CN104095141A - Method for coproducing collagen activity peptides and flavoured base materials with chicken bones - Google Patents
Method for coproducing collagen activity peptides and flavoured base materials with chicken bones Download PDFInfo
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Abstract
The invention discloses a method for coproducing collagen activity peptides and flavoured base materials with chicken bones. Through a two-step hot pressing and extracting method, and through slight pressure relief and pressure supplementation treatment, the yield of extracting the collagen activity peptides from the chicken bones is substantially increased. Through high-pressure homogenizing of osseins in extracting solutions before a step-by-step enzymolysis reaction of the osseins, the degree of hydrolysis of enzymolysis solutions is increased. In addition, two-step ultra filtration is performed on the enzymolysis solutions, and separation and purification of different products in the enzymolysis solutions are realized. Follow-up processing is performed aiming at the products of each molecule segment, different products of bone polysaccharide containing chondroitin sulfate, macromolecule polypeptide, micromolecule polypeptide and the like are obtained, making full use of ossein enzymolysis solutions is realized, and especially a Maillard reaction is performed on the macromolecule polypeptide, xylose, cysteine and thiamin to obtain the flavoured base material with a rich flavour.
Description
Technical field
The present invention relates to biological technical field, relate in particular to a kind of method of chicken bone coproduction collagen active peptide and local flavor base-material.
Background technology
In Jin10Nian Lai world meat production, chicken increase of production is very fast, and China's chicken output occupies second place of the world, and the annual production of chicken bone can, up to 7,000,000 tons, account for 25% of livestock and poultry gross weight alive.Chicken osteotrophy is quite abundant, and its protein is similar to equivalent fresh meat with fat content, is respectively 16.3% and 14.5%, and the mineral matter elements such as calcium, phosphorus, iron and zinc are the several times of fresh meat, and ratio is suitable, wherein calcium content is 3.95%, and phosphorus content is 2.04%.Chicken SPP1 is the soluble protein of full price comparatively, and biological value is high, is the protein sources of high-quality.In addition, in chicken bone, also contain the nutrient of a large amount of other needed by human, as each seed amino acid and vitamin A, D, B
1, B
12deng, especially contain the indispensable Phospholipids of brain and phosphoprotein, can prevent ossein and the chondroitin sulfate old and feeble and metabolism of reinforcement cortical cell and the egg ammonia enzyme that can promote liver function.As can be seen here, chicken bone has important comprehensive utilization and exploitation is worth, but for a long time due to the reason of the aspects such as fund, technology, a large amount of chicken bones are not reasonably utilized, and be processed to the product that added value is very low, as bone mud, feed or fertilizer etc., do not embody the nutritive value of chicken bone completely, the abandoned even having also causes serious environmental pollution.Along with the development of China's bird secondary industry, how to realize chicken bone higher value application, the added value of utilizing that improves chicken bone is the Tough questions facing at present.
At present, to the utilization of chicken bone, be mainly collagen or the chondroitin sulfate extracting in chicken bone, although the process adopting can be extracted the II collagen type in chicken bone cartilage, but very low or be only that chicken bone enzymolysis is obtained to chondroitin sulfate to accounting for the recovery rate of the type i collagen albumen in the os osseum of skeleton quality 60% left and right, enzymolysis liquid is as wastewater treatment, these methods not only utilization rate of chicken bone are low, also environment caused to larger pollution simultaneously.
Summary of the invention
A kind of method that the object of this invention is to provide chicken bone coproduction collagen active peptide and local flavor base-material, by adopting two sections of hot pressing method for extracting, and process through slight pressure release ftercompction, make chicken bone obtain abundant pyrolysis, type i collagen albumen in os osseum and the II collagen type in cartilage etc. are fully extracted, thereby increase substantially the productive rate that extracts collagen active peptide from chicken bone; In addition, high-pressure homogeneous by the ossein in extract was carried out before carrying out substep enzyme digestion reaction, promote that in ossein, collagen carries out physics depolymerization, thereby increase substantially the degree of hydrolysis of enzymolysis liquid.
Another object of the present invention is, by the ossein in extract is carried out to substep enzymolysis by compound protease and flavor protease, compound protein endonuclease capable increases restriction enzyme site on the one hand, thereby increase the effect of enzymolysis, obtain a large amount of polypeptide, flavor protease is modified peptide molecule chain end on the other hand, improves the product local flavor of collagen active peptide.
Another object of the present invention is, by enzymolysis liquid is carried out to two-stage ultrafiltration, realize the separation and purification of different products in enzymolysis liquid, and for each minute subsegment product, carry out following process and obtain comprising chondroitin sulfate at different products such as interior bone polysaccharide, large molecular polypeptide and micromolecule polypeptides, realization makes full use of ossein enzymolysis liquid, especially by large molecular polypeptide and wood sugar, cysteine and thiamine are carried out to Maillard reaction, obtain local flavor base-material with rich flavor.
