CN102406062B - Method for preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine - Google Patents
Method for preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine Download PDFInfo
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Abstract
The invention discloses a method for preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine, belonging to the field of food nutrition and the technical fields of microorganisms and feed. The method comprises the following steps: extracting Chinese herbal medicine by adopting asynchronous fermentation and enzymolysis technologies, namely adding water and nutritive salt in proportion to prepare a Chinese herbal medicine fermentation liquid after a plurality of Chinese herbal medicines are subjected to impurity removal and super-micro smashing; firstly inoculating Aspergillus niger for aerobic fermentation, thus cellulose and pectinase are prepared; and then inoculating enterococcus faecium and lactobacillus plantarum for anaerobic fermentation; and then carrying out enzymolysis on damage cell walls, and dissolving out active substances of saponin, alkaloid, phytosterol, enzymic preparations, organic acid and the like. A plate-and-frame filter press is adopted to filter the fermentation liquid, the filtration liquid is directly subjected to low-temperature spray drying to prepare antibiotic substituent powder A; a filtration cake is dried by adopting a roller drier to prepare antibiotic substituent powder B, and the power A and the powder B are mixed and formulated in proportion to replace fermented enzymolysis Chinese herbal medicine extracts of antibiotic for feed.
Description
Technical field
The invention belongs to food nutrition, microorganism and field of feed, be specifically related to the fermentation enzymolysis Chinese herbal medicine extract that the Chinese herbal medicine preparations such as aspergillus niger and lactic acid bacteria asynchronous fermentation camellia seedcake (dregs of rice) powder, folium cortex eucommiae, the Radix Astragali, hawthorn, pine needle meal, barrenwort, Radix Glycyrrhizae can replace feeding antibiotic.
Background technology
In recent years, food safety affair takes place frequently, and antibiotic harm receives much concern: the one, and antibiotic residues in the animal products such as meat, egg and milk easily, and harm humans is healthy; The 2nd, the livestock products local flavor is descended, and cause mankind itself's resistance to the action of a drug to increase.Therefore, some countries have prohibited use.1974, the European Community forbade using penicillin, tetracycline in feed, forbade using in feed the happy mycin of spiramvcin, Bacitracin Zinc, virginiamycin and peptide in 1998.2011, Korea S's feed aquaculture industry was forbidden antibiotic comprehensively.Therefore, to come substitute antibiotics be that one of current cultivation, feed circle need the extremely urgent important affair that solves to safe, the efficient and economic new bio feed of exploitation.
Since the eighties in 20th century, because the security of antibiotic, hormone and chemical synthesis additive is under suspicion gradually, people begin to enliven for the research and development of Chinese herbal medicine concentrated feed additive agent.A large amount of Chinese herbal medicine concentrated feeds that experimental results show that can not only improve production performance, and preventing disease theraping effect is fine, and some effect even is not second to antibiotic.Simultaneously, Chinese herbal medicine is also had little, residual low a, advantage such as security is high of toxic and side effect.As containing abundant saccharicter-penin in camellia seedcake (dregs of rice) powder, contain abundant chlorogenic acid in the folium cortex eucommiae, contain abundant astragalus polyose in the Radix Astragali, contain abundant pinke needle oil in the pine needle meal, contain abundant saponin in the barrenwort.Studies show that these saponins, alkaloid, phytosterol and organic acid isoreactivity material can obviously improve the immunity of livestock and poultry.
New Century, fermentation begin to attempt being applied to human Chinese herbal medicine with zymolysis technique and extract.Select appropriate zymotechnique and enzymolysis, not only can more leniently decompose plant tissue, accelerated release in vitro active ingredient, and can decompose and remove impurity affect liquid preparation such as starch, protein, pectin etc., promote the low liposoluble ingredient of some polarity to be converted into glucosides class soluble in water and be conducive to extraction.Human Chinese herbal medicine studies show that microbial fermentation can must discharge medium-height grass the effective elements of the medicine or active material to greatest extent, and active ingredient or the active material of Chinese herbal medicine are modified the new active material of generation.But the research of the fermentation of present feeding Chinese herbal medicine and enzymolysis is used in the ascendant, and a few feeding fermentedly extracts product with the enzymolysis Chinese herbal medicine or research remains in following 3 deficiencies both at home and abroad:
1., a few is feeding fermented with the research of enzymolysis and extraction Chinese herbal medicine or product all is single aerobic fermentation or anaerobic fermentation, namely or only adopts aerobic fermentation, or only adopts anaerobic fermentation, has no report or the product of aerobic and anaerobism asynchronous fermentation.2., prescription feeding fermented and the enzymolysis and extraction Chinese herbal medicine also is single mostly, lacks medium-height grass prescription drug compound fermented product or the research of science.3., the extraction of fermentation and enzymolysis Chinese herbal medicine and drying science not, the dregs of a decoction and liquid do not have separate drying.
