CN1392246A - Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process - Google Patents
Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process Download PDFInfo
- Publication number
- CN1392246A CN1392246A CN 02113142 CN02113142A CN1392246A CN 1392246 A CN1392246 A CN 1392246A CN 02113142 CN02113142 CN 02113142 CN 02113142 A CN02113142 A CN 02113142A CN 1392246 A CN1392246 A CN 1392246A
- Authority
- CN
- China
- Prior art keywords
- wsh
- pyruvic acid
- torulopsis glabrata
- vitamin
- strain
- 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
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention belongs to the field of bioengineering technology. The bacterium of the present invention is a kind of Torulopsis glabrata, WSH-, obtained with WSH-IP303 as starting strain and through NTG mutagenesis and breeding selection in culture medium with added acetic acid as replenishing carbon source. Compared with the starting strain, LQ307 has mush reduced pyruvate decarboxylase, and strong and stable pyruvate producing capacity. The yield of pyruvate reaches 46.2 g/L, 21% higher than that of starting strain when using acetic acid as replenishing carbon source and through shaking bottle culture for 48 hr; the yield of pyruvate may reach 68.7 g/L the glucose converting rate may reach 0.651 g/g through fermentation in a 5 L fermenting tank for 64 hr.
Description
Technical field
The present invention relates to a kind of acetate leakage type pyruvic acid high yield bacterium and selection thereof and, belong to technical field of bioengineering with this bacterium fermentative Production pyruvic acid.
Background technology
Pyruvic acid (Pyruvic acid) is the important intermediate product in the carbohydrate metabolism.Pyruvic acid is the important as precursors of synthetic multiple amino acids, VITAMIN and other useful matter, is widely used in industry such as medicine, daily use chemicals, agrochemicals, feed food.Though as a kind of Chemicals, pyruvic acid has been realized suitability for industrialized production already, but the pyruvic acid of chemical method production is still continued to use tartrate dehydration decarboxylation method, its main drawback is that the pyruvic acid productive rate is low, seriously polluted and because of expensive raw material price causes the pyruvic acid product price high, applies nature and also is restricted.The fermentative Production pyruvic acid has many advantages such as raw materials cost is low, wide material sources, product purity height, reaction conditions gentleness, has caused investigator's extensive interest.
" microorganism journal " 2000,40 (5): 528～534; " biotechnology journal " 2000,16 (2): 225～228; " industrial microorganism " 2001,31 (2): 10～13; Deng all report is arranged, in the former research, this laboratory (industrial biotechnology key lab of the Ministry of Education of Southern Yangtze University) obtains a strain torulopsis glabrata (Torulopsis glabrata) WSH-IP303 through repeatedly improveing, and is VitB1 (B
1), vitamin H (Bio), nicotinic acid (NA) and pyridoxol (B
6) auxotroph, promptly need these VITAMIN thalline just can keep the growth and accumulate high-caliber pyruvic acid, can be only nitrogen source with ammonium chloride.Nutritional condition, VITAMIN, oxygen-supply control mode etc. have also been studied to the influence of fermentation production of acetone acid and the metabolic characteristics of the excessive synthetic pyruvic acid of Torulopsis glabrata WSH-IP303.The pyruvic acid accumulating level is respectively 38.3g/l and 55.8g/l in shake flat experiment (48h) and 300L fermentor tank (68h), inversion rate of glucose is respectively 0.525g/g and 0.553g/g, but by product alcoholic acid content is higher and the pyruvic acid productive rate is not high in the fermented liquid, may be because the control pyruvic acid to the enzyme work of the pyruvic carboxylase (PDC) of the metabolism branch road of acetaldehyde than higher and make pyruvic acid more active to the metabolism of acetaldehyde, thereby cause the degraded and the generation of by product alcoholic acid of pyruvic acid.
Summary of the invention
The objective of the invention is to propose a kind of acetate leakage type pyruvic acid high yield bacterium and selection thereof and with this bacterium fermentative Production pyruvic acid
The present invention proposes a kind of title and is referred to as torulopsis glabrata (Torulopsis glabrata) WSH-LQ307, be deposited in Wuhan China typical culture collection center, be numbered CCTCC NO.M202019, this bacterial strain is to be starting strain with torulopsis glabrata (Torulops glabrata) WSH-IP303, after nitrosoguanidine (NTG) mutagenic treatment, in substratum, add acetate carbon source as a supplement again, get through cultivating screening and multiple sieve, it is carried out the checking of vitamin defective type, and conclusive evidence still is NA, B
1, B
6Auxotroph with four kinds of VITAMIN of Bio.It is the acetate seepage defect type pyruvic acid high productive mutant that pyruvic carboxylase (PDC) enzyme is lived and reduced.
