CN102578389A - Method for producing high-protein feed by using guanosine fermentation waste liquor - Google Patents
Method for producing high-protein feed by using guanosine fermentation waste liquor Download PDFInfo
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Abstract
The invention belongs to the field of environmental engineering and bioengineering, and particularly discloses a method for producing a protein feed by using a guanosine fermentation waste liquor. The method comprises the following steps of: 1, activating strains; 2, culturing seeds in a shake flask; 3, culturing seeds; 4, inoculating bacillus subtilis, bacillus licheniformis and saccharomycetes by taking the guanosine fermentation waste liquor as a raw material, and performing mixed fermentation; and 5, spray-drying, and crushing to obtain the protein feed. The total number of floras contained in the protein feed is 6.83*10<8> to 1.47*10<9>CFU/g, the content of crude protein is 49 to 51 percent, and the protein feed is rich in minerals and vitamins and is a high-quality protein poultry and livestock feed.
Description
Technical field
The invention belongs to environmental project and bioengineering field, is a kind of method of utilizing the guanosine fermentation waste water to produce protein feed specifically.
Background technology
Guanosine has another name called guanosine or 9-(β-D-ribofuranosyl) guanine.The purposes of guanosine very extensively; Can be used for fabricated food freshener-5`-Sodium guanylate, flavour nucleotide disodium and as the important intermediate of medical product; Can form a series of downstream medical product; Comprising ucleosides antiviral drugs such as Ribavirin, ACV etc., also is the primary raw material that is used to make acyclovir (Acyclovir), ribavirin (ATC), Sodium Guanosine Triphosphate medicines such as (GTP).At present, domestic market guanosine demand is greatly about 3 500 tons/year, and with annual 10% speed increment.
Fermentation method is adopted in guanosine production, and this method can produce bulk fermentation waste water.The residual sugar amount is about 3% in the guanosine fermentation waste water, and total nitrogen content is about 0.6%, and ammonia-nitrogen content is about 0.5%; Inorganic salt content is about 4.0%-5.0%, and waste liquor PH 7.0 is a kind of high BOD, the nutritious organic wastewater of high ammonia nitrogen; Directly discharging can cause the water body biology because of the anoxic mortality; Handling the back qualified discharge not only increases production cost, and the nutritional labeling in the waste water can not make full use of, and causes the wasting of resources.
Summary of the invention
The object of the invention is exactly to solve the guanosine fermented waste fluid directly to discharge the pollution problem to environment, and another purpose just provides a kind of method of utilizing the guanosine fermentation waste water to produce protein feed.
The present invention solves the problems of the technologies described above the technical scheme that is adopted: utilize the guanosine fermentation waste water to produce the method for protein feed, it is characterized in that including following steps:
1) in 1 L guanosine fermentation waste water, add sucrose 0.5 g-2.0 g, peptone 0.1 g-0.5 g, yeast extract 0.5 g-1.0 g obtains producing the fermentation medium of protein feed;
2) actication of culture; Picking bacillus subtilis, bacillus licheniformis, saccharomycete are inoculated on the corresponding slant medium respectively; In 30 ℃-37 ℃, cultivate 10 hr-20 hr, obtain bacillus subtilis, bacillus licheniformis, saccharomycetic inclined-plane seed respectively;
3) shake-flask seed is cultivated; With step 2) good bacillus subtilis, bacillus licheniformis, the saccharomycetic inclined-plane seed of activation that obtain be seeded to triangle respectively and shake in the bottle and cultivate, and obtains bacillus subtilis, bacillus licheniformis, saccharomycetic shake-flask seed;
4) fermentation seed is cultivated; With the bacillus subtilis of cultivating in the step 3), bacillus licheniformis, saccharomycetic shake-flask seed by volume the inoculum concentration of percentage 5%-10% be inoculated in the seeding tank respectively; In 30 ℃-37 ℃; Cultivate 20 hr-40 hr, obtain bacillus subtilis, bacillus licheniformis, saccharomycetic seed liquor;
5) mixing fermentation culture; With the bacillus subtilis of cultivating in the step 4), bacillus licheniformis, saccharomycetic seed liquor, adopt the described fermentation medium of step 1), be inoculated in the fermentation tank by total inoculum concentration of 5%-10%; In 34 ℃-36 ℃; Ventilating ratio is 0.13-0.15, cultivates 40 hr-48 hr, obtains mixed-culture medium;
6) spray-drying is carried out spray-drying, pulverizing with the mixed-culture medium of step 5), promptly gets protein feed.
