CN103829040B - A kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans and application - Google Patents
A kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans and application Download PDFInfo
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Abstract
The present invention discloses a kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans and application.The present invention utilizes Chen Xiangcha starter-making stage, and temperature controls, at 28 ~ 30 DEG C, to be beneficial to mildew bacterium, is conducive to the generation of protease, carbohydrase; Fermentation stage aerobic bacteria utilizes limited oxygen to grow, and it is strong that bacillus subtilis takes oxygen by force, and fermentation tank is in anaerobic state very soon, and mould and oxygen consumption bacterium cannot grow, and now saccharomycete, lactic acid bacteria metabolism are strengthened.Under enzyme participates in, anaerobic bacteria metabolism reduces ANFs content, improves organic acid and acid-soluble protein content, forms high-quality fermented bean dregs.By Chen Xiangcha compound bacteria and bacillus subtilis coordinative role, without the need to carrying out sterilization treatment to dregs of beans, sweat amphimicrobian and anaerobic fermentation, energy consumption is little, and suppress the growth of raw material pathogenic bacteria, dry loss is low.Technological process of the present invention is easy and simple to handle, and both applicable plant directly produced and throws something and feeds, and was also applicable to feed factory large-scale production.
Description
Technical field
The invention belongs to biotechnology fermentation field, be specifically related to a kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans and application.
Background technology
The protein content of dregs of beans is very high, be the base stock preparing livestock and poultry feed, but Direct-fed protein bio conversion ratio is lower.Fermented bean dregs is the solubility that improve dregs of beans albumen by beneficial microbe effect, is beneficial to digestion; Reduce the molecular weight of albumen in dregs of beans, a part has wherein reached little peptide level even amino acid levels, can directly be absorbed by animal; Fermented bean dregs has certain aromatic odor and delicate flavour, and have certain food calling effect, palatability is better; In dregs of beans, some non-starch polysaccharide obtain decomposition during the fermentation, are conducive to animal digestion.
The bacterial classification exploitation of fermented feed, directly will affect ferment effect, the various physical and chemical factors of different microorganisms to material require different, and are difficult to maintain some physico chemical factors in sweat, therefore, determine bean pulp fermentation jointly worked in coordination with by multiple-microorganism or carry out in order.The stability of fermented bean dregs product quality, generally, single bacterium is not as many bacterium, and pure culture (compatibility between the pure culture of multi-cultur es is not necessarily coordinated) is not as the sort of quyi (microorganism in the sort of quyi is natural composition, with strong complementarity each other).Therefore excellent fermented bacterium exploitation need to study further and grope.
Chen Xiangcha refers to the teas that fresh leaf is processed into through wet heap by long-time spontaneous fermentation or gross tea through complete, knead, dry (oven dry).Mould in Chen Xiangcha is mainly aspergillus niger, penicillium chrysogenum, Aspergillus glaucus and head mold, and wherein aspergillus niger is in the majority, accounts for the 80%(mass fraction of total number of molds).Bacterium mainly contains bacillus as bacillus coagulans (Bacillus coagulans), bacillus licheniformis (Bacilluslicheniformis), thermophilic bacillus amyloliquefacienses (Bacillus thermoamylovorans) etc., also have Pediococcus acidilactici (Pediococcus acidilactici) and Lactobacillus plantarum (Lactobacillus plantarum) in addition.Budding fungus mainly produces alcohol and produces the aroma-producing yeasts such as ester.As: Hansenula anomala (Hansenulaanomala), Lu Shi yeast (Zygosaccharomyces rouxii), torulopsis (Torulopsis Berlese), saccharomyces cerevisiae (Saccharomyces cerevisiae), brettanomyce belongs to (Brettanomyces sp.) etc.Chen Xiangcha is very complicated at later stage fermentation, and because the protein content in tea is few, make harmful bacteria be difficult to existence, great majority are beneficial bacteriums, and Chen Xiangcha fermentation is in faintly acid, and therefore the overwhelming majority is beneficial bacterium.Meanwhile, the polyphenols in tealeaves can suppress the growth of harmful bacteria.There are some researches show, the extract of Chen Xiang teas has certain antimutagenic effect, has potential antibacterial activity to Gram-positive staphylococcus and rod bacterium.Prove that the security of Chen Xiangcha is very high.
