CN102987057A - Production method of biological fish meal through microbial fermentation - Google Patents
Production method of biological fish meal through microbial fermentation Download PDFInfo
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- CN102987057A CN102987057A CN2012104946511A CN201210494651A CN102987057A CN 102987057 A CN102987057 A CN 102987057A CN 2012104946511 A CN2012104946511 A CN 2012104946511A CN 201210494651 A CN201210494651 A CN 201210494651A CN 102987057 A CN102987057 A CN 102987057A
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Abstract
The invention provides a production technology of biological fish meal by microbial fermentation method. The production method comprises the following steps of performing enzymolysis and scouring on corn gluten meal in a fermentation substrate to extract protein, and mixing the fermentation substrate with other raw materials in a certain proportion; carrying out digestion and sterilization and on the mixed fermentation substrate through a conditioner, adding water for reducing temperature, and adding purified fulvic acid which is 1 to 3 percent by weight of the fermentation substrate, and inoculating a mixed strain fermentation bacterium liquid consisting of 3 bacterium liquids; filling the inoculated fermentation substrate into a mobile fermentation track through a kick-out device, fermenting for several days indoors in the fermenting room; baking the fermented material at a low temperature, and crushing to obtain the biological fish bone finished product. The fermentation substrate consists of the following components in percentage by weight: 40 to 60 parts of corn gluten meal, 5 to 10 parts of rapeseed meal, 5 to 10 parts of alcohol meal, and 30 to 40 parts of vermicelli albumen powder. When the product is prepared to be supplied to a monogastric animal, the fish soluble condensed, which is 10% by weight of the fermentation substrate; and the three bacterium liquids are respectively monilia tropicalis, lactobacillus casei and bacterium liquid of glossy ganoderma.
Description
Technical field
The invention relates to a kind of method of producing biological fish meal by microbial fermentation, belong to the additive for microbe feedstuff field.
Background technology
Fish meal is of paramount importance dietary protein origin in the aquatic feeds, also is the material that accounts for maximum ratio in the aquatic feeds cost.It because of have the protein content height, be rich in the animal essential amino acid, digestibility is good, be called as easily " king of albumen " by advantages such as animal digest and assimilate.Along with signing Sino-Korean, Sino-Japan fisheries agreement, the reduction of world's fishery products amount of fishing, the imported fish meal price soars, and has brought stern challenge to feed industry.Seek the cheapness of part or all of Peru Fish Dietary and stable albumen, it is under-supply to alleviate fish meal on the one hand, also is the Important Action that reduces feed cost on the other hand.Therefore, how utilizing plant protein source or other cheap protein sources Peru Fish Dietary albumen is one of focus of Nutrition and Feed of Aquatic Animals research.At present, the effect of simple vegetable protein Peru Fish Dietary is not ideal, exists anti-nutritional factors in the vegetable protein source on the one hand; The plant protein source palatability is poor on the other hand, and amino acid forms uneven, lacks necessary amino acid and the contained UGF of fish meal of animal, and the relative fish meal of digestive utilization ratio is lower.Therefore, seek and develop the task of top priority that some new and high technologies that can improve vegetable protein utilization rate and conversion ratio become the practitioner.
Summary of the invention
The object of the invention is to, tackle above-mentioned form, a kind of method of producing biological fish meal by microbial fermentation is provided.
For achieving the above object, the present invention is by the following technical solutions:
1. technology that microbial fermentation processes is produced biological fish meal is characterized in that:
(1) corn protein powder in the fermentation substrate is carried out first enzymolysis and pickling and extract protein, and then mix with other raw materials by a certain percentage; Mixed fermentation substrate carries out boiling sterilization, adds water for cooling by quality-adjusting device, and then the ratio in fermentation substrate 1～3% adds the purification fulvic acid, inoculates at last the fermented by mixed bacterium bacterium liquid that is comprised of 3 kinds of bacterium liquid;
(2) postvaccinal fermentation substrate is filled in the portable fermenting vehicle by setting gauge, constant temperature bottom fermentation a couple of days in fermenting cellar;
(3) material that ferments is pulverized behind low temperature drying and is namely obtained described biological fish meal finished product;
In the step (1), the consisting of of described fermentation substrate: corn protein powder 40～60 weight portions, the dish dregs of rice 5～10 weight portions, the alcohol dregs of rice (DDGS) 5～10 weight portions, vermicelli protein powder 30～40 weight portions; When the nonruminant product was supplied with in preparation, the percentage by weight of also pressing fermentation substrate 10% added the molten slurry of fish;
In the step (1), described 3 kinds of bacterium liquid are respectively the bacterium liquid of candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei) and glossy ganoderma (Ganoderma lucidium).