Press such scheme, step 2) the used slant medium that is used for actication of culture is: the slant medium of bacillus subtilis consists of: glucose 0.5 g, peptone 0.6 g, yeast extract 0.7 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL; The slant medium of bacillus licheniformis consists of: glucose 0.5 g, peptone 0.7 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL; Saccharomycetic slant medium consists of: sucrose 2.0 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL.
Press such scheme; The shake-flask seed condition of culture does in the step 3), and bacillus subtilis is adopted following culture medium: in 100 mL guanosine fermentation waste waters, add yeast extract 1.0 g-3.0 g, medium pH 7.0; In 35 ℃-37 ℃, 20 hr-24 hr are cultivated in 200 rpm joltings; Bacillus licheniformis is adopted following culture medium: in 100 mL guanosine fermentation waste waters, add peptone 0.5 g-1.5 g, yeast extract 1.0 g-2.0 g, and medium pH 7.0, in 35 ℃-37 ℃, 26 hr-30 hr are cultivated in 180 rpm joltings; Saccharomycete adopts following culture medium: in 100 mL guanosine fermentation waste waters, add yeast extract 0.5 g-1.0 g, sucrose 0.5 g-2.0 g, in 28 ℃-32 ℃, 40 hr-48 hr are cultivated in 150 rpm joltings.
Press such scheme, the fermentation seed condition of culture is in the step 4): bacillus subtilis is adopted following culture medium: in 1 L guanosine fermentation waste water, add yeast extract 5 g-30 g, transfer pH7.0; Inoculum concentration is percent by volume 5%-7%, and in 35 ℃-37 ℃, ventilating ratio is 0.13-0.15; Cultivate 20 hr-24 hr, sampling is diluted 10 times; Survey cell density in 650 nm, make it to reach 1.46-1.52; Bacillus licheniformis is adopted following culture medium: in 1 L guanosine fermentation waste water, add peptone 5.0 g-15.0 g, yeast extract 10.0 g-20.0 g, transfer pH7.0.Inoculum concentration is 7%-10%, and in 35 ℃-37 ℃, ventilating ratio is 0.13-0.15, cultivates 26 hr-30 hr, and sampling is diluted 10 times, surveys cell density in 650 nm, makes it to reach 1.48-1.52; Saccharomycete adopts following culture medium: in 1 L guanosine fermentation waste water, add yeast extract 5.0 g-10.0 g, sucrose 5.0 g-20.0 g, inoculum concentration is 7%-10%, in 28 ℃-32 ℃; Ventilating ratio is 0.10-0.12; Cultivate 40 hr-48 hr, sampling is diluted 10 times; Survey cell density in 650 nm, make it to reach 1.44-1.53.
Press such scheme, the inoculum concentration of mixed culture fermentation is in the step 5): by bacillus subtilis inoculum concentration 1.0%-3.0%, bacillus licheniformis inoculum concentration 2.0%-4.0%, saccharomycete inoculum concentration 2.0%-3.0%.
Press such scheme, described protein feed is a pale yellow powder, flora sum 6.83 * 10
8-1.47 * 10
9CFU/g, crude protein content are 48%-51%.Being rich in mineral and vitamin, is a kind of fowl poultry protein feed of high-quality.
The present invention utilizes guanosine waste water to be raw material, fermentation production of protein feedstuff, and wherein flora adds up to 6.83 * 10
8-1.47 * 10
9CFU/g, crude protein content is 48%-51%, is rich in mineral and vitamin, is the albumen poultry and livestock feed of a high-quality.The present invention has not only solved the pollution of the discharge of wastewater of guanosine fermentation generation to environment, and has reduced the production cost of protein feed, accomplishes the maximum using of resource.
The specific embodiment
Below in conjunction with embodiment the present invention is done further detailed explanation, but can not be construed as limiting the invention.
1 one kinds of embodiment are following with guanosine fermentation waste water production protein feed method method:
Step 1, actication of culture, picking bacillus subtilis, bacillus licheniformis, saccharomycete are inoculated on the corresponding slant medium, and bacillus subtilis, bacillus licheniformis are cultivated 10 hr and 16 hr respectively in 37 ℃; Saccharomycete is cultivated 20 hr in 30 ℃.