Summary of the invention
Technical problem to be solved by this invention is: the research or the product that are intended to overcome a few feeding fermented extraction Chinese herbal medicine on the above-mentioned market all are single aerobic fermentation or anaerobic fermentations, and Chinese herbal medicine Chinese herbal medicine fermented product or research also be single mostly, lack scientific composition and the dregs of a decoction and liquid and do not have the problems such as separate drying, intend adopting asynchronous fermentation, zymolysis technique extracts Chinese herbal medicine, be about to several feeding Chinese herbal medicine impurity eliminations, after the ultramicro grinding, add water in proportion and nutritive salt is made Chinese herbal medicine fermented liquid, access first aspergillus niger and carry out aerobic fermentation, cellulase-producing and pectase, rear access lactoenterococcus and Lactobacillus plantarum are carried out anaerobic fermentation, the damaged cell membrane of enzymolysis again, the stripping saponin, alkaloid, sterol organic acid isoreactivity material.Zymotic fluid adopts plate and frame filter press to filter, the direct low temperature spray drying of filtrate is made antibiotic substitute A powder, filter cake adopts the roller drier oven dry to make antibiotic substitute B powder, A powder and B powder is mixed in proportion makes the fermentation enzymolysis Chinese herbal medicine extract that can replace feeding antibiotic.
The technical solution used in the present invention is as follows:
After Chinese herbal medicine impurity elimination, ultramicro grinding, be 1: 10: 0.005～10 mixing by Chinese herbal medicine, water, nutritive salt weight ratio, be warming up to 95～100 ℃ and soaked 2～4 hours, carry out the material lixiviate of Chinese medicine water-soluble active and sterilization, when treating that temperature drops to 38～40 ℃;
Described Chinese herbal medicine is comprised of camellia seed cake powder, folium cortex eucommiae, the Radix Astragali, hawthorn, pine needle meal, barrenwort, Radix Glycyrrhizae, and weight ratio is 1: 0.1～0.8: 0.1～0.8: 0.08～0.6: 0.08～0.6: 0.05～0.5: 0.05～0.5;
Be the aspergillus niger kind liquid of 10～30: 1 access cellulase-producing and pectase by Chinese herbal medicine fermented liquid, aspergillus niger kind liquid weight ratio, 6～96 hours cellulase-producings of aerobic fermentation and pectase;
Be 10～30: 1 access lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid by aerobic fermentation zymotic fluid, lactobacillus inoculation liquid weight ratio, anaerobic fermentation, enzymolysis 4～48 hours;
After the fermentation ends, 38～40 ℃ of enzymolysis of constant temperature 0.5～5 hour;
Use the plate and frame filter press filtering fermentating liquid;
The direct low temperature spray drying of filtrate is made antibiotic substitute A powder, and filter cake adopts the roller drier oven dry to make antibiotic substitute B powder, is that 1: 5～20 ratio is made the antibiotic substitute finished product by weight with A powder and B powder.
Preferably, the weight ratio of described Chinese herbal medicine is 1: 0.3: 0.3: 0.25: 0.25: 0.20: 0.20.
Further, described nutritive salt is by glucose, yeast extract, K
2HPO
4, NaCl, MgSO
4Form, weight ratio is 1: 0.15～0.3: 0.06～0.24: 0.03～0.09: 0.003～0.015.
Preferably, the weight ratio of described nutritive salt is 1: 0.24: 0.15: 0.06: 0.009.
Further, the weight ratio of the Chinese herbal medicine in the described Chinese herbal medicine fermented liquid, water, nutritive salt is 1: 10: 0.01.