Technical scheme of the present invention is:
A method for chicken bone coproduction collagen active peptide and local flavor base-material, is characterized in that,
Chicken bone is carried out to two sections of hot pressing extractings and obtain extract, and the ossein extracting from extract is carried out carrying out substep enzymolysis after high-pressure homogeneous processing, wherein said two sections of hot pressing method for extracting are: by chicken bone with water according to 1: 1-2 mass ratio mixes, carry out in turn one section of hot pressing extracting and two sections of hot pressing extractings, wherein one section of hot pressing extraction temperature is 110-120 ℃, extracting time 100-120min, two sections of hot pressing extraction temperatures are 130-140 ℃, extracting time 70-90min;
In addition, further comprising the steps of:
Step 1, substep enzyme digestion reaction, mix the ossein after high-pressure homogeneous processing to carry out substep enzyme digestion reaction with compound protease and flavor protease respectively according to sequencing, obtains enzymolysis liquid;
Step 2, two-stage ultrafiltering, adopt the milipore filter that molecular cut off is 9-10kDa to carry out one-level ultrafiltration gained enzymolysis liquid, obtain one-level trapped fluid and first-level filtering fluid, adopt the milipore filter that molecular cut off is 2-3kDa to carry out two-stage ultrafiltering first-level filtering fluid, obtain second entrapment liquid and secondary filter liquor;
The preparation of step 3, local flavor base-material, adopt ethanol precipitation to process gained one-level trapped fluid, staticly settle, get supernatant and second entrapment liquid is mixed to form Maillard reaction precursor, after being mixed with a certain amount of wood sugar, cysteine and thiamine, gained Maillard reaction precursor carries out hot pressure reaction, obtain maillard reaction product, by after gained maillard reaction product concentrate drying both described local flavor base-material;
The preparation of step 4, collagen active peptide, carries out concentrate drying by gained secondary filter liquor in step 2 and had both obtained collagen active peptide.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, described two sections of hot pressing extractive reactions carry out in hot pressure reaction tank, and after finishing, two sections of hot pressing extractive reactions carry out slight pressure release ftercompction, concrete grammar is: after two sections of hot pressing extractings finish, open the relief valve 8-10min of hot pressure reaction tank, make temperature in hot pressure reaction tank close relief valve 8-10min after reducing 5-10 ℃, and make temperature rising 5-10 ℃, according to this step, repeat 3-5 time.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the quality that adds compound protease and flavor protease in step 1 is respectively the 0.2-0.3% of ossein quality used, enzyme digestion reaction temperature is 51-53 ℃, each walks enzyme digestion reaction and carries out respectively 100-120min, and each walks enzyme digestion reaction and finishes to be warming up to respectively 90-100 ℃ and keep 20-30min to carry out enzyme-deactivating afterwards.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the concrete grammar of ethanol precipitation described in step 3 is: by described one-level trapped fluid with 90-100% ethanol according to 1: after the volume ratio of 2-3 is mixed, centrifugal through 8000-10000r/min, obtain the upper strata stillness of night and lower sediment.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, ethanol precipitation gained lower sediment is processed after 2-3 time with 90-100% ethanol dehydration, obtained after drying chondroitin sulfate.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the quality that adds wood sugar, cysteine and thiamine in step 3 is respectively the 0.4-0.6% of the front weight of Maillard reaction used, described hot pressure reaction temperature is 100-110 ℃, and the hot pressure reaction time is 80-100min.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the extracting method of described ossein is: after gained extract is screened out to bone slag, carry out stratification, both remove after upper oil phase to obtain described ossein;
In addition, ossein being carried out to high-pressure homogeneous method is: adopt high pressure homogenizer to carry out ossein high-pressure homogeneous, wherein set 70 ± 2 ℃ of homogenizing temperatures, pressure 30 ± 2MPa, rotating speed 700-800r/min, homogenizing time 5-8min.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, adopt vibratory sieve division to carry out bone ballast screen and remove, vibratory sieve used aperture is 40-50 order, and can be by bone slag remaining in the bone slag screening out and hot pressure reaction tank through pulverizing the dry bone meal that is prepared as.
Preferably, in described chicken bone coproduction collagen active peptide and the method for local flavor base-material, get the upper oil phase after stratification and upper oil phase is heated to 80 ℃ of left and right, adding volume is the 90-95 ℃ of hot water of upper oil phase 10-15%, 10-15r/min stirs 8-10min, after standing sedimentation 20-30min, remove water, remaining oil heat phase is obtained to animal oil at 30min with the interior 80-85 of being cooled to ℃ after 100-115 ℃.
