Method for manufacturing feedstuff protein by solid state fermentation of apple dregs liquid

[0033] Example 1:

[0034] Seed culture medium: 200g potato, 20g sucrose, and some water; its production method is: cut the potato into small pieces, add appropriate amount of tap water and boil for 20-30min, filter, take the filtrate and add tap water to 1000mL.

[0035] Cultivation of seed strains: Take 300mL seed culture medium into a 500mL Erlenmeyer flask and sterilize it at 121℃ for 20min. After cooling to room temperature, insert each slant strain and place it in an air bath shaker at 30-32℃, 120- Cultivate at 140rpm for 36-54h to obtain mature seed strains.

[0036] The composition of the liquid submerged fermentation medium is: dry apple pomace 90%, sucrose 5%, urea 2%, (NH 4 ) 2 SO 42%, KH 2 PO 4 1%. And add 10 times the solid material of water.

[0037] The connected mixed strains and ratio: Candida utilis: Trichoderma viride: Cider yeast = 4:3:1.5,

[0038] The composition of the solid medium is: dried apple pomace 75%, bran 20%, urea 2%, (NH 4 ) 2 SO 4 2%, KH 2 PO 4 1%, add water to adjust the initial water content to 60%.

[0039] After the fresh wet apple pomace is dried by a hot-air drying line at 75°C and crushed and sieved, 60 mesh raw materials are taken to prepare a liquid fermentation medium and placed in a large-scale fermentation tank, and at the same time, 10% (v/v) is connected to a seed tank for cultivation The mature mixed strains are fermented in a liquid state for 36 hours at 32°C, and then the submerged fermentation broth is poured into the prepared apple pomace solid medium at the amount of 30mL/100g of solid substrate, and the initial water content is 65% indestructible Under the condition of bacteria, the fermentation samples were taken out after 72 hours of solid culture at 32°C, dried, pelletized, and packaged to obtain the apple pomace feed protein product.

[0040] Comparative separate solid-state fermentation experiment: fresh wet apple pomace was dried and crushed on a hot-air drying line at 75°C and then mixed into a solid fermentation medium. Its composition was: dry apple pomace 75%, bran 20%, urea 2%, (NH 4 ) 2 SO 4 2%, KH 2 PO 4 1%. Insert 10% (v/v) into the seed tank to cultivate mature mixed strains (mixing ratio of Candida utilis: Trichoderma viride: Cider yeast = 4:3:1.5), with an initial water content of 65%, no Under sterilization conditions, the fermentation samples were taken out after 96 hours of solid culture at 32°C, dried, pelletized, and packaged to obtain the apple pomace feed protein product.

[0041] The main components, trace elements and amino acid composition determination results of the above two different fermentation forms are shown in Table 1, Table 2 and Table 3:

[0042] Table 1 Content of main ingredients (%)

[0043] Crude protein True protein Crude fat Crude fiber Ash

[0044] Fermentation raw materials 16.28 10.02 8.94 16.68 4.25

[0045] Individual solid-state fermentation products 28.43 26.59 13.47 10.84 6.52

[0046] Liquid and solid fermentation products 29.08 26.63 13.56 10.32 7.85

[0047] Table 2 Trace element content (mg/kg)

[0048] Ca(%) Mg(%) Fe Zn Mn Cu Se Pb Cd As

[0049] apple

[0050] 0.113 0.061 169.2 14.6 13.1 9.8 0.18 0.6 0.02 0.17

[0051] Slag

[0052] Ferment

[0053] 0.493 0.204 276.8 31.3 20.8 16.7 0.29 1.6 0.68 0.59

[0054] product

[0055]

[0056] test methods:

[0057] Determination of crude protein: adopt the national standard GB/T6432-86, namely the micro-Kjeldahl method.

[0058] Determination of true protein: using an improved micro-Kjeldahl method-weighing 5g of solid fermentation product, leaching with 40mL of 75% ethanol for 24h, fully stirring, suction filtration, and then washing the filter residue twice with 40mL of 75% ethanol. To wash away inorganic nitrogen and urea, while precipitating protein. Dry the precipitate at 65°C to a constant weight, accurately weigh about 1g for nitrification, and then use the micro-Kjeldahl method for determination, which is the above-mentioned national standard GB/T6432-86.

[0059] Determination of crude fat content: adopt the national standard GB/T6433-86, namely Soxhlet extraction method.

[0060] Determination of crude fiber content: adopt the national standard GB/T6434-86, that is, the acid-base washing method.

[0061] Determination of ash content: adopt the national standard GB/T6438-86, namely the ash method.

[0062] Determination of amino acids: determined by the 121-MB amino acid analyzer of the Analysis and Testing Center of Northwest Agriculture and Forestry University.

[0063] It can be seen from the above measurement results that with apple pomace as the main raw material, the main components of the product have changed significantly after solid-state fermentation of the mixed bacterial seed liquid, and the crude protein, true protein, crude fat, and ash content increased by 78.62%, 165.76%, respectively. 51.67% and 84.70%, while the crude fiber was reduced by 38.13%. After the analysis of 17 essential amino acids, its content increased sharply and was abundant and complete, especially serine, lysine and tyrosine, which are very important for the growth of poultry and livestock. The increase rates of acid and arginine compared with the fermentation raw materials were as high as 162.3%, 275.2%, 81.3% and 89.4%, respectively, and the nutritional value was significantly increased.

[0064] From the comparison of the main components and amino acid content of the two different types of fermentation products, it can be seen that the nutrient content of the liquid-solid fermentation is higher than that of the solid-state fermentation alone, which again shows that the liquid-solid-state fermentation is better than the solid-state fermentation alone.

[0065] It can be seen from the determination of trace elements that the content of beneficial elements is rich, and the content of heavy metal elements meets the requirements of the national sanitary standards for similar feed base materials. The fermented product was tested by the Feed Quality Supervision and Testing Center of the Ministry of Agriculture (Xi'an), an authoritative department, and no Salmonella was detected, aflatoxin B 1 0.0034mg/kg is far less than the requirements of the national feed hygiene standards (GB13078-91) for similar feed base materials such as bran and corn (0.05mg/kg). After the Shaanxi Provincial Center for Disease Control and Prevention conducted a toxicological test, the product is actually non-toxic and can be widely used as a protein feed in the field of breeding.

[0066] Example 2:

[0067] After the fresh wet apple pomace is dried on a hot-air drying line at 65°C and crushed and sieved, the 40 mesh raw materials are taken as liquid fermentation medium and placed in a large-scale fermentation tank. The composition of the liquid fermentation medium is: dried apple pomace 90%, sucrose 4% , Urea 2%, (NH 4 ) 2 SO 4 2%, KH 2 PO 4 2%, and add 12 times the moisture of the solid material. At the same time, 8% (v/v) is connected to the seed tank to cultivate the mature mixed strains, and the ratio of the connected strains: Candida utilis: Trichoderma viride: Wine yeast: Trichoderma koningii = 2:1.5 :6:3, at 33℃, deep fermentation in liquid state for 36h, and then pour the fermentation broth into the prepared apple pomace solid medium according to the amount of 20mL/100g solid substrate. The composition of the medium is: dried apple pomace 84% , Bran 10%, urea 3%, (NH 4 ) 2 SO 4 2%, KH 2 PO 4 1%. Under the condition of 70% water content and no sterilization, solid-state culture at 33°C for 78 hours, then take out the fermentation sample, dry, pellet, and package to obtain the apple pomace feed protein product. The crude protein content can reach 27.69%, and the true protein content is 25.21%.

[0068] The determination method is the same as above.