CN102125173A - Preparation and application of additive for reducing liver fat content of turbot - Google Patents
Preparation and application of additive for reducing liver fat content of turbot Download PDFInfo
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Abstract
The invention discloses a preparation of an additive for reducing the liver fat content of a turbot and the application of the additive in reducing the liver fat content of a turbot, wherein the preparation of the additive is implemented through the following steps: through taking minced pacific Pollock as raw materials, hashing the pacific Pollock, then raising the temperature of the obtained pacific Pollock for sterilization; after the temperature of the sterilized pacific Pollock raises to 90 DEG C, carrying out heat preservation on the pacific Pollock, then carrying out sterilization on the pacific Pollock for 20 minutes; adding water and a protease preparation into the obtained pacific Pollock and then mixing; carrying out enzymolysis on the obtained mixture under the condition of optimized enzymolysis process parameters; raising the temperature of the obtained enzymatic hydrolysate so as to carry out inactivation on proteases in the enzymatic hydrolysate; carrying out rough filtration on the enzymatic hydrolysate so as to separate spurs from the enzymatic hydrolysate; carrying out centrifugation on the obtained enzymatic hydrolysate so as to respectively obtain soluble hydrolytic fish protein powder; and carrying out ultrafiltration-purifying on the obtained soluble hydrolytic fish protein powder, then freezing and drying the obtained soluble hydrolytic fish protein powder so as to obtain hydrolytic fish protein powder, wherein the protease preparation refers to Alcalase + Flavourzyme; the enzymolysis process parameters are as follows: enzymolysis temperature is 50-60 DEG C; starting pH value is 8.5, ratio of material to water is 1:1, and enzymolysis time is 3 hours. Through the preparation disclosed by the invention, the hydrolytic fish protein powder (the molecular weight thereof is less than 100 Da and reaches 30%) can be obtained. When the hydrolytic fish protein powder is used for replacing 5% of fish protein concentrate in a turbot feed.
Description
Technical field:
The invention belongs to the preparation technique of turbot mixed feed, is a kind of preparation and application that reduces turbot fatty liver additive.
Background technology:
Fatty liver disease (Fatty Liver Syndrome) all has report various animals such as pig, chicken, ox, sheep, dog, fish, minks.Animal tallow hepatomegaly quantity research data shows that reasons such as heredity, nutrition, management, environment, hormone, noxious material all can cause the generation of fatty liver disease.Pathogenesis is relevant with fat metabolism, is commonly referred to be due to synthetic increase of liver cell fat and the oxidation minimizing, and nutrient balance has played considerable effect in this process.
The formation of fatty,fiss liver mainly is that its required nutrient imbalance and some lipotropic factor shortage cause.Also be subjected to the influence such as physiological metabolism characteristics, breeding environment, aquaculture model of fish simultaneously.Intensive culture, cultivation density strengthen, and the production cycle shortens, and the man-made feeds of nutritional imbalance substitute natural bait fully simultaneously, usually be difficult to satisfy the fish body fast, the needs of healthy growth, cause the Nutrition and Metabolism disorder of cultured fishes.Wherein liver fat metabolism disorder, deposition, infiltration, fat content raise, thereby cause fatty liver to take place.Fatty liver is one of nutritive disease common in the cultured fishes.Most of cultured fishes, especially the fish of sea-farming, as porgy, lefteye flounder, large yellow croaker, perch, black porgy etc., after ingest higher fatty acid, high protein or Hi CHO feed, poor appetite often appears, phenomenons such as unable, the poor growth of moving about and premunition reduction.Pathologic finding is found, ill fish liver hypertrophy, color is pale, fat is accumulated in a large number, liver fat is dripped increase, the hepatic tissue steatosis obviously and vacuolation, nucleus skew, even symptom such as hepatic tissue atrophy necrosis occurs.(Norway such as Lie, country nutrition and the Aquaculture of Food Research Inst. 1988) point out the fat of a large amount of accumulations in cod (Gadus morhua) liver, be mainly derived from fat in the feed in the fish body direct accumulation and feed in carbohydrate and the conversion of protein in the organism metabolism process synthesize.(France such as Deplano, research of agricultural science institute, Dis.Aquac.Org.1989) think and lack suitable mixed feed that promptly fat is excessive or trophic component is out of proportion and lack the major reason that anti-fattyliver substance is hyoid tooth perch (Dicentrarchus labrax) liver fat pathology in the feed.