The present invention has following beneficial effect: the method for a kind of chicken bone coproduction collagen active peptide of the present invention and local flavor base-material, by adopting two sections of hot pressing method for extracting, and process through slight pressure release ftercompction, make chicken bone obtain abundant pyrolysis, type i collagen albumen in os osseum and the collagens such as II collagen type in cartilage are fully extracted, make the content of collagen in extract reach 40mg/g left and right, especially for the extraction of the type i collagen albumen in os osseum, traditional atmospheric cooking method is difficult to extract the type i collagen albumen in os osseum, and the present invention has improved more than 3 times type i collagen protein extracting ratio in os osseum, type i collagen albumen in os osseum is utilized effectively, and increase substantially the productive rate that extracts collagen active peptide from chicken bone, the content of peptides of products therefrom reaches 91%, in addition, high-pressure homogeneous by the ossein in extract was carried out before carrying out substep enzyme digestion reaction, promote that in ossein, collagen carries out physics depolymerization, thereby increase substantially the degree of hydrolysis of enzymolysis liquid, degree of hydrolysis is reached more than 30%, and conventional method degree of hydrolysis only can reach 20% left and right.
In addition, by the ossein in extract is carried out to substep enzymolysis by compound protease and flavor protease, compound protein endonuclease capable increases restriction enzyme site on the one hand, fully enzymolysis is difficult for the former albumen of dissolved gum, thereby increases the effect of enzymolysis, obtains a large amount of polypeptide, flavor protease is modified peptide molecule chain end on the other hand, the bitter peptides content that enzymolysis is produced is controlled, and delicate flavour peptide content increases, and has greatly improved the local flavor of product.
Simultaneously, by enzymolysis liquid is carried out to two-stage ultrafiltration, realize the separation and purification of different products in enzymolysis liquid, and for each minute subsegment product, carry out following process and obtain comprising chondroitin sulfate at interior bone polysaccharide, the different products such as large molecular polypeptide and micromolecule polypeptide, wherein gained chondroitin sulfate cellulose content is higher than 92%, realization makes full use of ossein enzymolysis liquid, especially pass through large molecular polypeptide and wood sugar, cysteine and thiamine are carried out Maillard reaction, the intermediate product that in course of reaction, large molecular polypeptide and wood sugar generation carbonyl ammonia react generate reacts with the hot pressing product of cysteine and thiamine and generates Mei Lade product, obtain local flavor base-material with rich flavor, according to the interpretation of result of GC-MS, levels of pyrazine in local flavor base-material is approximately 10 times of enzymolysis liquid, aldoketones is approximately 2.5 times of enzymolysis liquid, proved the raw fragrant effect of Maillard reaction.When catering to the market demand, the present invention realizes the recycling of discarded object; and do not add any inorganic acid, alkali; technological process is compact; technological parameter is accurate, and artificial disturbance factor is few, is applicable to enterprise scale and produces; make full use of low value chicken bone and obtain the multiple product with market prospects; realize the high value added utilization of chicken bone, when reducing agricultural byproducts processing to the pollution of environment, improve Business Economic Benefit, for the comprehensive utilization of chicken bone provides technical support.
Accompanying drawing explanation
Fig. 1 is the method process chart of chicken bone coproduction collagen active peptide of the present invention and local flavor base-material;
Fig. 2 is hydrolysis degree change curve under the different enzymolysis times of substep enzyme digestion reaction of the present invention;
Tu3Wei Sigma company chondroitin sulfate standard items high-efficient liquid phase chromatogram;
Fig. 4 is the high-efficient liquid phase chromatogram of the chondroitin sulfate that extracts of the present invention;
Fig. 5 be under different hot pressing extracting conditions different material with the variation diagram of hot pressing extracting time;
Fig. 6 is that the one-tenth of bone meal of the present invention is grouped into table;
Fig. 7 is that the one-tenth of animal oil of the present invention is grouped into table;
Fig. 8 is bone polysaccharide physical and chemical index analysis result statistical form of the present invention;
Fig. 9 is each minute subsegment ultrafiltration product and enzymolysis liquid product non-oxidizability statistical form;
Figure 10 is maillard reaction product component list of the present invention.
The specific embodiment
Below in conjunction with accompanying drawing, 1-10 elaborates to the present invention, after making those of ordinary skills consult this description, can implement according to this.