Further, described aerobic fermentation is 20: 1 access aspergillus niger kind liquid by Chinese herbal medicine fermented liquid, aspergillus niger kind liquid weight ratio, aerobic fermentation 36 hours.
Further, described anaerobic fermentation is 20: 1 access lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid by aerobic fermentation zymotic fluid, lactobacillus inoculation liquid weight ratio, anaerobic fermentation, enzymolysis 16 hours.
Further, A powder and B powder are mixed in 1: 8 ratio and make antibiotic substitute.
The specific embodiment
Embodiment 1
Take by weighing camellia seed cake powder 1000kg, folium cortex eucommiae 300kg, Radix Astragali 300kg, hawthorn 250kg, pine needle meal 250kg, barrenwort 200kg, Radix Glycyrrhizae 200kg, after above-mentioned 8 kinds of Chinese herbal medicine impurity eliminations, being crushed to 80～120 orders (crossing the 0.15mm sieve) with micronizer after mixing, to make compound Chinese herb for subsequent use.
Take by weighing glucose 100kg, yeast extract 24kg, K
2HPO
415kg, NaCl 6kg, MgSO
40.9kg it is for subsequent use to make nutritive salt after the mixing.
With inclined-plane and MRS fluid nutrient medium with twice of aspergillus niger, lactoenterococcus and lactobacillus plantarum strain activation after, be inoculated in the 250mL triangular flask, every bottled liquid measure 100mL is after 39 ℃ of constant temperature leave standstill and cultivate 24h, by 2 * 10
7CFU/mL adds the concentration liquid seed fermentation tank of 5L that spreads cultivation, again by 1.5 * 10
7CFU/mL adds concentration and spreads cultivation in the liquid seed fermentation tank of 50L, and cultivation and fermentation 24h under 39 ℃ of conditions makes aspergillus niger strain liquid and lactoenterococcus, Lactobacillus plantarum mixed bacteria liquid is for subsequent use.
After taking by weighing above-mentioned compound Chinese herb 1000kg, nutritive salt 10kg and 10000kg water mixed dissolution, pass into steam and be warming up to 95～100 ℃ of immersions 3 hours, carry out the material lixiviate of Chinese medicine water-soluble active and sterilization.
When cooling to 38～40 ℃, inoculated aspergillus niger kind liquid 550kg passes into filtrated air fermentation 36 hours.
Inoculate lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid 500kg, anaerobic fermentation, enzymolysis 16 hours.
After the fermentation ends, 38～40 ℃ of enzymolysis of constant temperature 2 hours, use again the plate and frame filter press filtering fermentating liquid, the direct low temperature spray drying of filtrate is made antibiotic substitute A powder 118kg, filter cake adopts the roller drier oven dry to make antibiotic substitute B powder 980kg, and A powder and B powder are made Chinese herbal medicine fermentation and enzymatic hydrolyzed extract (antibiotic substitute finished product) in 1: 8 ratio.
Embodiment 2
Take by weighing camellia seed cake powder 500kg, folium cortex eucommiae 150kg, Radix Astragali 150kg, hawthorn 125kg, pine needle meal 125kg, barrenwort 100kg, Radix Glycyrrhizae 100kg, after above-mentioned 8 kinds of Chinese herbal medicine impurity eliminations, being crushed to 80～120 orders (crossing the 0.15mm sieve) with micronizer after mixing, to make compound Chinese herb for subsequent use.
Take by weighing glucose 50kg, yeast extract 12kg, K
2HPO
47.5kg, NaCl 3kg, MgSO
40.45kg it is for subsequent use to make nutritive salt after the mixing.
With inclined-plane and MRS fluid nutrient medium with twice of aspergillus niger, lactoenterococcus and lactobacillus plantarum strain activation after, be inoculated in the 250mL triangular flask, every bottled liquid measure 100mL is after 39 ℃ of constant temperature leave standstill and cultivate 24h, by 2 * 10
7CFU/mL adds the concentration liquid seed fermentation tank of 5L that spreads cultivation, again by 1.5 * 10
7CFU/mL adds concentration and spreads cultivation in the liquid seed fermentation tank of 50L, and cultivation and fermentation 24h under 39 ℃ of conditions makes aspergillus niger strain liquid and lactoenterococcus, Lactobacillus plantarum mixed bacteria liquid is for subsequent use.
