CN109749977A - By-product from sugarcane feed fermenting microbial inoculum and preparation method thereof - Google Patents
By-product from sugarcane feed fermenting microbial inoculum and preparation method thereof Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The invention discloses a kind of by-product from sugarcane feed fermenting microbial inoculums and preparation method thereof, the fermenting agent includes special domestication bacterium freeze-dried powder, and the freeze-dried powder of a variety of active ingredients such as candida utili, lactobacillus plantarum, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride, can effectively be degraded the by-product from sugarcane such as sugarcane end pin, bagasse, molasses, each mutually coordinated cooperation of strain, and fermentation is efficient, speed is fast, it consumes energy low, high conversion rate, stability is good.Domestication bacterium is from the Nature used in the present invention, it is obtained by enrichment, screening, domestication, breeding with super strength and adaptability, the cellulose in by-product from sugarcane can pointedly be decomposed, the polymeric carbohydrate that the animals such as crude fibre, lignin are difficult to sufficiently digest and assimilate can be converted to the absorbable lower-molecular substance utilized, the content for improving crude protein in fermented feed simultaneously, improves the trophism and palatability of feed.
Description
Technical field
The present invention relates to feed fermentation agent technical fields, specifically by-product from sugarcane feed fermenting microbial inoculum and its preparation
Method.
Background technique
Sugarcane (Saccharum of ficinarum L.) be a kind of perennial grass family (Gram ineae) chinese sorghum race
High stalk monocotyledon, tropical and subtropical region is planted extensively.Sugarcane belongs to C4 plant, has very high conversion of solar energy,
Yield per unit area is higher, is a kind of industrial crops for efficiently utilizing solar energy.Contain sufficient moisture content, sugar, albumen in sugarcane
Winter-spring season can be effectively relieved as a kind of important non-grain fodder crop in matter, fat, organic acid and various vitamins etc.
Save the problem of feed scarcity.Guangxi is sugarcane main producing region, there is sugar industry by-product abundant such as bagasse, sugarcane toppers leaf, molasses
Have that amount is big, concentrates and small by seasonal effect Deng, these by-products.It, not only can be with as can these sugar industry by product feeds
The difficult situation that people and animals strive grain is solved, considerable economic well-being of workers and staff is also brought for sugar refinery.
By-product from sugarcane (sugarcane toppers leaf, bagasse) has yield compared with other roughage raw material cornstalk, herbages etc.
Greatly, the advantages that place of production concentrates, is easy to purchase, cost is relatively low.Sugarcane toppers (leaf) are the by-products of Sugarcane Industry, are commonly called as sugarcane tail, entirely
About 20,000,000 tons of state's annual output, be a kind of cheap energy feed.Have the characteristics that yield is big, the place of production is concentrated, cost is relatively low.
Sugarcane toppers leaf contains protein, sugar, full of nutrition, is a kind of extraordinary livestock-raising feed resource.Bagasse is a kind of
Plant cellulose substance is the maximum by-product of cane sugar factory, its general yield accounts for about the 24~27% of raw material sugarcane, data
National about 10,000,000 tons of annual output of bagasse before statistical form improving eyesight, about 90% bagasse is used as fuel at present, for sugar refinery pot
Furnace power generation and supply steam, the bagasse of residue 10% or so are primarily used to papermaking and production animal feed etc..Although bagasse
Directly burning is the most succinct quick method of processing bagasse, but bagasse directly burns and not only wasted bagasse resource, but also can generate big
CO isothermal chamber gas is measured, serious pollution, therefore the rational exploitation and utilization problem of bagasse resource are caused to air and natural environment
It is urgently to be resolved.
There are also the open source literatures about fodder zymophyte agent in the prior art, but dedicated about by-product from sugarcane feed
Seldom there are no, such as following documents:
1, Chinese patent: a kind of crops straw stalk feed fermenting agent of highly effective and safe, its solid additive and preparation method；Shen
Please number: 201410208303.2；Applicant: Duan Yigao；Abstract: the invention discloses a kind of feedings of the agricultural crop straw of highly effective and safe
Expect fermenting agent, its solid additive and preparation method, selects one group of functional, it is the high fungi of combined effectiveness, actinomyces, thin
Nine strains of three classes of bacterium, through spawn rejuvenation, single bacterium kind expands culture, and mixing liquid is fermented, and concentrated mother liquor microbial inoculum is secondary
Expand culture；In preparation process, the concentrated mother liquor microbial inoculum has carried out well in the semi-open fermentor of solid straws feed
Draining and aeration condition under complete whole preparation process.Straw feed biosolids additive of the present invention is to various plant straw
After stalk carries out feed fermentation processing, the ability with efficient-decomposition softening stalk fibre can improve palatability, improve animal and disappear
The effects of change ability, raising straw utilization；Pesticide residue in the also degradable stalk of microbial bacterial agent simultaneously, chemical noxious substance
It infects, becomes the substitute products of antibiotic, improve the safety and health value of processing feedstuff with stalks, become Agro-ecology shape
Feed addictive.
2, a kind of Chinese patent: fermenting agent being used to prepare straw feed；Application number: 200810226318.6；Patent
Weigh people: China Agricultural University；Abstract: the invention discloses a kind of fermentations for being used to prepare straw feed
Microbial inoculum.Its active constituent includes lactobacillus fermenti (Lactobacillus fermentum), lactobacillus plantarum
(Lactobacillus plantarum) and lactobacillus paracasei (Lactobacillus paracasei).Use of the invention
Various lactic acid bacterias in the fermenting agent for preparing straw feed have the synergistic effect of height, form stable ecosystem self
System, can quickly breed in non-sterilized natural material, colonize and occupy advantage, generate a large amount of lactic acid rapidly, reduce fermentation
The pH value of material, improves the fermentation quality of feed, improves the nutritive value and palatability of feed, to the animal safety of feeding, and
It is low in cost using simple.