Bacillus subtilis (Bacillus subtilis) Ministry of Agriculture of base China allows one of the two kinds of bacillus as feed addictive, is more and more developed into microorganism formulation.Its preparation is nontoxic, noresidue, free of contamination " green " additive, and have vast potential for future development, now extensive use in animal husbandry, feedstuff industry, shows huge social benefit and ecological benefits.Bacillus subtilis has very strong protease, lipase, amylase isoreactivity, antibiotic can be produced, there is comparatively Johnson & Johnson's thing in animal intestinal and take oxygen ability by force, these characteristics to promoting zootrophicly to digest and assimilate, the feed conversion rate that improves animal and diseases prevention, growth promoting effects plays an important role.
Utilize the method with Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, not yet have report so far.
Summary of the invention
In order to the shortcoming overcoming prior art is with not enough, primary and foremost purpose of the present invention is to provide a kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans.The method overcome the problem of compatibility not science, the trade-off effect difference that composite ferment exists.
Another object of the present invention is to provide the composite fermentation obtained by above-mentioned preparation method dregs of beans.
Another object of the present invention is to provide the described application of composite fermentation dregs of beans on feed.
Object of the present invention is achieved through the following technical solutions: a kind of with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, comprises following steps: each component by mass percentage
(1) grind under Chen Xiangcha normal temperature, obtain Chen Xiangcha powder;
(2) sort of quyi preparation: the Chen Xiangcha powder obtained by 1% access step (1) of solid medium A total amount, mixing, cultivates; After mycelia is covered with solid medium A, shaking flask; Solid medium A surface covers with spore, and the sort of quyi has been cultivated;
(3) bent preparation is planted: by the sort of quyi that 5% ~ 15% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly, cultivate; After solid medium B surface covers with mycelia, start to turn over bent heat radiation; After whole solid medium B covers with spore, Chen Xiangcha microbial inoculum has been cultivated;
(4) bacillus subtilis formulation preparation: by the bacillus subtilis of 1% ~ 2% access activation of fluid nutrient medium total amount, cultivate, can bacillus subtilis formulation be obtained;
(5) by bacillus subtilis formulation 2% ~ 8%, molasses 2 ~ 10%, water 17% ~ 31% Homogeneous phase mixing that Chen Xiangcha microbial inoculum 10 ~ 20% obtained for step (3), step (4) obtain, then mix with 45 ~ 55% dregs of beans, obtain dregs of beans mixture;
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 3 ~ 6 days of 25 ~ 37 DEG C, the composite fermentation dregs of beans fermented is obtained;
(8) the composite fermentation dregs of beans fermented that obtains of step (7), can Direct-fed livestock and poultry.