Aforesaid method, preferably, pickling and the enzyme solution of the corn protein powder described in the step (1) are as follows:
A. enzymolysis: corn protein powder is added the water spice by material-water ratio 1:4～5, then use the salt acid for adjusting pH to 4.8 of 0.1～0.2mol/L～5.0, substrate liquid is warmed to 50～55 ℃, consumption by 20～40U/g and 5000～6000U/g adds cellulase and acid protease, constant temperature enzymolysis 30min～60min;
B. pickling: the salt acid for adjusting pH to 3.8 of substrate solution body and function 0.1～0.2mol/L that enzymolysis is good～4.0, under 30～32 ℃ of conditions, stir with 50～100r/min, keep 60min, centrifugation after the protein condenses precipitation, remove supernatant, get proteins precipitate, 75 ℃ of low temperature drying to moisture are 30%.
Aforesaid method preferably, the boiling sterilization described in the step (1), adds water for cooling and inoculation, and its concrete steps are as follows:
A. mixed fermentation substrate enters the ground floor of quenching and tempering device by the saturated vapor of 0.3～0.4MPa, carries out instantaneous boiling sterilization 1～2min under 140～150 ℃ of conditions;
B. the second layer that the fermentation substrate after sterilizing enters quenching and tempering device adds water, ventilation, cooling, and temperature of charge is down to 30～35 ℃, and moisture is adjusted between 40～55%;
C. inoculate for the 3rd layer that enters quenching and tempering device after the fermentation substrate cooling, mixed bacteria liquid is inoculated in the fermentation substrate by 5～10% percentage by weight, the ratio in fermentation substrate 3% adds the purification fulvic acid simultaneously.
Aforesaid method, preferably, the purity of described purification fulvic acid is greater than 95%.
Aforesaid method, preferably, described Candida tropicalis liquid is made by the following method:
Preparation first order seed culture medium: malt extract medium 130.1g and distilled water 1000mL mix, at 115～121 ℃ of lower sterilization 15～30min of temperature;
Preparation secondary seed medium and fermentation medium: cane molasses 120kg, glucose 10kg, Fructus Hordei Germinatus soak powder 10kg, yeast soaks powder 10kg, MgSO
47H
2O 1.5kg, KH
2PO
42.0kg, sterile pure water 1000L, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/80～100mL, under 28～30 ℃ with 100～130r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, is under the condition of 1:0.5～1, with 100～120r/min mechanical agitation at 28～30 ℃, ventilation volume ratio, cultivates 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, is under the condition of 1:0.5～1, with 100～120r/min mechanical agitation at 28～30 ℃, ventilation volume ratio, cultivates 24～48h.
Aforesaid method, preferably, described Lactobacillus casei bacterium liquid is made by the following method:
Preparation first order seed culture medium: casein peptone 10g, beef extract powder 10g, yeast soak powder 10g, glucose 5g, sodium acetate 5g, ammonium citrate 2g, Tween80 1g, K
2HPO
42g, MgSO
47H
2O 0.2g, MnSO
4H
2O 0.05g, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium and fermentation medium: whole milk powder 10kg, beef extract powder 10kg, yeast soak powder 10kg, glucose 5kg, sodium acetate 5kg, ammonium citrate 2kg, Tween80 1kg, K
2HPO
42kg, MgSO
47H
2O 0.2kg, MnSO
4H
2O 0.05kg, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed culture medium 1000mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/50～100mL, and 37～39 ℃ leave standstill cultivation 24～48h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, and 37～39 ℃ leave standstill and cultivate 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, and 37～39 ℃ leave standstill and cultivate 24～48h.
Aforesaid method, preferably, described lucidum bacteria liquid is made by the following method:
Preparation first order seed culture medium: peptone 5.0g, glucose 10g, NaCl 5.0g, CaCO
30.2g, distilled water 1.0L, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium and fermentation medium: glucose 10kg, Fructus Hordei Germinatus soak powder 5kg, peptone 5kg, corn flour 5kg, sterile pure water 1000L, and the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature.
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/50～100mL, under 23～25 ℃ with 100～120r/min shaken cultivation 24h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5%～10% weight ratio, is under the condition of 1:0.5～1, with 80～100r/min mechanical agitation at 23～25 ℃, ventilation volume ratio, cultivates 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5%～10% weight ratio, is under the condition of 1:0.5～1, with 50～80r/min mechanical agitation at 23～25 ℃, ventilation volume ratio, cultivates 24～48h.
