阅读说明：本技术 一种利用脆肉鲩内脏制备亚铁螯合肽及高油酸鱼油的生产方法 (Production method for preparing ferrous chelated peptide and high oleic acid fish oil by using crisped grass carp viscera ) 是由 李琳 潘子强 郑子懿 沈志华 于 2019-10-16 设计创作，主要内容包括：本发明公开了一种利用脆肉鲩内脏制备亚铁螯合肽及高油酸鱼油的生产方法,包括以下步骤:A、制备高油酸鱼油：将脆肉鲩内脏与纯净水混合搅拌破碎,加入合适的蛋白酶进行酶解,煮沸灭酶后,冷却至常温离心收集上层油相,得到的粗鱼油；精制得高油酸鱼油；B、制备亚铁螯合肽：下层酶解液继续酶解,煮沸灭酶后冷却至常温,离心去除上层油相后将酶解液过滤去除沉淀,调节酶解液pH值,加入活性炭静置,使用冷冻高速离心机离心处理,取离心之后的酶解液,加入氯化亚铁恒温震荡,然后置于旋转蒸发仪中真空浓缩后冷冻干燥,得到亚铁螯合酶解物。本发明通过生物酶解方式水解脆肉鲩内脏,确定适宜的蛋白酶及酶解条件,在释放品质较好鱼油的同时制备出具有螯合亚铁离子的酶解液。(The invention discloses a production method for preparing ferrous chelate peptide and high oleic acid fish oil by utilizing crisped grass carp viscera, which comprises the following steps: mixing and stirring crisped grass carp viscera and purified water, crushing, adding proper protease for enzymolysis, boiling to inactivate enzyme, cooling to normal temperature, centrifuging, and collecting an upper oil phase to obtain crude fish oil; refining to obtain high oleic acid fish oil; B. preparing ferrous chelate peptide: and continuously carrying out enzymolysis on the enzymolysis liquid on the lower layer, boiling to inactivate enzyme, cooling to normal temperature, centrifuging to remove the oil phase on the upper layer, filtering the enzymolysis liquid to remove precipitate, adjusting the pH value of the enzymolysis liquid, adding activated carbon for standing, carrying out centrifugal treatment by using a freezing high-speed centrifuge, taking the centrifuged enzymolysis liquid, adding ferrous chloride, carrying out constant-temperature oscillation, then placing in a rotary evaporator for vacuum concentration, and carrying out freeze drying to obtain the ferrous chelate enzymolysis product. According to the invention, crisped grass carp viscera is hydrolyzed in a biological enzymolysis mode, appropriate protease and enzymolysis conditions are determined, and the enzymatic hydrolysate with chelated ferrous ions is prepared while fish oil with better quality is released.)

1. A production method for preparing ferrous chelated peptide and high oleic acid fish oil by using crisped grass carp viscera is characterized by comprising the following steps:

A. preparing high oleic acid fish oil:

s1, cleaning the crisped grass carp viscera and draining off water, wherein 100 parts by weight of the crisped grass carp viscera are as follows: adding 100 parts of purified water in a ratio of 1:1, stirring, crushing, and uniformly mixing to obtain homogenate; heating the homogenate in an enzymolysis tank to 50-60 deg.C, adjusting pH, adding protease 1.5-3 parts, mixing, performing enzymolysis for 1-3 hr, boiling, and inactivating enzyme to obtain enzymolysis solution with amino acid content of 3.4-4.8 mg/ml;

s2, cooling the enzymolysis liquid to normal temperature, centrifuging at 6000-10000r/min and 1-6 ℃ for 10-20min, and collecting an upper oil phase to obtain crude fish oil;

s3, refining the crude fish oil obtained in the step S2 to obtain high oleic acid fish oil which is liquid at normal temperature and contains 44.52-45.92% of oleic acid;

B. preparing ferrous chelate peptide:

s-1, placing the lower layer enzymolysis liquid in the step S2 in the process of preparing the high oleic acid fish oil in an enzymolysis tank at 40-80 ℃ for continuous enzymolysis for 0.5-2h, and boiling to inactivate enzyme;

s-2, cooling the enzymatic hydrolysate in the step S-1 to normal temperature, centrifuging for 10-20min at 6000-10000r/min and 1-6 ℃, removing the upper oil phase, filtering the enzymatic hydrolysate to remove precipitates, adjusting the pH value of the enzymatic hydrolysate, taking 100 parts of the enzymatic hydrolysate by weight, adding 0.5-2 parts of activated carbon, standing for 10-30min to clarify the enzymatic hydrolysate from turbidity, using a freezing high-speed centrifuge, 10000-14000r/min and 2-6 ℃, centrifuging for 5-15 min, taking the centrifuged enzymatic hydrolysate, adding 1-3 parts of ferrous chloride, shaking for 20-90min at constant temperature of 20-60 ℃, placing in a rotary evaporator, concentrating in vacuum, and freeze-drying to obtain the ferrous chelate enzymatic hydrolysate.