In order to realize the present invention better:
Chen Xiangcha described in step (1) is preferably the black tea in Anhua, Hunan, Guangxi Liu Bao loose tea, yunnan puer tea or Guangdong great Ye green grass or young crops etc.; Be more preferably yunnan puer tea;
Normal temperature described in step (1) is preferably 15 ~ 25 DEG C;
Solid medium A described in step (2) is prepared preferably by following steps: each component by mass percentage: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min;
The temperature of the cultivation described in step (2) is preferably 28 ~ 30 DEG C;
Preferred every 8 hours shaking flasks 1 time after shaking flask described in step (2);
Dividing described in step (3) is preferably divided on bamboo sieve, and puts into kind of a bent room;
The thickness of described bamboo sieve is preferably 4 ~ 6cm;
The condition of the cultivation described in step (3) is preferably 28 ~ 30 DEG C, and humid control is more than 90%;
Solid medium B described in step (3) is prepared preferably by following steps: each component by mass percentage: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min;
The condition of the cultivation described in step (4) is preferably 37 DEG C of ventilate cavity 24 hours;
Fluid nutrient medium described in step (4) is prepared preferably by following steps: each component is by quality percent by volume (g/mL): 0.5% sodium chloride, 0.5% dusty yeast, and 1.0% peptone, adds water, 121 DEG C of sterilizing 15min;
Described Chen Xiangcha, wheat bran, molasses are commercially available prod;
Described composite fermentation dregs of beans is obtained by said method;
Described composite fermentation dregs of beans forms stronger wine flavour, band tart flavour; PH is preferably 3.08 ~ 4.35; Organic acid content is preferably 3.23% ~ 4.35%; Acid-soluble protein is preferably 10.76% ~ 15.29%; Urease activity preferably reduces by 80 ~ 94%; Bacillus globigii spores number is preferably 9.7 × 10
9more than cfu/g; Lactic acid bacteria number is preferably 8.3 × 10
8more than cfu/g;
The described application of composite fermentation dregs of beans on feed.
Described composite fermentation dregs of beans directly to be fed livestock and poultry as feed, or adds 4% premix mixing and to feed livestock and poultry.
Mechanism of the present invention is: utilize Chen Xiangcha starter-making stage, and temperature controls at 28 ~ 30 DEG C, is beneficial to mildew bacterium (aspergillus is main), is conducive to the generation of protease, carbohydrase.Fermentation stage aerobic bacteria utilizes limited oxygen to grow, and it is strong that bacillus subtilis takes oxygen by force, and fermentation tank is in anaerobic state very soon, and mould and oxygen consumption bacterium cannot grow, and now saccharomycete, lactic acid bacteria metabolism are strengthened.Under enzyme participates in, anaerobic bacteria metabolism reduces ANFs content, improves organic acid and acid-soluble protein content, forms high-quality fermented bean dregs.
The present invention, relative to prior art, has following advantage and effect:
(1) composite fermentation dregs of beans finished product is form the well reinforcing sort of quyi in Chen Xiangcha spontaneous fermentation maturation, and ecosystem is fermented, and Spawn incubation technique is simple.
(2) technological process of the present invention is easy and simple to handle, and both applicable plant directly produced and throws something and feeds, and was also applicable to feed factory large-scale production.
(3) by Chen Xiangcha compound bacteria and bacillus subtilis coordinative role, without the need to carrying out sterilization treatment to dregs of beans, sweat amphimicrobian and anaerobic fermentation, energy consumption is little, and suppress the growth of raw material pathogenic bacteria, dry loss is low.
(4) have strong sour fragrance, the most high energy of acid-soluble protein reaches 15.29%, and organic acid is up to 4.35%, and urease activity can reduce by 94%.Higher than market fermented bean dregs quality level.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment 1
Implement as follows: following component all by mass percentage
(1) grind under commercially available old scented tea (yunnan puer tea) normal temperature, obtain Chen Xiangcha powder;
(2) triangular flask sort of quyi preparation: be equipped with in 100g solid medium A500mL triangular flask, by culture medium 1% inoculum concentration access step (1) described Chen Xiangcha powder, mixing, 28 DEG C of cultivations.After mycelia is covered with culture medium, shaking flask, latter every 8 hours shaking flasks 1 time.Media surface covers with spore, and the triangular flask sort of quyi has been cultivated;
Described solid medium A mass percent: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
(3) bent preparation is planted: according to the triangular flask sort of quyi that 10% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly at bamboo sieve (commercially available prod) upper (thickness 6cm), put into kind of a bent room, 28 DEG C, humid control is cultivated more than 90%.After media surface covers with mycelia, start to turn over bent heat radiation.
After whole culture medium covers with spore, Chen Xiangcha microbial inoculum has been cultivated.