Aforesaid method, preferably, described mixed bacteria liquid is that the mixed proportion of 3 kinds of bacterium liquid by candida tropicalis 3～5%, Lactobacillus casei 2～3%, glossy ganoderma 4～5% is mixed to get, described mixed bacteria liquid is inoculated in the fermentation substrate by 5～10% percentage by weight, be filled in the portable fermenting vehicle by setting gauge after mixing, the control temperature is 28～35 ℃ in fermenting cellar, relative humidity 85～90% condition bottom fermentations 72～120h.
Aforesaid method, preferably, the temperature of the low temperature drying described in the step (3) is 70～80 ℃, the grinding particle size of material passes through 65 mesh sieves at least 95% by 80 mesh sieves and 100% after the described oven dry.
The biological fish meal for preparing of method as mentioned above.
The biological fish meal for preparing of method as mentioned above, wherein thick protein (DM) 〉=76.55%, crude fat (DM)＜5.00%, crude fibre (DM)＜0.12%, coarse ash (DM)＜3.03%, lysine (DM) 〉=3.60%, methionine (DM) 〉=1.54%, threonine (DM) 〉=2.84%, ganoderma polyoses content 〉=1.09 ㎎/100g, Lactobacillus casei (CFU/g) 〉=1.45 * 10
8, candida tropicalis (CFU/g) 〉=3.00 * 10
8, glucosinolate and meson alkali all do not detect.
Beneficial effect of the present invention is:
The present invention selects quality plant protein sources feed: corn protein powder, the dish dregs of rice, alcohol dregs of rice DDGS, the molten slurry of vermicelli protein powder and fish is raw material, adopt bacterial classification and the enzyme preparation of function admirable, extract protein by raw material at first being carried out enzymolysis and pickling, and then be mixed and made into fermentation substrate, through modified, ratio in fermentation substrate 1～3% after the sterilization adds the purification fulvic acid, inoculate at last by candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei), the mixed bacteria liquid that glossy ganoderma (Ganodermalucidium) forms carries out solid state fermentation, the complete oven dry of fermenting, the product of microorganism fermented forage that pulverizing makes.The advantage of the technology of the present invention is that crude protein content is high, amino acid forms better, digestibility is high, simultaneously animal is had good growth promotion and antioxidation.Do not contain ANFs in the product of the present invention, reasonable price, aboundresources, technique are simple, are a kind of best products of effective Peru Fish Dietary.
More particularly, the method for the biological fish meal of microbial fermentation production of utilizing provided by the invention has the following advantages:
1, the biological fish meal that utilizes the inventive method to obtain, its protein forms better, protein mainly is comprised of peptide albumen, vegetable protein and mycoprotein in the product, and crude protein content reaches more than 80%, and the amino acid balance principle meets the best amino acid pattern of growth of animal.
2, the ANFs such as the raw materials used middle glucosinolate of the present invention and meson alkali and crude fibre are degraded under the effect of multiple bacterium and enzyme and are transformed detoxification, and the high molecular weight protein degraded can be the small-molecule substances such as oligopeptides, Effective Raise the digestive utilization ratio of product.
3, the present invention makes unrighted acid and the useful micro-ecological bacterial group of containing the promoting animal growth factor in the product, contain simultaneously multiple digestive ferment such as cellulase and protease etc., can effectively improve the micro-ecological environment of animal intestinal, improve immunity of organisms, reduce the generation of disease.
4, the biological fish meal that adopts the inventive method to produce carries out detoxification, improved the taste of animal product, animal product meat, egg, milk etc. do not use the fishlike smell behind the fish meal, because the fermentation of microorganism is added in the product of the molten slurry of fish fishlike smell and is relatively desalinated, can metabolism, absorb in the animal product yet.
5, owing to the molten slurry of used fish in the inventive method has fish distinctive fish raw meat fragrance and special dietary composition, add the palatability and the absorptivity that have improved greatly animal in the product to.Also contain abundant amino acid, vitamin, mineral matter and UGF in the molten slurry of fish, replenish better and the perfect trophic structure of product.
6, adopt the inventive method to carry out the production of biological fish meal, can under the prerequisite that guarantees the cultivated animals normal growth, with the alternative expensive fish meal of the animal and plant protein source of cheapness, save feed cost, thereby reduce aquaculture cost.
7, adopt the inventive method to carry out the production of biological fish meal, can protect marine fishery resources, the output of restriction fish meal is guaranteed the sustainable development of sea fishery, and is protected halobiontic ecological diversity.