The slant medium of bacillus subtilis consists of: glucose 0.5 g, peptone 0.6 g, yeast extract 0.7 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL; The slant medium of bacillus licheniformis consists of: glucose 0.5 g, peptone 0.7 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL; Saccharomycetic slant medium consists of: sucrose 2.0 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL.
Step 2, shake-flask seed is cultivated, and the bacillus subtilis that activation in the step 1 is good is inoculated in to have sterilized and contains yeast extract 2.0 g, guanosine fermentation waste water 100 mL, and pH 7.0 shakes in the bottle, and in 37 ℃, 20 hr are cultivated in 200 rpm joltings; The bacillus licheniformis that activation in the step 2 is good is inoculated in to have sterilized and contains the shaking in the bottle of peptone 1.0 g, yeast extract 1.5 g, guanosine fermentation waste water 100 mL, pH7.0, and in 35 ℃, 24 hr are cultivated in 180 rpm joltings; The saccharomycete that activation in the step 2 is good is inoculated in to have sterilized and contains sucrose 1.0 g, yeast extract 0.7 g, and guanosine fermentation waste water 100 mL shake in the bottle, and in 30 ℃, 48 hr are cultivated in 150 rpm joltings.
Step 3, fermentation seed is cultivated.Be inoculated in respectively and carry out seed culture in the seeding tank shaking bacterium liquid that bottle cultivates in the step 2.
The bacillus subtilis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass, transfer pH7.0,121 ℃ of sterilization 30 min, cooling than the yeast extract that is 2.0%.With 5% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 37 ℃, ventilating ratio 0.13 is cultivated 20 hr, and the cell density that dilutes 10 times is 1.46 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 37 ℃, ventilating ratio 0.12 is cultivated 20 hr, and the cell density that dilutes 10 times is 1.48 at 650 nm places.
The bacillus licheniformis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 1.0% peptone, 1.5% yeast extract, transfer pH7.0.121 ℃ of sterilization 30 min, cooling.With 7% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 35 ℃, ventilating ratio 0.14 is cultivated 26 hr, and the cell density that dilutes 10 times is 1.49 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 35 ℃, ventilating ratio 0.14 is cultivated 26 hr, makes the cell density of 10 times of dilutions reach 1.48 at 650 nm places.
Saccharomycete seed liquid is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 1.0% sucrose, 0.7% yeast extract, the pH nature.121 ℃ of sterilization 30 min, cooling.With 10% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 30 ℃, ventilating ratio 0.10 is cultivated 48 hr, and the cell density that dilutes 10 times is 1.53 at 650 nm places.Be pressed into then in the seeding tank of 500 L, 30 ℃, ventilating ratio 0.10 is cultivated 48 hr, and the cell density that dilutes 10 times is 1.54 at 650 nm places.
Step 4, mixing fermentation culture.Get sucrose 18 kg, peptone 12 kg, yeast extract 18 kg, guanosine fermentation waste water 36 m
3, add 50 m behind the mixed dissolution
3Fermentation tank in, 121 ℃ the sterilization 30 min.Cooling, inoculation bacillus subtilis 1.2 m
3, bacillus licheniformis 1.4 m
3, saccharomycete 1.4 m
3(total inoculum concentration 10%), 36 ℃, ventilating ratio is 0.13, cultivates 40 hr, puts jar.
Step 5, spray-drying.Mixed culture fermentation liquid in the step 4 is carried out spray-drying, pulverizing, mycoprotein (being pale yellow powder), wherein to add up to be 1.07 * 10 to flora
9CFU/g, crude protein content are 50.6%.
2 one kinds of embodiment are following with guanosine fermentation waste water production protein feed method method:
Step 1 is carried out actication of culture with embodiment 1.