Described solid medium B mass percent: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
(4) bacillus subtilis formulation preparation: on liquid medium within, by the bacillus subtilis (Bacillus subtilis) (ATCC6633) (purchased from Guangdong Ying Wei feed corporation,Ltd) of 1% inoculum concentration access activation, 37 DEG C of ventilate cavity 24 hours, can obtain bacillus subtilis formulation.
The quality percent by volume (g/mL) of described fluid nutrient medium nutritional labeling: 0.5% sodium chloride, 0.5% dusty yeast, 1.0% peptone, adds water to 100mL, 121 DEG C of sterilizing 15min.
(5) by bacillus subtilis formulation 8%, molasses 6%, water 21% Homogeneous phase mixing that obtained for step (3) Chen Xiangcha microbial inoculum 10%, step (4) obtain, then mix with 55% dregs of beans, obtain dregs of beans mixture;
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 3 days of 37 DEG C, obtain the composite fermentation dregs of beans fermented, can directly directly use as finished feed.
The composite fermentation dregs of beans obtained as measurement result during finished feed in table 1.Wherein, the content of acid-soluble protein is according to QB/T2653-2004 method, and pH is recorded by pH meter, and organic acid is recorded by GB/T5009.39-2003 method, and urease activity is recorded by GB/T8622-2006.
The measurement result of table 1 finished feed
| Test item | Before fermentation | After fermentation | Ratio (before rear/fermentation of fermenting) |
| Color | Yellowish | Yellow | ------ |
| Particle | Particle is thicker | Particle is thin | ------ |
| Smell | Light beany flavour | Wine flavour and tart flavour | ------ |
| pH | 6.92 | 4.18 | ------ |
| Organic acid (%) | 0.62 | 4.14 | 6.68 |
| Acid-soluble protein (%) | 2.24 | 12.12 | 5.66 |
| Urease activity (U) | 0.402 | 0.081 | 20% |
Embodiment 2
Implement as follows: following component all by mass percentage
(1) grind under commercially available old scented tea (yunnan puer tea) normal temperature, obtain Chen Xiangcha powder;
(2) triangular flask sort of quyi preparation: be equipped with in 100g solid medium A500mL triangular flask, by culture medium 1% inoculum concentration access step (1) described Chen Xiangcha powder, mixing, 28 DEG C of cultivations.After mycelia is covered with culture medium, shaking flask, latter every 8 hours shaking flasks 1 time.Media surface covers with spore, and the triangular flask sort of quyi has been cultivated;
Described solid medium A mass percent: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
(3) bent preparation is planted: according to the triangular flask sort of quyi that 5%, 10%, 15% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly at bamboo sieve (commercially available prod) upper (thickness 5cm), put into kind of a bent room, 28 DEG C, humid control is cultivated more than 90%.After media surface covers with mycelia, start to turn over bent heat radiation.
After whole culture medium covers with spore, Chen Xiangcha microbial inoculum has been cultivated.
Described solid medium B mass percent: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
(4) bacillus subtilis formulation preparation: on liquid medium within, by the bacillus subtilis of 1%, 1.5%, 2% inoculum concentration access activation, 37 DEG C of ventilate cavity 24 hours, can obtain bacillus subtilis formulation.
The quality percent by volume (g/mL) of described fluid nutrient medium nutritional labeling: 0.5% sodium chloride, 0.5% dusty yeast, 1.0% peptone, adds water to 100mL, 121 DEG C of sterilizing 15min.
(5) by bacillus subtilis formulation 4%, molasses 4%, water 21% Homogeneous phase mixing that obtained for step (3) Chen Xiangcha microbial inoculum 10%, step (4) obtain, then mix with 55% dregs of beans, obtain dregs of beans mixture;
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 4 days of 25 DEG C, 30 DEG C, 37 DEG C, obtain the composite fermentation dregs of beans fermented, can directly directly use as finished feed.