The liver fat of fish is mainly from synthetic to the conversion of excessive protein and carbohydrate in direct absorption fatty in the feed and the feed.After these fat are transported to liver, if can not in time transport away, then can be piled up in and cause the liver metabolism disorder in the liver.Therefore, can realize prevention and treatment by regulating and control the source and the outlet of fat in the liver to fatty liver.Replenish these materials in the feed and help absorption and the utilization of fish to lipid, improve efficiency of feed utilization, reduce liver fat Study on content, Shang Weijian has the report of success.
Summary of the invention:
The objective of the invention is to propose a kind of Pacific Ocean wall pollack fish offal material that utilizes, the preparation liquid fish makes an addition in the turbot feed, substitutes 5% fish meal protein, can reduce turbot internal organ and liver index, reduce hepatic fat content, promotes growth.
The present invention finishes by following operating technology: comprise two parts: 1. the preparation of liquid fish; 2. protein hydrolysate reduces the application of turbot liver fat content.
1. the preparation of liquid fish: utilize Pacific Ocean wall pollack fish flakes, clean, rub the sterilization of heating of back input enzyme reactor through meat grinder, be warming up to 90 ℃ of insulations after the rubbing, water bath with thermostatic control 20min sterilization, adding water and protease enzyme preparation again mixes, enzymolysis under the enzymolysis process parameter condition of optimizing, enzymolysis liquid heats up and makes the protease deactivation then, isolated by filtration spur, the centrifugal soluble protein oligopeptides water that obtains respectively of the further three-phase of enzymolysis liquid, insoluble protein precipitation and fish oil, soluble protein oligopeptides water is purified through ultrafiltration, obtains liquid fish after the freeze drying.
Described protease enzyme preparation: adopt Alcalase+Flavourzyme complex enzyme zymohydrolysis method; Alcalase enzyme: E/S is 90AU, and Flavourzyme enzyme: E/S is 15000LAPU.
Described enzymolysis process parameter is: hydrolysis temperature: 50 ℃～60 ℃, initial pH=8.5, material-water ratio: 1: 1, and enzymolysis 3h.
The ultrafiltration of described protein oligopeptide is purified: adopted 8000rpm high speed centrifugation and the method that 100kDa milipore filter ultrafiltration purification combines, thoroughly held back macromolecular substances, obtained molecular weight and reach 30% protein oligopeptide less than 100Da.
2. protein hydrolysate reduces the suitable consumption of turbot liver fat content: substitute fish meal protein with 5% protein hydrolysate in the feed, can significantly improve the growth of turbot, reduce hepatic fat content.
The present invention and prior art contrast have following characteristics:
1. in feed, add protein hydrolysate and substitute fish meal protein,, reduce hepatic fat content for can significantly reducing turbot internal organ and liver index.
2. in feed, add additive of the present invention, the growth of turbot is had facilitation.
3, the present invention has easy to usely, has no side effect, and is pollution-free, characteristics such as noresidue.
Description of drawings:
Fig. 1: hydrolysis temperature is to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%); Enzymolysis parameter among the figure: A enzyme E/S=90AU, B enzyme E/S=15000LAPU, material-water ratio=1: 1, initial pH=7.0, enzymolysis time=6h.
Fig. 2: the initial pH level of enzymolysis is to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR), enzymolysis parameter among the figure: A enzyme E/S=90AU, B enzyme E/S=15000LAPU, material-water ratio=1: 1, temperature=50 ℃, enzymolysis time=6h.