As shown in Figure 1, a kind of method of chicken bone coproduction collagen active peptide and local flavor base-material, it is characterized in that, chicken bone is carried out to two sections of hot pressing extractings and obtain extract, after being removed to bone slag, extract drops into stratification in standing tank, at the bottom of utilizing tank, by-pass valve separation obtains ossein and upper oil phase, and the ossein of the extraction from extract is carried out carrying out substep enzymolysis after high-pressure homogeneous processing, wherein said two sections of hot pressing method for extracting are: chicken bone is rinsed with clear water, remove after surperficial bloodstain impurity, by chicken bone with water according to 1: 1-2 mass ratio mixes and adds in hot pressure reaction tank, carry out in turn one section of hot pressing extracting and two sections of hot pressing extractings, wherein one section of hot pressing extraction temperature is 110-120 ℃, extracting time 100-120min, two sections of hot pressing extraction temperatures are 130-140 ℃, extracting time 70-90min,
In addition, further comprising the steps of:
Step 1, substep enzyme digestion reaction, mix the ossein after high-pressure homogeneous processing to carry out substep enzyme digestion reaction with compound protease and flavor protease respectively according to sequencing, obtains enzymolysis liquid; Substep enzymolysis process carries out in reactor, and carries out stirring at low speed, and rotating speed is 20-25r/min, to increase the touch opportunity of enzyme and substrate, improves enzyme digestion reaction efficiency.In addition, described compound protease is a kind of enzyme being mixed by multiple biology enzyme, comprise bromelain, papain and neutral proteinase etc., described compound protease has a plurality of restriction enzyme sites, there are a plurality of protein loci cutting functions, and effective hydrophobic group in collagen protein enzymolysis, improve collagen solubility, fully enzymolysis is difficult for the former albumen of dissolved gum, obtain a large amount of polypeptide and amino acid, and under the effect of flavor protease, peptide molecule chain end is modified, thereby make in enzymolysis liquid bitter peptides content controlled, delicate flavour peptide content increases, improve the local flavor of product.
Step 2, two-stage ultrafiltration, adopt the milipore filter that molecular cut off is 9-10kDa to carry out one-level ultrafiltration gained enzymolysis liquid, obtain one-level trapped fluid and first-level filtering fluid, adopt the milipore filter that molecular cut off is 2-3kDa to carry out two-stage ultrafiltering first-level filtering fluid, obtain second entrapment liquid and secondary filter liquor; Enzymolysis liquid described in the present invention enters two-stage ultrafiltration coupling equipment after 200 order sieves filter, and ultrafiltration membrane elements molecular cut off used is 10kDa, 2kDa respectively, and ultrafiltration temperature is controlled at 45 ℃-50 ℃, operating pressure 0.5MPa.By enzymolysis liquid is carried out to two-stage ultrafiltration, realization is to having the separation and purification of different molecular weight product in enzymolysis liquid, obtain > 10kDa, 10kDa-2kDa and tri-sections of materials of < 2kDa, wherein > 10kDa main component is to comprise the bone polysaccharide of chondroitin sulfate and the precursor of part Maillard reaction; 10kDa-2kDa main component is large molecular polypeptide, is the precursor of main Maillard reaction; < 2kDa is mainly micromolecule polypeptide, is rich in biologically active, carries out following process obtain different product for the characteristic of each minute subsegment product, realizes making full use of ossein enzymolysis liquid.Each minute subsegment ultrafiltration product and enzymolysis liquid product non-oxidizability are shown in Fig. 9.
The preparation of step 3, local flavor base-material, adopt ethanol precipitation to process gained one-level trapped fluid, staticly settle, get supernatant and second entrapment liquid is mixed to form Maillard reaction precursor, after being mixed in hot pressure reaction still with a certain amount of wood sugar, cysteine and thiamine, gained Maillard reaction precursor carries out hot pressure reaction, in course of reaction, open stirring at low speed, rotating speed is 20-25r/min, obtains maillard reaction product; Gained maillard reaction product is carried out to after interlayer water-bath is cooled to 60-70 ℃, proceed to vacuum concentration pot and carry out Vacuum Concentration to Brix=40 °, vacuum is more than 0.08MPa, and with spray dryer by concentrate spray dry after both described local flavor base-material, wherein spray dryer feeding temperature is controlled as 180-190 ℃, it is 90 ℃ of left and right that drop temperature is controlled, and moisture content of finished products is not more than 5%.
