CN1724645A - Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine - Google Patents

Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine Download PDF

Info

Publication number
CN1724645A
CN1724645A CNA2005100138671A CN200510013867A CN1724645A CN 1724645 A CN1724645 A CN 1724645A CN A2005100138671 A CNA2005100138671 A CN A2005100138671A CN 200510013867 A CN200510013867 A CN 200510013867A CN 1724645 A CN1724645 A CN 1724645A
Authority
CN
China
Prior art keywords
histidine
mutant strain
resistance
glutamic acid
producing
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
Application number
CNA2005100138671A
Other languages
Chinese (zh)
Inventor
陈宁
刘辉
史楠
刘淑云
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tianjin University of Science and Technology
Original Assignee
Tianjin University of Science and Technology
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Tianjin University of Science and Technology filed Critical Tianjin University of Science and Technology
Priority to CNA2005100138671A priority Critical patent/CN1724645A/en
Publication of CN1724645A publication Critical patent/CN1724645A/en
Pending legal-status Critical Current

Links

Landscapes

  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to a glutamic acid bacilli mutant TL1105 that uses glutamic acid bacilli ATCC13032 as source fungus, selects 5-methyl tryptophan resistance, sulfaguanidine resistance, 5-fluorin tryptophane resistance, 8-azaguanine resistant, 6-meraptopurine resistant, and 2-thiazole alanine resistant genetic marker to cultivate TL1105. In the application in producing L-histidine by fermentation method, put the TL1105 into culture media for fermenting and cultivating. The PH value is 6.0-8.0, and the temperature is 20-40degree centigrade. Surging cultivating or fermenting cultivating for 2-5 days, and the inoculum size is 1%-10% (V/V). The invention could sharply increase the productivity of L-histidine.