3, it Chinese patent: solid fermentation microbial inoculum of chicken source bacillus subtilis and Pediococcus pentosaceus and preparation method thereof and answers
With；Application number: 201210428013.X；Patentee: Sichuan Agricultural University；Abstract: the invention discloses chicken source withered grass gemma
The preparation method of the solid fermentation microbial inoculum of bacillus and Pediococcus pentosaceus is selected bacillus subtilis (Bacillus subtilis)
SCS4562 bacterial strain and Pediococcus pentosaceus (Pediococcus pentosaceus) SCS4560 bacterial strain, bacillus subtilis
(Bacillus subtilis) SCS4562 bacterial strain deposit number: CGMCC NO.6565；Pediococcus pentosaceus (Pediococcus
Pentosaceus) SCS4560 bacterial strain deposit number: CGMCC NO.6566；Two bacterial strains obtain tunning through everfermentation
As solid fermentation microbial inoculum.Solid fermentation microbial inoculum of the invention is added directly into feed, maintains intestinal microecology health simultaneously
Nutritional ingredient in intestinal absorption feed is helped, chicken group's growth performance is improved.
In recent years, with Industrial Economic Development, China's ecological environment is continuous worsening.Due to overgrazing, estrepement denudation etc.
The generation of behavior causes grassland degeneration phenomenon serious, seriously threatens herbivorous stock raising sound development.Although China is sweet
Sugarcane plants big country, but the utilization mode of by-product from sugarcane is also relatively simple, is difficult to digest and assimilate containing a large amount of animal
Crude fibre, for the exploitation of by-product from sugarcane feed product, there are still biggish difficulty, it is difficult to be popularized in an all-round way, be caused big
The waste of by-product from sugarcane is measured, the bagasse after sugar cane crushing is only used for making by especially bagasse, many sugar enterprises
Paper or burning can not adequately utilize it, cause great waste.If can be scientific and reasonable to by-product
It is used, then can obtain huge economic, society and ecological benefits, it is therefore, by-product from sugarcane and food conversion is effective
Connect, develop the good dedicated hair of by-product from sugarcane feed of low a kind of efficient high-speed, energy consumption, high conversion rate, stability
Yeast-like fungi agent has good development prospect and social effect.
Summary of the invention
For the above the deficiencies in the prior art, the object of the present invention is to provide a kind of by-product from sugarcane feed fermenting bacterium
It is dedicated to develop efficient one kind, high speed, the good by-product from sugarcane feed of low, high conversion rate, the stability of consuming energy for agent and preparation method thereof
Fermenting agent.
The invention is realized by the following technical scheme:
By-product from sugarcane feed fermenting microbial inoculum, the raw material including following parts by weight: 15~30 parts of bacterium freeze-dried powder of domestication produces
8~10 parts of protein Candida freeze-dried powder, 6~8 parts of lactobacillus plantarum freeze-dried powder, 8~10 parts of bacillus subtilis freeze-dried powder, black song
4~6 parts of mould freeze-dried powder, 4~6 parts of aspergillus oryzae freeze-dried powder, 7~10 parts of Trichoderma viride freeze-dried powder, cellulase freeze-dried powder 18~20
Part；
The domestication bacterium freeze-dried powder the preparation method comprises the following steps: secondary domestication bacterium is taken to reach the bacterium solution of stationary phase when expanding culture, from
The heart takes bacterium mud to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out
Vacuum freeze drying is to get domestication bacterium freeze dried powder；
The candida utili freeze-dried powder the preparation method comprises the following steps: candida utili is inoculated in malt extract medium,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud nothing to stationary phase under the conditions of 36~38 DEG C, pH6.3~6.5
The washing of bacterium phosphate buffer, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and carry out vacuum freeze drying,
Up to candida utili freeze dried powder；
The lactobacillus plantarum freeze-dried powder the preparation method comprises the following steps: lactobacillus plantarum is inoculated in MRS culture medium, in 36~38
DEG C, culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud sterile phosphate to stationary phase under the conditions of pH6.3~6.5
Buffer washing, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and carry out vacuum freeze drying to get plant
Lactobacillus freeze dried powder；
The bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulose
Enzyme freeze-dried powder the preparation method comprises the following steps: each bacterium branch is inoculated in PCA fluid nutrient medium, in 36~38 DEG C, the condition of pH6.8~7.2
Lower culture takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is again to stationary phase
It is resuspended in sterile phosphate buffer, embedding medium is added, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-drying
Powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
The embedding medium is prepared from the following raw materials in parts by weight: 8~10 parts of maltodextrin, 6~8 parts of beta-cyclodextrin,
10~12 parts of starch, 8~20 parts of water.
The vacuum freeze drying design parameter are as follows: 65 DEG C of condenser temperature ﹣, 30~35 DEG C of heating temperature, vacuum degree≤
60pa, the dry moisture content to microbial inoculum are no more than 7%.
After domestication, the ability of decomposition of cellulose greatly promotes the domestication bacterium, especially decomposition bagasse
With the ability of sugarcane end pin, the survival ability in by-product from sugarcane is also improved；Domestication bacterium main component include:
Candida utili, lactic acid bacteria, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride.
The MRS culture medium constituent are as follows: 8~10 parts of peptone, 8~10 parts of beef extract, 4~6 parts of yeast extract, lemon
1.5~2.2 parts of diammonium of lemon acid hydrogen, 18~22 parts of glucose, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, dipotassium hydrogen phosphate
1.5~2.5 parts, 900~1000 parts of distilled water；The PCA fluid nutrient medium, constituent are as follows: 4~6 parts of tryptone, ferment
2~3.2 parts of female cream, 1~2 part of glucose, 950~1000 parts of distilled water.
It is described domestication bacterium acclimation method the following steps are included:
(1) it chooses bacterium mud: choosing the half a year above sugarcane field soil or sugarcane top stack retting mud or sugar refinery bagasse laydown area is stacked
Half a year sludge formed above seals at original bacterium mud scene in sterile glass vials up for safekeeping to get original bacterium mud, takes back laboratory work
It is cultivated for original flora；
These original bacterium muds, due to the influence of ecological environment, have gradually formed dominant bacteria in long-term stacking environment, survival
What is got off is the bacterium of decomposable by-product from sugarcane cellulose mostly, and such bacterium is easier to be tamed, the energy of degraded cellulose
Power can significantly improve, and have relative stability.