With triangular flask sort of quyi addition, bacillus subtilis inoculum concentration, cultivation temperature for Three factors-levels, the composite fermentation dregs of beans obtained as measurement result during finished feed in table 2.Wherein, the content of acid-soluble protein records according to QB/T2653-2004 method.
Table 2L
9(3
3) its acid-soluble protein content of orthogonal test table
From orthogonal arrage, bacterial classification production process, bacillus subtilis inoculum concentration, triangular flask sort of quyi amount are less on the impact of fermentation finished product acid-soluble protein.The impact of sweat temperature is larger.
Embodiment 3
Implement as follows: following component all by mass percentage
(1) grind under commercially available old scented tea (yunnan puer tea) normal temperature, obtain Chen Xiangcha powder;
(2) triangular flask sort of quyi preparation: be equipped with in 100g solid medium A500mL triangular flask, by culture medium 1% inoculum concentration access step (1) described Chen Xiangcha powder, mixing, 29 DEG C of cultivations.After mycelia is covered with culture medium, shaking flask, latter every 8 hours shaking flasks 1 time.Media surface covers with spore, and the triangular flask sort of quyi has been cultivated;
Described solid medium A mass percent: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
(3) bent preparation is planted: according to the triangular flask sort of quyi that 10% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly on bamboo sieve (thickness is about 5cm), put into kind of a bent room, 29 DEG C, humid control is cultivated more than 90%.After media surface covers with mycelia, start to turn over bent heat radiation.
After whole culture medium covers with spore, Chen Xiangcha microbial inoculum has been cultivated.
Described solid medium B mass percent: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
(4) bacillus subtilis formulation preparation: on liquid medium within, by the bacillus subtilis of 1% inoculum concentration access activation, 37 DEG C of ventilate cavity 24 hours, can obtain bacillus subtilis formulation.
The quality percent by volume (g/mL) of described fluid nutrient medium nutritional labeling: 0.5% sodium chloride, 0.5% dusty yeast, 1.0% peptone, adds water to 100mL, 121 DEG C of sterilizing 15min.
(5) by bacillus subtilis formulation 8%, molasses 10%, water 17% Homogeneous phase mixing that obtained for step (3) Chen Xiangcha microbial inoculum 20%, step (4) obtain, then mix with 45% dregs of beans, obtain dregs of beans mixture;
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 3,4,5,6 days of 30 DEG C, obtain the composite fermentation dregs of beans fermented, can directly directly use as finished feed.
Obtain the composite fermentation dregs of beans that ferments as measurement result during finished feed in table 3.The measuring method of organic acid, acid-soluble protein, urease activity is shown in embodiment 1.Amino-acid nitrogen records according to GB/T5009.39-2003.Urease activity reduction=(before fermentation the rear urease activity of urease activity-fermentation) front urease activity × 100% of/fermentation.
Table 3 ferments the measurement result of different number of days finished feed
Embodiment 4
Implement as follows: following component all by mass percentage
(1) grind under commercially available old scented tea (yunnan puer tea) normal temperature, obtain Chen Xiangcha powder;
(2) triangular flask sort of quyi preparation: be equipped with in 100g solid medium A500mL triangular flask, by culture medium 1% inoculum concentration access step (1) described Chen Xiangcha powder, mixing, 30 DEG C of cultivations.After mycelia is covered with culture medium, shaking flask, latter every 8 hours shaking flasks 1 time.Media surface covers with spore, and the triangular flask sort of quyi has been cultivated;
Described solid medium A mass percent: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
(3) bent preparation is planted: according to the triangular flask sort of quyi that 10% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly on bamboo sieve (thickness 6cm), put into kind of a bent room, 30 DEG C, humid control is cultivated more than 90%.After media surface covers with mycelia, start to turn over bent heat radiation.
After whole culture medium covers with spore, Chen Xiangcha microbial inoculum has been cultivated.
