CN102703339B - High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same - Google Patents
High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same Download PDFInfo
- Publication number
- CN102703339B CN102703339B CN 201110250427 CN201110250427A CN102703339B CN 102703339 B CN102703339 B CN 102703339B CN 201110250427 CN201110250427 CN 201110250427 CN 201110250427 A CN201110250427 A CN 201110250427A CN 102703339 B CN102703339 B CN 102703339B
- Authority
- CN
- China
- Prior art keywords
- citrulline
- parts
- glucose
- bacterial strain
- yield
- 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.)
- Active
Links
Abstract
The invention discloses a high-yield arginine deiminase bacterial strain and a method for producing L-citrulline by the same. The high-yield arginine deiminase bacterial strain of the invention is streptococcus faecalis BM-2 CGMCC No.4990. After small-scale fermentation of the bacterial strain in a 30-L fermentation tank, the L-citrulline yield is up to 98 g/L, which is increased by 50% when compared with the L-citrulline yield of the original strain; after continuous fermentation for 5 batches, the L-citrulline yield of the bacterial strain is stable, and the L-citrulline yield level is obviously higher than that of reported bacterial strains; and the bacterial strain of the invention has important industrial production application value.
Description
Technical field
The present invention relates to high yield arginine deiminase bacterial strain and produce the method for L-citrulline with it, belong to biological technical field.
Background technology
(Arginine deiminase ADI), is a kind ofly can decompose the arginic protein of L-to arginine deiminase.Scholars at first from mycoplasma Mycoplasma arginini extract purifying obtain ADI, discover that this enzyme can suppress the animal blood vessels endotheli ocytosis, correlative study further confirms, arginine deiminase can not only suppress the in-vitro multiplication of multiple malignant cell, can also suppress the hyperplasia of the malignant tumour in the body.Arginine deiminase has been subjected to more and more investigators' attention as a kind of new antitumoral material.
It is reported that arginine deiminase can be converted into the L-citrulline by catalysis L-arginine, this enzymatic reaction is a step only, can effectively avoid feedback regulation effect complicated in the complete synthesis approach of L-citrulline, makes the L-citrulline can run up to higher concentration.The L-citrulline has a lot of important physical functions, as the removing free radical, and vasorelaxation action, stabilizing blood pressure and diagnostics classes rheumatic arthritis etc. share in the treatment hyperammonemia with L-ornithine, L-arginine etc., and application prospect is very wide.This shows that the arginine deiminase effect is extensive, have very important exploitation and be worth.
The arginine deiminase of reporting before this comes from mycoplasma mostly, and mycoplasma is pathogenic micro-organism, causes disease easily, bigger danger is arranged, and mycoplasma is microparasite, and the cultivation difficulty is bigger, is unfavorable for the fermentative production of arginine deiminase.It is streptococcus faecium that the arginine deiminase of studying is herein produced bacterial strain, and this bacterial strain security is good, is easy to fermentative preparation, is conducive to scale operation and the clinical application of arginine deiminase.But present result of study does not reach gratifying level yet, mainly is because spawn activity is lower, is difficult to industrial applications.Mutagenic and breeding is to utilize physics or chemical factor to handle microorganism cells colony, impel genetic material (mainly the being DNA) structure in a few cell wherein to change, change thereby cause the microorganism hereditary proterties, manage from colony, to filter out the process of a small amount of good mutant strain then.The mutagenesis screening superior strain is the focus method that domestic and international researcher is studied in this respect always.
Ultraviolet mutagenesis is easy to drawing materials with respect to other several rays (X ray, β ray, gamma-rays etc.), and is also minimum to the injury of human body, is the most simple and convenient and highly effective a kind of physical mutagenesis method again.Chemomorphosis is to use the genetic material of chemical mutagen and organism to have an effect, and changes its structure, a kind of mutafacient system that its offspring is morphed.Chemical mutagen induced mutation rate height, and chromosome aberration is less, and light to handling material damage, the chemical mutagen that has is only limited to some privileged site of DNA.
Summary of the invention
The objective of the invention is to solve the defective that existing arginine deiminase is produced bacterial classification, solve than enzyme low problem alive.The invention provides a kind of method of ultraviolet ray, ethyl sulfate complex mutation of utilizing arginine deiminase production bacterial strain is carried out selection by mutation, screening has obtained the mutant strain of plant height product arginine deiminase, and this mutant strain is fermented, utilize arginine deiminase energy catalysis L-arginine to transform into the character of L-citrulline, by detecting the amount of newly-generated L-citrulline, calculate the vigor of arginine deiminase, find that the enzyme activity of mutant strain is greatly improved.
