CN109504713A - The method and application of high ester bond humic acid preparation are prepared using Clostridium beijerinckii - Google Patents
The method and application of high ester bond humic acid preparation are prepared using Clostridium beijerinckii Download PDFInfo
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Abstract
A kind of method and application preparing high ester bond humic acid preparation using Clostridium beijerinckii, it is that sodium humate is added in Clostridium beijerinckii fermentation system, esterification is occurred into for the Clostridium beijerinckii phenolic hydroxyl group in the butyric acid and sodium humate to be formed that ferments by lipase, butyric acid density in fermentation liquid is reduced, reduces butyric acid accumulation to the inhibiting effect of Clostridium beijerinckii fermentation process.The presence of sodium humate extends Clostridium beijerinckii fermentation time and product accumulation.The high ester bond humic acid that sodium humate and butyric acid are formed can be directly used as intestinal health Additive Product, enter enteron aisle and release humic acid and butyric acid under the action of pancreatic lipase later, promote intestinal health.
Description
Technical field
It the present invention relates to the use of Clostridium beijerinckii and prepare high ester bond humic acid preparation and application.
Background technique
Butyric acid is converted by carbon source in Clostridium beijerinckii fermentation process, with the accumulation of butyric acid, suppression can be generated to fermentation process
Production is used, and fermentation ends are caused.If can in time remove the butyric acid of generation in fermentation process, it can guarantee fermentation process
Stablize and carry out, improves fermentation process efficiency.
The one kind of butyric acid as volatile fatty acid plays a significant role in animal and bird intestines health, and butyric acid on the one hand can
Think that enteron aisle villus energizes, promote intestinal villus development, while harmful intestinal tract bacteria can be inhibited to grow, maintains intestinal microecology flat
Weighing apparatus.It will be appreciated that in use, a small amount of butyric acid added in feed can be absorbed by upper intestines, lead to fourth
Acid is hard to reach enteron aisle rear end and plays antibacterial and energy supply effect.Therefore number of patent application 201610999456.2 discloses a kind of micro-
Capsule is coated with sodium butyrate and preparation method thereof, is coated with using ethylene glycol to ingredients such as butyric acid.This method is by simple object
Coated method is managed, is wrapped up butyric acid etc. using coated fertilizer.The disadvantage is that release time and position of the coating object in enteron aisle
Set uncontrollable, being likely to appear in enteron aisle can not open in time, cause butyric acid etc. to be discharged with excrement, or because coating is not filled
Point, butyric acid etc. still can not arrive safe and sound enteron aisle back segment the problems such as.
Humic acid is the net high-polymer substance rich in phenolic hydroxyl group, not soluble in water and sour, but can be dissolved in alkali.Humic acid is in intestines
Astriction is played in road, promotes intestinal tract injury reparation.Phenolic hydroxyl group in butyric acid and humic acid is subjected to catalysis life using lipase
At ester bond, the inhibiting effect of butyric acid in fermentation liquid in Clostridium beijerinckii fermentation process, while butyric acid and sodium humate are on the one hand reduced
Occur only could slowly open ester bond under the action of pancreatic lipase in enteron aisle after esterification, butyric acid is enable to release
It puts；Due to ester bond open process be life entity normal physiological metabolism process, and very slowly, therefore high ester bond humic acid with
Chyme constantly moves and opens release humic acid and butyric acid by continuous, ensure that in entire enteron aisle with the presence of butyric acid, realizes fourth
The full enteron aisle of acid is antibacterial to be acted on energy supply.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, (such as Clostridium beijerinckii fermentation process can be by product fourth
Acid inhibits；Butyric acid can be absorbed after entering enteron aisle by enteron aisle upper section, lead to not arrival enteron aisle back segment and play a role；Tradition coating work
Skill not can guarantee the time that product is discharged in enteron aisle and position is controllable) and provide it is a kind of by Clostridium beijerinckii fermentation process plus
Enter sodium humate, under fatty enzyme effect, esterification is occurred into for butyric acid and humic acid, removes butyric acid in fermentation liquid, it is raw simultaneously
At high ester bond humic acid ensure that slow release of the butyric acid in enteron aisle, realize that the full enteron aisle of butyric acid is antibacterial and act on energy supply.