Fig. 3: Flavourzyme (B enzyme) E/S and enzymolysis time are to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%), enzymolysis parameter among the figure: the time dependent curvilinear equation of water-soluble protein hydrolysate nitrogen recovery (NR) % under A enzyme E/S=90AU, material-water ratio=1: 1, hydrolysis temperature=50 ℃, the initial pH=7.0 of enzymolysis, different B enzyme E/S condition, (◆=5000): NR (%)=11.39Ln (x)+49.55; (=10000): NR (%)=11.80Ln (x)+51.60; (▲=15000): NR (%)=13.71Ln (x)+51.98; (X=20000): NR (%)=13.36Ln (x)+53.54; (zero=30000): NR (%)=12.83Ln (x)+54.83.
Fig. 4: Alcalase (A enzyme) E/S and enzymolysis time are to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%), enzymolysis parameter among the figure: B enzyme E/S=15000LAPU, material-water ratio=1: 1, hydrolysis temperature=50 ℃, the initial pH=9.1 of enzymolysis, different A enzyme E/S be than the time dependent curvilinear equation of water-soluble protein hydrolysate nitrogen recovery (NR) % under the condition, (◆=30): NR (%)=10.75Ln (x)+54.69; (=60): NR (%)=6.98Ln (x)+66.88; (△=90): NR (%)=6.67Ln (x)+70.28; (X=120): NR (%)=6.49Ln (x)+71.26.
Fig. 5: material-water ratio is to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%), enzymolysis parameter among the figure: A enzyme E/S=90AU, B enzyme E/S=15000LAPU, temperature=50 ℃, initial pH=9.1, enzymolysis time=6h.
The specific embodiment:
Be described in detail preparation of the present invention and application process in conjunction with the accompanying drawings below by embodiment:
The present invention finishes by following operating technology: comprise two parts: one. the preparation of liquid fish; Two. protein hydrolysate reduces the application of turbot liver fat content.
One, the preparation of liquid fish:
The present invention utilizes Pacific Ocean wall pollack flesh of fish preparation protein hydrolysate powder, its enzymolysis process flow process is: select Pacific Ocean wall pollack fish flakes for use, be warming up to 90 ℃ of insulations, water bath with thermostatic control 20min sterilization after the rubbing, mix with quantitative protease preparation and water again, enzymolysis in the enzyme reactor of controllable parameters such as time, temperature, pH, after enzyme digestion reaction finishes, enzymolysis liquid is warming up to 90 ℃ of insulation 20min and carries out deactivation, centrifugation, film separation, each separates through freeze drying, has both obtained target product: water-soluble protein hydrolysate powder.
1, the hydrolysis result of two kinds of protease and combination thereof:
With reference to 2 kinds of commercially available protein enzyme preparation product specification parameters (table 1), designed the experimental program (table 2) of 4 groups of different protease and combination enzymolysis wall pollack fish flakes thereof.
The product description parameter of table 1, two kinds of commercially available protein enzyme preparations
The experimental program of table 2, two kinds of protease and the combination enzymolysis wall pollack flesh of fish thereof
Annotate: E/S: enzyme/substrate is than (enzyme activity unit/kg protein).
Experimental result shows (table 3), and the NR (65.3 ± 0.7%) when selecting single Alcalase enzyme for use is higher than the NR (41.6 ± 0.8%) of single use Flavourzyme enzyme far away, but the protein hydrolysate that obtains is obvious bitter taste; Adopt first Alcalase protease hydrolyzed, add the two step enzyme solutions that Flavourzyme protease continues enzymolysis after the deactivation again, can reach the effect to protein hydrolysate product debitterize, NR=65.2 ± 1.1% is not seen has further raising (P＞0.05); Experimental group 4 adopts the synchronous complex enzyme zymohydrolysis method of Alcalase+Flavourzyme enzyme, and NR (70.8 ± 0.7%) is significantly higher than other 3 experimental group (P＜0.05), and the product delicate flavour is obvious, no bitter taste.
Table 3, different protease and combination enzymolysis wall pollack thereof are put forward the effect (n=3) of the flesh of fish
Annotate: same column has and is designated as significant difference (P＜0.05) on the different English alphabets.