The present invention is starting strain with streptococcus faecium (streptococcus faecalis) ATCC 10180, utilize the method for ultraviolet ray, ethyl sulfate complex mutation to carry out selection by mutation, screening has obtained the mutant strain of plant height product arginine deiminase, i.e. streptococcus faecium (streptococcus faecalis) BM-2.This bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center and (has been called for short CGMCC on 06 27th, 2011, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica), preserving number is CGMCC No.4990.
Streptococcus faecium of the present invention (streptococcus faecalis) BM-2 CGMCC No.4990 has following Microbiological Characteristics: the thalline diameter is between 0.5-2 μ m, solid culture is mainly based on diplococcus, also have to be 3-5 the short chain of arranging on a small quantity, liquid culture is based on long-chain; Bacterium colony is circle, oyster white, and surperficial dimpling is moistening, and neat in edge is translucent, diameter 1-2mm.
Cultivation and the fermentation of streptococcus faecium of the present invention (streptococcus faecalis) BM-2CGMCC No.4990:
(1) solid medium and seed culture medium (latter does not contain agar) are made up of following components in part by weight:
1000 parts in casein peptone 5-50 part, extractum carnis 5-50 part, yeast extract 1-30 part, glucose 1-30 part, sodium acetate 1-30 part, dibasic ammonium citrate 1-30 part, tween 80 1-30 part, dipotassium hydrogen phosphate 1-30 part, magnesium sulfate heptahydrate 0.1-1 part, manganese sulfate monohydrate 0.01-0.1 part, agar 10-50 part and water;
Culture condition: pH 6.0-8.0, culture temperature is 25-40 ℃, incubation time 20-60h.
(2) fermention medium is formed (weight part): 1000 parts in glucose 5-50 part, corn steep liquor 2-50 part, peptone 5-50 part, extractum carnis 2-50 part, arginase 12-50 part, potassium primary phosphate 1-20 part, magnesium sulfate heptahydrate 0.1-1 part, sodium-chlor 1-20 part, manganese sulfate monohydrate 0.01-0.05 part and water.
Method so that streptococcus faecium (streptococcus faecalis) BM-2CGMCC No.4990 produces the L-citrulline is characterized in that,
(1) get a glycerine pipe of preserving streptococcus faecium (streptococcus faecalis) BM-2CGMCC No.4990, room temperature is placed to thawing, and absorption 0.5-1ml bacterium liquid is seeded to first order seed and shakes in the bottle, cultivates 20-60h for 25-40 ℃;
(2) be seeded to secondary seed according to 1% inoculum size and shake in the bottle, cultivate 20-60h for 25-40 ℃;
(3) be seeded in the fermentor tank according to the inoculum size of 1-5% then, cultivate 20-60h, culture condition: temperature 25-40 ℃, rotating speed 150-300r/min, pH 6.0-7.0; Flow feeding liquid in the fermenting process (glucose and ammoniacal liquor) makes that the content of glucose is not less than 0.5g/L in the fermented liquid, and pH maintains between the 6.0-7.0.
The invention has the beneficial effects as follows:
First, the arginine deiminase mutant strain that the present invention relates to is fermented through 30L fermentor tank lab scale, produce L-citrulline amount and reach 98g/L, produce L-citrulline amount than starting strain and improved 50%, and continuously ferment after 5 batches, it is stable to find that this bacterium produces the L-citrulline, produces L-citrulline level apparently higher than the bacterial strain of having reported, has important industrial production using value.
The second, a strain arginine deiminase high productive mutant provided by the invention produces acid amount height, and proterties is stable, has important economic benefit and social benefit, also lays a good foundation for making up genetic engineering bacterium.
Embodiment
The seed selection of embodiment 1 high yield arginine deiminase bacterial strain BM-2
(1) preparation of starting strain ATCC 10180 bacteria suspensions:
Streptococcus faecium (streptococcus faecalis) ATCC10180 bacterial strain with aseptic inoculation pin picking 1 ring inclined-plane is preserved carries out streak culture at solid medium.The activation back new fresh thalli of picking one ring dilutes with Sterile Saline, and bacterial concentration is diluted to 10
7Individual/ml, obtain bacteria suspension.