Step 2, shake-flask seed is cultivated, and the bacillus subtilis that activation in the step 1 is good is inoculated in to have sterilized and contains yeast extract 1.0 g, guanosine fermentation waste water 100 mL, and pH7.0 shakes in the bottle, and in 35 ℃, 24 hr are cultivated in 200 rpm joltings; The bacillus licheniformis that activation in the step 2 is good is inoculated in to have sterilized and contains the shaking in the bottle of peptone 0.5 g, yeast extract 2.0 g, guanosine fermentation waste water 100 mL, pH7.0, and in 36 ℃, 30 hr are cultivated in 180 rpm joltings; The saccharomycete that activation in the step 2 is good is inoculated in to have sterilized and contains sucrose 0.5 g, yeast extract 1.0 g, and guanosine fermentation waste water 100 mL shake in the bottle, and in 28 ℃, 44 hr are cultivated in 150 rpm joltings.
Step 3, fermentation seed is cultivated.Be inoculated in respectively and carry out seed culture in the seeding tank shaking bacterium liquid that bottle cultivates in the step 2.
The bacillus subtilis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass, transfer pH7.0,121 ℃ of sterilization 30 min, cooling than the yeast extract that is 1.0%.With 6% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 35 ℃, ventilating ratio 0.15 is cultivated 24 hr, and the cell density that dilutes 10 times is 1.51 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 35 ℃, ventilating ratio 0.15 is cultivated 24 hr, and the cell density that dilutes 10 times is 1.49 at 650 nm places.
The bacillus licheniformis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 0.5% peptone, 2.0% yeast extract, transfer pH7.0.121 ℃ of sterilization 30 min, cooling.With 9% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 36 ℃, ventilating ratio 0.13 is cultivated 30 hr, and the cell density that dilutes 10 times is 1.48 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 36 ℃, ventilating ratio 0.13 is cultivated 30 hr, and the cell density that dilutes 10 times is 1.49 at 650 nm places.
Saccharomycete seed liquid is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 0.5% sucrose, 1.0% yeast extract, the pH nature.121 ℃ of sterilization 30 min, cooling.With 7% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 28 ℃, ventilating ratio 0.12 is cultivated 44 hr, and the cell density that dilutes 10 times is 1.44 at 650 nm places.Be pressed into then in the seeding tank of 500 L, 28 ℃, ventilating ratio 0.12 is cultivated 44 hr, and the cell density that dilutes 10 times is 1.46 at 650 nm places.
Step 4, mixing fermentation culture.Get sucrose 40 kg, peptone 18 kg, yeast extract 24 kg, guanosine fermentation waste water 36 m
3, add 50 m behind the mixed dissolution
3Fermentation tank in, 121 ℃ the sterilization 30 min.Cooling, inoculation bacillus subtilis 0.8 m
3, bacillus licheniformis 1.1 m
3, saccharomycete 0.9 m
3(total inoculum concentration 7%), 34 ℃, ventilating ratio is 0.14, cultivates 45 hr, puts jar.
Step 5, spray-drying.Mixed culture fermentation liquid in the step 4 is carried out spray-drying, pulverizing, get mycoprotein (being pale yellow powder), wherein the flora sum 9.26 * 10
8CFU/g, crude protein content are 51.0%.
3 one kinds of embodiment are following with guanosine fermentation waste water production protein feed method method:
Step 1 is carried out actication of culture with embodiment 1.
Step 2, shake-flask seed is cultivated, and the bacillus subtilis that activation in the step 2 is good is inoculated in to have sterilized and contains yeast extract 3.0 g, guanosine fermentation waste water 100 mL, and pH7.0 shakes in the bottle, and in 36 ℃, 22 hr are cultivated in 200 rpm joltings; The bacillus licheniformis that activation in the step 1 is good is inoculated in to have sterilized and contains the shaking in the bottle of peptone 1.5 g, yeast extract 0.5 g, guanosine fermentation waste water 100 mL, pH7.0, and in 37 ℃, 28 hr are cultivated in 180 rpm joltings; The saccharomycete that activation in the step 2 is good is inoculated in to have sterilized and contains sucrose 2.0 g, yeast extract 0.5 g, and guanosine fermentation waste water 100 mL shake in the bottle, and in 32 ℃, 40 hr are cultivated in 150 rpm joltings.
Step 3, fermentation seed is cultivated.Be inoculated in respectively and carry out seed culture in the seeding tank shaking bacterium liquid that bottle cultivates in the step 2.