The preparation of step 4, collagen active peptide, carries out concentrate drying by gained secondary filter liquor in step 2 and had both obtained collagen active peptide.Method therefor of the present invention is for to carry out Vacuum Concentration to Brix=40 ° by secondary filter liquor, vacuum is more than 0.08MPa, and concentrate is sprayed dry with spray dryer, wherein spray dryer feeding temperature is controlled as 180-190 ℃, it is 90 ℃ of left and right that drop temperature is controlled, moisture content of finished products is not more than 5%, the content of peptides that obtains collagen active peptide is 91% left and right, the functions such as the anti-oxidant and ACE inhibition of little molecule oligopeptide are wherein remarkable, functional medicine with respect to manually synthesizing at present, has the features such as side effect is little, effect is remarkable.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, described two sections of hot pressing extractive reactions carry out in hot pressure reaction tank, and after finishing, two sections of hot pressing extractive reactions carry out slight pressure release ftercompction, concrete grammar is: after two sections of hot pressing extractings finish, open the relief valve 8-10min of hot pressure reaction tank, after making the interior temperature of hot pressure reaction tank reduce 5-10 ℃, close relief valve 8-10min, make 5-10 ℃ of temperature rising to former temperature, and repeat 3-5 time according to this step.Through slight pressure release ftercompction, process, can break the dynamic equilibrium in hot pressure reaction tank, promote the dynamic translation of material, improve the extract content of product, the type i collagen albumen of the present invention in can high efficiency extraction os osseum and the collagens such as II collagen type in cartilage, 100kg chicken bone can extract the ossein extract of 360kgBrix4% left and right, and wherein collagen content is 40mg/g left and right.
Total solable matter in extract under different hot pressing extracting time conditions, crude protein and collagen content changed with the hot pressing extracting time, wherein total solable matter is represented by TSS, chicken bone is the continuous stripping of material in it in hot pressing extractive process, wherein main material is protein and a small amount of solubility organic compound, as shown in Figure 5, total Soluble matter content is along with the raising of extracting time lengthening and extraction temperature has increase in various degree, between different extraction temperatures, total Soluble matter content changes from 0.9% to 5.2%, 1.9% to 5.6%, under different temperatures, total Soluble matter content is changed significantly, both P < 0.05, when extraction temperature is 120 ℃ and 130 ℃ between the different extracting time, total Soluble matter content is different, 90min before extracting particularly, total Soluble matter content has a big increase, 90min increases not remarkable later.Between different temperatures, crude protein content changes significant difference, P < 0.05, between different extraction temperatures, crude protein content is changed to from 0.54% to 3.54%, 1.23% to 3.82%, when wherein extraction temperature is 120 ℃, crude protein content amplification is maximum, when extraction temperature is 120 ℃ and 130 ℃, before extracting, 90min crude protein incremental amplitude is large, after 90min, amplification tends towards stability, when extraction temperature is 120 ℃, front 20min changes slowly, extracting 20min later its content sharply increases until also have the trend significantly increasing during 120min, illustrate and in chicken bone, also have many undissolved collagens.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the quality that adds compound protease and flavor protease in step 1 is respectively the 0.2-0.3% of ossein quality used, enzyme digestion reaction temperature is 51-53 ℃, each walks enzyme digestion reaction and carries out respectively 100-120min, and each walks enzyme digestion reaction and finishes to be warming up to respectively 90-100 ℃ and keep 20-30min to carry out enzyme-deactivating afterwards; In addition, in substep enzymolysis process, enzymolysis liquid is carried out to stirring at low speed.
The condition of described enzyme digestion reaction obtains through test of many times checking, extract is divided into the many parts of different biology enzymes of use and is hydrolyzed respectively 1h, 2h, 3h, 4h, 5h, and hydrolysis time on degree of hydrolysis (DH) impact as shown in Figure 2.Degree of hydrolysis is along with the prolongation of hydrolysis time constantly increases, and after hydrolysis surpasses certain hour, hydrolysis rate becomes slowly, and degree of hydrolysis ascensional range reduces, and is due to the prolongation with hydrolysis time, due to concentration of substrate reduces or enzyme activity declines.Papain, compound protein and neutral proteinase are before hydrolysis during 3h, and before the explanation of degree of hydrolysis rapid development, 3h hydrolysis is violent, and protein is degraded into free amino acid in a large number.And the growth trend of degree of hydrolysis becomes mild after 3h.Flavor protease before hydrolysis during 2h degree of hydrolysis increase slowly, after 2h, hydrolysis is violent, after hydrolysis 4h, degree of hydrolysis increases and is tending towards saturated.Although hydrolysis time extends the thorough hydrolysis that is conducive to albumen, hydrolysis surpasses after certain hour, and degree of hydrolysis growth trend is tending towards saturated, and the longer energy consumption of hydrolysis time is larger.Therefore, comprehensive degree of hydrolysis and energy consumption determine that flavor protease and the best enzymolysis time of compound protease are respectively 2h left and right.