It is a further object to provide a kind of high ester bond humic acids in feeding field with important application
Preparation.
A kind of high ester bond humic acid preparation, is added sodium humate according to a certain percentage in Clostridium beijerinckii fermentation liquid, fermentation
It is dense to reduce the butyric acid that dissociates in fermentation liquid through lipase-catalyzed formation ester bond for phenolic hydroxyl group in the butyric acid and sodium humate generated in the process
Degree, final humic acid and butyric acid generate the high ester bond humic acid preparation that ingredient is humic acid butyrate by esterification.Specifically
It is that esterification is occurred by the phenolic hydroxyl group that lipase ferments Clostridium beijerinckii in the butyric acid and sodium humate to be formed, reduces fermentation
Butyric acid density in liquid reduces butyric acid accumulation to the inhibiting effect of Clostridium beijerinckii fermentation process；The high ester that humic acid and butyric acid are formed
Key humic acid is further used as feed addictive, promotes animal and bird intestines health.
The purpose of invention is achieved in that a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
(1) seed liquor containing Clostridium beijerinckii is made, wherein bacterial concentration is 0.5 × 108-1.0×108CFU/ml；
Seed culture medium (TYA culture medium): 2 g of beef extract, 2 g of yeast powder, peptone 6 g, CH3COONH43 g,
KH2PO4 0.5 g, MgSO4·7H2O 0.2 g, FeSO4·7H26.5,121 DEG C of height of 0.01 g of O, distilled water 1000 mL, pH
Pressure 15 min of sterilizing, blow 5 min with sterile nitrogen, drive oxygen to the greatest extent.
(2) fermentation medium: K2HPO4 0.5 g、KH2PO4 0.5 g 、CH3COONH4 2.2 g、MgSO4·7H2O
0.2 g、MnSO4·H2O 0.01 g、NaCl 0.01 g、FeSO4·7H2O 0.01 g, thiamine (thiamines, vitamin
B1) 0.001g, biotin (biotin) 0.00001g, p-aminobenzoic acid (p- aminobenzoic acid)
0.001g adds water to 1000 mL.
(3) by sodium humate according to 4-10g/L, be added in fermentor.
(4) fermentation cylinder for fermentation medium sterilization terminates, and is cooled to room temperature, by lipase according to lipase and sodium humate
Fermentor is added in the ratio that mass ratio is 1:1-5.
(5) seed liquor containing Clostridium beijerinckii and fermentation medium will be obtained in step (1) to utilize according to 2% volume ratio
Peristaltic pump, which is added to, to be produced in butyric fermentation culture medium, and 37-42 DEG C, 50-120 revs/min of stir culture.
(6) 200g/L xylose utilization peristaltic pump is added in fermentor, maintaining xylose concentration in fermentation liquid is 20g/L,
Fermentation 96 hours.
(7) after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond humic acid for promoting enzymatic reaction to generate is heavy
Drop obtains high ester bond humic acid preparation through plate and frame filter press filters pressing, drying.
The specific feature of this programme is in addition, lipase is the Novozym 435 of Novozymes Company's production, enzyme activity 10000
PLU/g。
Clostridium beijerinckii (Clostridium beijerinckii) strain is Clostridium beijerinckii SE-2, culture presevation number is
CCTCC M2014384, preservation date on August 19th, 2014.Clostridium beijerinckii SE-2, Latin name Clostridium
beijerinckii SE-2；The bacterial strain was preserved in China typical culture collection center (CCTCC) on August 19th, 2014,
Address: Wuhan University, Wuhan, China city, postcode 430072, deposit number CCTCC No.M2014384.
A kind of application of high ester bond sodium humate preparation as described above, by high ester bond sodium humate preparation with 0.2% weight
The ratio of percentage is mixed with feed, Direct-fed livestock and poultry.