The selection of torulopsis glabrata (Torulopsis glabrata) WSH-LQ307 bacterial strain is characterized in that through nitrosoguanidine (NTG) mutagenic treatment, adds acetate carbon source as a supplement in the substratum.
Starting strain: torulopsis glabrata (Torulopsis glabrata) WSH-IP303 is nicotinic acid (NA), VitB1 (B
1), pyridoxol (B
6) and the auxotroph of four kinds of VITAMIN of vitamin H (Bio).
Substratum:
Inclined-plane and seed culture medium: glucose 30g, peptone 10g, KH
2PO
41g, MgSO
47H
2O 0.5g, agar 20g (inclined-plane is used), tap water is settled to 1L, pH5.5;
Fermention medium: glucose 100g, ammonium chloride 7g, KH
2PO
45g, MgSO
47H
2O 0.8g, KCl 5g, liquid microelement 5ml, vitamin 20 μ g, vitamin H 10 μ g, nicotinic acid 4mg, pyridoxine hydrochloride 100 μ g, riboflavin 50 μ g, CaCO
340g (shake bottle time add); The sodium acetate addition is that 0～10g decides on experiment, and optimum addition is 6g, and tap water is settled to 1L, pH5.0;
Minimum medium (MM): glucose 10g, NH
4Cl 3g, KH
2PO
41g, MgSO
47H
2O0.5g, liquid microelement 5ml, vitamin 20 μ g, vitamin H 10 μ g, nicotinic acid 4mg, pyridoxine hydrochloride 100 μ g, tap water is settled to 1L, pH5.0;
Screening culture medium (CM): on the basis of MM substratum, add the 6g sodium acetate.Claim perfect medium again.
Add 2% agar during preparation solid plate substratum.
Liquid microelement: CaCl
22H
2O 2g, FeSO
47H
2O 2g, CuSO
45H
2O 0.05g, ZnCl
20.5g, MnCl
24H
2O 0.2g is settled to 1L after the HCl dissolving of 2mol/L.
The mutagenesis of mutant strain and separation screening
Connect a ring WSH-IP303 bacterial classification from fresh inclined-plane and go into seed culture medium (50mL/500mL Erlenmeyer flask), after under 30 ℃, 200r/min, cultivating 12h, centrifugal collecting cell, after stroke-physiological saline solution and each washing once of 0.1mol/L potassium phosphate buffer (pH7.0), the cell suspension of breaing up is contained in the 0.1mol/L buffer solution of potassium phosphate (pH7.0) of 10g/L nitrosoguanidine (NTG) 30 ℃ of following oscillation treatment 1h in 20ml.The bacteria suspension of getting after 10mL handles dilutes 10 times of termination reactions with the 0.16mol/L hypo solution, centrifugal collecting cell, with cultivating 24h after the stroke-physiological saline solution washing in the middle of doing in its access seed culture medium, dilution then is coated with the CM flat board, cultivates 48h down for 30 ℃.The big bacterium colony of bacterium colony circle on the picking CM flat board, corresponding dibbling CM and MM flat board.Well-grown on the picking CM flat board and not growing on the MM flat board or the very weak bacterium colony of growing, the inoculation inclined-plane, behind 30 ℃ of cultivation 24h, every strain connects a ring and goes into fermention medium, determines to sieve again to use bacterial strain according to the height of output of pyruvic acid height and its pyruvic carboxylase vigor.Before carrying out multiple sieve, multiple sieve is confirmed with the vitamin defective type genetic marker of bacterial strain.When sieving again,, investigate every monobasic pyruvic acid synthesis capability (every strain connects 3 bottles of fermention mediums) with the continuous passage three times on the inclined-plane of every strain bacterium.According to output of pyruvic acid height, pyruvic carboxylase vigor size and product acid acceptance, screen definite bacterial strain.
By repeating mutagenic condition, can obtain identical WSH-LQ307 acetate leakage type pyruvic acid high yield bacterium.The test of the fermentative Production pyruvic acid by shaking a bottle level, 5L fermentor tank level and 300L fermentor tank level with this bacterium has industrial applicibility and is worth.