Described solid medium B mass percent: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
(4) bacillus subtilis formulation preparation: on liquid medium within, by the bacillus subtilis of 1% inoculum concentration access activation, 37 DEG C of ventilate cavity 24 hours, can obtain bacillus subtilis formulation.
The quality percent by volume (g/mL) of described fluid nutrient medium nutritional labeling: 0.5% sodium chloride, 0.5% dusty yeast, 1.0% peptone, adds water to 100mL, 121 DEG C of sterilizing 15min.
(5) by bacillus subtilis formulation 2%, molasses 2%, water 31% Homogeneous phase mixing that obtained for step (3) Chen Xiangcha microbial inoculum 15%, step (4) obtain, then mix with 50% dregs of beans, obtain dregs of beans mixture;
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 5 days of 25 DEG C, obtain the composite fermentation dregs of beans fermented, can directly directly use as finished feed.
Obtain the composite fermentation dregs of beans that ferments as measurement result during finished feed in table 4.Organic acid assay method is with embodiment 1.The measuring method of lactic acid bacteria number is with reference to GB4789.35-2010 method.Bacterial count of bacillus subtilis measuring method is with reference to GB4789.2-2010, and wherein sample treatment changes into and gets 25g sample and be placed in 225mL physiological saline and boil 15min.Urease activity reduction=(before fermentation the rear urease activity of urease activity-fermentation) front urease activity × 100% of/fermentation, measuring method is shown in embodiment 1.
Table 4 composite fermentation dregs of beans testing result
Embodiment 5
Implement as follows: following component all by mass percentage
(1) grind under commercially available old scented tea (yunnan puer tea) normal temperature, obtain Chen Xiangcha powder;
(2) triangular flask sort of quyi preparation: be equipped with in 100g solid medium A500mL triangular flask, by culture medium 1% inoculum concentration access step (1) described Chen Xiangcha powder, mixing, 30 DEG C of cultivations.After mycelia is covered with culture medium, shaking flask, latter every 8 hours shaking flasks 1 time.Media surface covers with spore, and the triangular flask sort of quyi has been cultivated;
Described solid medium A mass percent: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
(3) bent preparation is planted: according to the triangular flask sort of quyi that 10% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly on bamboo sieve (thickness is about 5cm), put into kind of a bent room, 30 DEG C, humid control is cultivated more than 90%.After media surface covers with mycelia, start to turn over bent heat radiation.
After whole culture medium covers with spore, Chen Xiangcha microbial inoculum has been cultivated.
Described solid medium B mass percent: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
(4) bacillus subtilis formulation preparation: on liquid medium within, by the bacillus subtilis of 1% inoculum concentration access activation, 37 DEG C of ventilate cavity 24 hours, can obtain bacillus subtilis formulation.
The quality percent by volume (g/mL) of described fluid nutrient medium nutritional labeling: 0.5% sodium chloride, 0.5% dusty yeast, 1.0% peptone, adds water to 100mL, 121 DEG C of sterilizing 15min.
(5) 1. obtained Chen Xiangcha microbial inoculum+2% molasses+step (4) of 20% step (3) obtains bacillus subtilis formulation 8%+25% water+45% dregs of beans respectively, stirs and obtains dregs of beans mixture; 2. obtained bacillus subtilis formulation 6%+25% water+45% dregs of beans of obtained Chen Xiangcha microbial inoculum+4% molasses+step (4) of 20% step (3), stirs and obtains dregs of beans mixture; 3. obtained bacillus subtilis formulation 4%+25% water+45% dregs of beans of obtained Chen Xiangcha microbial inoculum+6% molasses+step (4) of 20% step (3), stirs and obtains dregs of beans mixture; 4. obtained bacillus subtilis formulation 2%+25% water+45% dregs of beans of obtained Chen Xiangcha microbial inoculum+8% molasses+step (4) of 20% step (3), stirs and obtains dregs of beans mixture.
