CN101444252A - Method for preparing fermented virus-free cottonseed meal feed protein - Google Patents
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Abstract
The invention discloses a method for preparing fermented virus-free cottonseed meal feed protein. The invention relates to a production process which uses cottonseed meal as raw material and produces the virus-free cottonseed meal feed protein by microbial technology in a transformation way, in particular to a microbial fermenting process of using multi-strains mixed solid to ferment and prepare virus-free cottonseed meal. Fermentation cultures of one strain Saccharomyces cerevisiae which is cultured in an amplification way, one strain Candida tropicalis, one strain Bacillus ubtilis and one strain Lactobacillus are evenly mixed in the proportion of 2 to 1 to 1 to 1 and are inoculated in cottonseed meal (through 40 mesh sieve) with 5 percent of the inoculation amount, wherein, the water content of the cottonseed meal is 50 percent; simultaneously, substrate culture media (5 percent of molasses, 5 percent of bran, 0.5 percent of ammonium sulfate, and the initial pH value is 6.0) are added, evenly mixed, fermented for 72hours at 30 DEG C in aerobic way, dried and sampled at 60 DEG C; finally, the nutrient analysis is carried out on the cottonseed meal before and after fermentation; when weighed and packaged, finished products of the fermented cottonseed meal can be stored for 6 months at a cool and dry place. The fermented cottonseed meal can be used as feeds for beef cattle, chicken, tilapia, etc.
Description
Technical field
The present invention relates to the cottonseed dregs of rice is raw material, utilizes microbial technique to transform a kind of production technology of producing virus-free cottonseed meal feed protein, relates in particular to the microorganism fermentation process that a kind of many solid type mixed culture fermentations prepare virus-free cottonseed meal.
Background technology
In recent years, China's animal husbandry and feed industrial development are very rapid, and the contradiction that feed resource is in short supply becomes increasingly conspicuous.According to scholarly forecast, 2010～the year two thousand twenty, the difference of China's protein feeds is 2400～4,800 ten thousand tons, puffed soybean difference is 2,560 ten thousand tons.For a long time, the protein feed raw material dregs of beans that China mainly relies on, because of being subjected to influence of various factors such as output, weather, domestic and international market price fluctuation, supply day is becoming tight, and price irregularly goes up and fluctuates, and is bigger to the Feed Enterprise cost impact.Though and more existing its quantity of agricultural byproducts is abundant and price is cheap, because of containing ANFs or being subject to conventional machining process, has limited its effective utilization.At these problems, relevant researcher has carried out the developmental research of a lot of aspects about protein feeds both at home and abroad, and wherein effective utilization of the relevant cottonseed dregs of rice is exactly one of research emphasis.
The cottonseed dregs of rice are China's another kind of important puffed soybean protein feeds except that dregs of beans (cake), but because of containing to the virulent free gossypol of animal and some ANFs, its value and use amount are subjected to very big restriction, in order to enlarge its range of application, must carry out detoxification treatment earlier and be worth to improve applied nutrition.The main poison-removing method of the cottonseed dregs of rice has physics method, chemical method and bioanalysis at present.Though physics method and chemical detoxication method can significantly reduce free gossypol content in the cotton cake dregs, all have certain shortcoming.Handle the feeding value that the residual material in back and color and luster problem have often reduced cotton cake dregs with chemicals, organic solvent.Bioanalysis comprises enzymatic isolation method and microbe fermentation method, and the enzymatic isolation method detoxification efficiency is undesirable.Microbe fermentation method is a kind of new cotton cake dregs detoxification approach that at first grows up in China in the later stage eighties 20th century.This kind method not only detoxification efficiency is good, avoids the influence to the cottonseed protein functional character of chemical addition agent and physics method, also can improve palatability, increases protein content; The various digestive ferments that microbial fermentation produces, amino acid, vitamin etc. have also improved the nutritive value of cotton cake dregs greatly.Therefore, the cottonseed dregs of rice after microbial fermentation is handled, not only can be used as ruminant and aquatic animal feed, but also can partly replace protein feeds such as dregs of beans, fish meal in the livestock and poultry diet, or be used with other meal expeller and mealsolvent, both safe in utilization, can reduce cost again, increase economic efficiency, have good development potentiality.
The technology of the present invention is primarily aimed at the key issue of above-mentioned cottonseed dregs of rice detoxification, fermentation parameter by the many solid type mixed culture fermentations of system research, finally obtained a kind of microbial fermentation processes for preparing virus-free cottonseed meal feed protein, this all has important theory and realistic meaning for improving cottonseed dregs of rice microbial fermentation virus elimination rate and the aspects such as effective utilization of cottonseed cake protein in animal husbandry.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of fermented virus-free cottonseed meal feed protein, further promote the effective utilization of cotton cake dregs in animal husbandry, make it become one of approach that solves the present protein feed resource shortage of China.