Analytical procedure
The plastic centrifuge tube that will contain the 4mL fermented liquid places table model high speed centrifuge (TGL-16G), centrifugal 5min under 10000r/min, and it is standby to get supernatant liquor.
Pyruvic acid and glucose concn are used lactic dehydrogenase enzyme process and 3 respectively, and 5-dinitrosalicylic acid method is measured.
Pyruvic carboxylase (PDC) activity high-performance liquid chromatogram determination.
Acetaldehyde dehydrogenase (ADH) is measured with ultraviolet spectrophotometer.
Cell concentration OD
660Measure 1OD
660=0.23g dry mycelium (shakes bottle sample and also need add 2mL 2mol/L dissolving with hydrochloric acid CaCO
3).
Acetate high effective liquid chromatography for measuring in the substratum.
Protein measuring is measured with improved Folin-phenol method in the cell.
Shaking investigation original seed and the pyruvate fermentation performance of mutant strain and their PDC activity on bottle level, the results are shown in table 1.As seen from Table 1, the PDC activity of mutant strain is lower than starting strain, and output of pyruvic acid is high, and the PDC activity is low more, and output of pyruvic acid is high more, and productive rate is also high more.To the mutant strain experiment of going down to posterity, it is strong and stable to find that mutant strain WSH-LQ307 pyruvic acid in the process of going down to posterity produces ability.
The performance of table 1 original seed and mutant strain fermentation pyruvic acid is bacterium pyruvic carboxylase output of pyruvic acid glucose acid invert ratio cell concentration relatively
Active (g/L) be (g/L) (g/g)
(×10
-3μmol
pyr/min/mg/pro
Tein) the bacterium 15.38 38.3 0.46 9.1 strain LQ307 9.12 46.2 0.65 10.8 that set out
In minimum medium, add sodium acetate and investigate the influence of acetate starting strain and mutant strain growth.Mutant strain T.glabrata WSH-LQ307 bacterium minimum medium is dense than original seed low 28% as can be seen from Figure 1A, and from Figure 1B as can be seen, bacterium is dense in the perfect medium of WSH-LQ307 after having added the 6g/L sodium acetate exceeds 21% than original seed, and promptly the bacterium of LQ307 in perfect medium is dense exceeds 49% than the bacterium in minimum medium is dense.This shows, bacterial strain WSH-LQ307 can utilize acetate as a supplement carbon source grow, and growth must be faster than starting strain.Illustrate that acetate has promoted the growth of thalline as its supplementary carbon source.
Investigate PDC, ADH (ethanol dehydrogenase) activity of starting strain and mutant strain and the relation of output of pyruvic acid.Test result is as shown in table 2.The accumulation that shows pyruvic acid comes from the flat reduction of PDC enzyme running water, and the PDC activity is low more, and the accumulating level of pyruvic acid is high more.Change not quite and the enzyme running water of the ADH between starting strain and the mutant strain is flat, illustrate that ADH does not have influence on the further metabolism of pyruvic acid.Therefore, the raising of mutant strain WSH-LQ307 accumulation pyruvic acid level can only be the low result of PDC enzyme running water pancake, because the reduction that the PDC enzyme is lived makes pyruvic acid reduce to the flux of this metabolism branch road of acetaldehyde, pyruvic acid is accumulated.Mutant strain WSH-LQ307 can effectively utilize acetate as the energy, and can be pyruvic acid with conversion of glucose more efficiently than original seed, thereby realizes the high yield of pyruvic acid.
The PDC of table 2 original seed and mutant strain, ADH specific activity are than bacterial strain PDC ADH
(μ (μ
Mol/min/mg-protein) mol/min/mg-protein) starting strain 15.38 * 10
-31.41 * 10
-1WSH-LQ307 9.12 * 10
-31.34 * 10
-1
The experiment of going down to posterity of mutant strain WSH-LQ307
WSH-LQ307 goes down to posterity repeatedly to mutant strain, shaking ability and the PDC activity of measuring its accumulation pyruvic acid on bottle level, found that as shown in Figure 2.The heredity of WSH-LQ307, leavening property are stablized, and are bacterium of pyruvic acid generation preferably, and interpolation acetate carbon source accumulation output of pyruvic acid has as a supplement improved 21% than starting strain.