The present invention generates following good effect: 1, phenolic hydroxyl group in butyric acid and humic acid being carried out catalysis generation using lipase
Ester bond reduces the inhibiting effect of butyric acid in Clostridium beijerinckii fermentation process, guarantees going on smoothly for fermentation process, improves fermentation effect
Rate.
2, butyric acid and sodium humate occur only delay under the action of pancreatic lipase in enteron aisle after esterification
It is slow to open ester bond, it is released butyric acid；Since the process that ester bond is opened is very slowly, high ester bond humic acid is with chyme
Continuous movement simultaneously opens release humic acid and butyric acid by continuous, ensure that in entire enteron aisle with the presence of butyric acid, realizes butyric acid
Full enteron aisle is antibacterial to be acted on energy supply.
3, humic acid restrains intestinal tract injury, and butyric acid can not only be energized to enteron aisle villus, promotes intestinal villus development, and
Intestinal microecology balance can be improved, the synergistic effect of humic acid and butyric acid provides safeguard for intestinal health.
4, after fermentation, the characteristic using humic acid insoluble in acid and water promotes high ester bond humic acid by adjusting pH
Precipitating is formed in fermentation liquid, is convenient for later separation, saves separation costs.
On the one hand the present invention is promoted by the way that Clostridium beijerinckii tunning butyric acid and humic acid enzymatic are formed stable compound
Fermentation process is gone on smoothly, while high ester bond humic acid can help butyric acid slowly to discharge in enteron aisle again, realize full enteron aisle
Antibacterial and energy supply, enhances its effect on livestock and poultry cultivation, and product is made to have stronger competitiveness.
Specific embodiment
Embodiment 1: a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
The first step makes the seed liquor containing Clostridium beijerinckii, and wherein bacterial concentration is 0.5 × 108-1.0×108CFU/ml；It visits
Family name clostridium is deposit number CCTCC No.M2014384 Clostridium beijerinckii SE-2.
Seed culture medium (TYA culture medium): 40 g of glucose, 2 g of beef extract, 2 g of yeast powder, 6 g of peptone,
CH3COONH43 g, KH2PO4 0.5 g, MgSO4·7H2O 0.2 g, FeSO4·7H20.01 g of O, distilled water 1000 mL, pH
6.5,121 DEG C of 15 min of high pressure sterilization blow 5 min with sterile nitrogen, drive oxygen to the greatest extent.
Second step, production produce butyric fermentation culture medium, specifically: basal fermentation medium: K2HPO4 0.5 g、KH2PO4
0.5 g 、CH3COONH4 2.2 g、MgSO4·7H2O 0.2 g、MnSO4·H2O 0.01 g、NaCl 0.01 g、FeSO4·
7H2O 0.01 g, thiamine (thiamines, vitamin B1) 0.001g, biotin (biotin) 0.00001g, p-
Aminobenzoic acid (p- aminobenzoic acid) 0.001g, adds water to 1000 mL.
Third step, by sodium humate according to 4g/L, be added in fermentor.
4th step, fermentation cylinder for fermentation medium sterilization terminate, are cooled to room temperature, by lipase according to lipase and rotten plant
Fermentor is added in the ratio that sour sodium mass ratio is 1:5.
5th step, by the first step Clostridium beijerinckii seed liquor and fermentation medium according to 2% volume ratio utilize peristaltic pump
It is added in fermentation medium, 37 DEG C, 50 revs/min of stir cultures.
200g/L xylose utilization peristaltic pump is added in fermentor by the 6th step, and xylose concentration is in maintenance fermentation liquid
20g/L。
7th step, the 96th hour fermentation ends take fermentation liquid to analyze butyric acid density, butyric acid density 1.7g/L；It fermented
Xylose 118.7g/L is consumed in journey altogether.
8th step, after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond corruption for promoting enzymatic reaction to generate is planted
Acid deposition obtains high ester bond humic acid preparation through plate and frame filter press filters pressing, drying.
Embodiment 2: a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
The first step, second step, with embodiment 1.
Third step, by sodium humate according to 7 g/L, be added in fermentation medium.