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of high pressure sterilizations 10~
20min is added to prepared basal medium and is uniformly mixed after cooling, the weight ratio of sugarcane end pin and basal medium is
1.0~1.5:1 is to get primary domestication culture medium；
According to the difference of sugarcane end pin additional amount, be arranged 5~6 gradients (settable gradient is respectively as follows: 1.0:1,1.1:1,
1.2:1,1.3:1,1.4:1,1.5:1), the different primary domestication culture medium of several sugarcane end pin contents is prepared, it will be in step (1)
Original bacterium mud be inoculated into each culture medium, 35~40 DEG C, cultivate 20~30 days under the conditions of pH6.3~6.5, remembered every 2~4 days
The situation in a subculture is recorded, it is up to excellent with the percentage that sugarcane tail is decomposed, be so that bacterium solution is clarified, bacterium mud is not smelly
Excellent, in comprehensive each culture medium situation, selects one group of optimal culture medium, and the bacterium to survive in bacterium solution is primary domestication
Bacterium；
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains A
Material；It takes the bagasse powder after juicing to be broken to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains B material；It takes in sugar refinery
Sundries is removed in the sewage that bagasse shower water is formed, worry, obtains C material；
By above-mentioned A, B, C material carry out respectively combination of two or three's combination (according to the combination of different fiber material, can be formed AB,
Tetra- groups of AC, BC, ABC), it is added to basal medium, multiple secondary domestication culture mediums are made, it is primary in inoculation step (2) respectively
The bacterium solution for taming bacterium, is cultivated 15~30 days under the conditions of 35~42 DEG C, the situation in 2~4 days one subcultures of record, with
The percentage that sugarcane tail, bagasse decompose is up to excellent, with clarifying contaminated liquids, not smelly to be excellent, in comprehensive each culture medium
Situation, selects one group of optimal culture medium, and bacterium solution is secondary domestication bacterium；
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary domestication
Culture medium when culture medium optimal situation cultivates 40~50h；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
The basal medium is prepared from the following raw materials in parts by weight: 8~10 parts of peptone, 8~10 parts of beef extract,
4~6 parts of yeast extract, 5~10 parts of brewer's wort, 1.5~2.2 parts of diammonium hydrogen citrate, 18~22 parts of glucose, Tween 80 0.8~
1.2 parts, 4~6 parts of sodium acetate, 1.5~2.5 parts of dipotassium hydrogen phosphate, 4~6 parts of tryptone, 2~3.2 parts of yeast extract, glucose 1
~2 parts, 900~1000 parts of distilled water.
The by-product from sugarcane feed fermenting microbial inoculum is preparing the application in ruminant feed.
Candida utili of the present invention (Candida utilIs Torula utilis Henneb or edible torula) are called.Its
The content of protein and vitamin B is all higher than brewer's yeast, it can not be needed in the medium using urea and nitric acid as nitrogen source
Any growth factor, which is added, to be grown.It can utilize pentose and hexose, not produce alcohol under aerobic conditions, can utilize and make
The sulfite waste liquor of paper industry, moreover it is possible to produce the edible protein of people and animals using molasses, wood hydrolysis liquid etc..Candida utilis
The cell of yeast is rounded, oval or sausage shape, and size is (3.5~4.5) μ m (7~13) μm.Liquid Culture does not produce mould,
Tube bottom has bacterial sediment.In wort agar medium, bacterium colony milky, smoothly, and with or without gloss, neat in edge or mycelia
Shape.Animal intestinal micro-ecology balance is adjusted in candida utili, improves feed digestibility, enhances in its cell of animal body power
Rich in vitamin B and protein, moreover it is possible to part nutriment needed for providing animal.
Lactobacillus plantarum (Lactobacillus plantarum) be lactic acid bacteria one kind, nose circle straight-bar bacterium, usually
0.9~1.2vtm × 3.0~8.0 μm, anaerobism or amphimicrobian, strain be it is straight or curved rod-shaped, individually, sometimes in pairs or chaining
Shape, optimal pH 6.5 or so, belongs to homofermentative lactic bacteria.Compared with this bacterium is its viable count with the difference of other lactic acid bacterias
Height can largely produce acid, increase the pH stable in water not, and the acidic materials of its output can degrade heavy metal；It can hair
Ferment pentose or gluconate, 85% or more is lactic acid in final product；The energy distinctive lactobacillin of output in reproductive process,
Lactobacillin is a kind of preservative of bion, has certain antisepsis.
Bacillus subtilis (Bacillus subtilis) be bacillus one kind, individual cells 0.7~0.8 × 2
~3 microns, aerobic bacteria.Bacterium colony rough surface is opaque, dirty white or yellowish, when growing in liquid medium, is commonly formed
Wrinkle mould.Available protein, a variety of sugar and starch, decompose tryptophan and form indoles.During bacillus subtilis thalli growth
Subtilin, polymyxins, nystatin, the gramicidins isoreactivity substance of generation, these active materials are to pathogenic bacteria or interior
The conditioned pathogen of source sexuality dye has apparent inhibiting effect.
Aspergillus niger (Aspergillus niger), Deuteromycotina, Hyphomycetes, hyphomycetales, Moniliaceae, aspergillus is true
A Common Species in bacterium.It is distributed widely in grain all over the world, plant product and soil, is agriculturally used as production sugar
Change the strain of feed.Aspergillus niger is important fermentation industry strain, can produce amylase, acid protease, cellulase, pectin
Enzyme, glucose oxidase, citric acid, gluconic acid and gallic acid etc..
Aspergillus oryzae (Aspergillus oryzae ( Asp.oryzae)) category Deuteromycotina, Hyphomycetes, hyphomycetales, from
Geng Bao section, a Common Species in aspergillus fungi, 150~300 μm of diameter.Aspergillus oryzae can produce protease, amylase, saccharification
Enzyme, cellulase, phytase etc..Under the action of amylase, straight chain, the amylopectin in raw material are degraded to dextrin and various
Low molecule carbohydrate, such as maltose, glucose；Under the action of protease, stodgy macro-molecular protein is degraded to
Peptone, polypeptide and various amino acid, and the difficult mass degradation absorbed such as can make crude fibre in auxiliary material, phytic acid, improve battalion
Value, health-care efficacy and digestibility are supported, the fermentation industries such as food, feed, production kojic acid, wine brewing are widely used in.