Investigate the characteristic of WSH-LQ307 fermentation production of acetone acid
Sodium acetate concentration is to the influence of WSH-LQ307 accumulation pyruvic acid
Carbon content in the 5g sodium acetate is equivalent to the carbon content in the 5.5g glucose.The ability of bacterial strain WSH-LQ307 accumulation pyruvic acid and productive rate are as shown in Figure 3 when adding 0～10g sodium acetate in minimum medium.When adding the 6g/L sodium acetate, the WSH-LQ307 transforming glucose is the transformation efficiency of pyruvic acid the highest (0.586g/g), and (0.514g/g) exceeds 14% when not adding sodium acetate.So carbon source is better as a supplement for the sodium acetate of interpolation 6g/L as in fermention medium.
The experiment of WSH-LQ307 bacterial strain 5L fermentor tank
Fig. 4 is the pyruvate fermentation conditional curve of LQ307 in the 5L fermentor tank, and used substratum is the fermention medium that has added the 6g/L sodium acetate.As shown in Figure 4, the concentration of sodium acetate 6g/L from the outset reduces to 0 in the substratum behind fermentation 30h, illustrate that bacterial strain LQ-307 preferentially utilizes sodium acetate to grow, after the sodium acetate utilization is intact, utilize glucose to grow again, as can be seen from the figure after the sodium acetate completely consumed, the wear rate of glucose is obviously accelerated, and glucose major part except that the thalline that partly is used for growing then is the accumulation pyruvic acid.LQ-307 begins to accumulate pyruvic acid behind 16h, 20～56h pyruvic acid maintains more than the 0.6g/g substantially to the transformation efficiency of glucose, and 64h output of pyruvic acid (68.7g/L) and pyruvic acid exceed 24% and 17% than starting strain respectively to the transformation efficiency (0.651g/g) of glucose.So WSH-LQ307 is the strain excellent of fermentative Production pyruvic acid.
Advantage of the present invention is the acetate seepage defect type bacterial strain T.glabrata WSH-LQ307 from T.glabrata WSH-IP303 seed selection, show the ability of the high-level pyruvic acid of accumulation, this is owing to its PDC enzyme work with respect to starting strain has been lowered many, in substratum, added sodium acetate, be not sole carbon source, but add sodium acetate carbon source as a supplement with glucose.Mutant strain preferentially utilizes acetate to grow, and to have improved conversion of glucose be the transformation efficiency of pyruvic acid.By repeating mutagenic condition, can obtain identical WSH-LQ307 acetate leakage type pyruvic acid high yield bacterium.The test of the fermentative Production pyruvic acid by shaking a bottle level, 5L fermentor tank level and 300L fermentor tank level with this bacterium has industrial applicibility and is worth.
Description of drawings
Fig. 1 sodium acetate is to the influence (A is the MM substratum, and B is the CM substratum) of strain growth.
Fig. 2 passage number is to LQ307 output of pyruvic acid and the active influence of PDC.
Fig. 3 sodium acetate concentration is to the accumulation of pyruvic acid and the influence of productive rate.
The pyruvate fermentation conditional curve of Fig. 4 LQ-307 in the 5L fermentor tank.
Embodiment
Embodiment 1: with WSH-LQ307 fermentative Production pyruvic acid
Connect a ring LQ-307 bacterial classification from fresh inclined-plane and go into seed culture medium (50ml/500ml Erlenmeyer flask), at 30 ℃, 200rpm inserts fermention medium with 10% inoculum size (v/v) after cultivating 24h down.Seed culture medium and fermention medium are formed as described in this manual, add the 6g/L sodium acetate in the fermention medium.
Shake-flask culture: the liquid amount of fermention medium is 50ml in the 500ml Erlenmeyer flask, rotating speed 200rpm.Fermentation time is 48h, and output of pyruvic acid is 46.2g/L.
Embodiment 2: synthesize pyruvic acid with WSH-LQ307 at the 5L fermentor tank
Inclined-plane seed culture and fermentation inoculation condition are all with embodiment 1, and 5L fermentation cylinder for fermentation substratum (adding the 6g/L sodium acetate) volume is 3L.Air flow is 3L/min, 16h is 700rpm before the mixing speed, falls only 500rpm behind the 16h, controls pH about 5.0 with 5mol/LKOH or 5mol/L NaOH, leavening temperature is 30 ℃, and fermentation equipment is that the model that Korea S fermentor tank company produces is the automatic fermenter of KBT-5L.Fermentation time is 64h.Output of pyruvic acid is 68.7g/L, is 0.651g/g to the transformation efficiency of glucose.