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the concrete grammar of ethanol precipitation described in step 3 is: by described one-level trapped fluid with 90-100% ethanol according to 1: after the volume ratio of 2-3 is mixed, centrifugal through 8000-10000r/min, obtain the upper strata stillness of night and lower sediment.Ethanol precipitation gained lower sediment is processed after 2-3 time with 90-100% ethanol dehydration, obtained after drying the bone polysaccharide that contains 92% above chondroitin sulfate.Described bone polysaccharide physical and chemical index analysis result is shown in Fig. 8; Fig. 3 is the high-efficient liquid phase chromatogram of chondroitin sulfate standard items; Fig. 4 is the high-efficient liquid phase chromatogram of the chondroitin sulfate that extracts of the present invention, and in gained bone polyoses extract of the present invention, chondroitin sulfate cellulose content is 92.1% as calculated.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the quality that adds wood sugar, cysteine and thiamine in step 3 is respectively the 0.4-0.6% of the front weight of Maillard reaction used, described hot pressure reaction temperature is 100-110 ℃, and the hot pressure reaction time is 80-100min.The Maillard reaction precursor forming with second entrapment liquid and ethanol precipitation gained supernatant liquor is at wood sugar, under the participation of cysteine and thiamine, carry out Maillard reaction, wherein wood sugar provides carbonyl, cysteine and thiamine are main meat flavor precursors, the intermediate product that polypeptide in course of reaction in gained Maillard reaction precursor and wood sugar generation carbonyl ammonia react generate reacts with the thermal decomposition product of cysteine and thiamine and generates maillard reaction product with rich flavor, and obtain local flavor base-material through concentrate drying, according to the interpretation of result of GC-MS, acids and letones change less with enzymolysis time relative amount, levels of pyrazine in gained local flavor base-material is approximately 10 times of enzymolysis liquid, pyrazine class material mostly has barbecue taste, make enzymolysis product meat flavour strong, aldoketones is approximately 2.5 times of enzymolysis liquid, has further strengthened meat flavour and the fruital taste of enzymolysis liquid, and under the effect of the additives such as wood sugar, cysteine and thiamine, Maillard reaction has been strengthened the local flavor of enzymolysis liquid, makes product special flavour and function all good.The composition of described maillard reaction product forms as shown in figure 10, has proved the raw fragrant effect of Maillard reaction.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, the extracting method of described ossein is: after gained extract is screened out to bone slag, carry out stratification, both remove after upper oil phase to obtain described ossein; In addition, ossein being carried out to high-pressure homogeneous method is: adopt high pressure homogenizer to carry out ossein high-pressure homogeneous, wherein set 70 ± 2 ℃ of homogenizing temperatures, pressure 30 ± 2MPa, rotating speed 700-800r/min, homogenizing time 5-8min.The shearing of liquid stream, friction, centrifugal extruding and liquid strong, reciprocal in homogenizing process flow the comprehensive functions such as collision, promote collagen spiral physics depolymerization in ossein, increase its dissolubility and peptide chain gap, for follow-up substep enzymolysis provides condition, found that through high-pressure homogeneous ossein degree of hydrolysis after enzymolysis finishes be 23.4%, higher more than 6% than the ossein enzymolysis liquid degree of hydrolysis without excessively high-pressure homogeneous.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, adopt vibratory sieve division to carry out bone ballast screen and remove, in described bone slag, be rich in hydroxyapatite, vibratory sieve used aperture is 40-50 order.After hot pressing extracting finishes, most of bone slag is deposited in the bottom of hot pressing extracting tank, with ossein, the compression pump through hot pressing extracting tank pumps fraction bone slag, this part the bone slag being blended in so in ossein need to remove through Vibration Screen, and can will screen out bone slag remaining in resulting bone slag and hot pressure reaction tank together, through pulverizing to be dried, be prepared as bone meal, for animal feed or farmland inorganic fertilizer, wherein calcium, phosphorus content are respectively 62.3% and 29.4%.The composition of described bone meal forms as shown in Figure 6.
In described chicken bone coproduction collagen active peptide and the method for local flavor base-material, get the upper oil phase after stratification and upper oil phase is heated to 80 ℃ of left and right, adding volume is the 90-95 ℃ of hot water of upper oil phase 10-15%, 10-15r/min stirs 8-10min, after standing sedimentation 20-30min, remove water, by remaining oil heat phase to being cooled to rapidly 80-85 ℃ and obtaining animal oil with interior at 30min after 100-115 ℃.The composition of described animal oil forms as shown in Figure 7.As shown in Figure 7, the animal oil product of being made by chicken bone reaches edible blend oil standard substantially, and meat aromatic flavour, is of high nutritive value, and can be used for margarine and seasoning wet goods.