4th step, fermentation cylinder for fermentation medium sterilization terminate, are cooled to room temperature, by lipase according to lipase and rotten plant
Fermentor is added in the ratio that sour sodium mass ratio is 1:3.
5th step, by the first step Clostridium beijerinckii seed liquor and fermentation medium according to 2% volume ratio utilize peristaltic pump
It is added in fermentation medium, 40 DEG C, 80 revs/min of stir cultures.
200g/L xylose utilization peristaltic pump is added in fermentor by the 6th step, and xylose concentration is in maintenance fermentation liquid
20g/L。
7th step, the 96th hour fermentation ends take fermentation liquid to analyze butyric acid density, and butyric acid density is 0.46 g/L；Fermentation
Consume 143.3 g/L of xylose altogether in the process.
8th step, after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond corruption for promoting enzymatic reaction to generate is planted
Acid deposition obtains high ester bond humic acid preparation through plate and frame filter press filters pressing, drying.
Embodiment 3: a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
The first step, second step, with embodiment 1.
Third step, by sodium humate according to 10 g/L, be added in fermentation medium.
4th step, fermentation cylinder for fermentation medium sterilization terminate, are cooled to room temperature, by lipase according to lipase and rotten plant
Fermentor is added in the ratio that sour sodium mass ratio is 1:1.
5th step, by the first step Clostridium beijerinckii seed liquor and fermentation medium according to 2% volume ratio utilize peristaltic pump
It is added in fermentation medium, 42 DEG C, 100 revs/min of stir cultures.
200g/L xylose utilization peristaltic pump is added in fermentor by the 6th step, and xylose concentration is in maintenance fermentation liquid
20g/L。
7th step, the 96th hour fermentation ends take fermentation liquid to analyze butyric acid density, and butyric acid density is 0.90 g/L；Fermentation
Consume 129.8 g/L of xylose altogether in the process.
8th step, after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond corruption for promoting enzymatic reaction to generate is planted
Acid deposition obtains high ester bond humic acid preparation through plate and frame filter press filters pressing, drying.
Embodiment 4: a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
The first step, second step, with embodiment 1.
Third step, by sodium humate according to 7 g/L, be added in fermentation medium.
4th step, by the first step Clostridium beijerinckii seed liquor and fermentation medium according to 2% volume ratio utilize peristaltic pump
It is added in fermentation medium, 40 DEG C, 80 revs/min of stir cultures.
200g/L xylose utilization peristaltic pump is added in fermentor by the 5th step, and xylose concentration is in maintenance fermentation liquid
20g/L。
6th step, the 96th hour fermentation ends take fermentation liquid to analyze butyric acid density, and butyric acid density is 21.02 g/L；Fermentation
Consume 33.2 g/L of xylose altogether in the process.
7th step, after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond corruption for promoting enzymatic reaction to generate is planted
Acid deposition obtains high ester bond humic acid preparation through plate and frame filter press filters pressing, drying.
Embodiment 4 is only not added with lipase compared with Example 2, will be catalyzed butyric acid and humic acid due to lacking lipase
Esterification, therefore butyric acid accumulates in fermentation liquid, while xylose utilization amount is very low, only the 23.2% of embodiment 2, explanation
The presence of butyric acid causes fermentation process surprisingly to terminate.Meanwhile the two is calculated by xylose consumption, the fourth generated in whole process
It is very big that acid measures easy difference.
Embodiment 5: a kind of preparation method of high ester bond humic acid preparation, it includes the following steps:
The first step, second step, with embodiment 1.
Third step, by sodium humate according to 7 g/L, be added in fermentation medium.
4th step, by the first step Clostridium beijerinckii seed liquor and fermentation medium according to 2% volume ratio utilize peristaltic pump
It is added in fermentation medium, 40 DEG C, 80 revs/min of stir cultures.
200g/L xylose utilization peristaltic pump is added in fermentor by the 5th step, and xylose concentration is in maintenance fermentation liquid
20g/L。
6th step, the 96th hour fermentation ends take fermentation liquid to analyze butyric acid density, and butyric acid density is 21.02 g/L；Fermentation
Consume 33.2 g/L of xylose altogether in the process.