Embodiment 3: synthesize pyruvic acid with WSH-LQ307 at the 300L fermentor tank
Inclined-plane seed culture and fermentation inoculation condition are all with embodiment 1, and seed culture is the KF-30L automatic fermenter that Korea S fermentor tank company produces with 30L automatic fermenter (Changzhou Shuguang Chemical Factory).The reactor of fermentation usefulness is a 300L top mechanical agitator tank (Changzhou Shuguang Chemical Factory), and liquid amount is 200L, and air flow keeps 1L/Lmin, mixing speed 0～16h is 250rpm, be 220rpm behind the 16h, pH is 5.0 with industrial soda (mass concentration is about 300g/L) control, and leavening temperature is 30 ℃.Fermentation time is 65h.Output of pyruvic acid is 63.8g/L, is 0.588g/g to the transformation efficiency of glucose.
Claims (4)
1. a torulopsis glabrata (Torulopsis glabrata), it is characterized in that title is referred to as torulopsis glabrata (Torulopsis glabrata) WSH-LQ307 and (now has been preserved in Wuhan China typical culture collection center, numbering CCTCC NO.M202019), this bacterial strain is to be starting strain with torulopsis glabrata (Torulopsisglabrata) WSH-IP303, add through nitrosoguanidine (NTG) mutagenic treatment, in substratum acetate as a supplement the carbon source seed selection obtain, this bacterial strain remains nicotinic acid (NA), VitB1 (B
1), pyridoxol (B
6) and the auxotroph of four kinds of VITAMIN of vitamin H (Bio), be the acetate seepage defect type pyruvic acid high productive mutant that pyruvic carboxylase (PDC) enzyme is lived and reduced.
2. the selection of torulopsis glabrata (Torulopsis glabrata) WSH-LQ307 is characterized in that through nitrosoguanidine (NTG) mutagenic treatment, adds acetate carbon source as a supplement in substratum,
The A starting strain
Starting strain is torulopsis glabrata (Torulopsis glabrata) WSH-IP303, is nicotinic acid (NA), VitB1 (B
1), pyridoxol (B
6) and the auxotroph of four kinds of VITAMIN of vitamin H (Bio),
The B substratum
Inclined-plane and seed culture medium: glucose 30g, peptone 10g, KH
2PO
41g, MgSO
47H
2O 0.5g, agar 20g (inclined-plane is used), tap water is settled to 1L, pH5.5;
Fermention medium: glucose 100g, ammonium chloride 7g, KH
2PO
45g, MgSO
47H
2O0.8g, KCl 5g, liquid microelement 5ml, vitamin 20 μ g, vitamin H 10 μ g, nicotinic acid 4mg, pyridoxine hydrochloride 100 μ g, riboflavin 50 μ g, lime carbonate 40g (shake bottle time add), the sodium acetate addition is 0～10g, tap water is settled to 1L, pH5.0;
Minimum medium (MM): glucose 10g, ammonium chloride 3g, KH
2PO
41g, MgSO
47H
2O 0.5g, liquid microelement 5ml, vitamin 20 μ g, vitamin H 10 μ g, nicotinic acid 4mg, pyridoxine hydrochloride 100 μ g, tap water is settled to 1L, pH5.0;
Screening culture medium (CM): on the basis of MM substratum, add the 6g sodium acetate;
Add 2% agar during preparation solid plate substratum;
Liquid microelement: CaCl
22H
2O 2g, FeSO
47H
2O 2g, CuSO
45H
2O 0.05g, ZnCl
20.5g, MnCl
24H
2O 0.2g is settled to 1L after the HCl dissolving with 2mol/L;
The mutagenesis of C mutant strain and separation screening
Connect a ring WSH-IP303 bacterial classification from fresh inclined-plane and go into (50ml/500ml Erlenmeyer flask) the seed culture medium, at 30 ℃, after cultivating 12h under the condition of 200rpm, centrifugal collecting cell, respectively wash once with stroke-physiological saline solution and 0.1mol/L potassium phosphate buffer (pH7.0), the cell suspension broken up is contained in the buffer solution of potassium phosphate of 0.1mol/L of 10g/L nitrosoguanidine (NTG) in 20ml, 30 ℃ of following oscillation treatment 1h, the bacteria suspension of getting after 10ml handles dilutes 10 times of termination reactions with the 0.16mol/L hypo solution, centrifugal collecting cell, with in the middle of doing in its access seed culture medium, cultivating 24h after the stroke-physiological saline solution washing, and after dilution back coating CM flat board, cultivate 48h down at 30 ℃, the big bacterium colony of bacterium colony circle on the picking CM flat board, corresponding dibbling CM and MM flat board, well-grown on the picking CM flat board and the bacterium colony of not growing or growing faintlyer on the MM flat board.The inoculation slant medium, behind 30 ℃ of cultivation 24h, every strain connects a ring and goes into fermention medium, determine to sieve again the bacterial strain of usefulness according to the height of output of pyruvic acid and the active height of its pyruvic carboxylase (PDC), before the multiple sieve, to confirm with the vitamin defective type genetic marker of bacterial strain multiple sieve.