After taking by weighing above-mentioned compound Chinese herb 500kg, nutritive salt 8kg and 5000kg water mixed dissolution, pass into steam and be warming up to 95～100 ℃ of immersions 3 hours, carry out the material lixiviate of Chinese medicine water-soluble active and sterilization.
When cooling to 38～40 ℃, inoculated aspergillus niger kind liquid 260kg passes into filtrated air fermentation 36 hours.
Inoculate lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid 250kg, anaerobic fermentation, enzymolysis 16 hours.
After the fermentation ends, 38～40 ℃ of enzymolysis of constant temperature 2.5 hours, use again the plate and frame filter press filtering fermentating liquid, the direct low temperature spray drying of filtrate is made antibiotic substitute A powder 55kg, filter cake adopts the roller drier oven dry to make antibiotic substitute B powder 450kg, and A powder and B powder are made Chinese herbal medicine fermentation and enzymatic hydrolyzed extract in 1: 8.18 ratio.
Embodiment 3
Take by weighing camellia seed cake powder 5000kg, folium cortex eucommiae 1500kg, Radix Astragali 1500kg, hawthorn 1250kg, pine needle meal 1250kg, barrenwort 1000kg, Radix Glycyrrhizae 1000kg, after above-mentioned 8 kinds of Chinese herbal medicine impurity eliminations, being crushed to 80～120 orders (crossing the 0.15mm sieve) with micronizer after mixing, to make compound Chinese herb for subsequent use.
Take by weighing glucose 200kg, yeast extract 48kg, K
2HPO
430kg, NaCl 12kg, MgSO
41.8kg it is for subsequent use to make nutritive salt after the mixing.
With inclined-plane and MRS fluid nutrient medium with twice of aspergillus niger, lactoenterococcus and lactobacillus plantarum strain activation after, be inoculated in the 250mL triangular flask, every bottled liquid measure 100mL is after 39 ℃ of constant temperature leave standstill and cultivate 24h, by 2 * 10
7CFU/mL adds the concentration liquid seed fermentation tank of 5L that spreads cultivation, again by 1.5 * 10
7CFU/mL adds concentration and spreads cultivation in the liquid seed fermentation tank of 50L, and cultivation and fermentation 24h under 39 ℃ of conditions makes aspergillus niger strain liquid and lactoenterococcus, Lactobacillus plantarum mixed bacteria liquid is for subsequent use.
After taking by weighing above-mentioned compound Chinese herb 5000kg, nutritive salt 60kg and 50000kg water mixed dissolution, pass into steam and be warming up to 95～100 ℃ of immersions 3 hours, carry out the material lixiviate of Chinese medicine water-soluble active and sterilization.
When cooling to 38～40 ℃, inoculated aspergillus niger kind liquid 3000kg passes into filtrated air fermentation 36 hours again.
Inoculate lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid 2500kg, anaerobic fermentation, enzymolysis 16 hours.
After the fermentation ends, 38～40 ℃ of enzymolysis of constant temperature 1.5 hours, use again the plate and frame filter press filtering fermentating liquid, the direct low temperature spray drying of filtrate is made antibiotic substitute A powder 580kg, filter cake adopts the roller drier oven dry to make antibiotic substitute B powder 4800kg, and A powder and B powder are made Chinese herbal medicine fermentation and enzymatic hydrolyzed extract in 1: 8.26 ratio.
One, Chinese herbal medicine fermentation and enzymatic hydrolyzed extract activity substance content are as shown in table 1.
Activity substance content (butt) in table 1 aspergillus niger and the lactic acid bacteria asynchronous fermentation enzymolysis Chinese herbal medicine product
Annotate: main triterpenoid saponin (〉=30.0%) and the carbohydrate (total reducing sugar 〉=30.0%, wherein polysaccharide 〉=25.0%) of containing of saccharicter-penin (SHP)
Two, the application study of antibiotic substitute
1, on the impact of Production Performance of Weaning Pigs, as shown in table 2.
The fermentation of table 2 Chinese herbal medicine and the impact of enzymatic hydrolyzed extract on Production Performance of Weaning Pigs
2, on the impact of diarrhea of weaned piglets and intestinal microflora, as shown in table 3.