7th step, the ratio addition fermentor for being 1:3 according to lipase and sodium humate mass ratio by lipase, 40 DEG C,
80 revs/min are stirred to react 12 hours, and butyric acid density is 0.19 g/L in fermentation liquid.
8th step, after reaction by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond corruption for promoting enzymatic reaction to generate is planted
Acid deposition obtains high ester bond humic acid product through plate and frame filter press filters pressing, drying.
Embodiment 5 is adding lipase compared with Example 4 after fermentation, promotes the butyric acid of sodium humate and generation
Esterification occurs, forms high ester bond humic acid.
Embodiment 6: a kind of preparation method of humic acid preparation, it includes the following steps:
The first step is same as Example 5 to the 6th step；
7th step, after reaction by the way that fermentation liquid pH is adjusted to 2.5, the humic acid sedimentation for promoting enzymatic reaction to generate, warp
Plate and frame filter press filters pressing, drying obtain humic acid preparation.
21.02 g/L butyric acid are added according to fermentation volume into the humic acid of the 7th step acquisition in 8th step, obtain final produce
Product.
Embodiment 6 and the difference of embodiment 5 are that the butyric acid and humic acid in embodiment 5 are connected with ester bond, and embodiment
Butyric acid is with butyric acid molecule individualism in 6.
Embodiment 7: a kind of application of high ester bond sodium humate preparation as described above, the high ester bond that embodiment 2 is obtained
Humic acid preparation is mixed with feed by 0.2% weight percent, direct feeding.
Test result:
Experimental subjects is the birth piglet of certain market pig.
Experimental group: the feed of high ester bond humic acid preparation is added in newborn piglet feeding, without injection of antibiotic agent medication
The methods of prevention and control, other feeding managements by pig farm conventional method carry out.
Control group: injection of antibiotics and other medicines after diarrhea occur for piglet, and the Case management on pig farm is combined to give one
The treatment of fixed other drugs, daily ration, nutritional ingredient and feeding management etc. are completely identical as experimental group.Test result such as table 1:
Table 1
The result shows that: compared with the control group, which can effectively prevent the diarrhea of piglet to experimental group, and
Reduce drug cost.
Embodiment 8: a kind of application of high ester bond sodium humate preparation as described above, the humic acid that embodiment 4 is obtained
Preparation is mixed with feed by 0.2% weight percent, direct feeding.
Test result:
Experimental subjects is the birth piglet of certain market pig.
Experimental group: the feed of high ester bond humic acid preparation is added in newborn piglet feeding, without injection of antibiotic agent medication
The methods of prevention and control, other feeding managements by pig farm conventional method carry out.
Control group: injection of antibiotics and other medicines after diarrhea occur for piglet, and the Case management on pig farm is combined to give one
The treatment of fixed other drugs, daily ration, nutritional ingredient and feeding management etc. are completely identical as experimental group.Test result such as table 2:
Table 2
The result shows that: experimental group compared with the control group, can prevent the diarrhea of piglet, but compared with Example 7, embodiment 8
In product (embodiment 4) used due to being not added with lipase, sodium humate certain effect is finally only played in enteron aisle,
Fermentation process generation can not be acted on；And butyric acid in final humic acid preparation and does not contain because being dissolved in fermentation liquid
Butyric acid, final embodiment 8(embodiment 4) although with embodiment 7(embodiment 2) there was only one Xiang Butong of lipase, product pair
The prevention effect of grice diarrhoea is but very different.
Embodiment 9: a kind of application of high ester bond sodium humate preparation as described above, the humic acid that embodiment 5 is obtained
Preparation is mixed with feed by 0.2% weight percent, direct feeding.
Test result:
Experimental subjects is the birth piglet of certain market pig.
Experimental group: the feed of high ester bond humic acid preparation is added in newborn piglet feeding, without injection of antibiotic agent medication
The methods of prevention and control, other feeding managements by pig farm conventional method carry out.