3. the purposes of torulopsis glabrata (Torulopsis glabrata) WSH-LQ307 is characterized in that the bacterial strain as the fermentative Production pyruvic acid with WSH-LQ307, and the sodium acetate that adds 0～10g/L in fermention medium is carbon source as a supplement.
4. the purposes of torulopsis glabrata as claimed in claim 3 (Torulopsis glabrata) WSH-LQ307 is characterized in that the bacterial strain as the fermentative Production pyruvic acid, and carbon source is better as a supplement to add the 6g/L sodium acetate in fermention medium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021131422A CN1176205C (en) | 2002-06-08 | 2002-06-08 | Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB021131422A CN1176205C (en) | 2002-06-08 | 2002-06-08 | Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1392246A true CN1392246A (en) | 2003-01-22 |
| CN1176205C CN1176205C (en) | 2004-11-17 |
Family
ID=4742469
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB021131422A Expired - Fee Related CN1176205C (en) | 2002-06-08 | 2002-06-08 | Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1176205C (en) |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1297652C (en) * | 2003-04-03 | 2007-01-31 | 中国科学院微生物研究所 | Metabolic engineering microzyme for producing pyruvic acid |
| CN101157941B (en) * | 2007-09-19 | 2010-06-09 | 江南大学 | Method for controlling dextrose and vitamine concentration for improving fermentation and acetonic acid yield |
| CN101225410B (en) * | 2008-01-29 | 2010-08-25 | 江南大学 | Method for promoting acetonic acid excess accumulation by adding proline |
| CN1962851B (en) * | 2006-11-29 | 2010-09-15 | 华东理工大学 | Culture medium formulation for fermentation production of lactic acid |
| WO2011035492A1 (en) * | 2009-09-24 | 2011-03-31 | 上海天伟生物制药有限公司 | High yield antibiotics producing fungus strain, preparation method and use thereof |
| CN101319235B (en) * | 2008-07-01 | 2011-07-20 | 江南大学 | Method for improving production volume of pyruvic acid preparation of zymotechnics with additive gluconic acid sodium salt |
| CN101157942B (en) * | 2007-09-19 | 2011-09-07 | 江南大学 | Method for controlling culture system temperature and improving fermentation acetonic acid yield |
| CN101691545B (en) * | 2009-09-09 | 2011-10-05 | 江南大学 | Structuring of pyruvic acid-producing recombinant bacteria strain and method for enhancing synthesis rate of pyruvic acid by pyruvic acid-producing recombinant bacteria |
| CN102286550A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Method for improving pyruvic acid yield via fermentation process by using urea as nitrogen source and application |
| CN102622533A (en) * | 2012-04-06 | 2012-08-01 | 江南大学 | Construction and application technology of Torulopsis glabrata genome metabolism model |
-
2002
- 2002-06-08 CN CNB021131422A patent/CN1176205C/en not_active Expired - Fee Related
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1297652C (en) * | 2003-04-03 | 2007-01-31 | 中国科学院微生物研究所 | Metabolic engineering microzyme for producing pyruvic acid |
| CN1962851B (en) * | 2006-11-29 | 2010-09-15 | 华东理工大学 | Culture medium formulation for fermentation production of lactic acid |
| CN101157941B (en) * | 2007-09-19 | 2010-06-09 | 江南大学 | Method for controlling dextrose and vitamine concentration for improving fermentation and acetonic acid yield |
| CN101157942B (en) * | 2007-09-19 | 2011-09-07 | 江南大学 | Method for controlling culture system temperature and improving fermentation acetonic acid yield |
| CN101225410B (en) * | 2008-01-29 | 2010-08-25 | 江南大学 | Method for promoting acetonic acid excess accumulation by adding proline |
| CN101319235B (en) * | 2008-07-01 | 2011-07-20 | 江南大学 | Method for improving production volume of pyruvic acid preparation of zymotechnics with additive gluconic acid sodium salt |
| CN101691545B (en) * | 2009-09-09 | 2011-10-05 | 江南大学 | Structuring of pyruvic acid-producing recombinant bacteria strain and method for enhancing synthesis rate of pyruvic acid by pyruvic acid-producing recombinant bacteria |