Trichoderma viride (Trichoderma viride) be Trichoderma one kind, it is extensive in distributed in nature, often it is saprophytic in
On timber, seed and plant residue.Trichoderma glues spore mushroom in Deuteromycotina, Hyphomycetes, hyphomycetales.Trichoderma viride energy
Generate one of a variety of biologically active enzyme systems, the highest bacterial strain of institute's cellulase-producing activity, generated cellulase pair
Crop has degradation.
Cellulase (cellulase), also known as β-Isosorbide-5-Nitrae-glucan -4- glucan hydrolysis, is that degraded cellulose generates Portugal
The general name of one group of enzyme of grape sugar, is a kind of complex enzyme, mainly by circumscribed 1,4 beta-glucanase, Endo-β-glucanase and β-glucose
The composition such as glycosides enzyme, there are also the zytases of very high vigor.Biocatalysis from decomposition of cellulose, can be by cellulose point
Solution is mainly used in feed, alcohol, weaving and food etc. at oligosaccharides or the protein of monosaccharide.
Beneficial effects of the present invention:
1, after domestication, the ability of decomposition of cellulose is greatly promoted domestication bacterium of the invention, especially decomposition bagasse
With the ability of sugarcane end pin, the survival ability in by-product from sugarcane is also improved.In conjunction with candida utili, plant
Object lactobacillus, bacillus subtilis, aspergillus niger, aspergillus oryzae, Trichoderma viride, cellulase etc., addition embedding medium carry out at embedding
Reason, through vacuum freeze drying, is finally mixed to form microbial inoculum powder according to specific ratio.It is special suitable for by-product from sugarcane fodder
It is not the feed for preparing the ruminants such as ox, sheep, deer, the palatability and nutritive value of feed can be improved, and to sugarcane pair
The good approach that product is comprehensively utilized has good development prospect and social effect.
2, by-product from sugarcane feed fermenting microbial inoculum of the present invention, active constituent include candida utili,
Lactobacillus plantarum, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride etc., each mutually coordinated cooperation of strain, fermentation are high
Effect, speed is fast, and consume energy low, high conversion rate, and stability is good, can make up the deficiency of single culture fermentation, can also solve nature
Fermenting microbe excessively complexity, the problem of fermentation process controllability difference.
3, during livestock and poultry feeding large-scale development, the long-term drug resistance that antibiotic is used continuously and causes animal body, thus
So that mankind pathogen is also generated drug resistance, endangers the health of the mankind.Fermenting agent of the invention during the fermentation, passes through micro- life
Object and its a variety of physiological metabolism substances of generation, can effectively kill various pathogens, to some nuisances original in feed
Matter has carried out degradation and harmless processing, avoids the use of antibiotic.
4, the domestication bacterium used in the present invention obtains from the Nature by enrichment, screening, domestication, with super strength
Breeding and adaptability, can pointedly decompose the cellulose in the by-product from sugarcane such as bagasse, sugarcane end pin, can be by thick fibre
The polymeric carbohydrate that the animals such as dimension, lignin are difficult to sufficiently digest and assimilate is converted to the absorbable low-molecular material utilized
Matter, and the content of crude protein in fermented feed is improved simultaneously, improve the trophism and palatability of feed.
Specific embodiment
In order to keep technical solution of the present invention clearer, below with reference to the embodiment of the present invention, to technology of the invention
Scheme is clearly and completely described.
Embodiment 1
Tame bacterium acclimation method the following steps are included:
(1) it chooses bacterium mud: choosing sugarcane top stack retting mud more than half a year, it is formed above that sugar refinery bagasse laydown area stacks half a year
Sludge mixes to get original bacterium mud according to weight ratio 1:1, original bacterium mud scene is sealed up for safekeeping in sterile glass vials, experiment is taken back
It is cultivated as original flora room；
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 200 mesh, in 121 DEG C of high pressure sterilization 15min, is added to after cooling
Prepared basal medium is uniformly mixed, and obtains once taming culture medium, the weight ratio of sugarcane end pin and basal medium is
1.0~1.5:1；
According to the differences of sugarcane end pin additional amount, 6 gradients are set and are respectively as follows: 1.0:1,1.1:1,1.2:1,1.3:1,1.4:1,
The different primary domestication culture medium of 6 kinds of sugarcane end pin contents is made in 1.5:1, and one group of culture medium is arranged in each gradient, and every group has
3 parallel test culture mediums；Original bacterium mud in step (1) is inoculated into each culture medium, 38 DEG C, cultivate under the conditions of pH6.4
30 days, the situation in 2~4 days one subcultures of record, culture first 12 days primary every 4 days records, and the 18th~21 day every
Primary every 3 days records, the 21st~30 day primary every 2 days records, is scored according to the case where each record.
During being primary domestication such as the following table 1, according to the difference of sugarcane end pin additional amount, in five groups of culture mediums of record
Situation grade form.Up to excellent with the percentage that sugarcane tail is decomposed, full marks are 10 points；It is so that bacterium solution is clarified, bacterium mud is not smelly
Excellent, full marks are 10 points, and the relevant professional person of Feed Manufacturing research is artificially engaged in scoring, and 50 people of number makes even and respectively (retains two
Bit digital) it is final score；According to the same judgment criteria, the case where culture medium of record, score as follows:
Table 1 once tames record sheet
| ①1.0:1 | ②1.1:1 | ③1.2:1 | ④1.3:1 | ⑤1.4:1 | ⑥1.5:1 | |
| 4th day | 1.8 | 2.0 | 2.1 | 2.2 | 2.2 | 2.1 |
| 8th day | 2.7 | 2.8 | 2.8 | 3.0 | 2.9 | 2.9 |
| 12nd day | 3.8 | 3.8 | 3.9 | 4.1 | 4.2 | 4.2 |
| 15th day | 5.5 | 5.7 | 6.2 | 6.5 | 6.5 | 6.4 |
| 18th day | 6.9 | 7.0 | 7.6 | 8.2 | 8.1 | 8.0 |
| 21st day | 8.8 | 8.9 | 9.0 | 9.4 | 9.3 | 9.2 |
| 24th day | 9.0 | 9.2 | 9.3 | 9.4 | 9.4 | 9.3 |
| 26th day | 9.4 | 9.6 | 9.3 | 9.7 | 9.5 | 9.6 |
| 28th day | 9.5 | 9.6 | 9.7 | 9.9 | 9.8 | 9.8 |
| 30th day | 9.5 | 9.7 | 9.8 | 9.9 | 9.9 | 9.8 |
| Total score | 66.9 | 68.3 | 69.7 | 72.3 | 71.8 | 71.3 |
As seen from the above table, total score best result is the 3. group, and has reached 8.2 points when the 18th day, is reached within the 21st day
To 9.4 points, reaches best result 9.9 within the 28th day and divide, be considered as optimal set.