8, used fulvic acid has hemostasis, anti-inflammatory, convergence, absorption, antiallergy, urgees to secrete, remove necrosis and promote granulation, adjust gastrointestinal function in the inventive method, improves the effects such as immunity of organisms.Fulvic acid is nutriment, is again class growth hormone, to improving the price of deed, promoting growth of animal, increases resistance against diseases and the resistance to oxidation of body, and treatment viral infectious aspect has very significant effect.
9, adopt in the biological fish meal of the inventive method production, thick protein (DM) 〉=76.55%, crude fat (DM)＜5.00%, crude fibre (DM)＜0.12%, coarse ash (DM)＜3.03%, lysine (DM) 〉=3.60%, methionine (DM) 〉=1.54%, threonine (DM) 〉=2.84%, ganoderma polyoses content 〉=1.09 ㎎/100g, Lactobacillus casei (CFU/g) 〉=1.45 * 10
8, candida tropicalis (CFU/g) 〉=3.00 * 10
8, glucosinolate and meson alkali all do not detect.
Description of drawings
Fig. 1 is the schematic flow sheet of one of them embodiment of the inventive method.
Fig. 2 is the schematic flow sheet of another embodiment of the inventive method.
The specific embodiment
Below describe technology of the present invention and characteristics in detail by specific embodiment, but these embodiment limit protection scope of the present invention.
Candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei) and glossy ganoderma (Ganoderma lucidium) used in following examples of the present invention are for to buy in the wild-type strain of Chinese industrial microorganism fungus kind preservation administrative center (CICC).Preserving number is respectively: CICC 1254, CICC 6105, CICC 14042.
Embodiment 1
Referring to Fig. 1, prepare in accordance with the following methods the biological fish meal of present embodiment:
1. take by weighing corn protein powder 500kg, dish dregs of rice 50kg, alcohol dregs of rice DDGS 50kg, vermicelli protein powder 400kg.
2. corn protein powder is carried out pickling and enzymolysis, concrete grammar is as follows:
A. enzymolysis: the 600kg corn protein powder that weighs up is poured in the 5000L enzymatic vessel, add the water spice by material-water ratio 1:5, then use the salt acid for adjusting pH to 4.8 of 0.2mol/L, substrate liquid is warmed to 52 ℃, the consumption of pressing 30U/g and 5000U/g adds cellulase and acid protease, constant temperature enzymolysis 60min;
B. pickling: the salt acid for adjusting pH to 3.8 of the substrate solution body and function 0.2mol/L that enzymolysis is good, under 31 ℃ of conditions, stir 80r/min, keep 60min, supernatant is removed in centrifugation after the protein condenses precipitation, get proteins precipitate, 75 ℃ of low temperature dryings are to moisture 30%.
3. the corn protein powder of enzymolysis, pickling processes being crossed mixes with other raw materials, then carries out boiling sterilization, adds water for cooling and inoculation through quenching and tempering device, and idiographic flow and method are as follows:
A. fermentation substrate enters the ground floor of quenching and tempering device by the saturated vapor of 0.4MPa, carries out instantaneous boiling sterilization 1min under 150 ℃ of conditions.
B. the second layer that the fermentation substrate after sterilizing enters quenching and tempering device adds water, the cooling of ventilating, and temperature of charge is down to 31 ℃, and moisture is adjusted to 45%.
C. inoculate for the 3rd layer that enters quenching and tempering device after the fermentation substrate cooling, fermented by mixed bacterium liquid is inoculated in the fermentation substrate by 10% percentage by weight, the ratio in fermentation substrate 3% adds the purification fulvic acid simultaneously.
4. described mixed bacteria liquid is to mix after candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei) and glossy ganoderma (Ganoderma lucidium) are increased respectively bacterium cultivation and liquid state fermentation by the following method, specifically:
(1) candida tropicalis (Candida tropicalis)
A. prepare the first order seed culture medium: malt extract medium 130.1g, distilled water 1000mL mixes, at 115 ℃ of lower sterilization 15min of temperature.
B. prepare secondary seed medium and fermentation medium: cane molasses 120kg, glucose 10kg, Fructus Hordei Germinatus soak powder 10kg, yeast soaks powder 10kg, MgSO
47H
2O 1.5kg, KH
2PO
42.0kg, sterile pure water 1000L, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature.
C. condition of culture:
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, inoculate strain inclined plane in the first order seed culture medium in the ratio of ring/100mL 28 ℃, 130r/min, shaken cultivation 24h.
Secondary seed is cultivated: the 10L automated seed canned fermentation medium 5L that ferments, then the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 10% weight ratio, 28 ℃, ventilation volume ratio 1:1, mechanical agitation 100r/min, cultivates 24h.
Liquid fermentation and culture: 50L Fermentation dress fermentation medium 25L, then the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, 28～30 ℃, ventilation volume ratio 1:1, mechanical agitation 100r/min, cultivates 24h.