Purpose of the present invention is achieved by the following technical programs:
The preparation method of a kind of fermented virus-free cottonseed meal feed protein provided by the invention the steps include:
A. belong to picking one ring thalline bacterium and the Blastocystis fungi from original bacterial classification hay bacillus, be inoculated into respectively in LB seed culture medium and the brewer's wort seed culture medium, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table is cultivated after 16-24 hour, up to OD
600=0.7-0.9 stops to cultivate, as seed liquor.Picking one ring thalline is inoculated in the genus lactubacillus culture medium from original bacterial classification genus lactubacillus bacterium, and static cultivation is 40-48 hour under 30 ℃ of-35 ℃ of temperature, as seed liquor.
B. in liquid enlarging LB culture medium (hay bacillus belongs to bacterium) and malt extract medium (Blastocystis fungi), inoculate seed liquor with 1-5% inoculum concentration, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table was cultivated 20-28 hours.With 1-5% inoculum concentration inoculation seed liquor, static cultivation is 48 hours under 30 ℃ of-35 ℃ of temperature in liquid expansion genus lactubacillus culture medium (genus lactubacillus bacterium).The various strain fermentation nutrient solutions of saccharomyces cerevisiae, candida tropicalis, bacillus subtilis, lactic acid bacteria are mixed in the 2:1:1:1 ratio, as cottonseed dregs of rice fermented bacterium liquid.
C. cottonseed dregs of rice fermented bacterium liquid among the b being inoculated into water content with 5% inoculum concentration is in 50% the cottonseed dregs of rice (crossing 40 mesh sieves), add substrate cultivation base (molasses 5% simultaneously, wheat bran 5%, ammonium sulfate 0.5%, initial pH value is 6.0) and mix, aerobic fermentation is 72 hours under 30 ℃ of temperature, then in 60 ℃ of dry sample preparations down, at last the cottonseed dregs of rice before and after the fermentation is carried out the nutritional labeling analysis.
D. the cottonseed dregs of rice finished product that ferments is put in shady and cool dry place after weighing, packing, at room temperature can deposit 6 months.
E. the cottonseed dregs of rice after the fermentation promptly can be used as the feed of beef cattle, chicken, Tilapia mossambica etc.
Bacterial classification seed LB culture medium described in the step a of the present invention contains 0.3%-0.5% sodium chloride, 0.3%-0.5% peptone and 0.5%-1.0% yeast extract by weight percentage, and the genus lactubacillus culture medium contains the glucose, 0.3% KH of NaCl, 0.2%-0.4% of tryptone, the 0.3%-0.5% of 0.2%-0.4%
2PO
4, 0.3% K
2HPO
4, 0.02% MgCl
2, 0.005% CaSO
4With 0.2% starch, malt extract medium contains 10% malt extract powder and 0.01% chloramphenicol.
Bacterial classification described in the step a of the present invention is bacillus subtilis (Bacillus subtilis) 1 strain, lactic acid bacteria (Lactobacillus) 1 strain, saccharomyces cerevisiae (Saccharomyces crevisiae) 1 strain and candida tropicalis (Candida tropicalis) 1 strain.
The prescription of three kinds of fluid enlargement culture bases described in the step b of the present invention is identical with step a.
The present invention has following beneficial effect: the present invention adopts the microbial fermentation processing cottonseed dregs of rice, and not only detoxification efficiency is good, has avoided the influence to the cottonseed protein functional character of chemical addition agent and physics method, also can improve palatability, increases protein content; The various digestive ferments that sweat produces, amino acid, vitamin etc. have also improved the nutritive value of the cottonseed dregs of rice greatly.Therefore, the cotton cake dregs after microbial fermentation is handled is even also can partly replace protein feeds such as dregs of beans, fish meal in livestock and poultry diet, or be used with other meal expeller and mealsolvent, both safe in utilization, can reduce cost again, increase economic efficiency, have good development potentiality.
The specific embodiment
The preparation method's of a kind of fermented virus-free cottonseed meal feed protein provided by the invention embodiment is as follows:
The used bacterial classification of present embodiment is bacillus subtilis (Bacillus subtilis) 1 strain, lactic acid bacteria (Lactobacillus) 1 strain, saccharomyces cerevisiae (Saccharomyces crevisiae) 1 strain and candida tropicalis (Candida tropicalis) 1 strain.Its concrete steps are:
A. the LB fluid nutrient medium of being made up of 5 gram peptones, 5 gram yeast extracts, 3 gram sodium chloride and 1 premium on currency (pH7.0) is poured in the aseptic triangular flask, makes bacillus subtilis seed fluid nutrient medium and fluid enlargement culture base; By 2 gram peptones, 5 gram NaCl, 2 gram glucose, 3 gram KH
2PO
4, 3 the gram K
2HPO
4, 0.2 the gram MgCl
2, 0.05 the gram CaSO
4, the genus lactubacillus culture medium formed of 2 gram starch and 1 premium on currency (pH6.5),, pour in the aseptic triangular flask, make lactobacillus solution body culture medium and fluid enlargement culture base; By the malt extract medium that 100 gram malt extract powder, 0.01 gram chloramphenicol and 1 premium on currency (pH6.0) are formed, pour in the aseptic triangular flask, make saccharomycete seed fluid nutrient medium and fluid enlargement culture base.