Description

Glutamic acid rod bacillus mutant strain TL 1105 and the application in fermentation method of producing L-histidine
[technical field]: the invention belongs to the biochemical engineering field, relate to a kind of Corynebacterium (Corynebacteriumglutamicum) bacterial classification and use the method that this strain fermentation is produced the L-Histidine.
[background technology]: the chemistry of L-Histidine (L-Histidine) is called L-pantonine-imidazolylpropionic acid, is the basic aminoacids that contains imidazole nucleus in the molecule.The L-Histidine has different physiological roles, is widely used in medicine, feed and food service industry, and especially the effect in medical research comes into one's own day by day.
At present, the production method of L-Histidine mainly contains two kinds:
The one, the proteolysis method, being about to animal blood meal or hair or hoof first is raw material, at first carries out acid hydrolysis, with hydrolyzed solution concentrating under reduced pressure, activated carbon decolorizing, upper prop, after refining Histidine.The main drawback of this method is the cost height, and productive rate is low, and is seriously polluted, and no commercial exploitation is worth.
The 2nd, direct fermentation.Be to have self needed amino acid whose ability of synthesizing by means of microorganism, by mutagenic treatment to specified microorganisms, select auxotroph or analog resistant strain, with feedback inhibition and the feedback repression in the releasing metabolic regulation, thereby reach certain amino acid whose purpose of excess accumulation.At present, Histidine is carbon source mostly with glucose, obtains resistant mutant strain direct fermentation with brevibacterium flavum, Corynebacterium glutamicum, clayey Sai Shi bacillus etc. through inducing.It is low that this method has a raw materials cost, wide material sources, and the product purity height, advantages such as reaction conditions gentleness have the potentiality of suitability for industrialized production.
At present at home, the microorganism direct fermentation of L-Histidine still is in the strain improvement stage, and output all is lower than 10g/L.Up to now, the domestic patent application of still not having L-Histidine microorganism direct fermentation.
[summary of the invention]: one of the object of the invention provides a kind of Corynebacterium glutamicum (Corynebacterium glutamicum) of Corynebacterium, and this bacterium has the ability that improves L-Histidine productive rate.
Two of the technical issues that need to address of the present invention provide the production method that adopts above-mentioned strain fermentation to produce the L-Histidine.
Corynebacterium glutamicum provided by the invention (Corynebacterium glutamicum) mutant strain TL 1105, Corynebacterium glutamicum ATCC13032 (being preserved in Chinese industrial microbial strains preservation administrative center deposit number is CICC10226) with Corynebacterium is the bacterial classification that sets out, and has 5-methyl tryptophan resistance (5-MT by seed selection progressively r), Sulphaguanidine resistance (SG r), 5-fluorotryptophan resistance (5-FT r), 8-azaguanine resistance (8-AG r), Ismipur resistance (6-MP r), 2-thiazole L-Ala resistance (2-TA r) mutant strain TL 1105 of genetic marker, make it have the biosynthesis ability of L-Histidine.
The application of Corynebacterium glutamicum (Corynebacterium glutamicum) mutant strain TL 1105 in fermentation method of producing L-histidine:
This method adds fermentation culture in the fermention medium with glutamic acid rod bacillus mutant strain TL 1105, wherein, carbon source content should be 1%～30% in the fermention medium, carbon-nitrogen ratio N: C should be between 5: 100～5 (between 0: 100, inorganic salt content should be 0.0001～10%, the initial p H6.0 of fermention medium ~ 8.0,20 ℃ ~ 40 ℃ of culture temperature, shaking culture or fermentor cultivation 2 ~ 5 days, inoculum size are 1% ~ 10% (V/V); In the fermenting process, add lime carbonate and stream and add PH6.0 ~ 8.0 that urea, ammoniacal liquor or sodium hydroxide are regulated fermention medium.
Carbon source can be utilized sugar, glucose, sucrose, fructose, maltose, seminose, starch and syrup; Or utilize sugar alcohol, glycerine; Or organic acid, acetic acid; Or low mass molecule alcohol, ethanol; Carbon source content the best is 10%～20%.
Nitrogenous source can utilize ammonia or ammonium salt: ammonium sulfate, ammonium acetate, ammonium nitrate, ammonium phosphate, ammonium acetate or ammonium chloride; Also can utilize organonitrogen: peptone, extractum carnis, corn steep liquor, or soya-bean cake hydrolyzed solution.