(2) Lactobacillus casei (Lactobacillus casei)
A. prepare the first order seed culture medium: casein peptone 10g, beef extract powder 10g, yeast soak powder 10g, glucose 5g, sodium acetate 5g, ammonium citrate 2g, Tween80 1g, K
2HPO
42g, MgSO
47H
2O0.2g, MnSO
4H
2O 0.05g, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature.
B. prepare secondary seed medium and fermentation medium: whole milk powder 10kg, beef extract powder 10kg, yeast soak powder 10kg, glucose 5kg, sodium acetate 5kg, ammonium citrate 2kg, Tween80 1kg, K
2HPO
42kg, MgSO
47H
2O 0.2kg, MnSO
4H
2O 0.05kg, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature.
C. condition of culture:
First order seed is cultivated: dress first order seed culture medium 1000mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/50mL, and 37 ℃ leave standstill cultivation 24h.
Secondary seed is cultivated: the 10L automated seed canned fermentation medium 5L that ferments, and the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 10% weight ratio, and 37 ℃ leave standstill and cultivate 24h.
Liquid fermentation and culture: the 50L automated seed canned fermentation medium 25L that ferments, the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 10% weight ratio, and 37 ℃ leave standstill and cultivate 24h.
(3) glossy ganoderma (Ganoderma lucidium)
A. prepare the first order seed culture medium: peptone 5.0g, glucose 10g, NaCl 5.0g, CaCO
30.2g, distilled water 1.0L, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20～30min of temperature.
B. prepare secondary seed medium and fermentation medium: glucose 10kg, Fructus Hordei Germinatus soak powder 5kg, peptone 5kg, corn flour 5kg, sterile pure water 1000L, and the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature.
C. condition of culture:
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, inoculate strain inclined plane in the first order seed culture medium in the ratio of ring/50mL 25 ℃, 100r/min, shaken cultivation 24h.
Secondary seed is cultivated: the 10L automated seed canned fermentation medium 5L that ferments, then the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 10% weight ratio, 23 ℃, ventilation volume ratio 1:0.5, mechanical agitation 80r/min, cultivates 24h.
Liquid fermentation and culture: the 50L automated seed canned fermentation medium 25L that ferments, then the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, 25 ℃, volume ratio ventilation 1:0.5, mechanical agitation 50r/min, cultivates 24h.
5. in the described mixed bacteria liquid, the mixed proportion of candida tropicalis, Lactobacillus casei and lucidum strain liquid is: candida tropicalis 3%, Lactobacillus casei 2%, glossy ganoderma 5%.
Described mixed bacteria liquid is inoculated in the fermentation substrate by 10% percentage by weight, is filled in the portable fermenting vehicle by setting gauge after mixing, and the control temperature is 32 ℃ in fermenting cellar, relative humidity 85～90% condition bottom fermentation 120h.
6. the material that ferments carries out behind 70 ℃ of low temperature dryings that fine powder is broken to get product, grinding particle size at least 95% by 80 mesh sieves, 100% by 65 mesh sieves.
7. the biological fish meal for preparing according to above method, record thick protein (DM) 〉=80.07%, crude fat (DM)＜5.00%, crude fibre (DM)＜0.05%, coarse ash (DM)＜1.35%, lysine (DM) 〉=3.75%, methionine (DM) 〉=1.68%, threonine (DM) 〉=3.00%, ganoderma polyoses content 1.20 ㎎/100g, Lactobacillus casei (CFU/g) 〉=1.50 * 10
8, candida tropicalis (CFU/g) 〉=3.00 * 10
8, glucosinolate and meson alkali all do not detect.
Embodiment 2
Referring to Fig. 1, prepare in accordance with the following methods the biological fish meal of present embodiment:
1. take by weighing corn protein powder 500kg, dish dregs of rice 100kg, alcohol dregs of rice DDGS100kg, vermicelli protein powder 400kg.
2. corn protein powder is carried out pickling and enzymolysis, concrete grammar is as follows:
A. enzymolysis: the 500kg corn protein powder that weighs up is poured in the 5000L enzymatic vessel, add the water spice by material-water ratio 1:5, then use the salt acid for adjusting pH to 4.8 of 0.2mol/L, substrate liquid is warmed to 52 ℃, the consumption of pressing 30U/g and 5000U/g adds cellulase and acid protease, constant temperature enzymolysis 60min;
B. pickling: the salt acid for adjusting pH to 3.8 of the substrate solution body and function 0.2mol/L that enzymolysis is good, under 31 ℃ of conditions, stir 80r/min, keep 60min, supernatant is removed in centrifugation after the protein condenses precipitation, get proteins precipitate, 75 ℃ of low temperature dryings are to moisture 30%.