2:Alcalase (A the enzyme)+technology characteristic of Flavourzyme (B enzyme) complex enzyme solution under different enzymolysis parameter levels
The Alcalase enzyme is a kind of efficient enzyme preparation that is widely used in all kinds of protein digestions, but there is tangible bitter taste in its enzymolysis product, has limited its range of application.Macro-molecular protein itself is tasteless, and works as protein by enzymolysis, and peptide chain is stretched, cuts off, and exposes the former inner group that is wrapped in, and the bitter peptides that wherein specific intermediate molecular weight has the hydrophobic grouping end is the principal element that produces bitter taste.The Flavourzyme enzyme is a kind of albumen/peptide enzyme preparation that has the circumscribed and inscribe activity of peptide chain simultaneously, it is lower to the enzymolysis efficiency of complete macro-molecular protein, its characteristic is the further protein peptides of enzymolysis intermediate molecular weight, improve the enzymolysis degree (the highest DH value can reach 60%) of enzymolysis product, the molecular structure that simultaneously can the modified protein peptide or the hydrophobic grouping of decomposition bitter peptides.By the synergy of Alcalase+Flavourzyme enzyme, the mellow nature of enzymolysis product local flavor of acquisition, no bitter taste can reach and removes the bitter taste effect.
General knowledge according to the protease hydrolyzed dynamics research, the optimum organization of material-water ratio, hydrolysis temperature, pH value, enzymolysis time, protease preparation and protease preparation consumption E/S (enzyme activity unit/kg protein) can make nitrogen recovery, the albumen oligomeric peptide physical characteristic of water-solubility protein oligomeric peptide, the overall targets such as energy resource consumption of technical process reach optimum efficiency.Therefore on above-mentioned experimental result basis, further the technological parameter to A enzyme+B enzyme complex enzyme solution has carried out optimizing experiment.
Shown in Figure 1: hydrolysis temperature is to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%): when the enzymolysis parameter: at A enzyme E/S=90AU, B enzyme E/S=15000LAPU, material-water ratio=1: 1, initial pH=7.0, enzymolysis time=6h, under different hydrolysis temperature conditions, the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR) is that 67.7 ± 1.7% (70 ℃) are to 82.4 ± 1.9% (60 ℃).NR under 50 ℃, 55 ℃, 60 ℃ hydrolysis temperature conditions is apparently higher than other hydrolysis temperature, when hydrolysis temperature when 45 ℃ are brought up to 50 ℃, the NR absolute value has improved 6.4%, when 50 ℃ are brought up to 60 ℃, the absolute value of NR has only improved 3.9%, when continue improving hydrolysis temperature to 65 ℃, the absolute value of NR has reduced by 7.4% on the contrary.So the best hydrolysis temperature condition of Alcalase+Flavourzyme complex enzyme conforms to substantially with the enzyme preparation product specification between 50 ℃～60 ℃, hydrolysis temperature is crossed to exceed to hang down all can influence protease activities.
Shown in Figure 2: the initial pH level of enzymolysis is to the influence (n=2) of the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR): when the enzymolysis parameter: when A enzyme E/S=90AU, B enzyme E/S=15000LAPU, material-water ratio=1: 1, temperature=50 ℃, enzymolysis time=6h.The natural pH of wall pollack meat mincing is 6.6 ± 0.2 approaching neutrality, when adjusting initial pH to 5.5, NR significantly descends, when initial pH value is adjusted to 8.5,9.0,9.5,10, corresponding N R brings up to 84.6 ± 0.5% by 81.1 ± 0.1%, though the NR the highest (84.6 ± 0.5%) during initial pH=10, this moment enzymolysis liquid mouthfeel and smell variation, continue to improve initial pH value, the anti-reduction trend that is of NR.