Solid medium and seed culture medium (latter does not contain agar) are made up of weight components: 1000 parts in 10 parts of casein peptones, 20 parts of extractum carniss, 20 parts of yeast extracts, 20 parts of glucose, 10 parts of sodium acetates, 5 parts of dibasic ammonium citrates, 2 parts of tween 80s, 2 parts of dipotassium hydrogen phosphates, 0.1 part of magnesium sulfate heptahydrate, 0.05 part of manganese sulfate monohydrate, 20 parts in agar and water:
The processing condition pH 7.0 for preparing dull and stereotyped bacterial classification, culture temperature is 37 ℃, incubation time 24h.
(2) starting strain ultraviolet mutagenesis:
Absorption 0.1ml dilutes good bacteria suspension and is coated on the solid plate substratum, and behind ultraviolet lamp (power 20W, irradiation distance 35cm) the irradiation certain hour (2min), 30 ℃ of following lucifuges are cultivated 24h, observations.Select edge-smoothing, the of uniform size bacterium colony of lethality rate on the flat board of 70-80%, carry out the test of fermentation shake flask primary dcreening operation.
(3) the fermentation shake flask primary dcreening operation of ultraviolet mutation bacterial strain:
After the mutagenesis, select the well-grown inoculation of 200 strains to slant medium from flat board and cultivate.Bacterial strain on the picking one ring slant medium inserts in the 100ml seed culture medium, and 37 ℃, 200r/min cultivates 24h.Cultivation finishes, and seed liquor is seeded in the 200ml fermention medium according to 1% ratio, and 37 ℃, 200r/min cultivates 24h.Fermented liquid is centrifugal, abandon precipitation, measure the L-citrulline content in the supernatant liquor.From 200 plant mutant strains, screen the bacterial strain that 20 strain L-citrulline output improve, to the test of going down to posterity of 20 strain bacterium, when reaching for the 10th generation, carry out the 30L jar and test.
Fermention medium is made up of following parts by weight of component: 1000 parts in 40 parts of glucose, 20 parts of corn steep liquors, 5 parts of peptones, 10 parts in beef, 10 parts of arginine, 5 parts of potassium primary phosphates, 0.5 part of magnesium sulfate heptahydrate, 2 parts in sodium-chlor, 0.02 part of manganese sulfate monohydrate and water.
(4) the multiple sieve of the 30L jar of ultraviolet mutation bacterial strain fermentation:
The 20 plant mutant strains that obtain behind the ultraviolet primary dcreening operation are activated at flat board, and the single bacterium colony on the plate of making even is connected in the 200ml seed culture medium, and 37 ℃, 200r/min cultivates 24h.In the 30L fermentor tank, inoculum size by 5% inserts in the fermention medium, cultivate 24h, culture condition: 37 ℃, 200r/min, pH7.0,5 cycles of fermentation to the begin flow feeding liquid, make that the content of glucose is not less than 0.5g/L in the fermented liquid, and the pH value maintains between the 6.5-7.0 (by the end of fermentation ends, generally need add glucose 500g, add 20% ammoniacal liquor 500ml).After the fermentation ends, measure L-citrulline output in the fermented liquid.The bacterial strain that improves with L-citrulline output is that starting strain carries out ultraviolet mutagenesis again, primary dcreening operation, multiple sieve.Through the three-wheel mutagenesis screening, finally obtain the highest mutant strain ATCC 10180-A of a strain L-citrulline output, its L-citrulline output reaches 70g/L.
(5) preparation of ultraviolet mutagenesis mutant strain bacteria suspension, ethyl sulfate mutagenesis:
Ultraviolet mutagenesis mutant strain ATCC 10180-A with aseptic inoculation pin picking 1 ring inclined-plane is preserved carries out streak culture at solid medium.The activation back new fresh thalli of picking one ring dilutes with Sterile Saline, and bacterial concentration is diluted to 10
7Individual/ml, obtain bacteria suspension.
Get the 2mL bacteria suspension, add to the phosphoric acid buffer of 15mL pH6.5, to wherein adding the 0.5mL ethyl sulfate, handle 40min for 35 ℃.Get the 0.1mL treatment solution and coat on the solid plate, behind 37 ℃ of cultivation 24h, therefrom select well-grown bacterium colony 300 strains, insert the inclined-plane, 4 ℃ of refrigerators are preserved.