B. belong to picking one ring thalline bacterium and the Blastocystis fungi from original bacterial classification hay bacillus, be inoculated into respectively in LB seed culture medium and the brewer's wort seed culture medium, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table is cultivated after 16-24 hour, up to OD
600=0.7-0.9 stops to cultivate, as seed liquor.Picking one ring thalline is inoculated in the genus lactubacillus culture medium from original bacterial classification genus lactubacillus bacterium, and static cultivation is 40-48 hour under 30 ℃ of-35 ℃ of temperature, as seed liquor.
C. in liquid enlarging LB culture medium (hay bacillus belongs to bacterium) and malt extract medium (Blastocystis fungi), inoculate seed liquor with 1-5% inoculum concentration, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table was cultivated 20-28 hours.With 1-5% inoculum concentration inoculation seed liquor, static cultivation is 48 hours under 30 ℃ of-35 ℃ of temperature in liquid expansion genus lactubacillus culture medium (genus lactubacillus bacterium).The strain fermentation nutrient solution of saccharomyces cerevisiae, candida tropicalis, bacillus subtilis, lactic acid bacteria is mixed in the 2:1:1:1 ratio, as cottonseed dregs of rice fermented bacterium liquid.
D. cottonseed dregs of rice fermented bacterium liquid being inoculated into water content with 5% inoculum concentration is in 50% the cottonseed dregs of rice (crossing 40 mesh sieves), add substrate cultivation base (molasses 5% simultaneously, wheat bran 5%, ammonium sulfate 0.5%, initial pH value is 6.0) and mix, aerobic fermentation is 72 hours under 30 ℃ of temperature, then in 60 ℃ of dry sample preparations down.
E. the cottonseed dregs of rice before and after the fermentation are carried out the nutritional labeling analysis.
F. the cottonseed dregs of rice finished product that ferments is put in shady and cool dry place after weighing, packing, at room temperature can deposit 6 months.
G. the cottonseed dregs of rice after fermenting among the step f promptly can be used as the feed of beef cattle, chicken, Tilapia mossambica etc.
The ANFs of the cottonseed dregs of rice finished product of microbe fermentation method preparation obviously reduces in above-mentioned example, crude protein and soluble protein content are increased, middle small molecular weight protein showed increased, improved palatability greatly, amino acid content tends to balance, vitamin is abundant, and contains beneficial bacterium and UGF, and nutritive value improves greatly.Can improve use amount and the scope of application of the cottonseed dregs of rice on animal produces more significantly.Its effect is as follows:
Select yellow fryer 480 plumages in the healthy 21 active age in days commodity south of the Five Ridges, adopt the single factor experiment design, establish 4 groups, establish 4 repetitions for every group, each repeats 30 plumages.Control group is for adding the typical daily ration of the 3% common cottonseed dregs of rice, and the common cottonseed dregs of rice and part dregs of beans that test group 1,2,3 is added respectively in the alternative basal diet of 5%, the 8% and 10% fermentation cottonseed dregs of rice are formulated.26 ages in days enter formal test, 22 days experimental periods.The growth performance index of last determination test fryer, butcher performance, alimentary canal ponderal index, immune indexes, blood parameters.The result is:
(1) adds 5%, 8% and 10% the fermentation cottonseed dregs of rice in the daily ration of broiler respectively to all not remarkable (p〉0.05) (table 1) of average daily ingestion amount, average daily gain and the feed conversion rate difference of fryer; (2) add 5%, 8% and 10% the fermentation cottonseed dregs of rice in the daily ration of broiler respectively to complete clean thorax rate, half clean thorax rate, chest muscle rate and the leg flesh rate difference of fryer all not significantly (p〉0.05), meat production is good.Add 10% the fermentation cottonseed dregs of rice in the daily ration, dressing percentage is significantly higher than and adds 5% the fermentation cottonseed dregs of rice, and along with the rising of adding fermentation cottonseed dregs of rice ratio in the daily ration, dressing percentage and half clean thorax rate have trend of rising (table 2); (3) add in the daily ration 5% the fermentation cottonseed dregs of rice extremely significantly (p＜0.01) reduced the abdomen fat rate of fryer, but along with the rising of fermentation cottonseed dregs of rice ratio in the daily ration, fryer abdomen fat rate has trend of rising (table 2); (4) add 5%, 8% and 10% the fermentation cottonseed dregs of rice in the daily ration of broiler respectively alimentary canal, immune organ and the blood parameters aspect of fryer are not all had negative effect (table 3-5); (5) the cottonseed dregs of rice that adopt this experimental condition and mode to ferment, adding proportion can reach 10% in daily ration of broiler, but best with the fermentation cottonseed dregs of rice feeding effect that adds 5% ratio.