Known in the protein digestion process, fracture along with peptide chain, protein peptides terminal carboxyl group amount increases, the pH value is continuous reduction trend, and the pH that keep whole enzymolysis process is constant, need constantly add the alkali neutralization, this can cause the final products ash content higher, also make technology controlling and process too complicated, usual way is by adjusting initial pH value, the pH of enzymolysis process being in the suitable relatively scope.In this experiment, when initial pH value was adjusted into 5.5 to 11.5, the endpoint pH of respectively organizing enzymolysis liquid behind the 6h was 5.5～7.8, and the optimum pH of Alcalase and Flavourzyme product description is respectively 6.5～8.5 and 5.5～7.5.This prompting difference of water-solubility protein oligopeptides NR under different pH condition, except with the influence factor of pH value to enzymatic activity, also may be relevant in the solubility under the different pH condition with proteins/peptides.In this experiment, when initial pH＞10, terminal point pH＞7 of wall pollack enzymolysis liquid behind the enzymolysis 6h.
Shown in Figure 3: Flavourzyme (B enzyme) E/S and the influence (n=2) of enzymolysis time to the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%): under A enzyme E/S=90AU, material-water ratio=1: 1, temperature=50 ℃, initial pH=7.0, different Flavourzyme (B enzyme) E/S condition, the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR) is seen Fig. 3 with the change curve of enzymolysis and extraction time lengthening.When B enzyme E/S is respectively 5000,10000,15000,20000 and during 30000LAPU, NR behind the enzymolysis 2h is respectively 57.5 ± 0.7,60.1 ± 0.9,61.5 ± 0.3,63.0 ± 1.2 and 63.7 ± 0.5%, NR is respectively 65.3 ± 0.5,67.2 ± 1.0,70.9 ± 0.4,71.7 ± 0.4 and 72.6 ± 0.3% behind the enzymolysis 4h, and NR is respectively 74.7 ± 1.0,77.6 ± 0.6,82.1 ± 0.1,83.0 ± 0.5 and 82.9 ± 1.2% behind the enzymolysis 9h.Along with bringing up to 10000,10000 to 15000,15000 to 20000,20000 by 5000, B enzyme E/S brings up to 30000LAPU again, at enzymolysis time is under the identical enzymatic hydrolysis condition of 9h, NR increases thereupon, the every increase of E/S 1000LAPU, NR absolute value average mark you can well imagine high 0.58,0.9,0.18 and-0.01%.
Fig. 4 is the influence (n=2) to the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%) of Alcalase (A enzyme) E/S and enzymolysis time, at B enzyme E/S=15000LAPU, material-water ratio=1: 1, temperature=50 ℃, initial pH=9.1, under different al calase (A enzyme) the E/S level conditions, wall pollack is oppressed the nitrogen recovery (NR) of water-soluble protein hydrolysate.When A enzyme E/S from 30AU be increased to 60,90,120AU, NR increases with E/S and improves, (enzymolysis time 4h) NR is respectively 69.6 ± 0.6,76.6 ± 1.1,80.7 ± 0.5 and 80.7 ± 0.5% under the identical enzymatic hydrolysis condition, the average every raising of E/S 10AU, the NR absolute value improves 2.3,1.37% and 0% respectively.
Fig. 3, the prompting of 4 experimental results, in the enzymolysis process of the wall pollack flesh of fish, Alcalase enzyme and Flavourzyme enzyme E/S level and NR proportional relation in certain scope, tend towards stability then, when the E/S of Alcalase enzyme and Flavourzyme enzyme is higher than 90AU and 15000LAPU respectively, near or during the value of reaching capacity, NR will be tending towards a plateau value, no longer further obviously improve with the increase of E/S, this is consistent with existing correlative study report.
In the two enzyme enzymatic hydrolysis systems of this research Alcalase+Flavourzyme, water-soluble protein hydrolysate NR of wall pollack and the enzymolysis and extraction time relation of being proportionate (Fig. 3,4), along with enzymolysis time prolongs, the growth of NR mainly occurs in the incipient stage, tends towards stability then.(Fig. 4 after during enzymolysis 2h, △=90) NR is up to 74.6 ± 0.6%, 0～2h NR absolute value on average improves 0.62%/min, enzymolysis 2～4h, 4～6h, 6～9h, that NR absolute value average mark be you can well imagine is high by 0.05%, 0.01 and 0.01%/min, behind the enzymolysis 14h, NR is up to 89.5 ± 0.8%, and the NR absolute value on average improves 0.02%/min during 9～14h.Results suggest NR behind enzymolysis 4h still can constantly increase with the prolongation of enzymolysis time, and the raising of NR absolute value is tending towards fixed value and is approximately 0.01%/min.
