, potassium dihydrogen phosphate0.1g/L, magnesium sulfate 0.1g/L, ferrous sulfate 0.1g/L, regulates pH to 2. In 250ml triangular flask, fill liquidAmount is 50ml, 120 DEG C, sterilizing in 20 minutes. Get ProteusvulgarisATCC25933 glycerine pipe seedLiquid 0.5mL inoculation, 35 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, zymotic fluid is centrifugal(rotating speed 10000rpm, time 10min), removes supernatant and obtains thalline, and the thalline of centrifugal acquisition is with asepticWater cleans twice. Get 0.1g wet thallus and put into the 2ml solution that L-Phe concentration is 2g/L, and adjustJoint pH to 2, mixes. Static conversion 24 hours in 37 DEG C of anaerobic culture boxes, filtering with microporous membraneConversion fluid, in liquid-phase chromatographic analysis conversion fluid, D-phenyllactic acid concentration is 0.2g/L and L-Phe residual quantityFor 1.6g/L.
Embodiment 9
Culture medium consists of: glucose 1g/L, peptone 1g/L, potassium dihydrogen phosphate 0.1g/L, magnesium sulfate0.1g/L, ferrous sulfate 0.1g/L, L-Phe 20g/L regulates pH to 8. In 250ml triangular flask, fillLiquid measure is 50ml, 120 DEG C, and sterilizing in 20 minutes. Get ProteusmirabilisATCC33946 glycerine pipe kindSub-liquid 0.5mL inoculation, 35 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, miillpore filter mistakeFilter conversion fluid, in liquid-phase chromatographic analysis conversion fluid, D-phenyllactic acid concentration is that 10.2g/L and L-Phe are residualAmount is 6.3g/L.
Embodiment 10
Culture medium consists of: glucose 1g/L, urea 1g/L, diammonium hydrogen phosphate 1g/L, magnesium sulfate 0.1G/L, ferrous sulfate 0.1g/L, regulates pH to 8. In 500ml triangular flask, liquid amount is 100ml, 120 DEG C,Sterilizing in 20 minutes, totally 10 bottles. Respectively get ProteuspenneriATCC33519 glycerine pipe seed liquor 0.5mLInoculation, 35 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, by centrifugal zymotic fluid (rotating speed10000rpm, time 10min), remove supernatant and obtain thalline, the thalline sterile water wash of centrifugal acquisitionTwice. Get 5g wet thallus and put into the 10ml solution that L-Phe concentration is 10g/L, and regulate pHTo 5, mix. Vibrate (100rpm) after 10 hours in 35 DEG C of shaking tables, filtering with microporous membrane conversion fluid,In liquid-phase chromatographic analysis conversion fluid, D-phenyllactic acid concentration is that 7.5g/L and L-Phe residual quantity are 0.23g/L.
Embodiment 11
Culture medium consists of: glucose 1g/L, peptone 1g/L, diammonium hydrogen phosphate 1g/L, biphosphatePotassium 0.1g/L, magnesium sulfate 0.1g/L, ferrous sulfate 0.1g/L. Regulate pH to 5. In 250ml triangular flask, fillLiquid measure is 50ml, 120 DEG C, and sterilizing in 20 minutes. Get ProteusmirabilisATCC33946 glycerine pipe kindSub-liquid 0.5mL inoculation, 37 DEG C of fermentation temperatures, shaking speed 200rpm. After cultivating 24, miillpore filter mistakeFilter conversion fluid, in liquid-phase chromatographic analysis zymotic fluid, D-phenyllactic acid concentration is 0.2g/L.
Claims (4)
1. Proteus microbial conversion L-Phe is produced R-(+)-PLA, concreteMicroorganism has: proteus vulgaris, proteus mirabilis, the glutinous proteus of product, Pan Shi deformed rodBacterium and Hao Shi proteus.
2. according to the bacterial classification described in right 1, its incubation can also can be entered at good oxygen condition at anaerobic stateOK.
3. the thalline obtaining according to the cultivation described in right 2, it transforms L-Phe can be at anaerobic stateAlso can carry out at good oxygen condition.