(6) the fermentation shake flask primary dcreening operation of ultraviolet mutagenesis mutant strain:
The bacterial strain that picking one ring inclined-plane is preserved inserts in the 100mL seed culture medium, and 37 ℃, 200r/min shaking culture spend the night.By 1% inoculum size seed culture medium is accessed in the 200mL fermention medium, 37 ℃, 200r/min are cultivated 24h.Fermented liquid is centrifugal, abandon precipitation, measure the content of L-citrulline in the supernatant liquor.From 300 plant mutant strains, screen the bacterial strain that 10 strain L-citrulline output improve, to the test of going down to posterity of this 10 strain bacterium, when reaching for the 10th generation, carry out the 30L jar and test.
(7) the 30L ferment tank of ultraviolet mutagenesis mutant strain sieves again:
10 plant mutant strains are activated at fresh solid plate, and the single bacterium colony on the picking flat board is connected in the 200mL seed culture medium, cultivates 24h for 37 ℃.In the 30L fermentor tank, insert in the fermention medium by 5% inoculum size, cultivate 24h, culture condition: 37 ℃ of temperature, rotating speed 200r/min, pH7.0.8 cycles of fermentation to the begin flow feeding liquid (by the end of fermentation ends, generally need add glucose 800g, add 20% ammoniacal liquor 800ml).After the fermentation ends, measure L-citrulline output in the fermented liquid.Through screening, obtain the highest mutant strain BM-2 of a strain L-citrulline output, its L-citrulline output reaches 98g/L.
(8) with this mutant strain, carry out the glycerine pipe and preserve:
Earlier this bacterial strain is applied in the eggplant bottle, behind 30 ℃ of cultivation 30h, the glycerine with 30% is stored in it in deep cooling pipe, places in-70 ℃ of refrigerators.
(9) mutant strain Microbiological Characteristics: the thalline diameter is between 0.5-2 μ m, and solid culture also has to be 3-5 the short chain of arranging on a small quantity mainly based on diplococcus, and liquid culture is based on long-chain; Bacterium colony is circle, oyster white, and surperficial dimpling is moistening, and neat in edge is translucent, diameter 1-2mm.
Embodiment 2L-citrulline production method
Seed culture medium is formed (weight part): 1000 parts in 7.5 parts of casein peptones, 10 parts of extractum carniss, 5 parts of yeast extracts, 10 parts of glucose, 5 parts of sodium acetates, 2 parts of dibasic ammonium citrates, 1 part of tween 80,4 parts of dipotassium hydrogen phosphates, 0.3 part of magnesium sulfate heptahydrate, 0.03 part of manganese sulfate monohydrate and water;
Fermention medium is formed (weight part): 1000 parts in 5 parts of glucose, 2 parts of corn steep liquors, 10 parts of peptones, 30 parts of extractum carniss, 0 part of arginase 12,5 parts of potassium primary phosphates, 0.5 part of magnesium sulfate heptahydrate, 3 parts in sodium-chlor, 0.03 part of manganese sulfate monohydrate and water.
(1) get a glycerine pipe, room temperature is placed to thawing, and absorption lml bacterium liquid is seeded to first order seed and shakes in the bottle, behind 30 ℃ of cultivation 30h;
(2) be seeded to secondary seed according to 1% inoculum size and shake in the bottle, cultivate 30h for 30 ℃;
(3) be seeded in the 30L fermentor tank according to 5% inoculum size then, cultivate 35h, culture condition: 30 ℃ of temperature, rotating speed 150r/min, pH 6.5-7.0.10 cycles of fermentation to the begin flow feeding liquid (glucose and ammoniacal liquor), make that the content of glucose is not less than 0.5g/L in the fermented liquid, and pH maintains between the 6.5-7.0, by the end of fermentation ends, adds glucose 800g altogether, adds 20% ammoniacal liquor 800ml.After the fermentation ends, L-citrulline content can reach 98.5g/L in the mensuration fermented liquid.
Embodiment 3
Seed culture medium is formed (weight part): 1000 parts in 20 parts of casein peptones, 5 parts of extractum carniss, 10 parts of yeast extracts, 20 parts of glucose, 10 parts of sodium acetates, 10 parts of dibasic ammonium citrates, 10 parts of tween 80s, 10 parts of dipotassium hydrogen phosphates, 0.5 part of magnesium sulfate heptahydrate, 0.05 part of manganese sulfate monohydrate and water;
Fermention medium is formed (weight part): 1000 parts in 20 parts of glucose, 10 parts of corn steep liquors, 20 parts of peptones, 20 parts of extractum carniss, 30 parts of arginine, 10 parts of potassium primary phosphates, 0.8 part of magnesium sulfate heptahydrate, 10 parts in sodium-chlor, 0.02 part of manganese sulfate monohydrate and water.