After during Alcalase+Flavourzyme enzymolysis wall pollack flesh of fish 2h (Fig. 4, △=90), NR is up to 74.6 ± 0.6%, conforms to the result of experiment 1.Adopt hyperfiltration process to measure the molecular weight distribution (Fig. 4, △=90, enzymolysis time 6h) that wall pollack is oppressed water-soluble protein hydrolysate, molecular weight＜5000Da accounts for 97 ± 2%, and molecular weight＜3000Da accounts for 89 ± 3%.
Improve nitrogen recovery (NR), shortening enzymolysis time that protease preparation consumption E/S helps improving water-soluble protein hydrolysate, production cost then can increase because of the raising of protease consumption on the other hand; Prolong enzymolysis time and can reduce the enzyme preparation consumption in right amount and improve water-soluble protein hydrolysate NR, then reduced the turnover utilization rate of equipment on the other hand.The selection of best E/S ratio and enzymolysis time needs comprehensive every economic and technical norms to determine.
Fig. 5 is the influence (n=2) of material-water ratio to the water-soluble protein hydrolysate nitrogen recovery of wall pollack (NR%), when the enzymolysis parameter: when A enzyme E/S=90AU, B enzyme E/S=15000LAPU, temperature=50 ℃, initial pH=9.1, enzymolysis time=6h, the result shows, under different material-water ratio level 1.25,1.00,0.67,0.50 conditions, the nitrogen recovery (NR) of wall pollack solubility protein hydrolysate is 81.5 ± 0.5%～81.9 ± 0.2%, does not have significant difference.This and existing single enzyme enzymolysis research are reported different, generally, too high substrate and production concentration in the enzymatic hydrolysis system, the activated centre of meeting inhibitory enzyme molecule and the separation process that reaches with product that combines of substrate, reduce the catalytic efficiency of enzyme, in the complex enzyme hydrolysis system of this experiment A enzyme+B enzyme, macromolecular wall pollack fish protein is at first by A enzyme institute enzymolysis, its enzymolysis product is simultaneously again by B enzyme institute enzymolysis, thereby reduced the relative concentration of substrate and product, this may be in the material-water ratio scope that this experiment is selected, the one of the main reasons that different material-water ratios have no significant effect NR.Improve material-water ratio and help improving NR usually, but also increased follow-uply concentrate, the energy resource consumption of drying process, need comprehensive the measurement during selection.
3, separate:
Centrifugal and milipore filter combined method: Pacific Ocean wall pollack meat mincing fishy smell composition adopts the method for 8000rpm high speed centrifugation and 100kDa milipore filter combining ultrafiltration mainly from fat, can hold back big molecule fishy smell composition and anaphylactogen and heat source substance more up hill and dale.
This case process process is as follows:
Two, protein hydrolysate reduces the application of turbot liver fat content
Substitute 5%, 10%, 15%, 20%, 25% fish meal protein respectively respectively with liquid fish, control group is not for adding the basal feed of liquid fish.Feedstuff is crossed 80 mesh sieves through ultramicro grinding, and each raw material mixes after quantitatively by proportioning, adds an amount of adhesive then and kneads, and squeezes out the feed that diameter is respectively two kinds of particle diameters of 1.5mm through the twin-screw banded extruder.After drying, cut into the long particle of 3mm and be stored in-20 ℃ of refrigerators stand-by.Experiment feed formula such as table 4.
Table 4, experiment feed formula
1Vitamin mixtures (mg or g/kg feed): thiamine, 25mg; Riboflavin, 45mg; Puridoxine hydrochloride, 20mg; Cobalamin, 0.1mg; Prokeyvit, 10mg; Inositol, 800mg; Pantothenic acid, 60mg; Nicotinic acid, 200mg; Folic acid, 20mg; Biotin, 1.20mg; Vitamin A, 32mg; Vitamin D, 5mg; Vitamin E, 120mg; Inferior powder 18.67g.