Inorganic salt can utilize potassium phosphate salt, sal epsom, lime carbonate, ferrous sulfate, or manganous sulfate.
Can add nutrition in the fermention medium, amino acid and VITAMIN.
Advantage of the present invention and positively effect: the present invention utilizes Corynebacterium glutamicum (Corynebacteriumglutamicum) mutant strain fermentative production L-Histidine, can improve the productive rate of L-Histidine greatly, and its output all is higher than 12g/L.
Utilize purine or pyrimidine structure analogue such as 8-azaguanine resistance (8-AG r) mutant strain, Ismipur resistance (6-MP r) mutant strain, utilize tryptophane analog such as 5-methyl tryptophan resistance (5-MT r) mutant strain, 5-fluorotryptophan resistance (5-FT r) mutant strain, can remove it to the feedback inhibition of key enzyme phosphoribose pyrophosphokinase and check, strengthen HMP metabolism distributions, increase the biosynthesizing of precursor PRPP, help the further accumulation of Histidine;
Utilize Histidine analog such as 2-thiazole L-Ala resistance (2-TA r) mutant strain can remove the feedback regulation of Histidine to its route of synthesis, excessive synthetic Histidine effectively;
Sulfa drugs is the competitive inhibitor of the necessary metabolite para-amino benzoic acid of bacterial growth (PABA).They combine with dihydrofolate synthetase competitively, stop or have replaced para-amino benzoic acid to be incorporated in the folate molecule and to go, thereby blocked the synthetic of bacterial cell important component folic acid.Utilize sulfa drugs such as Sulphaguanidine resistance (SG r) mutant strain, thereby the restraining effect of releasing Sulphaguanidine, and cause a large amount of synthetic of tetrahydrofolic acid (THFA).Tetrahydrofolic acid (THFA) is the coenzyme of plurality of enzymes, plays a carbon back transferance, thereby very important to the eubolism of keeping thalline.
[description of drawings]:
Fig. 1 is that Corynebacterium glutamicum TL1105 is at 5L jar batch fermentation conditional curve figure.
[embodiment]:
Embodiment 1:
Provide among the present invention, that be used to produce the L-Histidine is a kind of Corynebacterium glutamicum (Corynebacterium glutamicum) TL1105 (hereinafter to be referred as TL1105) that belongs to Corynebacterium, and this bacterial classification original strain is ATCC13032 (being preserved in Chinese industrial microbial strains preservation administrative center).Pass through many ethyl sulfates of ATCC13032 (DES) mutagenic treatment, and adopt the filter paper method to select the interior single bacterium colony of inhibition zone, have 5-methyl tryptophan resistance (5-MT by shaking bottle primary dcreening operation and multiple sieve, progressively selecting r), Sulphaguanidine resistance (SG r), 5-fluorotryptophan resistance (5-FT r) mutant strain of genetic marker, 8-azaguanine resistance (8-AG r) mutant strain of genetic marker, Ismipur resistance (6-MP r) mutant strain of genetic marker, 2-thiazole L-Ala resistance (2-TA r) etc. the mutant strain of genetic marker, finally just can obtain TL1105 through separation and purification.
The L-Histidine is produced the seed selection of bacterial strain:
The first step, mutagenesis primary dcreening operation:
Will be after the conventional mutagenesis of DES bacterial strain, make bacteria suspension through centrifuge washing, coat minimum medium flat board (glucose 20g/L, (NH after the dilution 4) 2SO 410g/L, KH 2PO 43H 2O 1g/L, MgSO 47H 2O 0.4g/L, FeSO 47H 2O 0.01g/L, MnSO 4H 2O 0.01g/L, vitamin H 100 μ g/L, VB 1100 μ g/L, agar powder 20g/L, PH7.0～7.2), in every flat board, place an aseptic filter paper sheet that speckles with the resistance soup again, 30 ℃ of constant temperature leave standstill to be cultivated 4～6 days, selected the single bacterium colony in the inhibition zone around the filter paper, and preserved in the inclined-plane and wait to sieve.
Second step, multiple sieve
Shake on bottle level former bacterial strain and each mutant strain are produced L-Histidine performance relatively
Picking one encircles former bacterial strain and each mutant strain lawn (glucose 25g/L, (NH in seed culture medium respectively from fresh inclined-plane 4) 2SO 45g/L, corn steep liquor 40mL, KH 2PO 43H 2O 1.0g/L, MgSO 47H 2O 0.5g/L, CaCO 310g/L, pH7.0～7.2), 500mL triangular flask liquid amount 40mL, 9 layers of gauze seal.Place (170r/min) on the rotary shaking table 30 ℃ of shaking culture 11 hours.With inoculum size 10%, initial pH7.0 inserts (glucose 150g/L, (NH in the fermention medium 4) 2SO 435g/L, corn steep liquor 10mL, KH 2PO 43H 2O 1.0g/L, MgSO 47H 2O 0.5g/L, CaCO 330g/L), 500mL triangular flask liquid amount 25mL.30 ℃ of shaking culture 72 hours, rotating speed is 200r/min, intermittent flow adds NaOH control pH about 7.0.Former bacterial strain and each mutant strain product L-Histidine performance are relatively seen Table 1.