Control group: injection of antibiotics and other medicines after diarrhea occur for piglet, and the Case management on pig farm is combined to give one
The treatment of fixed other drugs, daily ration, nutritional ingredient and feeding management etc. are completely identical as experimental group.Test result such as table 3:
Table 3
Embodiment 10: a kind of application of sodium humate preparation as described above, the humic acid preparation and feed that embodiment 6 is obtained
It is mixed by 0.2% weight percent, direct feeding.
Test result:
Experimental subjects is the birth piglet of certain market pig.
Experimental group: the feed of high ester bond humic acid preparation is added in newborn piglet feeding, without injection of antibiotic agent medication
The methods of prevention and control, other feeding managements by pig farm conventional method carry out.
Control group: injection of antibiotics and other medicines after diarrhea occur for piglet, and the Case management on pig farm is combined to give one
The treatment of fixed other drugs, daily ration, nutritional ingredient and feeding management etc. are completely identical as experimental group.
Test result such as table 4:
Table 4
The result shows that: experimental group compared with the control group, can prevent the diarrhea of piglet, although in 6 product of embodiment 5 and embodiment
Humic acid is identical as the content of butyric acid, but butyric acid is connected with humic acid with ester bond in embodiment 5, and the butyric acid in embodiment 6 is with fourth
Acid molecule form exists, therefore butyric acid reaches complete because of the such slow release of butyric acid in no image of Buddha embodiment 9 in embodiment 10
Enteron aisle is antibacterial with the effect for promoting intestinal villus development, and the grice diarrhoea rate in embodiment 10 that eventually leads to is higher than embodiment 9.The knot
Fruit further illustrates, by adding sodium humate, connect butyric acid with humic acid enzymatic using lipase, exists for improving product
Effect in enteron aisle is necessary.
Claims (5)
1. a kind of high ester bond humic acid preparation, it is characterized in that humic acid is added according to a certain percentage in Clostridium beijerinckii fermentation liquid
Sodium, phenolic hydroxyl group reduces fermentation liquid middle reaches through lipase-catalyzed formation ester bond in the butyric acid and sodium humate generated in fermentation process
From butyric acid density, final humic acid and butyric acid generate the high ester bond humic acid system that ingredient is humic acid butyrate by esterification
Agent.
2. a kind of preparation method of high ester bond humic acid preparation, it is characterized in that it includes the following steps:
(1) seed liquor containing Clostridium beijerinckii is made, wherein bacterial concentration is 0.5 × 108-1.0×108CFU/ml；
Seed culture medium: 2 g of beef extract, 2 g of yeast powder, peptone 6 g, CH3COONH43 g, KH2PO4 0.5 g,
MgSO4·7H2O 0.2 g, FeSO4·7H26.5,121 DEG C of high pressure sterilizations 15 of 0.01 g of O, distilled water 1000 mL, pH
Min blows 5 min with sterile nitrogen, drives oxygen to the greatest extent；
(2) fermentation medium: K2HPO4 0.5 g、KH2PO4 0.5 g 、CH3COONH4 2.2 g、MgSO4·7H2O 0.2 g、
MnSO4·H2O 0.01 g、NaCl 0.01 g、FeSO4·7H20.01 g of O, thiamines vitamin B1 0.001g, biotin
0.00001g, p- aminobenzoic acid 0.001g, add water to 1000 mL；
(3) by sodium humate according to 4-10g/L, be added in fermentor；
(4) fermentation cylinder for fermentation medium sterilization terminates, and is cooled to room temperature, by lipase according to lipase and sodium humate quality
Than fermentor is added for the ratio of 1:1-5；
(5) seed liquor containing Clostridium beijerinckii and fermentation medium will be obtained in step (1) according to 2% volume ratio using wriggling
Pump is added in fermentor, and 37-42 DEG C, 50-120 revs/min of stir culture；
(6) 200g/L xylose utilization peristaltic pump is added in fermentor, maintaining xylose concentration in fermentation liquid is 20g/L, fermentation
96 hours；
(7) after fermentation by the way that fermentation liquid pH is adjusted to 2.5, the high ester bond humic acid sedimentation for promoting enzymatic reaction to generate,
High ester bond humic acid preparation is obtained through plate and frame filter press filters pressing, drying.