The bacillus subtilis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass, transfer pH7.0,121 ℃ of sterilization 30 min, cooling than the yeast extract that is 3.0%.With 7% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 36 ℃, ventilating ratio 0.14 is cultivated 22 hr, and the cell density that dilutes 10 times is 1.49 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 36 ℃, ventilating ratio 0.14 is cultivated 22 hr, and the cell density that dilutes 10 times is 1.52 at 650 nm places.
The bacillus licheniformis seed liquor is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 1.5% peptone, 0.5% yeast extract, transfer pH7.0.121 ℃ of sterilization 30 min, cooling.With 10% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 37 ℃ of ventilating ratios 0.15 are cultivated 28 hr, and the cell density that dilutes 10 times is 1.52 at 650 nm places.Be pressed into then in the seeding tank of 500 L, in 37 ℃, ventilating ratio 0.15 is cultivated 28 hr, and the cell density that dilutes 10 times is 1.51 at 650 nm places.
Saccharomycete seed liquid is cultivated.Medium preparation: in the guanosine fermentation waste water, add volume mass than being 2.0% sucrose, 0.5% yeast extract, the pH nature.121 ℃ of sterilization 30 min, cooling.With 9% inoculum concentration, with the shake-flask seed of cultivating in the step 2, be inoculated in the seeding tank of 10 L, 32 ℃, 150 rpm, ventilating ratio 0.11 is cultivated 40 hr, and the cell density that dilutes 10 times is 1.45 at 650 nm places.Be pressed into then in the seeding tank of 500 L, 32 ℃, 150 rpm, ventilating ratio 0.11 is cultivated 40 hr, and the cell density that dilutes 10 times is 1.46 at 650 nm places.
Step 4, mixing fermentation culture.Get sucrose 72 kg, peptone 3.6 kg, yeast extract 36 kg, guanosine fermentation waste water 36 m
3, add 50 m behind the mixed dissolution
3Fermentation tank in, 121 ℃ the sterilization 30 min.Cooling, inoculation bacillus subtilis 0.5 m
3, bacillus licheniformis 0.8 m
3, saccharomycete 0.7 m
3(total inoculum concentration 5%), 35 ℃, ventilating ratio is 0.15, cultivates 48 hr, puts jar.
Step 5, spray-drying.Mixed culture fermentation liquid in the step 4 is carried out spray-drying, pulverizing, get mycoprotein (being pale yellow powder), wherein the flora sum 6.97 * 10
8CFU/g, crude protein content are 48.6%.
Claims (6)
1. utilize the guanosine fermentation waste water to produce the method for protein feed, it is characterized in that including following steps:
1) in 1 L guanosine fermentation waste water, add sucrose 0.5 g-2.0 g, peptone 0.1 g-0.5 g, yeast extract 0.5 g-1.0 g obtains producing the fermentation medium of protein feed;
2) actication of culture; Picking bacillus subtilis, bacillus licheniformis, saccharomycete are inoculated on the corresponding slant medium respectively; In 30 ℃-37 ℃, cultivate 10 hr-20 hr, obtain bacillus subtilis, bacillus licheniformis, saccharomycetic inclined-plane seed respectively;
3) shake-flask seed is cultivated; With step 2) good bacillus subtilis, bacillus licheniformis, the saccharomycetic inclined-plane seed of activation that obtain be seeded to triangle respectively and shake in the bottle and cultivate, and obtains bacillus subtilis, bacillus licheniformis, saccharomycetic shake-flask seed;
4) fermentation seed is cultivated; With the bacillus subtilis of cultivating in the step 3), bacillus licheniformis, saccharomycetic shake-flask seed by volume the inoculum concentration of percentage 5%-10% be inoculated in the seeding tank respectively; In 30 ℃-37 ℃; Cultivate 20 hr-40 hr, obtain bacillus subtilis, bacillus licheniformis, saccharomycetic seed liquor;
5) mixing fermentation culture; With the bacillus subtilis of cultivating in the step 4), bacillus licheniformis, saccharomycetic seed liquor, adopt the described fermentation medium of step 1), be inoculated in the fermentation tank by total inoculum concentration of 5%-10%; In 34 ℃-36 ℃; Ventilating ratio is 0.13-0.15, cultivates 40 hr-48 hr, obtains mixed-culture medium;
6) spray-drying is carried out spray-drying, pulverizing with the mixed-culture medium of step 5), promptly gets protein feed.