In summary the situation in each culture medium, selects one group of optimal culture medium, i.e. sugarcane end pin and basis is cultivated
The primary domestication bacterium that the weight ratio of base is tamed when being 1.3:1, for the qualified primary domestication bacterium of optimal a batch of domestication.
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100 mesh, in 121 DEG C of high pressure sterilization 15min, obtains A material；It takes
Bagasse powder after juicing is broken to 200 mesh, in 121 DEG C of high pressure sterilization 15min, obtains B material；Take bagasse shower water shape in sugar refinery
At sewage, worry remove sundries, obtain C material；
Above-mentioned A, B, C material are subjected to combination of two and three's combination respectively, each material combine be etc. weight combination, and total weight
It is identical, tetra- groups of AB, AC, BC, ABC are formed, the basal medium of material total weight a quarter is added separately to, is made multiple two
Secondary domestication culture medium, every group of culture medium for having 3 parallel tests；Sugarcane end pin and basis are cultivated in inoculation step (2) respectively
The primary domestication bacterium bacterium solution that the weight ratio of base is tamed when being 1.3:1, is cultivated 20 days under the conditions of 40 DEG C, is remembered every 2~3 days
The situation in a subculture is recorded, culture first 12 days are primary every 3 days records, and the 12nd~20 day is primary every 2 days records, according to
The case where record, scores every time.
Situation during being secondary domestication such as the following table 2, according to the combination of different fiber material, in four groups of culture mediums of record
Grade form.Up to excellent with the percentage that sugarcane tail is decomposed, full marks are 10 points；It is not smelly to be excellent with bacterium solution clarification, bacterium mud, it is full
It is divided into 10 points；The relevant professional person of Feed Manufacturing research is artificially engaged in scoring, and 50 people of number makes even and respectively (retains double figures
Word) it is final score；According to the same judgment criteria, the case where culture medium of record, score as follows:
2 two domestication record sheets of table
| AB | AC | BC | ABC | |
| 3rd day | 6.8 | 6.6 | 6.5 | 6.6 |
| 6th day | 7.5 | 7.4 | 7.3 | 7.5 |
| 9th day | 8.2 | 8.1 | 8.0 | 8.6 |
| 12nd day | 8.5 | 8.5 | 8.6 | 8.9 |
| 14th day | 9.0 | 8.9 | 9.0 | 9.4 |
| 16th day | 9.3 | 9.1 | 9.4 | 9.7 |
| 18th day | 9.6 | 9.5 | 9.6 | 9.9 |
| 20th day | 9.9 | 9.9 | 9.8 | 10.0 |
| Total score | 68.8 | 68.0 | 68.2 | 70.6 |
As seen from the above table, total score best result is ABC group, has reached 9.4 points when the 14th day, can reach most within the 20th day
High score 10.0 divides, and is considered as optimal set.
In summary the situation in each culture medium selects one group of optimal culture medium, i.e., ABC group, bacterium solution are two
Secondary domestication bacterium.
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary
Culture medium when culture medium optimal situation is tamed, 45h is cultivated；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
The basal medium is prepared from the following raw materials in parts by weight: 10 parts of peptone, 10 parts of beef extract, and yeast extract
5 parts, 10 parts of brewer's wort, 2 parts of diammonium hydrogen citrate, 20 parts of glucose, 1 part of Tween 80,5 parts of sodium acetate, 2 parts of dipotassium hydrogen phosphate,
5 parts of tryptone, 2.5 parts of yeast extract, 1.8 parts of glucose, 1000 parts of distilled water.
After domestication, the ability of decomposition of cellulose greatly promotes the domestication bacterium, especially decomposition bagasse
With the ability of sugarcane end pin, the survival ability in by-product from sugarcane is also improved；Domestication bacterium main component include:
Candida utili, lactic acid bacteria, bacillus subtilis, aspergillus niger, aspergillus oryzae and Trichoderma viride.
According to acclimation method described in the embodiment of the present invention 1, the domestication bacterium being prepared is for following example 2~4
By-product from sugarcane feed fermenting microbial inoculum preparation.
Embodiment 2
By-product from sugarcane feed fermenting microbial inoculum, the raw material including following parts by weight: it is false to produce protein for 30 parts of bacterium freeze-dried powder of domestication
8 parts of freeze-dried powder of yeast of silk, 6 parts of lactobacillus plantarum freeze-dried powder, 8 parts of bacillus subtilis freeze-dried powder, 4 parts of aspergillus niger freeze-dried powder, rice
4 parts of aspergillus freeze-dried powder, 7 parts of Trichoderma viride freeze-dried powder, 18 parts of cellulase freeze-dried powder；
The domestication bacterium freeze-dried powder the preparation method comprises the following steps: secondary domestication bacterium is taken to reach the bacterium solution of stationary phase when expanding culture, from
The heart takes bacterium mud to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out
Vacuum freeze drying is to get domestication bacterium freeze dried powder；
The candida utili freeze-dried powder the preparation method comprises the following steps: candida utili is inoculated in malt extract medium,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud sterile phosphate slow to stationary phase under the conditions of 37 DEG C, pH6.4
Fliud flushing washing, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and it is false to get protein is produced to carry out vacuum freeze drying
Silk yeast freeze dried powder；
The lactobacillus plantarum freeze-dried powder the preparation method comprises the following steps: lactobacillus plantarum is inoculated in MRS culture medium, in 37 DEG C,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer to stationary phase under the conditions of pH6.4
It washs, centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get lactobacillus plantarum jelly
Dry powder doses；
The bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulose
Enzyme freeze-dried powder the preparation method comprises the following steps: each bacterium branch is inoculated in PCA fluid nutrient medium, culture is to steady under the conditions of 37 DEG C, pH6.8
Periodically, it takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile again
Embedding medium is added in phosphate buffer, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger
Freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
The embedding medium is prepared from the following raw materials in parts by weight: 8 parts of maltodextrin, 6 parts of beta-cyclodextrin, and starch 10
Part, 8 parts of water.