2Inorganic salt mixt (mg or g/kg feed): sodium fluoride, 2mg; KI, 0.8mg; Cobalt chloride, 50mg; Copper sulphate, 10mg; Ferric sulfate, 80mg; Zinc sulfate, 50mg; Magnesium sulfate, 1200mg; Calcium dihydrogen phosphate, 3000mg; Common salt, 100mg; Zeolite powder, 15.45g.
Adopt the mixed feed that above-mentioned prescription makes to feed 10 weeks of turbot, observe its influence, result such as table 5 turbot growth and internal organ and liver index, hepatic fat content.
The result is cultured in table 5, each group test
Annotate: have significant difference (P＜0.05) with expression between line data subscript letter a, the b difference person.
Culture experiment finishes each test group turbot growth performance performance of back well, and survival rate is 100%, whole opisthosoma weight, rate of body weight gain, specific growth rate significant difference between each group of experiment.Substitute 5% group of fish meal protein, its rate of body weight gain is significantly higher than control group, substitutes fish meal protein 15%, substitutes fish meal protein 20% and substitutes 25% group of fish meal (P＜0.05), but close with alternative fish meal protein 10%, both there was no significant differences (P＞0.05).The specific growth rate that substitutes fish meal protein 15% is 1.1 times of control group.Control group, alternative fish meal protein 15%, alternative fish meal protein 20% and alternative 25% group of (P＜0.05) there was no significant difference of fish meal (P＞0.05).
Table 6, the different influence (%) that substitutes the liquid fish of level to turbot internal organ index, liver index, tripe tallow ratio, richness
Annotate: have significant difference (P＜0.05) with expression between line data subscript letter a, the b difference person.
As shown in Table 6, the different influences that substitute the liquid fish of level to turbot internal organ index, liver index, tripe tallow ratio, richness, the influence of the liquid fish liver index of different alternative levels, tripe tallow ratio and richness is less, control group internal organ index, liver index, tripe tallow are slightly higher than, fullness level, but with all the other each group differences not significantly (P＞0.05).Substitute fish meal protein 5% and substitute fish meal protein 10% internal organ than significantly being lower than other each experimental group (P＜0.05).
The different influences (%) that substitute the liquid fish of level to the biochemical composition of the full fish of turbot, muscle, liver in table 7, the feed
Annotate: have significant difference (P＜0.05) with expression between the different persons of line data subscript letter;
The liquid fish of different alternative levels sees Table 6 to the biochemical composition of the full fish of turbot, muscle, liver, and the biochemistry group Chengdu of full fish, dorsal muscles and the liver of each group of experiment is subjected to the influences of the liquid fish of different alternative levels to a certain extent.Wherein, the moisture that substitute fish meal protein 10%, substitutes fish meal protein 20% is the highest, with control group and 25% group of otherness of alternative fish meal protein significantly (P＜0.05), does not have significant difference (P＞0.05) but compare for 5% group with alternative fish meal protein.The protein of the full fish of control group, fat content and variant alternative fish meal protein group difference is remarkable (P＞0.05) not; Different liquid fish dorsal muscles moisture and the fatty there was no significant differences (P＜0.05) that substitute level, the protein content that substitute fish meal protein 5%, substitutes fish meal protein 10% is significantly higher than control group, substitutes fish meal protein 15%, substitutes fish meal protein 20%, substitutes fish meal protein 25% (P＜0.05), and control group, substitutes fish meal protein 15%, substitutes fish meal protein 20%, substitutes fish meal protein 25% there was no significant difference (P＞0.05).Substitute fish meal protein 5% for the liver moisture and significantly be lower than other each group (P＜0.05), protein content control group, alternative fish meal protein 5%, alternative fish meal protein are significantly higher than and substitute fish meal protein 10%, substitute fish meal protein 15% and alternative fish meal protein 25% for 20% group; It is minimum to substitute fish meal protein 5% for hepatic fat content, significantly be lower than each experimental group (P＜0.05), alternative fish meal protein 10%, alternative fish meal protein 15% significantly are lower than control group, substitute fish meal protein 20% and substitute 25% group of fish meal protein (P＜0.05).
