CN102757929B - Method for promoting multiplication of clostridium butyricum - Google Patents
Method for promoting multiplication of clostridium butyricum Download PDFInfo
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- CN102757929B CN102757929B CN 201210244458 CN201210244458A CN102757929B CN 102757929 B CN102757929 B CN 102757929B CN 201210244458 CN201210244458 CN 201210244458 CN 201210244458 A CN201210244458 A CN 201210244458A CN 102757929 B CN102757929 B CN 102757929B
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Abstract
The invention discloses a method for promoting multiplication of clostridium butyricum. The method comprises the steps of: adding acid oligosaccharide into a culture medium containing clostridium butyricum or an animal. The method has the advantage that the plant oligosaccharide and the clostridium butyricum can obviously promote the multiplication of clostridium butyricum under the in vitro or in vivo coexistence condition.
Description
Technical field
The present invention relates to a kind of method that promotes clostridium butyricum propagation.
Background technology
In recent years, the enteric microorganism flora is illustrated gradually to the influence of human body and animal health, and the reproduction balance of enteric microorganism flora also is familiar with by people gradually to the significance of health.Wherein clostridium butyricum is in recent years by a kind of animal intestinal probiotic bacterium of people's common concern.Clostridium butyricum is a kind of anaerobism bacillocin, has heat-resisting acidproofly and common antibiotics had characteristics such as resistance, has the intestinal mucosa of reparation, recovers the intestinal microflora balance, improves immunizing power, anti-inflammatory presses down many-sided physiological actions such as cancer.Have research report to confirm, clostridium butyricum has the effect of the propagation of the clostridieum welchii that suppresses to cause pathogenic bacterium such as vibrio cholerae, Shigellae, Salmonellas, pathogenic colon bacillus, the Enterohemorrhagic E.coli of dysentery and cause pseudomembranous enteritis.The clostridium butyricum preparation is used for the treatment of bacterial diarrhea, viral diarrhea and mycotic diarrhoea, the untoward reaction of the suitable and antibiotic-free of its curative ratio and microbiotic.Clostridium butyricum can promote the growth and breeding of beneficial bacteria of intestinal tract such as bifidus bacillus, milk-acid bacteria and streptococcus faecium.In addition, the meta-bolites butyric acid of clostridium butyricum also have activation and repair animal intestinal epithelial cell, treatment inflammatory enteritis, promote animal intestinal to the absorption of moisture and nutritive substance, suppress the effects such as generation of corrupt substance in the intestines.In food, feed, pharmaceuticals, has purposes widely.But, characteristic such as it is low that clostridium butyricum also has the growth and breeding amount, and the vitro culture difficulty is big.Up to the present, also do not report the material and the method that can effectively promote clostridium butyricum propagation both at home and abroad.
Summary of the invention
Technical problem to be solved by this invention is: a kind of method that promotes clostridium butyricum propagation efficiently is provided.
In order to solve the problems of the technologies described above, the technical solution used in the present invention is: promote the method for clostridium butyricum propagation, comprising: acidic oligosaccharide is joined in the substratum or animal body that contains clostridium butyricum.
In order to solve the problems of the technologies described above better, the further technical scheme that the present invention adopts is: described acidic oligosaccharide can be powder, tablet or liquid.
In order to solve the problems of the technologies described above better, the further technical scheme that the present invention adopts is: described acidic oligosaccharide can be used with auxiliary material, and described auxiliary material can be: the mixing of one or more of carbohydrate, protein, VITAMIN or mineral element.
Advantage of the present invention is: the propagation that can obviously promote clostridium butyricum in stripped or animal body under the condition that plant oligose and clostridium butyricum coexist.
Description of drawings
Fig. 1 acidic oligosaccharide is to the influence of clostridium butyricum growth
Fig. 2 acidic oligosaccharide is to the influence of rat enteron aisle clostridium butyricum increment
Fig. 3 acidic oligosaccharide is to the influence of chicken enteron aisle clostridium butyricum increment
Fig. 4 acidic oligosaccharide is to the influence of intestine of young pigs clostridium butyricum increment.
Embodiment
Describe particular content of the present invention in detail below in conjunction with the drawings and specific embodiments.