| WO2011035492A1 (en) * | 2009-09-24 | 2011-03-31 | 上海天伟生物制药有限公司 | High yield antibiotics producing fungus strain, preparation method and use thereof |
| US8431383B2 (en) | 2009-09-24 | 2013-04-30 | Shanghai Techwell Biopharmaceutical Co., Ltd. | Mutagenized strain of Glarea lozoyensis and a method of preparing a compound from the mutagenized strain |
| CN102286550A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Method for improving pyruvic acid yield via fermentation process by using urea as nitrogen source and application |
| CN102622533A (en) * | 2012-04-06 | 2012-08-01 | 江南大学 | Construction and application technology of Torulopsis glabrata genome metabolism model |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1176205C (en) | 2004-11-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ge et al. | A shortcut to the production of high ethanol concentration from Jerusalem artichoke tubers | |
| CN1216150C (en) | Method for producing alcohol by solid fermentation of stalks | |
| CN103756939B (en) | Sporolactobacillus terrae and application thereof | |
| CN102719371B (en) | Clostridium beijerinckii and method for preparing biological butanol through fermentation of xylose residue serving as raw material thereof | |
| AU2009262334A1 (en) | Method of producing yeast biomass | |
| Wang et al. | Efficient L-lactic acid production from sweet sorghum bagasse by open simultaneous saccharification and fermentation | |
| CN1176205C (en) | Acetic acid leakage type high-yield pyruvate bacterium and its breeding method and producing pyruvic acid by said bacterium via fermentation process | |
| CN101869181B (en) | Preparation method of bacillus polymyxa raw powder with 100 billion live spores per gram | |
| CN100338221C (en) | Preparation of lactic acid from a pentose-containing substrate | |
| CN1824767A (en) | Method of producing xylosic alcohol and its special baterial strain | |
| CN102690764B (en) | Bacillus coagulans used to preparing L-lactic acid and application method thereof | |
| CN102925365B (en) | Trichoderma atroviride strain and application thereof in preparation of cellulase | |
| CN113621674B (en) | Method for producing L-lactic acid by using liquor distiller grains | |
| CN1840653A (en) | Mutant strain of lactobacillus casei, its preparation method and fermentation production of L-lactic acid | |
| CN101792753B (en) | Construction method for production of recombinant yeast of ethanol by fermented xylose | |
| Garcia-Kirchner et al. | Mixed submerged fermentation with two filamentous fungi for cellulolytic and xylanolytic enzyme production | |
| CN1117854C (en) | Inulinase generating saccharomycetes strain and its application in producing high fructure syrup | |
| CN112481316A (en) | Cathode electric fermentation method for preparing butyric acid by fermenting straw through enhanced anaerobic mixed flora | |
| CN115197977A (en) | Method for synthesizing lactic acid from lignocellulose by using fungus-bacterium mixed system with complementary functions | |
| CN109536565A (en) | Method for producing succinic acid by utilizing mixed fermentation of high-temperature anaerobic bacteria for pyrolyzing sugar and actinobacillus succinogenes | |
| Vinche et al. | Chitosan: A valuable byproduct of ethanolic fermentation by Rhizopus oryzae | |
| CN103275886A (en) | Bacterium for stable and high yielding of 2,3-butylene glycol, and method for utilizing low-temperature plasma and diethyl sulfate compound mutation | |
| CN111139211B (en) | Gluconobacter oxydans adaptive evolution method for efficiently utilizing non-glucose carbon source and application thereof | |
| CN108587923B (en) | Method for improving malic acid fermentation performance | |
| He et al. | Ethanol production by mixed-cultures of Paenibacillus sp. and Zymomonas mobilis using the raw starchy material from sweet potato |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| 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 | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |