CN103749957A - Preparation method for blue-green algae single-cell protein feed - Google Patents
Preparation method for blue-green algae single-cell protein feed Download PDFInfo
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Abstract
The invention provides a preparation method for a blue-green algae single-cell protein feed. According to the preparation method, aspergillus niger with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.8640 is used for carrying out solid-state fermentation on blue-green algae. The preparation method comprises the specific steps: (1) dehydrating the blue-green algae and carrying out filter pressing to prepare a blue-green algae dehydrated filtering block; (2) inoculating aspergillus niger liquid into a solid-state culture medium and culturing to obtain solid-state aspergillus niger; (3) adding the solid-state aspergillus niger into the blue-green algae dehydrated filtering block and mixing and fermenting to obtain a blue-green algae fermentation product; (4) drying the blue-green algae fermentation product by hot air, and crushing and sieving to obtain the blue-green algae single-cell protein feed. According to the preparation method for the blue-green algae single-cell protein feed, waste resources are utilized, the dehydration is energy-saving and the environmental pollution is not caused; the detoxification of micro-capsule blue-green algae toxin and the synchronous production of the single-cell protein feed are realized.
Description
Technical field
The present invention relates to a kind of waste resource fodder and utilize technology, specifically a kind of preparation method of blue-green algae single-cell protein feed.
Technical background
Along with industrial and agricultural production continues develop rapidly, the frequent aggravation of mankind's activity, has caused inland-lake pool pollution condition to be on the rise, and the rudimentary plant blue-green algae in lake can overgrow in 6 annual-October, is referred to as " wawter bloom "." wawter bloom " takes place frequently, and to local society economic development, resident's life and life and health have been brought adverse influence, it multiple, and continuation and seriousness have caused the very big concern of government and society all circles.
Blue-green algae is planted at Chinese frequent species nearly more than 300, wherein dominant species name is called microcystic aeruginosa (Microcystis aeruyinvsa), when in the inland lakes such as Taihu Lake, Chaohu and Dian Chi, " wawter bloom " occurs, occupation rate is about more than 95%, and almost full lake is all this absolute predominance population.The Microcystin (Microcystin is called for short MC) that micro-capsule blue-green algae produces is that one has hepatotoxicity wind agitation and short carcinous cyclic amino acid peptide matters.Report according to the study, Microcystin MC can cause that fish are poisoning, and in heavy dose of situation, severe patient can lethally be died, and Microcystin MC can cause hepatic injury to mammal small white mouse, causes hepatomegaly and liver tissue lesions.But also there is research to point out, Microcystin MC does not show MA to mouse bone marrow cells chromosome, does not find its aberration inducing effect yet, at least can say, Microcystin MC, to mammal hereditary material DNA, also lacks the abundant foundation of Mutagenicity and MA.
The annual blue-green algae that micro-capsule cyanobacterial bloom produces in lake, biomass is huge.Only jiangsu wuxi one city's statistics, the blue-green algae liquid (wherein containing blue-green algae dry 1-2%) of salvaging from Taihu Lake out from 2011 ~ in June, 2013-October has nearly 1,000,000 tons more than every year, and this is only a part for blue-green algae generation in lake.How good these blue algae resources of processing and utilizing, turn waste into wealth it, have great Social Ecology benefit and economic implications.
By the analysis to Taihu Lake Microcystis aeruginosa resource, wherein nitrogenous approximately 10% left and right, phosphorously approach 1%, the crude protein content that measure and calculation obtains is estimated in 50% left and right, blue algae polysaccharide 5%, crude fat approaches 0.3%, also contains natural colouring matter, unrighted acid and cyanophycean toxin, contains in addition varied physiologically active components such as antibacterial, antiviral and anticancer.Micro-capsule blue-green algae nutrition rich connotation, physiologically active presents many-sided function, and resources development and utilization is well worth doing.
At present the exploitation of micro-capsule blue-green algae are not yet entered to the industrialization compared with wide-range, except by can producing marsh gas power generation to the anaerobic fermentation of blue-green algae and producing organic fertilizer, mostly also in the experimental exploring stage, also do not reach plant-scale extensive utilization.Applicant thinks, blue-green algae is made to protein feed, and significant is paid close attention to, significant:
The first, micro-capsule blue-green algae is produced to protein feed, the resources advantage of itself can farthest be brought into play, and its nitrogenous height, cyanophycin enrich, and kind is varied.Once had report, dry blue-green algae rear grinds, by blue algae powder, with other high-quality protein resource comparisons, blue algae powder albumen is 1.34 ~ 1.46 times of soybean protein, is 3.5 ~ 4.1 times of milk powder, is eel, soft-shelled turtle powder 4.5 ~ 5.8 times.
Second, blue-green algae is remarkable to the feeding effect of livestock and poultry and aquatic livestock, from blue algae powder, make an addition to the effect of carrying out feeding trial feed, 4 ~ 6% blue algae powders in pork pig basal feed, have been added, can increase weight 18 ~ 40%, 0.1 ~ 1% blue algae powder is added in layer diets, and laying rate can improve 3.5% ~ 5.3%.Blue-green algae is applied to aquaculture, with containing 5% blue algae powder, makes bait and breeds fish, and compared with the control, can make carp weightening finish 20.1%, crucian weightening finish 18.3%, bream weightening finish 10.6%.Separately have bibliographical information, the broiler chicken of the blue algae powder of feeding, can significantly improve the appearance luster of chicken leg and sternal rib, has improved outward appearance and the class of product.
The 3rd, the shortage of protein feed is the development bottleneck of feed industry and aquaculture for many years.According to statistics, the whole nation provides the artifical compound feeds such as livestock and poultry and aquatic livestock to reach 1.3 hundred million tons/year, is equivalent to 1/4 ~ 1/5 of National Grain.If making blue-green algae " without grain protein feed ", realization can save considerable grain.The innovative development of this feed technology and popularization, contribute to alleviate the problem of current China protein feed deficiency.
But the micro-capsule blue-green algae that utilizes lake outburst " wawter bloom " to produce is made cyanophycin feed, mainly contains two large technical problems: the one, how effectively to utilize nutrition composition in micro-capsule blue-green algae, and transform and produce cyanophycin feed; The 2nd, how effectively to remove micro-capsule cyanophycean toxin MC.
Existing technology, also in laboratory scale experimental stage, is difficult to form industrialization.Substantially be all first to pass through physics or chemical method, at the lower cyanophycean toxin of removing of strong condition (high temperature, strong acid, highly basic), then make feed.For example, adopting physical method (Chinese invention patent " processing method of blue-green algae ", publication number CN102150747A) is by 150 ~ 300 ℃, 10 ~ 60min high-temperature process detoxification element, by≤60 ℃, be dried and obtain blue-green algae dry powder, it is said and can be used as feed addictive.This method major defect is that treatment temperature is too high, and protein peptide bond structure is had to destruction, can affect the quality of forage protein; Meanwhile, adopt high like this temperature to remove cyanophycean toxin also undesirable, 240 ℃ of processing of high temperature 1 hour, MC-LR also remains and has 53.19ug/kg.Again for example, adopt chemical method (Chinese invention patent " a kind of preparation method of detoxicated algae ", publication number CN1268317A) first blue-green algae is made to dry algae powder, then adopt 1 ~ 3% high-concentration ozone water to carry out cyanophycean toxin detoxification, or by ozone-containing air (O
3content is 20%) carry out cyanophycean toxin detoxification.But, the method to these two kinds of ozone detoxifications that propose herein, whether concrete effect is how, does not disclose concrete data in file, as for the product after detoxification, can, for feed adopts, also not be described further.Again for example, blue-green algae hydrolysis is obtained to amino acid whose chemical method (Chinese invention patent " bloom blue algae hydrolysis obtains the method for free amino acid ", publication number CN101133778) rich blue algae water (water content 75 ~ 97%) is added to the concentrated sulfuric acid (or concentrated hydrochloric acid) in reaction pot, first hydrolysis 5 ~ 8 hours in 100 ~ 120 ℃, add again zinc salt or mantoquita as catalyst, hydrolysis 2 ~ 5 hours again, after cooling, add alkali neutralization, filter and remove precipitation, supernatant hydrolyzate adopts the dry or vacuum drying of spraying or freeze drying to obtain amino-acid powder.But, in strong acid, high temperature, hydrolysis can destroy the tryptophan in protein composition, also make cystine and the oxidation of methionine generating portion, and by the hydrolyzate convection drying after neutralization, volume is very large, not only the salt content of product is high, and production cost is very high, the popularization beyond affordability of feed circle and aquaculture.
Summary of the invention
For above-mentioned technical barrier, the invention provides a kind of microorganism solid fermentation method, synchronously realize the detoxification of micro-capsule cyanophycean toxin and form the two-way object of single-cell protein feed, the object of the present invention is achieved like this:
A preparation method for blue-green algae single-cell protein feed, is characterized in that, black-koji mould take preserving number as CGMCC No.8640 (
aspergillus niger) for zymophyte, blue-green algae is carried out to solid state fermentation, concrete steps are as follows:
1) blue algae dewatering, press filtration are made to the blue algae dewatering filter block that water content is 79%-87.2%;
2) by concentration, be 1 × 10
6-1 × 10
8the black-koji mould liquid of cfu/ml by volume mass ratio 3%-5% is inoculated in solid medium, in 28-35 ℃, under relative humidity 85-90 ﹪ environment, cultivates 36-48h, obtains solid-state black-koji mould;
Described solid medium formula is that mass ratio is wheat bran and the water of 1-1.5;
3 ﹚ are to adding auxiliary material in blue algae dewatering filter block described in step 1, the blue-green algae culture medium that formation water content is 60%, in blue-green algae culture medium, add the solid-state black-koji mould in step 2, form fermentation medium, the mass ratio of described solid-state black-koji mould and blue-green algae culture medium is 4%-6%; Fermentation medium is mixed and is placed in koji tray, in 30-37 ℃, the environment bottom fermentation 36-48h of relative humidity 85-90%, obtain blue algae fermentation product;
The blue algae fermentation product that 4 ﹚ obtain step 3, by the heated-air drying of 80-100 ℃, makes its water content < 10%, then pulverizes and sieves, and obtains blue-green algae single-cell protein feed.
In the present invention, described black-koji mould liquid is to obtain like this:
Black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, then streak inoculation is in Kolle flask Czapek's medium, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then add sterilized water scraping inclined-plane bacterium colony and make bacteria suspension, as black-koji mould liquid;
Or black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then add sterilized water scraping inclined-plane bacterium colony and make after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, as black-koji mould liquid;
Or black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then adding sterilized water scraping inclined-plane bacterium colony makes after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium, 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, with volume ratio 5-10 ﹪ shaking table bacterium liquid, be inoculated in seeding tank fluid nutrient medium, logical filtrated air stirs, at 30-32 ℃ of fermented and cultured 36-48h, obtain zymocyte liquid, as black-koji mould liquid,
Wherein, Czapek's medium formula is: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o, 30g sucrose and 15-20g agar, adjusting pH is 6.0-6.5;
Triangle shaking flask fluid nutrient medium and seeding tank Liquid Culture based formulas are: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o and 30g sucrose, adjusting pH is 6.0-6.5.
In the present invention, described auxiliary material is wheat bran, or blue algae powder, or powder of straw or the combination between them.
The present invention adopts microorganism solid fermentation method, has particularly adopted unsterilised Solid Fermentation of Uncooked Material technology, and synchronously removing cyanophycean toxin MC and transforming blue-green algae is the technical process of two aspects of single-cell protein feed.The biological oxidation, the metabolism assimilation that by fermentation, produce on the one hand, and further the produced catalyticing decomposition action of enzymatic production, the a series of resultant effects such as open loop, decomposition and utilization to micro-capsule cyanophycean toxin have been formed, cause algae toxin be constantly decomposed during the fermentation, destroy and utilize, constantly removed; On the other hand simultaneously, fermentation also makes the diversified nutrition composition in blue-green algae obtain reconfiguring and recycling, and zymophyte self is bred rapidly, unicellular organism thalline and precursor thereof that quantity of formation is huge, become blue-green algae single cell protein, thereby realize the fodder of blue-green algae.
The invention has the advantages that: 1, solid state fermentation conditions nature, ecological, gentle, tunning is abundant, without micro-capsule cyanophycean toxin MC, except results mycoprotein, also can produce multiple digestive ferment, organic acid, vitamin and natural antimicrobial substance, and the somatomedin of multiple the unknown, the blue-green algae forage protein nutrition that forms is good, inherent quality is high; 2, in solid state fermentation production process, present the dehydration energy saving effect of highly significant, and non-environmental-pollution, be again to utilize waste resource, make low cost product, be conducive to apply in various cultivation industries; 3, solid state fermentation ratio is easier to realize industrial large-scale production, can utilize in a large number blue algae resource.
The specific embodiment
In conjunction with specific embodiments, further illustrate the present invention, should understand these embodiment and only for the present invention is described, is not used in and limits the scope of the invention.
The preparation of embodiment 1 black-koji mould
The black-koji mould that the application adopts derives from Institute of Microorganism, Academia Sinica, applicant is by carrying out this bacterial strain after blue-green algae domestication adaptation, what the strain that obtains was new has a lifting single cell protein, improve the black-koji mould of fermentation dehydration rate and anti-living contaminants, applicant is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 27th, 2013 by this new Aspergillus niger strain, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101, preserving number is CGMCC No.8640.
The preparation of embodiment 2 blue algae dewatering filter blocks
Water content is carried out to pressure floatation air at 98% above blue algae collecting liquid by pressurized dissolved air flotation machine, then use sludge filter press press filtration, making water content is the blue algae dewatering filter block of 79%-87.2%.
Embodiment 3 blue-green algae solid state fermentations are prepared single-cell protein feed
The culture medium that the present embodiment is related:
Solid medium formula is that mass ratio is wheat bran and the water of 1-1.5;
Czapek's medium formula: add 3g NaNO in 1L distilled water
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o, 30g sucrose and 15-20g agar, adjusting pH is 6.0-6.5.
Get embodiment 1 obtains black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated 3-5 days, after inclined-plane covers with bacterium colony, picking colony, streak inoculation in Kolle flask Czapek's medium, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, treat that inclined-plane covers with bacterium colony, adding sterilized water scraping inclined-plane bacterium colony and make bacteria suspension, as black-koji mould liquid, is 1 × 10 by 3ml concentration
6-1 × 10
8the black-koji mould liquid of cfu/ml is inoculated in (volume mass ratio is 3%) in 100g solid medium, under 28-35 ℃, relative humidity 85-90 ﹪ environment, cultivates 36-48h, obtains solid-state black-koji mould.
Take method described in embodiment 2, obtain water content as 87.2% dehydration filter block 500g, add 312g wheat bran, make blue-green algae moisture content in medium adjust to 60% left and right, add again the solid-state black-koji mould of 32.5g (mass ratio is about 4%), mix, obtain being granular fermentation medium, fermentation medium is layered in ventilative koji tray, thickness is 4cm, 35 ℃ of temperature, relative humidity 86%, natural ventilation oxygen environment bottom fermentation 48h, fermentation medium can all form the tunning of mycelium enclosed mass, tunning is pulverized and sieved after by the heated-air drying of 80-100 ℃, obtain blue-green algae single-cell protein feed 299g, by Kjeldahl's method, record crude protein content 27.6%.
Embodiment 4 blue-green algae solid state fermentations are removed algae toxin
The culture medium that the present embodiment is related:
Solid medium formula is that mass ratio is wheat bran and the water of 1-1.5;
Czapek's medium formula: add 3g NaNO in 1L distilled water
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o, 30g sucrose and 15-20g agar, adjusting pH is 6.0-6.5;
Triangle shaking flask Liquid Culture based formulas is: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o and 30g sucrose, adjusting pH is 6.0-6.5.
Get the aspergillus niger strain streak inoculation that embodiment 1 obtains activates on Cha Shi slant medium, 28-30 ℃ of lower inclined plane cultivated 3-5 days, picking colony after inclined-plane covers with bacterium colony, streak inoculation is the 28-30 ℃ of 3-4 days that spreads cultivation on Kolle flask inclined-plane, treat that inclined-plane covers with bacterium colony, add sterilized water scraping inclined-plane bacterium colony and make bacteria suspension, with the about 3-5% of volume ratio, be inoculated in triangle shaking flask fluid nutrient medium, 30-32 ℃, 150-180rpm. shaker fermentation cultivation 40-50h obtains the streak inoculation of shaking table strain liquid black-koji mould and activates on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then adding sterilized water scraping inclined-plane bacterium colony makes after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium, 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, as black-koji mould liquid, by 4ml concentration, be 1 × 10
6-1 × 10
8the black-koji mould liquid of cfu/ml is inoculated in (volume mass ratio is 4%) in 100g solid medium, under 28-35 ℃, relative humidity 85-90 ﹪ environment, cultivates 36-48h, obtains solid-state black-koji mould.
With method described in embodiment 2, obtain the blue algae dewatering filter block of 1000g water content 80.8%, add wherein powder of straw 100g, wheat bran 304g, its water content is to 60% left and right, then after adding 70.2g solid black Aspergillus (mass ratio is about 5%) fully to mix, obtain fermentation medium, the fermentation medium that takes out half amount is dried immediately as fermentation medium dry powder, is celadon (as the contrast before fermentation).Second half fermentation medium adopts the standing solid state fermentation of tray, be paved into thickness 4cm, 35 ℃ of temperature, relative humidity is more than 86%, fermentation 48h, fermentation medium forms the tunning of mycelia enclosed mass, and tunning is obtained to tunning dry powder 285g by the heated-air drying of 80-100 ℃, be yellowish-brown, by Kjeldahl's method, record crude protein content 38.7%.Send in the lump Southern Yangtze University's Food Science and the test of analysis center of technology National Key Laboratory to detect the content of algae toxin (MC-LR, MC-YR and MC-RR) in fermentation medium dry powder (sample before fermentation) and tunning dry powder (sample after fermentation), test data is as shown in table 1: the virus elimination rate (%) that calculates toxin:
Sample content of toxins contrast before and after table 1 fermentation
From table 1 result, the virus elimination rate of MC-LR and two kinds of toxin kinds of MC-RR is all more than 70%, and MC-YR tunning does not detect, and virus elimination rate reaches 100%.Applicant thinks, the application adopts black-koji mould solid state fermentation remarkable to the detoxification efficiency of micro-capsule blue-green algae.
The impact of the blue algae dewatering filter block of embodiment 5 different moisture contents on tunning
The culture medium that the present embodiment is related:
Solid medium formula is that mass ratio is wheat bran and the water of 1-1.5;
Czapek's medium formula: add 3g NaNO in 1L distilled water
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o, 30g sucrose and 15-20g agar, adjusting pH is 6.0-6.5.
Triangle shaking flask fluid nutrient medium and seeding tank Liquid Culture based formulas are: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o and 30g sucrose, adjusting pH is 6.0-6.5.
Get the black-koji mould streak inoculation that embodiment 1 obtains activates on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then adding sterilized water scraping inclined-plane bacterium colony makes after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium, 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, with volume ratio 5-10 ﹪ shaking table bacterium liquid, be inoculated in seeding tank fluid nutrient medium, logical filtrated air stirs, at 30-32 ℃ of fermented and cultured 36-48h, obtain zymocyte liquid, as black-koji mould liquid, by 5ml concentration, be 1 × 10
6-1 × 10
8the black-koji mould liquid of cfu/ml is inoculated in (volume mass ratio is 5%) in 100g solid medium, under 28-35 ℃, relative humidity 85-90 ﹪ environment, cultivates 36-48h, obtains solid-state black-koji mould.
With method described in embodiment 2, obtain water content and be respectively 87.2%, 86.5%, 84.4%, 79%, 81.5%, 80.8% the each 500g of dehydration filter block, number respectively 1-6 group, adding respectively wherein wheat bran is 312g, 300g, 277g, 215g, 244g and 236g, make moisture content in medium adjust to 60% left and right, add respectively again 48.7g, 48.0g, 46.6g, 42.9g, the solid-state aspergillus niger (mass ratio is about 6%) of 44.6g and 44.2g, mix, obtain being granular fermentation medium, fermentation medium is layered in ventilative koji tray, thickness is 4cm, 35 ℃ of temperature, relative humidity 86%, natural ventilation oxygen environment bottom fermentation 48h, fermentation medium can all form the tunning of mycelium enclosed mass, tunning is pulverized and sieved after by the heated-air drying of 80-100 ℃, by Kjeldahl's method, record the crude protein content of tunning, result is as shown in table 2:
The blue algae dewatering filter block fermentation of table 2 different moisture content after crude protein content
From table 2 result, the protein content of tunning becomes negative correlation with the water content of blue algae dewatering filter block before fermentation, by reducing the water content of filter block, can improve the protein content of product after solid state fermentation.
The above is only the preferred embodiment of the present invention, it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention, can also make some improvement, and these improve and also should be considered as protection scope of the present invention.
Claims (3)
1. a preparation method for blue-green algae single-cell protein feed, is characterized in that, black-koji mould take preserving number as CGMCC No.8640 (
aspergillus niger) for zymophyte, blue-green algae is carried out to solid state fermentation, concrete steps are as follows:
Blue algae dewatering, press filtration are made to the blue algae dewatering filter block that water content is 79%-87.2%;
By concentration, be 1 × 10
6-1 × 10
8the black-koji mould liquid of cfu/ml by volume mass ratio 3%-5% is inoculated in solid medium, in 28-35 ℃, under relative humidity 85-90 ﹪ environment, cultivates 36-48h, obtains solid-state black-koji mould;
Described solid medium formula is that mass ratio is wheat bran and the water of 1-1.5;
3 ﹚ are to adding auxiliary material in blue algae dewatering filter block described in step 1, the blue-green algae culture medium that formation water content is 60%, in blue-green algae culture medium, add the solid-state black-koji mould in step 2, form fermentation medium, the mass ratio of described solid-state black-koji mould and blue-green algae culture medium is 4%-6%; Fermentation medium is mixed and is placed in koji tray, in 30-37 ℃, the environment bottom fermentation 36-48h of relative humidity 85-90%, obtain blue algae fermentation product;
The blue algae fermentation product that 4 ﹚ obtain step 3, by the heated-air drying of 80-100 ℃, makes its water content < 10%, then pulverizes and sieves, and obtains blue-green algae single-cell protein feed.
2. the preparation method of blue-green algae single-cell protein feed according to claim 1, is characterized in that described black-koji mould liquid is to obtain like this:
Black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, then streak inoculation is in Kolle flask Czapek's medium, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then add sterilized water scraping inclined-plane bacterium colony and make bacteria suspension, as black-koji mould liquid;
Or black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then add sterilized water scraping inclined-plane bacterium colony and make after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, as black-koji mould liquid;
Or black-koji mould streak inoculation is activated on Czapek's medium, 28-30 ℃ of lower inclined plane cultivated picking colony after 3-5 days, streak inoculation is in Kolle flask Czapek's medium again, the 28-30 ℃ of lower inclined plane 3-4 days that spreads cultivation, then adding sterilized water scraping inclined-plane bacterium colony makes after bacteria suspension, with volume ratio 3-5%, be inoculated in triangle shaking flask fluid nutrient medium, 30-32 ℃, 150-180rpm. shaker fermentation is cultivated 40-50h and is obtained shaking table bacterium liquid, with volume ratio 5-10 ﹪ shaking table bacterium liquid, be inoculated in seeding tank fluid nutrient medium, logical filtrated air stirs, at 30-32 ℃ of fermented and cultured 36-48h, obtain zymocyte liquid, as black-koji mould liquid,
Wherein, Czapek's medium formula is: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o, 30g sucrose and 15-20g agar, adjusting pH is 6.0-6.5;
Triangle shaking flask fluid nutrient medium and seeding tank Liquid Culture based formulas are: in 1L distilled water, add 3g NaNO
3, 1g K
2hPO
4, 0.5g KCL, 0.5g MgSO
4﹒ 7H
2o, 0.01g FeSO
4﹒ 4H
2o and 30g sucrose, adjusting pH is 6.0-6.5.
3. according to the preparation method of blue-green algae single-cell protein feed described in claim 1 or 2, it is characterized in that, described auxiliary material is wheat bran, or blue algae powder, or powder of straw, or the combination between them.
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| CN106617043A (en) * | 2016-09-22 | 2017-05-10 | 苏州太阳都生化技术有限公司 | Preparation method and application of bloom cyanobacteria nutrient solution |
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| CN101849618A (en) * | 2010-04-13 | 2010-10-06 | 浙江大学 | Method for producing complex enzyme for feed |
| CN102132762A (en) * | 2011-03-29 | 2011-07-27 | 南开大学 | Straw feed and preparation method thereof |
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| CN101215539A (en) * | 2008-01-07 | 2008-07-09 | 周文彩 | Microorganism bacterium composition, blue algae biological bacterium leaven containing the same, nutriment and biological organic fertilizer, and preparing method thereof |
| CN101317625A (en) * | 2008-06-27 | 2008-12-10 | 刘新虎 | Comprehensive treatment method for pollution of livestock and poultry aquaculture, special feedstuff bridging agent, biological edible warm pad and biological organic fertilizer |
| CN101653189A (en) * | 2009-08-26 | 2010-02-24 | 四川国凤生物科技有限公司 | Natural plant small molecular group substance concentrate and production method thereof |
| CN101849618A (en) * | 2010-04-13 | 2010-10-06 | 浙江大学 | Method for producing complex enzyme for feed |
| CN102132762A (en) * | 2011-03-29 | 2011-07-27 | 南开大学 | Straw feed and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106617043A (en) * | 2016-09-22 | 2017-05-10 | 苏州太阳都生化技术有限公司 | Preparation method and application of bloom cyanobacteria nutrient solution |
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Denomination of invention: A preparation method of cyanobacteria single cell protein feed Effective date of registration: 20200812 Granted publication date: 20150520 Pledgee: Wuxi rural commercial bank Limited by Share Ltd. Yixing branch Pledgor: SKYSTONE FEED (YIXING) Co.,Ltd. Registration number: Y2020990000944 |