(6) the dregs of beans mixture obtained in step (5) is directly seated in fermentation cylinder for fermentation respectively, sealing prevents moisture from distributing;
(7) at the condition bottom fermentation 4 days of 37 DEG C, obtain the composite fermentation dregs of beans fermented respectively, can directly directly use as finished feed.
Obtain the composite fermentation dregs of beans that ferments as measurement result during finished feed in table 5.Organic acid assay method is with embodiment 1, and bacterial count of bacillus subtilis assay method is shown in embodiment 3, and acid-soluble protein measuring method is shown in embodiment 1.
Table 5 composite fermentation dregs of beans testing result
Application Example 1
The application of composite fermentation dregs of beans of the present invention: added mass fraction 4% grower pigs compound premix and throw something and feed, carries out pig and feeds growth performance test.
The temperature of pig house and humidity: duration of test in every day 10:00,14:00 measure temperature and the humidity of pig house, get its mean value as the temperature on pig house same day and humidity; After measured, in duration of test pig house, mean temperature is 30.52 ± 2.03 DEG C, medial humidity is 69.21 ± 2.53%.
Animal used as test: body weight is about Du × length × large hybridization galt 100 of 60kg;
Animal used as test divides into groups: 100 pigs are divided into control group and test group at random, and often organize 5 repetitions, each repetition 10 pigs, each repetition is 1 hurdle, and single hurdle area is 4.5m × 5.0m.
Feed: the conventional daily ration of control group fed, add mass fraction 4% grower pigs compound premix (buying from profit Bioisystech Co., Ltd of Beijing agricultural university), daily ration used adopts corn-soybean meal, with reference to NRC (1998) swine rearing standard preparation; Test group is fed the composite fermentation dregs of beans that embodiment 1 prepares, and adds mass fraction 4% grower pigs compound premix.
Every day respectively throws something and feeds 1 time sooner or later, until stop searching for food; The nursing phase is 30 days; Experimental animal is pressed hurdle and is raised, and freely drink water, other feedings and managements are carried out routinely.
After the nursing phase terminates, the growth performance index of determination experiment animal.Assay method is as follows:
Growth performance index determining: on-test and at the end of fasting 1 night, weigh to by head animal, the independent record of feed feed intake of each repetition, calculates adding weight, daily ingestion amount, feed-weight ratio.
Experimental result, in table 6
Table 6 fermented feed is on the impact of Performance of Finishing Pigs
P<0.01, shows that difference is extremely remarkable; P<0.05, shows significant difference; P>0.05, shows that difference is not remarkable.
Application Example 2
The application of composite fermentation dregs of beans of the present invention: it directly thrown something and fed, carries out chicken feeding trial.
Animal used as test: AA meat sold on the market baby chick, 1000.
Animal used as test divides into groups: 1000 chickens are divided into control group and test group at random, and often organize 5 repetitions, each repetition 100 chickens, each repetition is 1 hurdle.
Feed: test group is fed the composite fermentation dregs of beans that embodiment 1 prepares, and control group is identical with test group nutritional labeling, and feeding and management method is identical.
The nursing phase is 55 days, after the nursing phase terminates, and the growth performance index of determination experiment animal.In table 7.
Table 7 composite fermentation dregs of beans is on the impact of growth of meat chicken performance
| Group | On average every quality (kg) | Feedstuff-meat ratio | Survival rate (%) |
| Control group | 2.08 | 2.57 | 94.1 |
| Test group | 2.86 | 2.26 | 98.3 |
After composite fermentation dregs of beans prepared by the present invention directly feeds chicken, hair color light, health is better, and the hair color of control group is obviously poor.In feeding process, the conversion ratio of feed is high, decreases the generation of disease, has high survival rate.