The vacuum freeze drying design parameter are as follows: 65 DEG C of condenser temperature ﹣, 30 DEG C of heating temperature, vacuum degree 60pa, do
The dry moisture content to microbial inoculum is no more than 7%.
The MRS culture medium constituent are as follows: 8 parts of peptone, 10 parts of beef extract, 6 parts of yeast extract, diammonium hydrogen citrate
2.2 parts, 18 parts of glucose, 0.8 part of Tween 80,4 parts of sodium acetate, 2.5 parts of dipotassium hydrogen phosphate, 1000 parts of distilled water；Described
PCA fluid nutrient medium, constituent are as follows: 6 parts of tryptone, 3.2 parts of yeast extract, 1 part of glucose, 1000 parts of distilled water.
Embodiment 3
By-product from sugarcane feed fermenting microbial inoculum, the raw material including following parts by weight: it is false to produce protein for 22 parts of bacterium freeze-dried powder of domestication
9 parts of freeze-dried powder of yeast of silk, 7 parts of lactobacillus plantarum freeze-dried powder, 9 parts of bacillus subtilis freeze-dried powder, 5 parts of aspergillus niger freeze-dried powder, rice
5 parts of aspergillus freeze-dried powder, 8 parts of Trichoderma viride freeze-dried powder, 19 parts of cellulase freeze-dried powder；
The domestication bacterium freeze-dried powder the preparation method comprises the following steps: secondary domestication bacterium is taken to reach the bacterium solution of stationary phase when expanding culture, from
The heart takes bacterium mud to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out
Vacuum freeze drying is to get domestication bacterium freeze dried powder；
The candida utili freeze-dried powder the preparation method comprises the following steps: candida utili is inoculated in malt extract medium,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud sterile phosphate slow to stationary phase under the conditions of 38 DEG C, pH6.4
Fliud flushing washing, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and it is false to get protein is produced to carry out vacuum freeze drying
Silk yeast freeze dried powder；
The lactobacillus plantarum freeze-dried powder the preparation method comprises the following steps: lactobacillus plantarum is inoculated in MRS culture medium, in 38 DEG C,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer to stationary phase under the conditions of pH6.4
It washs, centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get lactobacillus plantarum jelly
Dry powder doses；
The bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulose
Enzyme freeze-dried powder the preparation method comprises the following steps: each bacterium branch is inoculated in PCA fluid nutrient medium, culture is to steady under the conditions of 38 DEG C, pH7.0
Periodically, it takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile again
Embedding medium is added in phosphate buffer, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger
Freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
The embedding medium is prepared from the following raw materials in parts by weight: 9 parts of maltodextrin, 7 parts of beta-cyclodextrin, and starch 11
Part, 15 parts of water.
The vacuum freeze drying design parameter are as follows: 65 DEG C of condenser temperature ﹣, 33 DEG C of heating temperature, vacuum degree 55pa, do
The dry moisture content to microbial inoculum is no more than 7%.
The MRS culture medium constituent are as follows: 9 parts of peptone, 9 parts of beef extract, 5 parts of yeast extract, diammonium hydrogen citrate 2
Part, 20 parts of glucose, 1 part of Tween 80,5 parts of sodium acetate, 1.8 parts of dipotassium hydrogen phosphate, 950 parts of distilled water；The PCA liquid
Culture medium, constituent are as follows: 5 parts of tryptone, 2 parts of yeast extract, 1 part of glucose, 950 parts of distilled water.
Embodiment 4
By-product from sugarcane feed fermenting microbial inoculum, the raw material including following parts by weight: it is false to produce protein for 15 parts of bacterium freeze-dried powder of domestication
10 parts of freeze-dried powder of yeast of silk, 8 parts of lactobacillus plantarum freeze-dried powder, 10 parts of bacillus subtilis freeze-dried powder, 6 parts of aspergillus niger freeze-dried powder,
6 parts of aspergillus oryzae freeze-dried powder, 10 parts of Trichoderma viride freeze-dried powder, 20 parts of cellulase freeze-dried powder；
The domestication bacterium freeze-dried powder the preparation method comprises the following steps: secondary domestication bacterium is taken to reach the bacterium solution of stationary phase when expanding culture, from
The heart takes bacterium mud to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out
Vacuum freeze drying is to get domestication bacterium freeze dried powder；
The candida utili freeze-dried powder the preparation method comprises the following steps: candida utili is inoculated in malt extract medium,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud sterile phosphate slow to stationary phase under the conditions of 37 DEG C, pH6.4
Fliud flushing washing, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and it is false to get protein is produced to carry out vacuum freeze drying
Silk yeast freeze dried powder；
The lactobacillus plantarum freeze-dried powder the preparation method comprises the following steps: lactobacillus plantarum is inoculated in MRS culture medium, in 37 DEG C,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer to stationary phase under the conditions of pH6.4
It washs, centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out vacuum freeze drying to get lactobacillus plantarum jelly
Dry powder doses；
The bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulose
Enzyme freeze-dried powder the preparation method comprises the following steps: each bacterium branch is inoculated in PCA fluid nutrient medium, culture is to steady under the conditions of 37 DEG C, pH7.1
Periodically, it takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is resuspended in sterile again
Embedding medium is added in phosphate buffer, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-dried powder, aspergillus niger
Freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
The embedding medium is prepared from the following raw materials in parts by weight: 10 parts of maltodextrin, 8 parts of beta-cyclodextrin, and starch 12
Part, 20 parts of water.