The screening of table 1 Histidine superior strain and shake flask fermentation result
Bacterial strain genetic marker output (g/L)
ATCC13032 trace
TQ2223 (5-MT r+SG r+5-FT r) 1.03
S037 (5-MT r+SG r+5-FT r) 1.25
A021 (5-MT r+SG r+5-FT r+8-AG r) 3.08
A118 (5-MT r+SG r+5-FT r+8-AG r) 4.16
M011 (5-MT r+SG r+5-FT r+8-AG r+6-MP r) 6.81
M018 (5-MT r+SG r+5-FT r+8-AG r+6-MP r) 7.19
T007 (5-MT r+SG r+5-FT r+8-AG r+6-MP r+2-TA r) 9.27
T110 (5-MT r+SG r+5-FT r+8-AG r+6-MP r+2-TA r) 10.80
TL1105 (5-MT r+SG r+5-FT r+8-AG r+6-MP r+2-TA r) 12.02
By table 1 as seen, the product L-Histidine performance of mutant strain TL 1105 is the strongest, and T007 and T110 take second place.
Embodiment 2:
Utilize bacterial classification of the present invention to produce the L-Histidine:
Accumulate the L-Histidine by in substratum, cultivating, can effectively produce the L-Histidine as the above-mentioned Corynebacterium glutamicum that obtains with in substratum.
In order to utilize Corynebacterium glutamicum of the present invention to produce the L-Histidine, can utilize and contain carbon source, nitrogenous source and inorganic salt and other nutritive substance of trace such as the substratum of amino acid and VITAMIN are cultivated.
Different carbon-nitrogen ratios (N: C) to the influence of fermentation and acid
In seed culture medium, inclined-plane seed culture and seed and fermentation culture conditions are all with embodiment 1 from the lawn of fresh inclined-plane picking one ring TL1105 bacterial strain, and the seed culture based component comprises: glucose 30g/L, (NH 4) 2SO 43g/L, corn steep liquor 50mL, soya-bean cake hydrolyzed solution 20mL, KH 2PO 43H 2O 1.0g/L, MgSO 47H 2O0.4g/L, MnSO 4H 2O 0.01g/L, FeSO 47H 2O 0.0 1G/L, VB 1300 μ/L, VH200 μ g/L, CaCO 33g/L, pH7.2.The fermentation culture based component comprises: glucose 150g/L, corn steep liquor 10mL, soya-bean cake hydrolyzed solution 25mL, KH 2PO 43H 2O 1.0g/L, MgSO 47H 2O 0.4g/L, MnSO 4H 2O 0.01g/L, FeSO 47H 2O 0.01g/L, VB 1100 μ g/L, VH 50 μ g/L, CaCO 330g/L, stream adds NaOH adjusting PH7.2 in the fermenting process.(the NH of different content 4) 2SO 4Be added in the fermention medium, different Histidine output are listed in the table 2.
As shown in Table 2, (N: during C) less than 5: 100, Histidine output is relatively low when carbon-nitrogen ratio.On the other hand, (N: during C) greater than 5: 100, Histidine output is higher when carbon-nitrogen ratio.
The bottle that shakes of table 2 different nitrogen contents produces sour result relatively
(NH 4) 2SO 4% N% C: N output (g/L)
0.5 0.11 100∶1.8 5.9
1.0 0.21 100∶3.5 6.2
2.0 0.43 100∶7.2 8.5
3.0 0.64 100∶10.7 14.2
4.0 0.85 100∶14.2 14.5
5.0 1.05 100∶17.5 14.0
6.0 1.26 100∶21.0 14.8
7.0 1.47 100∶24.5 15.4
8.0 1.68 100∶28.0 17.0
9.0 1.89 100∶31.5 16.5
10.0 2.10 100∶35.0 16.6
Embodiment 3
In seed culture medium, inclined-plane seed culture and seed culture condition are all with embodiment 1 from the lawn of fresh inclined-plane picking one ring TL1105 bacterial strain, and seed culture medium inserts in the fermention medium with 5% inoculum size with embodiment 2.The fermentation culture based component comprises: glucose 128g/L, (NH 4) 2SO 4100g/L, corn steep liquor 10mL, the dense 20mL of beans, KH 2PO 43H 2O 1.2g/L, MgSO 47H 2O 0.5g/L, MnSO 4H 2O 0.02g/L, FeSO 47H 2O 0.02g/L, VB 1100 μ g/L, VH 50 μ g/L, CaCO 330g/L., 30 ℃ of shaking culture 72 hours, rotating speed is 200r/min, stream adds NaOH and regulates PH7.2 in the fermenting process.Producing the L-Histidine after the fermentation ends is 20.91g/L.
Embodiment 4
Encircle the lawn of TL1105 bacterial strain in seed culture medium from fresh inclined-plane picking one, inclined-plane seed culture and seed and fermention medium are all with embodiment 1, by 10% inoculum size seed liquor 250mL is inserted in the 5L fermentor tank, initial loading liquid measure 2.5L, ventilation 500L/h, mixing speed 600r/min, 30 ℃ of culture temperature are by auto-feeding ammonia soln control pH7.0 ± 0.05.Between yeast phase, survey residual sugar every the 2h sampling, reducing to 2～3g/L when residual sugar is pre-feed supplement.Determine the feed supplement amount according to sugar consumption rate, keep glucose concn 2～6g/L.Feed supplement finishes, and residual sugar is lower than 2g/L and finishes fermentation.The TL1105 bacterial strain enters behind the 15h in fermentation and produces the acid phase, reaches at about 60h and produces sour peak period, and its high acid amount reaches 14.8g/L.
Referring to Fig. 1, TL1105 is at 5L jar batch fermentation conditional curve.