The fermentation of table 3 Chinese herbal medicine and the impact of enzymatic hydrolyzed extract on diarrhea of weaned piglets and intestinal microflora
Annotate: 1. severe diarrhea finger to finger test piglet ight soil outward appearance is aqueous, and not moulding, and liquid dung has segregation phenomenon;
2. intestinal microflora unit: Log
10The cfu/g intestinal contents.
3, on the impact of meat chicken production performance, as shown in table 4.
The fermentation of table 4 Chinese herbal medicine and the impact of enzymatic hydrolyzed extract on meat chicken production performance
Can find out from above table,
The antibiotic substitute of the present invention's preparation all has good production performance to various animals, can substitute preferably original antibiotic goods.
Above content only is to structure example of the present invention and explanation; under those skilled in the art described specific embodiment is made various modifications or replenish or adopt similar mode to substitute; only otherwise depart from the structure of invention or surmount this scope as defined in the claims, all should belong to protection scope of the present invention.
Claims (8)
1. the method for a preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine is characterized in that:
After Chinese herbal medicine impurity elimination, ultramicro grinding, be 1: 10: 0.005～10 mixing by Chinese herbal medicine, water, nutritive salt weight ratio, be warming up to 95～100 ℃ and soaked 2～4 hours, carry out the material lixiviate of Chinese medicine water-soluble active and sterilization, when treating that temperature drops to 38～40 ℃;
Described Chinese herbal medicine is comprised of camellia seed cake powder, folium cortex eucommiae, the Radix Astragali, hawthorn, pine needle meal, barrenwort, Radix Glycyrrhizae, and weight ratio is 1: 0.1～0.8: 0.1～0.8: 0.08～0.6: 0.08～0.6: 0.05～0.5: 0.05～0.5;
Be the aspergillus niger kind liquid of 10～30: 1 access cellulase-producing and pectase by Chinese herbal medicine fermented liquid, aspergillus niger kind liquid weight ratio, 6～96 hours cellulase-producings of aerobic fermentation and pectase;
Be 10～30: 1 access lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid by aerobic fermentation zymotic fluid, lactobacillus inoculation liquid weight ratio, anaerobic fermentation, enzymolysis 4～48 hours;
After the fermentation ends, 38～40 ℃ of enzymolysis of constant temperature 0.5～5 hour;
Use the plate and frame filter press filtering fermentating liquid;
The direct low temperature spray drying of filtrate is made antibiotic substitute A powder, and filter cake adopts the roller drier oven dry to make antibiotic substitute B powder, is that 1: 5～20 ratio is made the antibiotic substitute finished product by weight with A powder and B powder.
2. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1 is characterized in that, the weight ratio of described Chinese herbal medicine is 1: 0.3: 0.3: 0.25: 0.25: 0.20: 0.20.
3. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1 is characterized in that, described nutritive salt is by glucose, yeast extract, K
2HPO
4, NaCl, MgSO
4Form, weight ratio is 1: 0.15～0.3: 0.06～0.24: 0.03～0.09: 0.003～0.015.
4. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 3 is characterized in that, the weight ratio of described nutritive salt is 1: 0.24: 0.15: 0.06: 0.009.
5. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1 is characterized in that, the weight ratio of the Chinese herbal medicine in the described Chinese herbal medicine fermented liquid, water, nutritive salt is 1: 10: 0.01.
6. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1, it is characterized in that, described aerobic fermentation is 20: 1 access aspergillus niger kind liquid by Chinese herbal medicine fermented liquid, aspergillus niger kind liquid weight ratio, aerobic fermentation 36 hours.
7. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1, it is characterized in that, described anaerobic fermentation is 20: 1 access lactoenterococcus and Lactobacillus plantarum mixed bacteria liquid by aerobic fermentation zymotic fluid, lactobacillus inoculation liquid weight ratio, anaerobic fermentation, enzymolysis 16 hours.
8. the method for a kind of preparing antibiotic substituent for feed by probiotic asynchronous fermentation of Chinese herbal medicine according to claim 1 is characterized in that, A powder and B powder are mixed in 1: 8 ratio and make antibiotic substitute.
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