The vacuum freeze drying design parameter are as follows: 65 DEG C of condenser temperature ﹣, 35 DEG C of heating temperature, vacuum degree 50pa, do
The dry moisture content to microbial inoculum is no more than 7%.
The MRS culture medium constituent are as follows: 8 parts of peptone, 8 parts of beef extract, 6 parts of yeast extract, diammonium hydrogen citrate
1.5 parts, 22 parts of glucose, 0.8 part of Tween 80,4 parts of sodium acetate, 2.5 parts of dipotassium hydrogen phosphate, 1000 parts of distilled water；Described
PCA fluid nutrient medium, constituent are as follows: 4 parts of tryptone, 2 parts of yeast extract, 2 parts of glucose, 1000 parts of distilled water.
Application Example
By-product from sugarcane feed fermenting microbial inoculum prepared by the embodiment of the present invention 3 is applied to bagasse, dregs of beans be main
In the Feed Manufacturing processing of raw material, the additional amount of fermenting agent is that 0.2% fermentation material feed nutrient of raw material weight changes such as
Shown in the following table 3:
The nutritional ingredient of 3 fermentation material of table changes
| Initial feed | Ferment 7d | Ferment 14d | Ferment 21d | Ferment 28d | |
| Protein (%) | 17.79 | 18.38 | 18.67 | 19.54 | 20.08 |
| Viable count (CFU/g × 1010) | 0.137 | 1.90 | 2.00 | 0.395 | 0.390 |
| Moisture (%) | 60.00 | 61.06 | 63.01 | 65.27 | 65.32 |
| pH | 4.54 | 4.29 | 3.89 | 3.76 | 3.71 |
| Neutral detergent fiber (%) | 49.63 | 49.03 | 36.20 | 35.03 | 34.84 |
| Acidic cleaning lignin (%) | 4.55 | 4.63 | 4.08 | 3.53 | 3.48 |
| Crude fat (%) | 9.01 | 12.46 | 11.61 | 10.02 | 9.98 |
| Coarse ash (%) | 12.30 | 11.85 | 11.24 | 10.83 | 10.78 |
Seen from table 3, indices are relatively stable after fermenting 21 days, and gas production at this time has been decreased obviously, feed aromatic flavour,
Quality is stablized, and can be used for animal feeding, palatability is excellent.
By by-product from sugarcane feed fermenting microbial inoculum prepared by the present invention, with common two kinds of microbial inoculums on the market, it is used for
Fermentation produces protein feed using bagasse as the same materials of primary raw material, and comparing result is as shown in table 4 below.
The nutritional ingredient variation of the different bacteria fermentation production feeds of table 4
Wherein, D microbial inoculum is by-product from sugarcane feed fermenting microbial inoculum prepared by the present invention, and N microbial inoculum is that biological section is preced in Guangzhou agriculture
The feed special bacteria agent of skill Co., Ltd production, H microbial inoculum are the fermenting agent of Yichun Qiangwei Biotechnology Co., Ltd.'s production.
As can be seen from Table 4, fermenting agent of the invention, it is every under fermentation condition and the identical situation of fermentation time
The test result of index is superior to remaining two kinds of microbial inoculum.Fermenting agent of the invention is for neutral detergent fiber and acidic cleaning
The degradation capability of lignin is substantially better than N microbial inoculum and H microbial inoculum, and crude ash content is minimum after fermentation, and crude protein content obtains
Biggish raising illustrates that the ferment effect of microbial inoculum of the present invention is optimal.
Claims (8)
1. by-product from sugarcane feed fermenting microbial inoculum, it is characterised in that: the raw material including following parts by weight: domestication bacterium freeze-drying
15~30 parts of powder, 8~10 parts of candida utili freeze-dried powder, 6~8 parts of lactobacillus plantarum freeze-dried powder, bacillus subtilis freeze-drying
8~10 parts of powder, 4~6 parts of aspergillus niger freeze-dried powder, 4~6 parts of aspergillus oryzae freeze-dried powder, 7~10 parts of Trichoderma viride freeze-dried powder, cellulose
18~20 parts of enzyme freeze-dried powder；
The domestication bacterium freeze-dried powder the preparation method comprises the following steps: secondary domestication bacterium is taken to reach the bacterium solution of stationary phase when expanding culture, from
The heart takes bacterium mud to be washed with sterile phosphate buffer, and centrifugation is resuspended in sterile phosphate buffer again, and embedding medium is added, and carries out
Vacuum freeze drying is to get domestication bacterium freeze dried powder；
The candida utili freeze-dried powder the preparation method comprises the following steps: candida utili is inoculated in malt extract medium,
Culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud nothing to stationary phase under the conditions of 36~38 DEG C, pH6.3~6.5
The washing of bacterium phosphate buffer, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and carry out vacuum freeze drying,
Up to candida utili freeze dried powder；
The lactobacillus plantarum freeze-dried powder the preparation method comprises the following steps: lactobacillus plantarum is inoculated in MRS culture medium, in 36~38
DEG C, culture takes bacterium solution to be centrifuged, discards supernatant liquid, take bacterium mud sterile phosphate to stationary phase under the conditions of pH6.3~6.5
Buffer washing, centrifugation are resuspended in sterile phosphate buffer again, and embedding medium is added, and carry out vacuum freeze drying to get plant
Lactobacillus freeze dried powder；
The bacillus subtilis freeze-dried powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulose
Enzyme freeze-dried powder the preparation method comprises the following steps: each bacterium branch is inoculated in PCA fluid nutrient medium, in 36~38 DEG C, the condition of pH6.8~7.2
Lower culture takes bacterium solution to be centrifuged, discards supernatant liquid, bacterium mud is taken to be washed with sterile phosphate buffer, centrifugation is again to stationary phase
It is resuspended in sterile phosphate buffer, embedding medium is added, carries out vacuum freeze drying, that is, respectively obtains bacillus subtilis freeze-drying
Powder, aspergillus niger freeze-dried powder, aspergillus oryzae freeze-dried powder, Trichoderma viride freeze-dried powder, cellulase freeze-dried powder.