Claims (5)

1, a kind of Corynebacterium glutamicum (Corynebacterium glutamicum) mutant strain TL 1105, Corynebacterium glutamicum ATCC13032 with Corynebacterium (is preserved in Chinese industrial microbial strains preservation administrative center, deposit number is CICC10226) be the bacterial classification that sets out, have 5-methyl tryptophan resistance (5-MT by seed selection progressively r), Sulphaguanidine resistance (SG r), 5-fluorotryptophan resistance (5-FT r), 8-azaguanine resistance (8-AG r), Ismipur resistance (6-MP r), 2-thiazole L-Ala resistance (2-TA r) mutant strain TL 1105 of genetic marker, make it have the biosynthesis ability of L-Histidine.
2, the application of a kind of Corynebacterium glutamicum (Corynebacterium glutamicum) mutant strain TL 1105 in fermentation method of producing L-histidine, it is characterized in that: glutamic acid rod bacillus mutant strain TL 1105 is added fermentation culture in the fermention medium, wherein, carbon source content should be 1%～30% in the fermention medium, carbon-nitrogen ratio N: C should be between 5: 100～50: 100, inorganic salt content should be 0.0001～10%, the initial p H6.0 of fermention medium～8.0,20 ℃～40 ℃ of culture temperature, shaking culture or fermentor cultivation 2～5 days, inoculum size are 1%～10% (V/V); In the fermenting process, add lime carbonate and stream and add PH6.0～8.0 that urea, ammoniacal liquor or sodium hydroxide are regulated fermention medium.
3, the application of glutamic acid rod bacillus mutant strain TL 1105 according to claim 2 in fermentation method of producing L-histidine is characterized in that the carbon source in the fermention medium can be utilized sugar, glucose, sucrose, fructose, maltose, seminose, starch and syrup; Or utilize sugar alcohol, glycerine; Or organic acid, acetic acid; Or low mass molecule alcohol, ethanol; Carbon source content the best is 10%～20%.
4, according to claim 2 or 3 application of described glutamic acid rod bacillus mutant strain TL 1105 in fermentation method of producing L-histidine, it is characterized in that the nitrogenous source in the fermention medium can utilize ammonia or ammonium salt: ammonium sulfate, ammonium acetate, ammonium nitrate, ammonium phosphate, ammonium acetate or ammonium chloride; Also can utilize organonitrogen: peptone, extractum carnis, corn steep liquor, or soya-bean cake hydrolyzed solution.
5, the application of glutamic acid rod bacillus mutant strain TL 1105 according to claim 2 in fermentation method of producing L-histidine is characterized in that inorganic salt, can utilize potassium phosphate salt, sal epsom, lime carbonate, ferrous sulfate, or manganous sulfate.
CNA2005100138671A 2005-06-22 2005-06-22 Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine Pending CN1724645A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CNA2005100138671A CN1724645A (en) 2005-06-22 2005-06-22 Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CNA2005100138671A CN1724645A (en) 2005-06-22 2005-06-22 Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine

Publications (1)

Publication Number Publication Date
CN1724645A true CN1724645A (en) 2006-01-25

Family

ID=35924275

Family Applications (1)

Application Number Title Priority Date Filing Date
CNA2005100138671A Pending CN1724645A (en) 2005-06-22 2005-06-22 Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine

Country Status (1)