Promote the method for clostridium butyricum propagation, comprising: acidic oligosaccharide is joined in the substratum or animal body that contains clostridium butyricum.Described acidic oligosaccharide is the patent No.: 200910012598.5 patent names: have fat-reducing with the acidic oligosaccharide of lipid-lowering effect and the acidic oligosaccharide of being mentioned in using.Described acidic oligosaccharide can be powder, tablet or liquid.Described acidic oligosaccharide can be used with auxiliary material, and described auxiliary material can be: the mixing of one or more of carbohydrate, protein, VITAMIN or mineral element.
Isolated experiment:
Join the acidic oligosaccharide powder in the substratum (glucose 1%, yeast powder 1%, NaCl0.5%) of clostridium butyricum in 0.25% ratio, with the substratum that do not add acidic oligosaccharide in contrast, insert a certain amount of clostridium butyricum after sterilising treatment, 37 ℃ are carried out anaerobism and cultivate behind the 36h absorbancy at its nutrient solution of 660 nm places detection.Judge the promoter action of acidic oligosaccharide with the variation of absorbancy to the clostridium butyricum growth.The result as shown in Figure 1, after the co-cultivation, the bacterium number of its reproductive growth is significantly more than the bacterium number that does not add the plant oligosaccharides under isolated condition for acidic oligosaccharide and clostridium butyricum.
The experiment of rat caecum:
Rat is divided into 2 groups of control group and sample sets, 6 every group at random.Every rat of sample sets is irritated the acidic oligosaccharide of gastric juice attitude every day by 150 mg/kg body weight dosage, control group is irritated stomach distilled water every day, once a day, 2 week of continuous irrigation stomach, dissected the back, and the ight soil separation and Culture (substratum: peptone 15g/L, yeast powder 2g/L, glucose 20g/L, Zulkovsky starch 0. 5g/L, NaCl 5g/L, 5% halfcystine 10ml/L) of getting the caecum part is investigated the changing conditions of its clostridium butyricum colony number.The result as shown in Figure 2, acidic oligosaccharide can increase the quantity of the probiotic bacterium-clostridium butyricum in the rat enteron aisle significantly, plays the effect of regulating intestinal colony.
Chicken manure is just tested:
Adopt randomized block design, select 45 of the chick of same kind, be divided into 3 groups at random, 15 every group.The 1st group is control group (Yu Mi – dregs of beans type basal diet), the 2nd group is added clostridium butyricum preparation (1 * 10 in basal diet
8Cfu/g, 5g/kg), the 3rd group is added clostridium butyricum preparation (1 * 10 in basal diet
8Cfu/g, 5g/kg) and acidic oligosaccharide (2.25g/kg).Each is organized every day and all regularly feeds 3 times, does not limit food consumption, freely drinks water.Fed continuously 14 days, in taking chicken manure just to aseptic after being tried the last 24h of thing, be inoculated into the last 37 ℃ of anaerobism of selective medium (peptone 15g/L, yeast powder 2g/L, glucose 20g/L, Zulkovsky starch 0. 5g/L, NaCl 5g/L, 5% halfcystine 10ml/L) through the isolate processing and cultivate 48h, to investigate the quantity of clostridium butyricum in the ight soil.The result reaches with contrast as shown in Figure 3
Clostridium butyricum uses separately and compares, and acidic oligosaccharide can significantly promote the propagation of clostridium butyricum in the chicken enteron aisle.
The swine excrement experiment
Select 30 of the weanling pigs of same kind, be divided into 3 groups at random, 10 every group.The 1st group is control group (Yu Mi – dregs of beans type basal diet), the 2nd group is added clostridium butyricum preparation (1 * 10 in basal diet
8Cfu/g, 5g/kg), the 3rd group is added clostridium butyricum preparation (1 * 10 in basal diet
8Cfu/g, 5g/kg) and acidic oligosaccharide (4g/kg).Tested 14 days by a definite date, each is organized and regularly feeds every day 3 times, does not limit food consumption, freely drinks water.Fed continuously 14 days, in to the aseptic swine excrement of taking after being tried the last 24h of thing, be inoculated into the last 37 ℃ of anaerobism of selective medium (peptone 15g/L, yeast powder 2g/L, glucose 20g/L, Zulkovsky starch 0. 5g/L, NaCl 5g/L, 5% halfcystine 10ml/L) through the isolate processing and cultivate 48h to investigate the quantity of clostridium butyricum in the ight soil.The result compares with contrast and independent use of clostridium butyricum as shown in Figure 4, and acidic oligosaccharide can significantly promote the propagation of clostridium butyricum in the chitling road.