2. according to the right 1 described method of utilizing the guanosine fermentation waste water to produce protein feed; It is characterized in that step 2) the used slant medium that is used for actication of culture is: the slant medium of bacillus subtilis consists of: glucose 0.5 g; Peptone 0.6 g, yeast extract 0.7 g, sodium chloride 0.5 g; Agar 2.0 g, deionized water 100 mL; The slant medium of bacillus licheniformis consists of: glucose 0.5 g, peptone 0.7 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL; Saccharomycetic slant medium consists of: sucrose 2.0 g, yeast extract 0.8 g, sodium chloride 0.5 g, agar 2.0 g, deionized water 100 mL.
3. according to the right 1 described method of utilizing the guanosine fermentation waste water to produce protein feed; It is characterized in that the shake-flask seed condition of culture does in the step 3); Bacillus subtilis is adopted following culture medium: in 100 mL guanosine fermentation waste waters, add yeast extract 1.0 g-3.0 g; Medium pH 7.0, in 35 ℃-37 ℃, 20 hr-24 hr are cultivated in 200 rpm joltings; Bacillus licheniformis is adopted following culture medium: in 100 mL guanosine fermentation waste waters, add peptone 0.5 g-1.5 g, yeast extract 1.0 g-2.0 g, and medium pH 7.0, in 35 ℃-37 ℃, 26 hr-30 hr are cultivated in 180 rpm joltings; Saccharomycete adopts following culture medium: in 100 mL guanosine fermentation waste waters, add yeast extract 0.5 g-1.0 g, sucrose 0.5 g-2.0 g, in 28 ℃-32 ℃, 40 hr-48 hr are cultivated in 150 rpm joltings.
4. utilize in the method that the guanosine fermentation waste water produces protein feed according to right 1 is described, it is characterized in that the fermentation seed condition of culture is in the step 4): bacillus subtilis is adopted following culture medium: in 1 L guanosine fermentation waste water, add yeast extract 5 g-30 g, transfer pH7.0; Inoculum concentration is percent by volume 5%-7%, and in 35 ℃-37 ℃, ventilating ratio is 0.13-0.15; Cultivate 20 hr-24 hr, sampling is diluted 10 times; Survey cell density in 650 nm, make it to reach 1.46-1.52; Bacillus licheniformis is adopted following culture medium: in 1 L guanosine fermentation waste water, add peptone 5.0 g-15.0 g, yeast extract 10.0 g-20.0 g, transfer pH7.0, inoculum concentration is 7%-10%; In 35 ℃-37 ℃, ventilating ratio is 0.13-0.15, cultivates 26 hr-30 hr; Sampling; Dilute 10 times, survey cell density, make it to reach 1.48-1.52 in 650 nm; Saccharomycete adopts following culture medium: in 1 L guanosine fermentation waste water, add yeast extract 5.0 g-10.0 g, sucrose 5.0 g-20.0 g, inoculum concentration is 7%-10%, in 28 ℃-32 ℃; Ventilating ratio is 0.10-0.12; Cultivate 40 hr-48 hr, sampling is diluted 10 times; Survey cell density in 650 nm, make it to reach 1.44-1.53.
5. utilize in the method that the guanosine fermentation waste water produces protein feed according to right 1 is described, it is characterized in that the inoculum concentration of mixed culture fermentation in the step 5) is: inoculate by bacillus subtilis inoculum concentration 1.0%-3.0%, bacillus licheniformis inoculum concentration 2.0%-4.0%, saccharomycete inoculum concentration 2.0%-3.0%.
6. utilize in the method that the guanosine fermentation waste water produces protein feed according to right 1 is described, it is characterized in that described protein feed is a pale yellow powder, flora sum 6.83 * 10
8-1.47 * 10
9CFU/g, crude protein content are 48%-51%.
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Cited By (3)
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| CN103120253A (en) * | 2013-03-06 | 2013-05-29 | 左英龙 | Method for producing nutritional feed through utilizing L-arginine extraction waste fluid |
| CN103636921A (en) * | 2013-12-18 | 2014-03-19 | 海南椰国热带水果食品加工有限公司 | Microorganism feed additive and preparation method thereof |
| CN112592946A (en) * | 2020-12-31 | 2021-04-02 | 河南巨龙生物工程股份有限公司 | Environment-friendly high-yield guanosine production process method |
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