3. two kinds of raw materials of the corn protein powder of enzymolysis, pickling processes being crossed and other mix, and then carry out boiling sterilization, add water for cooling and inoculation through quenching and tempering device, and idiographic flow and method are as follows:
A. fermentation substrate enters the ground floor of quenching and tempering device by the saturated vapor of 0.4MPa, carries out instantaneous boiling sterilization 1min under 150 ℃ of conditions.
B. the second layer that the fermentation substrate after sterilizing enters quenching and tempering device adds water, the cooling of ventilating, and temperature of charge is down to 31 ℃, and moisture is adjusted to 45%.
C. inoculate for the 3rd layer that enters quenching and tempering device after the fermentation substrate cooling, mixed bacteria liquid is inoculated in the fermentation substrate by 10% percentage by weight, the ratio in fermentation substrate 3% adds the purification fulvic acid simultaneously.
Described mixed bacteria liquid be candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei) and glossy ganoderma (Ganoderma lucidium) are increased by method as described in Example 1 that bacterium is cultivated respectively and liquid state fermentation after mix.
ML5. in the described mixed bacteria liquid, the mixed proportion of candida tropicalis, Lactobacillus casei and lucidum strain liquid is: candida tropicalis 4%, Lactobacillus casei 2%, glossy ganoderma 4%.
Mixed bacteria liquid is inoculated in the fermentation substrate by 10% percentage by weight, is filled in the portable fermenting vehicle by setting gauge after mixing, and the control temperature is 32 ℃ in fermenting cellar, relative humidity 85～90% condition bottom fermentation 120h.
6. the material that ferments carries out behind 70 ℃ of low temperature dryings that fine powder is broken to get product, grinding particle size at least 95% by 80 mesh sieves, 100% by 65 mesh sieves.
7. the biological fish meal for preparing according to above method, record thick protein (DM) 〉=76.55%, crude fat (DM)＜4.7%, crude fibre (DM)＜0.12%, coarse ash (DM)＜1.78%, lysine (DM) 〉=3.60%, methionine (DM) 〉=1.54%, threonine (DM) 〉=2.84%, ganoderma polyoses content 〉=1.09 ㎎/100g, Lactobacillus casei (CFU/g) 〉=1.45 * 10
8, candida tropicalis (CFU/g) 〉=3.85 * 10
8, glucosinolate and meson alkali all do not detect.
Embodiment 3
Referring to Fig. 2, prepare in accordance with the following methods the biological fish meal of present embodiment:
1. take by weighing corn protein powder 600kg, dish dregs of rice 50kg, alcohol dregs of rice DDGS 50kg, vermicelli protein powder 400kg, the molten slurry of fish 100kg.
2. corn protein powder is carried out pickling and enzymolysis, concrete grammar is as follows:
A. enzymolysis: the 600kg corn protein powder that weighs up is poured in the 5000L enzymatic vessel, add the water spice by material-water ratio 1:5, then use the salt acid for adjusting pH to 4.8 of 0.2mol/L, substrate liquid is warmed to 52 ℃, the consumption of pressing 30U/g and 5000U/g adds cellulase and acid protease, constant temperature enzymolysis 60min;
B. pickling: the salt acid for adjusting pH to 3.8 of the substrate solution body and function 0.2mol/L that enzymolysis is good, under 31 ℃ of conditions, stir 80r/min, keep 60min, supernatant is removed in centrifugation after the protein condenses precipitation, get proteins precipitate, 75 ℃ of low temperature dryings are to moisture 30%.
3. the corn protein powder of enzymolysis, pickling processes being crossed mixes with other raw materials, then carries out boiling sterilization, adds water for cooling and inoculation through quenching and tempering device, and idiographic flow and method are as follows:
A. fermentation substrate enters the ground floor of quenching and tempering device by the saturated vapor of 0.4MPa, carries out instantaneous boiling sterilization 1min under 150 ℃ of conditions.
B. the second layer that the fermentation substrate after sterilizing enters quenching and tempering device adds water, the cooling of ventilating, and temperature of charge is down to 31 ℃, and moisture is adjusted to 45%.
C. inoculate for the 3rd layer that enters quenching and tempering device after the fermentation substrate cooling, mixed bacteria liquid is inoculated in the fermentation substrate by 10% percentage by weight, the ratio in fermentation substrate 3% adds the purification fulvic acid simultaneously.