Brief summary:
Can significantly improve the specific growth rate of turbot when substituting fish meal protein with 5% protein hydrolysate in the feed, to its survival rate to influence difference not remarkable; When the alternative fish meal protein amount of liquid fish in the feed reaches 5%, can significantly improve the growth performance of turbot, significantly reduce turbot liver fat content.
This experiment is to carry out in Yantai Tian Yuan breed company, has at first carried out bench-scale testing, on this basis, has carried out pilot scale, can effectively improve the growth performance of turbot, reduces turbot liver fat content, and application prospect is extensive.
Claims (2)
1. preparation that reduces turbot liver fat content additive, the preparation method who it is characterized in that it is: utilize Pacific Ocean wall pollack fish flakes, clean, rub the sterilization of heating of back input enzyme reactor through meat grinder, be warming up to 90 ℃ of insulations after the rubbing, water bath with thermostatic control 20min sterilization, adding water and protease enzyme preparation again mixes, enzymolysis under the enzymolysis process parameter condition of optimizing, heat up then and make the protease deactivation, coarse filtration is separated spur, the centrifugal soluble protein oligopeptides water that obtains respectively of the further three-phase of enzymolysis liquid, insoluble protein precipitation and fish oil, solubility protein hydrolysate is purified through ultrafiltration, obtains the protein hydrolysate powder after the freeze drying.
Described protease enzyme preparation: adopt Alcalase+Flavourzyme complex enzyme zymohydrolysis method; Alcalase enzyme: E/S is 90AU, and Flavourzyme enzyme: E/S is 15000LAPU.
Described enzymolysis process parameter is: hydrolysis temperature: 50 ℃～60 ℃, initial pH=8.5, material-water ratio: 1: 1, and enzymolysis time: 3h.
The ultrafiltration of described protein oligopeptide is purified: adopted 8000rpm high speed centrifugation and the method that 100kDa milipore filter ultrafiltration purification combines, thoroughly held back macromolecular substances, obtained molecular weight and reach 30% liquid fish less than 100Da.
2. one kind is reduced turbot liver fat content Application of Additives, it is characterized in that its suitable consumption is: substitute fish meal protein with 5% protein hydrolysate in the feed.
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| CN2010105523286A Pending CN102125173A (en) | 2010-11-10 | 2010-11-10 | Preparation and application of additive for reducing liver fat content of turbot |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103340315A (en) * | 2013-07-12 | 2013-10-09 | 河北师范大学 | Low-fish-meal turbot compound feed and preparation method thereof |
| CN103494039A (en) * | 2013-10-10 | 2014-01-08 | 中国水产科学研究院黄海水产研究所 | Additive for reducing visceral fat deposition of weever and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1845748A (en) * | 2003-07-04 | 2006-10-11 | 伯格生物医药有限公司 | Fish protein hydrolyzate |
| CN101703155A (en) * | 2009-11-17 | 2010-05-12 | 中国水产科学研究院黄海水产研究所 | Preparation method and application of prawn non-specific immunity reinforcing agent oligopeptide |
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2010
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1845748A (en) * | 2003-07-04 | 2006-10-11 | 伯格生物医药有限公司 | Fish protein hydrolyzate |
| CN101703155A (en) * | 2009-11-17 | 2010-05-12 | 中国水产科学研究院黄海水产研究所 | Preparation method and application of prawn non-specific immunity reinforcing agent oligopeptide |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103340315A (en) * | 2013-07-12 | 2013-10-09 | 河北师范大学 | Low-fish-meal turbot compound feed and preparation method thereof |
| CN103494039A (en) * | 2013-10-10 | 2014-01-08 | 中国水产科学研究院黄海水产研究所 | Additive for reducing visceral fat deposition of weever and application thereof |
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Application publication date: 20110720 |