Claims (3)
1. promote the method for clostridium butyricum propagation, it is characterized in that: comprising: acidic oligosaccharide is joined the substratum that contains clostridium butyricum in 0.25% ratio, the preparation method of described acidic oligosaccharide is as follows: (1) gets the pectin aqueous solution that mass percentage concentration is 1%-2%, accent pH is 2-4, at 90-120 ℃, decomposition reaction 15 minutes-24 hours; Or get the pectin aqueous solution that mass percentage concentration is 1%-2%, the ratio in every gram pectin adding 0.01-1.0Unit enzyme unit alive adds pectin hydrolase or pectin lyase, at 30-50 ℃, pH is 3.5-4.5, and decomposition reaction 30 minutes-24 hours obtains pectin decomposer; (2) pectin decomposer that obtains is handled with hollow-fibre membrane or ceramic filter membrane, after reclaiming the following oligosaccharide mixture of molecular weight 6000 dalton, with the oligosaccharide mixture of nanofiltration membrane recovery molecular weight 200-6000, obtain the acidity oligosaccharide of pectin that overlap ratio is 2-20 with Bio-gel P-2 gel column or the separation of Sephadex G-25 gel filtration chromatography; Maybe the pectin decomposer that obtains is handled with hollow-fibre membrane or ceramic filter membrane, after reclaiming the following oligosaccharide mixture of molecular weight 6000 dalton, reclaim the oligosaccharide mixture of molecular weight 200-6000 again with nanofiltration membrane, cross DEAE-Sephadex A-25 anionite-exchange resin, inorganic salt solution gradient elution with suitable bed volume 5-10 0.05-1M doubly obtains the acidity oligosaccharide of pectin that overlap ratio is 2-20.
2. according to the method for the described promotion clostridium butyricum propagation of claim 1, it is characterized in that: described acidic oligosaccharide is powder, tablet.
3. according to the method for claim 1 or 2 described promotion clostridium butyricums propagation, it is characterized in that: described acidic oligosaccharide can be used with auxiliary material, and described auxiliary material is: the mixing of one or more of carbohydrate, protein, VITAMIN or mineral element.
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| CN105861411B (en) * | 2016-05-26 | 2019-10-25 | 湖北工业大学 | A method of improving Miyarisan Fermentative growth efficiency |
| CN105950538B (en) * | 2016-05-26 | 2019-10-11 | 湖北工业大学 | A method of promoting Miyarisan Fermentative growth |
| CN110055207B (en) * | 2019-03-27 | 2020-12-29 | 贺州普唯尔生命科技有限公司 | New application of N-acetylglucosamine |
| CN111671778A (en) * | 2020-05-26 | 2020-09-18 | 深圳市东荣生物科技有限责任公司 | Compound microbial preparation for protecting liver and stomach |
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| CN101624611A (en) * | 2009-08-13 | 2010-01-13 | 浙江大学 | Preparation method of inulase zymolyte for promoting clostridium butyricum to grow |
| CN102404990A (en) * | 2009-02-24 | 2012-04-04 | 里特制药股份有限公司 | Prebiotic formulations and methods of use |
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| CN102404990A (en) * | 2009-02-24 | 2012-04-04 | 里特制药股份有限公司 | Prebiotic formulations and methods of use |
| CN101624611A (en) * | 2009-08-13 | 2010-01-13 | 浙江大学 | Preparation method of inulase zymolyte for promoting clostridium butyricum to grow |
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Denomination of invention: Methods to promote the proliferation of Clostridium butyricum Effective date of registration: 20221115 Granted publication date: 20130724 Pledgee: Industrial Bank Co.,Ltd. Yancheng branch Pledgor: JIANGSU YUANSHAN BIOLOGY TECHNOLOGY CO.,LTD. Registration number: Y2022320000705 |
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