Described mixed bacteria liquid be candida tropicalis (Candida tropicalis), Lactobacillus casei (Lactobacillus casei) and glossy ganoderma (Ganoderma lucidium) are increased by method as described in Example 1 that bacterium is cultivated respectively and liquid state fermentation after mix.
5. in the described mixed bacteria liquid, the mixed proportion of candida tropicalis, Lactobacillus casei and lucidum strain liquid is: candida tropicalis 3%, Lactobacillus casei 2%, glossy ganoderma 5%.
Mixed bacteria liquid is inoculated in the fermentation substrate by 10% percentage by weight, is filled in the portable fermenting vehicle by setting gauge after mixing, and the control temperature is 32 ℃ in fermenting cellar, relative humidity 85～90% condition bottom fermentation 120h.
6. the material that ferments carries out behind 70 ℃ of low temperature dryings that fine powder is broken to get product, grinding particle size at least 95% by 80 mesh sieves, 100% by 65 mesh sieves.
7. the biological fish meal for preparing according to above method, record thick protein (DM) 〉=82.51%, crude fat (DM)＜5.00%, crude fibre (DM)＜0.06%, coarse ash (DM)＜3.03%, lysine (DM) 〉=4.00%, methionine (DM) 〉=1.93%, threonine (DM) 〉=3.28%, ganoderma polyoses content 〉=1.17 ㎎/100g, Lactobacillus casei (CFU/g) 〉=1.85 * 10
8, candida tropicalis (CFU/g) 〉=4.50 * 10
8, glucosinolate and meson alkali all do not detect.
Claims (10)
1. method of producing biological fish meal by microbial fermentation is characterized in that:
(1) corn protein powder in the fermentation substrate is carried out first enzymolysis and pickling and extract protein, and then mix with other raw materials by a certain percentage; Mixed fermentation substrate carries out boiling sterilization, adds water for cooling by quality-adjusting device, and then the ratio in fermentation substrate 1～3% adds the purification fulvic acid, inoculates at last the fermented by mixed bacterium bacterium liquid that is comprised of 3 kinds of bacterium liquid;
(2) postvaccinal fermentation substrate is filled in the portable fermenting vehicle by setting gauge, constant temperature bottom fermentation a couple of days in fermenting cellar;
(3) material that ferments is pulverized behind low temperature drying and is namely obtained described biological fish meal finished product;
In the step (1), the consisting of of described fermentation substrate: corn protein powder 40～60 weight portions, the dish dregs of rice 5～10 weight portions, the alcohol dregs of rice 5～10 weight portions, vermicelli protein powder 30～40 weight portions; When the nonruminant product was supplied with in preparation, the percentage by weight of also pressing fermentation substrate 10% added the molten slurry of fish;
In the step (1), described 3 kinds of bacterium liquid are respectively the bacterium liquid of candida tropicalis, Lactobacillus casei and glossy ganoderma.
2. method according to claim 1 is characterized in that, pickling and the enzyme solution of the corn protein powder described in the step (1) are as follows:
A. enzymolysis: corn protein powder is added the water spice by material-water ratio 1:4～5, then use the salt acid for adjusting pH to 4.8 of 0.1～0.2mol/L～5.0, substrate liquid is warmed to 50～55 ℃, consumption by 20～40U/g and 5000～6000U/g adds cellulase and acid protease, constant temperature enzymolysis 30min～60min;
B. pickling: the salt acid for adjusting pH to 3.8 of substrate solution body and function 0.1～0.2mol/L that enzymolysis is good～4.0, under 30～32 ℃ of conditions, stir with 50～100r/min, keep 60min, centrifugation after the protein condenses precipitation, remove supernatant, get proteins precipitate, 75 ℃ of low temperature drying to moisture are 30%.
3. method according to claim 1 is characterized in that, the boiling sterilization described in the step (1), adds water for cooling and inoculation, and its concrete steps are as follows:
A. mixed fermentation substrate enters the ground floor of quenching and tempering device by the saturated vapor of 0.3～0.4MPa, carries out instantaneous boiling sterilization 1～2min under 140～150 ℃ of conditions;
B. the second layer that the fermentation substrate after sterilizing enters quenching and tempering device adds water, ventilation, cooling, and temperature of charge is down to 30～35 ℃, and moisture is adjusted between 40～55%;
C. inoculate for the 3rd layer that enters quenching and tempering device after the fermentation substrate cooling, mixed bacteria liquid is inoculated in the fermentation substrate by 5～10% percentage by weight, the ratio in fermentation substrate 3% adds the purification fulvic acid simultaneously.
4. method according to claim 1 is characterized in that, the purity of described purification fulvic acid is greater than 95%.