3. the preparation method of high ester bond humic acid preparation according to claim 2, it is characterized in that lipase is that Novi's letter is public
The Novozym 435 of production is taken charge of, enzyme activity is 10000 PLU/g.
4. the preparation method of high ester bond humic acid preparation according to claim 2, it is characterized in that Clostridium beijerinckii strain is to visit
Family name clostridium SE-2, Latin name Clostridium beijerinckii SE-2；The bacterial strain was protected on August 19th, 2014
It is hidden in China typical culture collection center (CCTCC), address: Wuhan, China Wuhan University, deposit number CCTCC
No.M2014384。
5. a kind of application of high ester bond sodium humate preparation, it is characterized in that by high ester bond humic acid preparation with 0.2% weight percent
It is mixed with feed, Direct-fed livestock and poultry.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115521885A (en) * | 2022-08-24 | 2022-12-27 | 中国农业科学院饲料研究所 | Clostridium beijerinckii for producing butyric acid and application thereof |
| WO2024009036A1 (en) * | 2022-07-05 | 2024-01-11 | Adisseo France S.A.S. | Additive for animal nutrition |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000016785A2 (en) * | 1998-09-23 | 2000-03-30 | Enerkom (Proprietary) Limited | Humic acid and its use in the treatment of various conditions |
| WO2007036363A2 (en) * | 2005-09-28 | 2007-04-05 | Südzucker Aktiengesellschaft Mannheim/Ochsenfurt | Compositions containing butyric acid ester of carbohydrates and carbohydrate polyols |
| CN104673712A (en) * | 2015-01-16 | 2015-06-03 | 山东省科学院能源研究所 | Bacterial strain for producing alcohol fuels by synchronously utilizing glucose and xylose and application of bacterial strain |
-
2018
- 2018-12-04 CN CN201811473511.XA patent/CN109504713B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000016785A2 (en) * | 1998-09-23 | 2000-03-30 | Enerkom (Proprietary) Limited | Humic acid and its use in the treatment of various conditions |
| WO2007036363A2 (en) * | 2005-09-28 | 2007-04-05 | Südzucker Aktiengesellschaft Mannheim/Ochsenfurt | Compositions containing butyric acid ester of carbohydrates and carbohydrate polyols |
| CN101316598A (en) * | 2005-09-28 | 2008-12-03 | 甜糖（曼海姆/奥克森富特）股份公司 | Butyrate esters of carbohydrates and carbohydrate polyols |
| CN104673712A (en) * | 2015-01-16 | 2015-06-03 | 山东省科学院能源研究所 | Bacterial strain for producing alcohol fuels by synchronously utilizing glucose and xylose and application of bacterial strain |
Non-Patent Citations (4)
| Title |
|---|
| T. E. WEBER等: "Effects of dietary humic and butyric acid on growth performance and response to lipopolysaccharide in young pigs", 《J. ANIM. SCI.》 * |
| ZHONG-TIAN ZHANG等: "In Situ Esterification and Extractive Fermentation for Butyl Butyrate Production with Clostridium tyrobutyricum", 《BIOTECHNOLOGY AND BIOENGINEERING》 * |
| 余东游等: "三丁酸甘油酯的生物学功能及其在畜牧生产中的应用", 《中国畜牧杂志》 * |
| 韦良开等: "腐植酸类物质的生物学功能及其在畜禽养殖中的应用", 《中国饲料》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024009036A1 (en) * | 2022-07-05 | 2024-01-11 | Adisseo France S.A.S. | Additive for animal nutrition |
| FR3137537A1 (en) * | 2022-07-05 | 2024-01-12 | Adisseo France S.A.S. | Additive for animal nutrition |
| CN115521885A (en) * | 2022-08-24 | 2022-12-27 | 中国农业科学院饲料研究所 | Clostridium beijerinckii for producing butyric acid and application thereof |
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| CN109504713B (en) | 2022-05-17 |
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