Table 1 adds the fermentation cottonseed dregs of rice of different proportion to the growth of meat chicken Effect on Performance
Annotate: the different differences extremely significantly (p＜0.01) of representing of colleague's shoulder motes capitalization, the different significant differences (p＜0.05) of representing of colleague's shoulder motes lowercase, colleague's shoulder motes identical lowercase represent difference not significantly (p〉0.05), and the mark person does not represent difference not remarkable, down with.
The fermentation cottonseed dregs of rice that add different proportion in table 2 daily ration are butchered the influence of index to fryer
Table 3 adds the influence of the fermentation cottonseed dregs of rice of different proportion to fryer alimentary canal ponderal index
Table 4 adds the influence of the fermentation cottonseed dregs of rice of different proportion to the fryer immune organ
Table 5 adds the influence of the fermentation cottonseed dregs of rice of different proportion to the fryer blood parameters
Claims (3)
1, a kind of preparation method of fermented virus-free cottonseed meal feed protein the steps include:
A. belong to picking one ring thalline bacterium and the Blastocystis fungi from original bacterial classification hay bacillus, be inoculated into respectively in LB seed culture medium and the brewer's wort seed culture medium, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table is cultivated after 16-24 hour, up to OD
600=0.7-0.9 stops to cultivate, as seed liquor.Picking one ring thalline is inoculated in the genus lactubacillus culture medium from original bacterial classification genus lactubacillus bacterium, and static cultivation is 40-48 hour under 30 ℃ of-35 ℃ of temperature, as seed liquor.
B. in liquid enlarging LB culture medium (hay bacillus belongs to bacterium) and malt extract medium (Blastocystis fungi), inoculate seed liquor with 1-5% inoculum concentration, under 30 ℃ of-37 ℃ of temperature, 150-170rpm, shaking table was cultivated 20-28 hours.With 1-5% inoculum concentration inoculation seed liquor, static cultivation is 48 hours under 30 ℃ of-35 ℃ of temperature in liquid expansion genus lactubacillus culture medium (genus lactubacillus bacterium).The various strain fermentation nutrient solutions of saccharomyces cerevisiae, candida tropicalis, bacillus subtilis, lactic acid bacteria are mixed in the 2:1:1:1 ratio, as cottonseed dregs of rice fermented bacterium liquid.
C. cottonseed dregs of rice fermented bacterium liquid among the b being inoculated into water content with 5% inoculum concentration is in 50% the cottonseed dregs of rice (crossing 40 mesh sieves), add substrate cultivation base (molasses 5% simultaneously, wheat bran 5%, ammonium sulfate 0.5%, initial pH value is 6.0) and mix, aerobic fermentation is 72 hours under 30 ℃ of temperature, then in 60 ℃ of dry sample preparations down, at last the cottonseed dregs of rice before and after the fermentation is carried out the nutritional labeling analysis.
D. the cottonseed dregs of rice finished product that ferments is put in shady and cool dry place after weighing, packing, at room temperature can deposit 6 months.
E. the cottonseed dregs of rice after the fermentation promptly can be used as the feed of beef cattle, chicken, Tilapia mossambica etc.
2, the preparation method of fermented virus-free cottonseed meal feed protein according to claim 1, bacterial classification seed LB culture medium described in its step a contains 0.3%-0.5% sodium chloride, 0.3%-0.5% peptone and 0.5%-1.0% yeast extract by weight percentage, and the genus lactubacillus culture medium contains the glucose, 0.3% KH of NaCl, 0.2%-0.4% of tryptone, the 0.3%-0.5% of 0.2%-0.4%
2PO
4, 0.3% K
2HPO
4, 0.02% MgCl
2, 0.005% CaSO
4With 0.2% starch, malt extract medium contains 10% malt extract powder and 0.01% chloramphenicol.
3, the preparation method of fermented virus-free cottonseed meal feed protein according to claim 1, the bacterial classification described in its step a is bacillus subtilis (Bacillus subtilis) 1 strain, lactic acid bacteria (Lactobacillus) 1 strain, saccharomyces cerevisiae (Saccharomycescrevisiae) 1 strain and candida tropicalis (Candida tropicalis) 1 strain.
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|---|---|
| CN101444252B (en) | 2011-06-08 |
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