5. method according to claim 1 is characterized in that, described Candida tropicalis liquid is made by the following method:
Preparation first order seed culture medium: malt extract medium 130.1g and distilled water 1000mL mix, at 115～121 ℃ of lower sterilization 15～30min of temperature;
Preparation secondary seed medium and fermentation medium: cane molasses 120kg, glucose 10kg, Fructus Hordei Germinatus soak powder 10kg, yeast soaks powder 10kg, MgSO
47H
2O 1.5kg, KH
2PO
42.0kg, sterile pure water 1000L, the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/80～100mL, under 28～30 ℃ with 100～130r/min shaken cultivation, 24～48h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, is under the condition of 1:0.5～1, with 100～120r/min mechanical agitation at 28～30 ℃, ventilation volume ratio, cultivates 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, is under the condition of 1:0.5～1, with 100～120r/min mechanical agitation at 28～30 ℃, ventilation volume ratio, cultivates 24～48h.
6. method according to claim 1 is characterized in that, described Lactobacillus casei bacterium liquid is made by the following method:
Preparation first order seed culture medium: casein peptone 10g, beef extract powder 10g, yeast soak powder 10g, glucose 5g, sodium acetate 5g, ammonium citrate 2g, Tween80 1g, K
2HPO
42g, MgSO
47H
2O 0.2g, MnSO
4H
2O 0.05g, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium and fermentation medium: whole milk powder 10kg, beef extract powder 10kg, yeast soak powder 10kg, glucose 5kg, sodium acetate 5kg, ammonium citrate 2kg, Tween80 1kg, K
2HPO
42kg, MgSO
47H
2O 0.2kg, MnSO
4H
2O 0.05kg, distilled water 1000mL mixes, and pH6.8 is at 115～121 ℃ of lower sterilization 20～30min of temperature;
First order seed is cultivated: dress first order seed culture medium 1000mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/50～100mL, and 37～39 ℃ leave standstill cultivation 24～48h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5～10% weight ratio, and 37～39 ℃ leave standstill and cultivate 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5～10% weight ratio, and 37～39 ℃ leave standstill and cultivate 24～48h.
7. method according to claim 1 is characterized in that, described lucidum bacteria liquid is made by the following method:
Preparation first order seed culture medium: peptone 5.0g, glucose 10g, NaCl 5.0g, CaCO
30.2g, distilled water 1.0L, pH7.2～7.4 are at 115～121 ℃ of lower sterilization 20～30min of temperature;
Preparation secondary seed medium and fermentation medium: glucose 10kg, Fructus Hordei Germinatus soak powder 5kg, peptone 5kg, corn flour 5kg, sterile pure water 1000L, and the pH nature is at 115～121 ℃ of lower sterilization 20～30min of temperature.
First order seed is cultivated: dress first order seed culture medium 500mL in the 1000mL triangular flask, strain inclined plane is inoculated in the first order seed culture medium in the ratio of ring/50～100mL, under 23～25 ℃ with 100～120r/min shaken cultivation 24h;
Secondary seed is cultivated: the bacterium liquid that will cultivate through described first order seed is inoculated in the secondary seed tank according to 5%～10% weight ratio, is under the condition of 1:0.5～1, with 80～100r/min mechanical agitation at 23～25 ℃, ventilation volume ratio, cultivates 24～48h;
Liquid fermentation and culture: the bacterium liquid that will cultivate through described secondary seed is inoculated in the fermentation tank according to 5%～10% weight ratio, is under the condition of 1:0.5～1, with 50～80r/min mechanical agitation at 23～25 ℃, ventilation volume ratio, cultivates 24～48h.
8. method according to claim 1, it is characterized in that, described mixed bacteria liquid is that the mixed proportion of 3 kinds of bacterium liquid by candida tropicalis 3～5%, Lactobacillus casei 2～3%, glossy ganoderma 4～5% is mixed to get, described mixed bacteria liquid is inoculated in the fermentation substrate by 5～10% percentage by weight, be filled in the portable fermenting vehicle by setting gauge after mixing, the control temperature is 28～35 ℃ in fermenting cellar, relative humidity 85～90% condition bottom fermentations 72～120h.
9. method according to claim 1 is characterized in that, the temperature of the low temperature drying described in the step (3) is 70～80 ℃, and the grinding particle size of material passes through 65 mesh sieves at least 95% by 80 mesh sieves and 100% after the described oven dry.
10. the biological fish meal for preparing according to each described method in the claim 1～9.
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| CN104920791A (en) * | 2015-05-29 | 2015-09-23 | 东莞市林氏生物技术股份有限公司 | A fermented fish meal with high digestibility rate and a preparation method thereof |
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