2. by-product from sugarcane feed fermenting microbial inoculum according to claim 1, it is characterised in that: the embedding medium by
The raw material of following parts by weight is made: 8~10 parts of maltodextrin, 6~8 parts of beta-cyclodextrin, 10~12 parts of starch, 8~20 parts of water.
3. by-product from sugarcane feed fermenting microbial inoculum according to claim 1, it is characterised in that: the domestication bacterium is tamed and dociled
Change method the following steps are included:
(1) it chooses bacterium mud: choosing the half a year above sugarcane field soil or sugarcane top stack retting mud or sugar refinery bagasse laydown area is stacked
Half a year sludge formed above seals at original bacterium mud scene in sterile glass vials up for safekeeping to get original bacterium mud, takes back laboratory work
It is cultivated for original flora；
(2) primary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, cold in 121 DEG C of high pressure sterilization 10~20min
But it is added to prepared basal medium afterwards to be uniformly mixed, the weight ratio of sugarcane end pin and basal medium is 1.0~1.5:
1 to get primary domestication culture medium；
According to the difference of sugarcane end pin additional amount, 5~6 gradients are set, prepare different primary tame and docile of several sugarcane end pin contents
Change culture medium, the original bacterium mud in step (1) is inoculated into each culture medium, 35~40 DEG C, cultivate under the conditions of pH6.3~6.5
20~30 days, the situation in 2~4 days one subcultures of record was up to excellent with the percentage that sugarcane tail is decomposed, with bacterium
Liquid clarification, not smelly bacterium mud are excellent, in comprehensive each culture medium situation, select one group of optimal culture medium, deposit in bacterium solution
Bacterium living is primary domestication bacterium；
(3) secondary domestication: taking fresh sugarcane end pin to be crushed to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains A
Material；It takes the bagasse powder after juicing to be broken to 100~200 mesh, in 121 DEG C of 10~20min of high pressure sterilization, obtains B material；It takes in sugar refinery
Sundries is removed in the sewage that bagasse shower water is formed, worry, obtains C material；
Above-mentioned A, B, C material are subjected to combination of two or three's combination respectively, are added to basal medium, is made multiple secondary tame and docile
Change culture medium, once tames the bacterium solution of bacterium in inoculation step (2) respectively, cultivated 15~30 days under the conditions of 35~42 DEG C, every 2
~4 days record one subculture in situation, with sugarcane tail, bagasse decompose percentage it is up to excellent, with clarifying contaminated liquids,
Not smelly is excellent, in comprehensive each culture medium situation, selects one group of optimal culture medium, bacterium solution is secondary domestication bacterium；
(4) expand culture: the bacterium solution for the secondary domestication bacterium for taking step (3) to obtain expands culture, and culture medium is secondary domestication
Culture medium when culture medium optimal situation cultivates 40~50h；
(5) it dispenses, the sub- liquid of domesticated strain can be carried out continuing to expand culture, be formed a large amount of by preservation to get the sub- liquid of domesticated strain
Dominant bacteria, to adapt to industrialized production.
4. by-product from sugarcane feed fermenting microbial inoculum according to claim 2, it is characterised in that: the basis culture
Base is prepared from the following raw materials in parts by weight: 8~10 parts of peptone, 8~10 parts of beef extract, and 4~6 parts of yeast extract, brewer's wort 5~
10 parts, 1.5~2.2 parts of diammonium hydrogen citrate, 18~22 parts of glucose, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, phosphorus
1.5~2.5 parts of sour hydrogen dipotassium, 4~6 parts of tryptone, 2~3.2 parts of yeast extract, 1~2 part of glucose, distilled water 900~1000
Part.
5. by-product from sugarcane feed fermenting microbial inoculum according to claim 1, it is characterised in that: the vacuum refrigeration
Dry design parameter are as follows: 65 DEG C of condenser temperature ﹣, 30~35 DEG C of heating temperature, vacuum degree≤60pa, the dry moisture to microbial inoculum contains
Amount is no more than 7%.
6. by-product from sugarcane feed fermenting microbial inoculum according to claim 1, it is characterised in that: the domestication bacterium warp
It crosses after domestication, the ability of decomposition of cellulose greatly promotes, and the ability of bagasse and sugarcane end pin is especially decomposed, sweet
Survival ability in sugarcane by-product is also improved；The main component of domestication bacterium includes: candida utili, lactic acid bacteria, withered
Careless bacillus, aspergillus niger, aspergillus oryzae and Trichoderma viride.
7. by-product from sugarcane feed fermenting microbial inoculum according to claim 1, it is characterised in that: the MRS culture
Base constituent are as follows: 8~10 parts of peptone, 8~10 parts of beef extract, 4~6 parts of yeast extract, 1.5~2.2 parts of diammonium hydrogen citrate,
18~22 parts of glucose, 0.8~1.2 part of Tween 80,4~6 parts of sodium acetate, 1.5~2.5 parts of dipotassium hydrogen phosphate, distilled water 900
~1000 parts；The PCA fluid nutrient medium, constituent are as follows: 4~6 parts of tryptone, 2~3.2 parts of yeast extract, glucose 1
~2 parts, 950~1000 parts of distilled water.
8. by-product from sugarcane feed fermenting microbial inoculum described in claim 1 is preparing the application in ruminant feed.
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| CN114181979A (en) * | 2020-09-14 | 2022-03-15 | 中国科学院大连化学物理研究所 | A kind of method for non-enzymatic biomass hydrolysis saccharification liquid to avoid detoxification and efficiently produce lactic acid |
| CN114958628A (en) * | 2021-02-20 | 2022-08-30 | 柳州市德缘科技开发有限责任公司 | Preparation method of anaerobic active biological leavening agent special for cane mud |
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| CN114958628A (en) * | 2021-02-20 | 2022-08-30 | 柳州市德缘科技开发有限责任公司 | Preparation method of anaerobic active biological leavening agent special for cane mud |
| CN114958628B (en) * | 2021-02-20 | 2024-02-20 | 柳州市德缘科技开发有限责任公司 | Preparation method of special anaerobic active biological starter for bagasse |
| CN113076698A (en) * | 2021-04-20 | 2021-07-06 | 广西大学 | Dynamic multi-target collaborative optimization method and system based on workshop big data |
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