Embodiment 1
Step 1, 300kg chicken bone is rinsed with clear water, remove surperficial bloodstain impurity, being divided into three batches drops into respectively in hot pressure reaction tank and adds quantitative clear water according to the mass ratio of 1: 2, carry out two sections of hot pressing extractings, one section of hot pressing extraction temperature is 120 ℃, extracting time 120min, two sections of hot pressing extraction temperatures are 130 ℃, extracting time 90min, slight pressure release/ftercompction during this time improves product and puies forward rate, method is: after two sections of hot pressing extractings finish, open the relief valve 10min of hot pressure reaction tank, make temperature in hot pressure reaction tank close relief valve 10min after reducing by 5 ℃, and make 5 ℃ of temperature risings return back to former temperature, according to this step, repeat 3 times.
Step 2, adopt vibratory sieve division to remove bone slag extract, vibratory sieve aperture is 40 orders, enter standing tank standing, utilize the separated ossein of flow divider and upper oil phase, ossein enters enzyme digestion reaction still under 30 ± 2MPa condition after homogeneous depolymerization through high pressure homogenizer.
Step 3, adopt substep enzymatic isolation method to carry out enzymolysis ossein, by 0.2% of substrate quality, successively add compound protease and flavor protease, two step enzyme digestion reaction temperature are 51-53 ℃, two step enzyme digestion reactions carry out respectively 120min, two step enzyme digestion reactions finish to be warming up to respectively 90 ℃ and keep 20min to carry out enzyme-deactivating afterwards, in enzymolysis process, in reactor, open stirring at low speed, rotating speed 20～25r/min.
Step 3, enzymolysis liquid enter two-stage ultrafiltration coupling equipment after 200 order sieves filter, and ultrafiltration membrane elements molecular cut off is 10kDa, 2kDa respectively, and ultrafiltration temperature is controlled at 45 ℃-50 ℃, operating pressure 0.5MPa.
Step 4, described one-level trapped fluid is mixed according to the volume ratio of 1: 3 with 95% ethanol after, centrifugal through 10000r/min, obtain the upper strata stillness of night and lower sediment, lower sediment continues with after 95% ethanolic solution dehydration 2 times, the dry bone polysaccharide finished product that contains chondroitin sulfate of obtaining, supernatant liquor adds second entrapment liquid to form Maillard reaction precursor.
Step 5, Maillard reaction precursor is moved into hot pressure reaction still, the wood sugar, cysteine and the thiamine that add respectively Maillard reaction precursor quality 0.5%, hot pressure reaction 90min at 105 ℃, open stirring at low speed, the Mei Lade product obtaining, interlayer water-bath proceeds to vacuum concentration pot and carries out Vacuum Concentration to Brix=40 ° after being cooled to 60-70 ℃, and vacuum is more than 0.08MPa, with spray dryer, concentrate is dry, obtain local flavor base-material.
Step 6, by the dry collagen active peptide that obtains of secondary filter liquor Vacuum Concentration; Bone slag remaining in the bone slag screening out in extract and hot pressure reaction tank is made bone meal after ultramicro grinding is dry, can be used for animal feed and farmland inorganic fertilizer; Animal oil is transferred to animal oil holding vessel by by-pass valve, to 90 ℃ of hot water that add 15% in the animal oil of 80 ℃ of left and right temperature, 10-15r/min stirs 10min, standing sedimentation 30min discards waste water, animal oil is heated to 115 ℃ and removes moisture, with interlayer water-bath, be cooled to rapidly 80-85 ℃, temperature fall time is controlled in 30min, obtains finished chicken animal oil.
Although embodiment of the present invention are open as above, but it is not restricted to listed utilization in description and embodiment, it can be applied to various applicable the field of the invention completely, for those skilled in the art, can easily realize other modification, therefore do not deviating under the universal that claim and equivalency range limit, the present invention is not limited to specific details and illustrates here and the legend of describing.