(1) get a glycerine pipe, room temperature is placed to thawing, and absorption 0.5ml bacterium liquid is seeded to first order seed and shakes in the bottle, behind 37 ℃ of cultivation 24h;
(2) be seeded to secondary seed according to 1% inoculum size and shake in the bottle, cultivate 24h for 37 ℃;
(3) be seeded in the 30L fermentor tank according to 3% inoculum size then, cultivate 30h, culture condition: 37 ℃ of temperature, rotating speed 170r/min, pH 6.5-7.0.8 cycles of fermentation to the begin flow feeding liquid (glucose and ammoniacal liquor), make that the content of glucose is not less than 0.5g/L in the fermented liquid, and pH maintains between the 6.5-7.0, by the end of fermentation ends, adds glucose 700g altogether, adds 20% ammoniacal liquor 650ml.L-citrulline in the fermented liquid is measured, and content reaches 99.5g/L.
Claims (3)
1. a streptococcus faecium (streptococcus faecalis) BM-2 is characterized in that deposit number is CGMCC No.4990.
2. the method for producing the L-citrulline with the described streptococcus faecium BM-2 of claim 1 is characterized in that,
(1) get a glycerine pipe of preserving streptococcus faecium BM-2, room temperature is placed to thawing, and absorption 0.5-1ml bacterium liquid is seeded to first order seed and shakes in the bottle, cultivates 20-60h for 25-40 ℃;
(2) be seeded to secondary seed according to 1% inoculum size and shake in the bottle, cultivate 20-60h for 25-40 ℃;
(3) be seeded in the fermentor tank according to the inoculum size of 1-5% then, cultivate 20-60h, culture condition: temperature 25-40 ℃, rotating speed 150-300r/min, pH6.0-7.0; Stream adds and mends glucose and ammoniacal liquor in the fermenting process, makes that the content of glucose is not less than 0.5g/L in the fermented liquid, and pH maintains between the 6.0-7.0.
3. the method for production as claimed in claim 2 L-citrulline, it is characterized in that the shake-flask seed substratum of described step (1) and (2) is made up of following components by part by weight: 1000 parts in casein peptone 5-50 part, extractum carnis 5-50 part, yeast extract 1-30 part, glucose 1-30 part, sodium acetate 1-30 part, dibasic ammonium citrate 1-30 part, tween 80 1-30 part, dipotassium hydrogen phosphate 1-30 part, magnesium sulfate heptahydrate 0.1-1 part, manganese sulfate monohydrate 0.01-0.1 part and water; The fermention medium of described step (3) is made up of following components by part by weight: 1000 parts in glucose 5-50 part, corn steep liquor 2-50 part, peptone 5-50 part, extractum carnis 2-50 part, arginase 12-50 part, potassium primary phosphate 1-20 part, magnesium sulfate heptahydrate 0.1-1 part, sodium-chlor 1-20 part, manganese sulfate monohydrate 0.01-0.05 part and water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110250427 CN102703339B (en) | 2011-08-29 | 2011-08-29 | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201110250427 CN102703339B (en) | 2011-08-29 | 2011-08-29 | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102703339A CN102703339A (en) | 2012-10-03 |
| CN102703339B true CN102703339B (en) | 2013-08-14 |
Family
ID=46896394
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201110250427 Active CN102703339B (en) | 2011-08-29 | 2011-08-29 | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102703339B (en) |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104805144B (en) * | 2015-05-07 | 2018-05-04 | 江南大学 | A kind of method of efficiently production L-citrulline |
| CN105177075A (en) * | 2015-07-01 | 2015-12-23 | 滨州市生物技术研究院有限责任公司 | Method for preparation of L-citrulline with arginine as raw material |
| CN105176954B (en) * | 2015-07-01 | 2018-06-26 | 山东民强生物科技股份有限公司 | A kind of method that arginine deiminase is prepared using bacterial strain MQO-153 |
| CN105176859B (en) * | 2015-07-01 | 2018-06-26 | 山东民强生物科技股份有限公司 | The bacterial strain MQO-153 of one plant of production arginine deiminase |
| CN105017086B (en) * | 2015-07-01 | 2017-05-03 | 滨州市生物技术研究院有限责任公司 | Separation and purification method for L-citrulline |
| CN106591270B (en) | 2017-01-23 | 2018-09-21 | 江南大学 | One plant of Fixedpoint mutation modified genetic engineering arginine deiminase |