Above-described embodiment is the present invention's preferably embodiment; but embodiments of the present invention are not restricted to the described embodiments; change, the modification done under other any does not deviate from Spirit Essence of the present invention and principle, substitute, combine, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1., with a method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that comprising following steps: each component by mass percentage
(1) grind under Chen Xiangcha normal temperature, obtain Chen Xiangcha powder;
(2) sort of quyi preparation: the Chen Xiangcha powder obtained by 1% access step (1) of solid medium A total amount, mixing, cultivates; After mycelia is covered with solid medium A, shaking flask; Solid medium A surface covers with spore, and the sort of quyi has been cultivated;
(3) bent preparation is planted: by the sort of quyi that 5% ~ 15% access step (2) of solid medium B total amount is obtained, divide after mixing thoroughly, cultivate; After solid medium B surface covers with mycelia, start to turn over bent heat radiation; After whole solid medium B covers with spore, Chen Xiangcha microbial inoculum has been cultivated;
(4) bacillus subtilis formulation preparation: by the bacillus subtilis of 1% ~ 2% access activation of fluid nutrient medium total amount, cultivate, obtain bacillus subtilis formulation;
(5) bacillus subtilis formulation 2% ~ 8%, molasses 2 ~ 10%, water 17% ~ 31% Homogeneous phase mixing that the Chen Xiangcha microbial inoculum 10 ~ 20% step (3) obtained, step (4) are obtained, mix with 45 ~ 55% dregs of beans again, obtain dregs of beans mixture;
(6) after the dregs of beans mixture obtained in step (5) being directly seated in fermentation cylinder for fermentation sealing, at the condition bottom fermentation 3 ~ 6 days of 25 ~ 37 DEG C;
(7) the composite fermentation dregs of beans fermented is obtained.
2. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the Chen Xiangcha described in step (1) is that the black tea in Anhua, Hunan, Guangxi Liu Bao loose tea, yunnan puer tea or Guangdong great Ye are blue or green.
3. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the solid medium A described in step (2) is prepared as follows: each component by mass percentage: 5% dregs of beans, 45% wheat bran, 50% water, 121 DEG C of sterilizing 30min.
4. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the solid medium B described in step (3) is prepared as follows: each component by mass percentage: 10% molasses, 60% dregs of beans, 30% water, 121 DEG C of sterilizing 30min.
5. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the temperature of the cultivation described in step (2) is 28 ~ 30 DEG C;
The condition of the cultivation described in step (3) is 28 ~ 30 DEG C, and humid control is more than 90%.
6. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the shaking flask described in step (2) is every 8 hours shaking flasks 1 time;
Dividing as dividing on bamboo sieve described in step (3), and put into kind of a bent room.
7. according to claim 1 with the method for Chen Xiangcha and bacillus subtilis composite fermentation dregs of beans, it is characterized in that: the condition of the cultivation described in step (4) is 37 DEG C of ventilate cavity 24 hours;
Fluid nutrient medium described in step (4) is prepared as follows: each component is by quality percent by volume: 0.5% sodium chloride, 0.5% dusty yeast, and 1.0% peptone, adds water, 121 DEG C of sterilizing 15min.
8. a composite fermentation dregs of beans, is obtained by the method described in any one of claim 1 ~ 7.
9. composite fermentation dregs of beans according to claim 8, is characterized in that: described composite fermentation dregs of beans forms stronger wine flavour, band tart flavour; PH is 3.08 ~ 4.35; Organic acid content is 3.23% ~ 4.35%; Acid-soluble protein is 10.76% ~ 15.29%; Urease activity reduces by 80 ~ 94%; Bacillus globigii spores number is 9.7 × 10
9more than cfu/g; Lactic acid bacteria number is 8.3 × 10
8more than cfu/g.
10. the application of composite fermentation dregs of beans on feed described in claim 8 or 9.
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| CN105685479A (en) * | 2016-03-09 | 2016-06-22 | 广东海洋大学 | Preparation method of fermented soybean meal |
| CN107788301A (en) * | 2016-03-18 | 2018-03-13 | 海口椰彩食品有限公司 | A kind of compound fermented beverage of walnut coconut and its preparation method and application |
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