Country Link
CN (1) CN1724645A (en)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100455670C (en) * 2006-11-21 2009-01-28 江南大学 Strain for production of L-serine and method for production of L-serine by using same
CN102391977A (en) * 2011-12-01 2012-03-28 天津科技大学 Corynebacterium glutamicum and production method of alpha-ketoglutarate through fermentation thereof
CN110396493A (en) * 2019-09-09 2019-11-01 廊坊梅花生物技术开发有限公司 The method of culture medium combination and production isoleucine
CN112118747A (en) * 2017-12-08 2020-12-22 驰若莫塞尔公司 Tryptophan derivatives as sweeteners

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100455670C (en) * 2006-11-21 2009-01-28 江南大学 Strain for production of L-serine and method for production of L-serine by using same
CN102391977A (en) * 2011-12-01 2012-03-28 天津科技大学 Corynebacterium glutamicum and production method of alpha-ketoglutarate through fermentation thereof
CN102391977B (en) * 2011-12-01 2013-03-20 天津科技大学 Corynebacterium glutamicum and production method of alpha-ketoglutarate through fermentation thereof
CN112118747A (en) * 2017-12-08 2020-12-22 驰若莫塞尔公司 Tryptophan derivatives as sweeteners
EP3720295A4 (en) * 2017-12-08 2021-09-08 Chromocell Corporation Tryptophan derivatives as sweeteners
CN110396493A (en) * 2019-09-09 2019-11-01 廊坊梅花生物技术开发有限公司 The method of culture medium combination and production isoleucine
CN110396493B (en) * 2019-09-09 2021-09-07 廊坊梅花生物技术开发有限公司 Culture medium composition and method for producing isoleucine

Similar Documents

Publication Publication Date Title
CN100999756B (en) Process of preparing gamma-polyglutamic acid by bacillus subtilis and glutamic acid bacillus mixed cultivating system
CN1169946C (en) L-arginine producing strain and its mutation method and usage in producing L-arginine
CN1884565B (en) Process for producing D-alanine using microbe
CN1844356A (en) Yellow bacillus brevis mutant and process for fermentation production of L-valine by using same
CN1181191C (en) Microorganism producing 5'-inosinic acid and process for producing 5'-inosinic acid using the same
CN1724645A (en) Glutamic acid rod bacillus mutant strain TL1105 and application in fermentation method of producing L-histidine
Razak et al. Optimization of fermentation upstream parameters and immobilization of Corynebacterium glutamicum MH 20-22 B cells to enhance the production of L-lysine
CN1834227A (en) Yellow graminic mutant strain and prodn. technique for producing L-leucine by fermentation process
CN101235402B (en) Fermentation technique for producing stability isotope 15N marking L-leucine
CN1834228A (en) Yellow graminic mutant strain and its application in prodn. of L-isoleucine by fementation process
CN100455670C (en) Strain for production of L-serine and method for production of L-serine by using same
CN1140761A (en) Process for producing L-leucine
CN1225552C (en) Microorganism for producing riboflavin and method for producing riboflavin using the same
CN1869244A (en) Production method of 15N stability isotop labeling L-arginine
JP3245881B2 (en) Simultaneous fermentation of basic and acidic amino acids
CN117757859A (en) Method for co-producing L-isoleucine and L-valine through microbial fermentation
CN1977048B (en) Fermentation of L-lysine using acid-treated product of soybean meal
CN1218043C (en) Method for producing L-glutamic acid by continuous fermentation
CN1117869C (en) Process for producing L-lysine by fermentation
JPS5886094A (en) Preparation of l-lysine by fermentation
JPS5828292A (en) Preparation of l-lysine by fermentation
KR0181297B1 (en) Process for producing l-lysine by fermentation
JPH01199589A (en) Production of l-lysine by fermentation method
US20060270004A1 (en) Fermentation processes with low concentrations of carbon-and nitrogen-containing nutrients
Irshad et al. Optimization of physico-chemical parameters for hyperproduction of lysine by mutated strain of Brevibacterium flavum.

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C12 Rejection of a patent application after its publication
RJ01 Rejection of invention patent application after publication