Claims (9)
1. a method for chicken bone coproduction collagen active peptide and local flavor base-material, is characterized in that,
Chicken bone is carried out to two sections of hot pressing extractings and obtain extract, and the ossein extracting from extract is carried out carrying out substep enzymolysis after high-pressure homogeneous processing, wherein said two sections of hot pressing method for extracting are: by chicken bone with water according to 1: 1-2 mass ratio mixes, carry out in turn one section of hot pressing extracting and two sections of hot pressing extractings, wherein one section of hot pressing extraction temperature is 110-120 ℃, extracting time 100-120min, two sections of hot pressing extraction temperatures are 130-140 ℃, extracting time 70-90min;
In addition, further comprising the steps of:
Step 1, substep enzyme digestion reaction, mix the ossein after high-pressure homogeneous processing to carry out substep enzyme digestion reaction with compound protease and flavor protease respectively according to sequencing, obtains enzymolysis liquid;
Step 2, two-stage ultrafiltering, adopt the milipore filter that molecular cut off is 9-10kDa to carry out one-level ultrafiltration gained enzymolysis liquid, obtain one-level trapped fluid and first-level filtering fluid, adopt the milipore filter that molecular cut off is 2-3kDa to carry out two-stage ultrafiltering first-level filtering fluid, obtain second entrapment liquid and secondary filter liquor;
The preparation of step 3, local flavor base-material, adopt ethanol precipitation to process gained one-level trapped fluid, staticly settle, get supernatant and second entrapment liquid is mixed to form Maillard reaction precursor, after being mixed with a certain amount of wood sugar, cysteine and thiamine, gained Maillard reaction precursor carries out hot pressure reaction, obtain maillard reaction product, by after gained maillard reaction product concentrate drying both described local flavor base-material;
The preparation of step 4, collagen active peptide, carries out concentrate drying by gained secondary filter liquor in step 2 and had both obtained collagen active peptide.
2. the method for chicken bone coproduction collagen active peptide as claimed in claim 1 and local flavor base-material, it is characterized in that, described two sections of hot pressing extractive reactions carry out in hot pressure reaction tank, and after finishing, two sections of hot pressing extractive reactions carry out slight pressure release ftercompction, concrete grammar is: after two sections of hot pressing extractings finish, open the relief valve 8-10min of hot pressure reaction tank, make temperature in hot pressure reaction tank close relief valve 8-10min after reducing 5-10 ℃, and make temperature rising 5-10 ℃, according to this step, repeat 3-5 time.
3. the method for chicken bone coproduction collagen active peptide as claimed in claim 1 and local flavor base-material, it is characterized in that, the quality that adds compound protease and flavor protease in step 1 is respectively the 0.2-0.3% of ossein quality used, enzyme digestion reaction temperature is 51-53 ℃, each walks enzyme digestion reaction and carries out respectively 100-120min, and each walks enzyme digestion reaction and finishes to be warming up to respectively 90-100 ℃ and keep 20-30min to carry out enzyme-deactivating afterwards.
4. the method for chicken bone coproduction collagen active peptide as claimed in claim 1 and local flavor base-material, it is characterized in that, the concrete grammar of ethanol precipitation described in step 3 is: by described one-level trapped fluid with 90-100% ethanol according to 1: after the volume ratio of 2-3 is mixed, centrifugal through 8000-10000r/min, obtain the upper strata stillness of night and lower sediment.
5. the method for chicken bone coproduction collagen active peptide as claimed in claim 4 and local flavor base-material, is characterized in that, ethanol precipitation gained lower sediment is processed after 2-3 time with 90-100% ethanol dehydration, obtains after drying chondroitin sulfate.
6. the method for chicken bone coproduction collagen active peptide as claimed in claim 1 and local flavor base-material, it is characterized in that, the quality that adds wood sugar, cysteine and thiamine in step 3 is respectively the 0.4-0.6% of the front weight of Maillard reaction used, described hot pressure reaction temperature is 100-110 ℃, and the hot pressure reaction time is 80-100min.
7. the method for chicken bone coproduction collagen active peptide as claimed in claim 1 and local flavor base-material, is characterized in that, the extracting method of described ossein is: after gained extract is screened out to bone slag, carry out stratification, both remove after upper oil phase to obtain described ossein;
In addition, ossein being carried out to high-pressure homogeneous method is: adopt high pressure homogenizer to carry out ossein high-pressure homogeneous, wherein set 70 ± 2 ℃ of homogenizing temperatures, pressure 30 ± 2MPa, rotating speed 700-800r/min, homogenizing time 5-8min.
8. the method for chicken bone coproduction collagen active peptide as claimed in claim 7 and local flavor base-material, it is characterized in that, adopting vibratory sieve division to carry out bone ballast screen to extract removes, vibratory sieve used aperture is 40-50 order, and can be by bone slag remaining in the bone slag screening out and hot pressure reaction tank through pulverizing the dry bone meal that is prepared as.
9. the method for chicken bone coproduction collagen active peptide as claimed in claim 8 and local flavor base-material, it is characterized in that, get the upper oil phase after stratification and upper oil phase is heated to 80 ℃ of left and right, adding volume is the 90-95 ℃ of hot water of upper oil phase 10-15%, 10-15r/min stirs 8-10min, after standing sedimentation 20-30min, remove water, remaining oil heat phase is obtained to animal oil at 30min with the interior 80-85 of being cooled to ℃ after 100-115 ℃.
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