| CN107828682A (en) * | 2017-11-06 | 2018-03-23 | 四川恒通动保生物科技有限公司 | A kind of fermentation medium of alctasin and application |
| CN107916282B (en) * | 2017-12-13 | 2021-06-04 | 湖北新生源生物工程有限公司 | Method for preparing L-citrulline and L-ornithine by biological method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1518595A (en) * | 2000-11-28 | 2004-08-04 | 凤凰药理学公司 | Modified arginine deiminase |
-
2011
- 2011-08-29 CN CN 201110250427 patent/CN102703339B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1518595A (en) * | 2000-11-28 | 2004-08-04 | 凤凰药理学公司 | Modified arginine deiminase |
Non-Patent Citations (4)
| Title |
|---|
| "Control of Enzyme Synthesis in the Arginine Deiminase Pathway of Streptococcus faecalis";JEAN-PAUL SIMON 等;《JOURNAL OF BACTERIOLOGY》;19820630;第150卷(第3期);第1085-1090页 * |
| "粪链球菌转化合成L-瓜氨酸的研究";姚海峰 等;《食品工业科技》;20080630;第29卷(第6期);第256-258页 * |
| JEAN-PAUL SIMON 等."Control of Enzyme Synthesis in the Arginine Deiminase Pathway of Streptococcus faecalis".《JOURNAL OF BACTERIOLOGY》.1982,第150卷(第3期),第1085-1090页. |
| 姚海峰 等."粪链球菌转化合成L-瓜氨酸的研究".《食品工业科技》.2008,第29卷(第6期),第256-258页. |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102703339A (en) | 2012-10-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102703339B (en) | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same | |
| CN102174433B (en) | Clostridium beijerinckii with high stress resistance and application thereof | |
| CN102643770B (en) | Colibacillus capable of generating succinic acid by anaerobic growth in synthetic medium pure and application thereof | |
| CN102597251A (en) | A process for integrated production of ethanol and seaweed sap from kappaphycus alvarezii | |
| CN108624524A (en) | The bacterial strain and its separating screening method of one plant of production bacteria cellulose | |
| CN105132331A (en) | Komagataeibacter nataicola and application thereof | |
| CN109536409A (en) | A kind of resistance is high and using the Pedicoccus acidilacticii strain of several kinds of carbon source and the method for producing lactic acid using the bacterial strain | |
| CN107201315A (en) | A kind of Paecilomyces thermaphila mutagenic strain and its method of mutagenesis and application | |
| CN107760623B (en) | The A Shi bacillus of the neutral uncooked amylum enzyme of one plant of production | |
| CN105754925B (en) | A method of improving Pichia kudriavezii thermo-tolerance | |
| CN103571772A (en) | Novel strain for producing butanol and method for producing butanol | |
| CN102864113B (en) | Strain capable of producing succinic acid, method for producing succinic acid and application thereof | |
| CN106811438A (en) | A kind of straw degradative acidifying microbial inoculum and preparation method thereof | |
| CN102634460B (en) | Rhizopus oryzae RH1-5 and separating and culturing method thereof | |
| CN112239738A (en) | Escherichia coli capable of producing succinic acid and application thereof | |
| CN105969702B (en) | Serratia marcescens RZ 21-C6 and its application | |
| CN103421850A (en) | Method used for producing bioethanol with Scenedesmusabundans | |
| CN100415870C (en) | Nie's brevibacterium new strain and process for preparing enzyme by it | |
| CN110129225A (en) | γ~polyglutamic acid producing strains and breeding prepare γ~polyglutamic acid method | |
| CN110241043A (en) | The bacterial strain of one plant height temperature fermenting lactic acid and the method for producing lactic acid | |
| CN110205250A (en) | One plant of cellulase high-yield and its screening technique and application | |
| CN105838652B (en) | One plant of bacterial strain for strengthening glycerol metabolism and its application | |
| CN102268379B (en) | Trametes trogii and method for producing cellulase by using same | |
| CN104745554B (en) | Bacillus produces the fermentation medium and fermentation process of protease and gemma | |
| CN109161507A (en) | A kind of Corynebacterium glutamicum of high yield L-Orn and its application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same Effective date of registration: 20180428 Granted publication date: 20130814 Pledgee: Zouping Pudong Development Bank and Limited by Share Ltd Pledgor: Shandong NB Biological Engineering Co.,Ltd. Registration number: 2018370000082 |