4. according to can directly adding L-Phe length of side thalline limit in the thalline incubation described in right 2Transform and produce R-(+)-PLA.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610086443.6A CN105603009B (en) | 2016-02-15 | 2016-02-15 | A kind of method of microorganism conversion |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610086443.6A CN105603009B (en) | 2016-02-15 | 2016-02-15 | A kind of method of microorganism conversion |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105603009A true CN105603009A (en) | 2016-05-25 |
| CN105603009B CN105603009B (en) | 2019-03-15 |
Family
ID=55983350
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610086443.6A Active CN105603009B (en) | 2016-02-15 | 2016-02-15 | A kind of method of microorganism conversion |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105603009B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925535A (en) * | 2012-11-26 | 2013-02-13 | 厦门大学 | Screening and identification method of electricigen enzyme |
| CN103045514A (en) * | 2012-12-27 | 2013-04-17 | 江南大学 | Proteus mirabilis for synthetizing acidic ethyl urethane hydrolytic enzyme and application thereof |
-
2016
- 2016-02-15 CN CN201610086443.6A patent/CN105603009B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925535A (en) * | 2012-11-26 | 2013-02-13 | 厦门大学 | Screening and identification method of electricigen enzyme |
| CN103045514A (en) * | 2012-12-27 | 2013-04-17 | 江南大学 | Proteus mirabilis for synthetizing acidic ethyl urethane hydrolytic enzyme and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| HENRIKSEN S D等: "A comparison of the phenylpyruvic acid reaction and the urease test in the differentiation of Proteus from other enteric organisms", 《JOURNAL OF BACTERIOLOGY》 * |
| LABOURÉ A M等: "Regulation of Phenylalanine Oxidase Synthesis in Proteus", 《JOURNAL OF BACTERIOLOGY》 * |
| MÜLLER H E等: "Production and degradation of indole by gram-negative bacteria", 《ZENTRALBLATT FÜR BAKTERIOLOGIE MIKROBIOLOGIE UND HYGIENE》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105603009B (en) | 2019-03-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103865855A (en) | Bacillus subtilis strain and applications thereof | |
| CN103146786A (en) | Method for producing adenosine by sequentially controlling fermentation with gradient pH | |
| CN105316257A (en) | lysinibacillus xylanilyticus and method for preparing alpha-ketoacid with same | |
| CN103146595B (en) | Bacillus subtilis and method for fermentation production of D- ribose | |
| CN105543292A (en) | Microbial transformation method | |
| CN106479923B (en) | The lactobacillus fermenti of one plant of degrade simultaneously arginine and urea | |
| CN105603006A (en) | Microorganism converting method | |
| CN105543290A (en) | Microbial transformation method | |
| CN102433288A (en) | Strain for producing ornithine and method for biologically synthesizing ornithine with same | |
| CN104087535A (en) | Method for preparing theanine by biological process | |
| CN103409486A (en) | Method for improving adenosine fermentation output through feeding hypoxanthine | |
| CN101270346B (en) | Butachlor degradation bacterium and application thereof | |
| CN102433289B (en) | Strain for producing citrulline and method for biologically synthesizing citrulline with same | |
| CN102268389B (en) | Bacillusmethylotrophicus and method using same for producing gamma-polyglutamic acid by fermentation | |
| CN105670975A (en) | Pseudomonas for mass production of exopolysaccharides and application thereof | |
| CN105779363B (en) | A kind of method of liquefied Serratia and its Synthesis heliotropin | |
| CN105603009A (en) | Microorganism converting method | |
| CN102433290B (en) | Strain for producing citrulline and method for biologically synthesizing citrulline with same | |
| CN106479922B (en) | The lactobacillus plantarum of one plant of degrade simultaneously arginine and urea | |
| CN100467587C (en) | Pseudomonas zjwp-14 and its uses in asymmetrical hydrolyzed preparation of L-cysteine | |
| CN105543291A (en) | Microbial transformation method | |
| CN100564515C (en) | One strain Bordetella and the application in preparation rCO and courage steroid-4-alkene-3-ketone thereof | |
| CN105603007A (en) | Microorganism converting method | |
| CN105624217A (en) | Microbial transformation method | |
| CN100510090C (en) | Process for producing theanine |
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 | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230329 Address after: Floor 20, Unit 2, Building 1, Jinlan West Jingyuan, No. 56, Shinan Road, Science Avenue, High-tech Industrial Development Zone, Zhengzhou City, Henan Province, 450000 Patentee after: Zhuohong Chaoyuan Biotechnology (Zhengzhou) Co.,Ltd. Address before: No. 1800 road 214122 Jiangsu Lihu Binhu District City of Wuxi Province Patentee before: Jiangnan University |