2. The method for producing ferrous chelated peptide and high oleic acid fish oil from grass carp viscera as claimed in claim 1, wherein the crude fish oil is refined in step S3 by degumming, neutralization, decolorization and deodorization.

3. The method for producing ferrous chelated peptide and high oleic acid fish oil with grass carp viscera as claimed in claim 1, wherein the pH value of the enzymolysis in step S1 is 7.8-8.2.

4. The method for producing ferrous chelated peptide and high oleic acid fish oil with grass carp viscera as claimed in claim 1, wherein the enzyme is boiled and deactivated in step S1 for 10-30 min.

5. The method for producing ferrous chelated peptide and high oleic acid fish oil by using grass carp viscera as claimed in claim 1, wherein the pH of the enzymatic hydrolysate is adjusted to 6.9-7.1 in step S-2.

6. The method for producing ferrous chelated peptide and high oleic acid fish oil from grass carp viscera as claimed in claim 1, wherein the protease in step S1 is a mixture of more than two of lcalse, papain, neutral protease, pepsin and trypsin.

7. The method for producing ferrous chelated peptide and high oleic acid fish oil from crusted grass carp viscera as claimed in claim 6, wherein the protease is a mixture of alcale, papain, neutral protease, pepsin and trypsin at a mass ratio of 1:3:0.5:2.5: 1.5.

8. The method for producing ferrous chelated peptide and high oleic acid fish oil by using grass carp viscera as claimed in claim 1, wherein the vacuum concentration in step S-2 is performed at 20-40 deg.C for 2-4 h; freeze drying at-50 deg.C to-30 deg.C for 10-18 h.

9. The method for producing ferrous chelated peptide and high oleic acid fish oil by using grass carp viscera as claimed in claim 1, wherein the molar concentration of ferrous chloride in step S-2 is 0.5-1.5 mol/L.

10. The method for preparing ferrous chelated peptide and high oleic acid fish oil from grass carp viscera as claimed in claim 1, wherein the extraction rate of the crude fish oil in step S2 is 56.50-67.27%, and the iodine value is 70.47-87.6gI₂100g of; the acid value is 7.11-13.62g/100 g; the peroxide value is 3.84-4.67milliequivalent/1000 g.

Technical Field

The invention belongs to the technical field of comprehensive utilization of aquatic product wastes, and relates to a method for treating crusian fishbone viscera and simultaneously preparing ferrous chelated peptide and fish oil by utilizing an enzymolysis technology.

Background

Grass carp becomes a new breeding object, namely crisped grass carp, after brittle breeding, has the characteristics of crystal color and luster, crisp and compact meat quality, difficult fish meat drawing, no rot after long-term cooking, sweet and delicious taste, high protein, low cholesterol, rich nutrition and the like, is popular among fishermen and consumers, and becomes a characteristic fish species in south China. With the deep processing and remote sale of crisped grass carp, a great deal of leftovers including fish head, fish scales, viscera and the like are produced. If the leftovers are discarded at will, not only is the resource waste caused, but also the environment is polluted. Crisped grass carp is large in size, generally about 6kg-10kg, large in internal organs, mostly 300 g-800 g, rich in fat and protein. Through detection, the water content of the crisped grass carp viscera is 11.58 +/-0.53%, the ash content is 1.25 +/-0.21%, the protein content is 6.94 +/-0.76%, and the fat content is 78.67 +/-2.53%, so that the crisped grass carp viscera is a good source of animal fat.

The protease can destroy the connection between fat cells and fish visceral tissues, the fat cells are released, the enzymolysis condition is mild, the quality of the produced fish oil is high, and if the enzymolysis process can be controlled (the type and the enzymolysis condition of the enzyme), the protein in the tissues is enzymolyzed while the fish oil is released, so that amino acid or short peptide with high nutritive value and physiological activity is produced. The common iron element in food exists mainly in trivalent form and can not be directly absorbed and utilized by the body, so that iron deficiency becomes one of the most common nutritional deficiency problems in the world at present. The currently used iron supplement preparation mainly comprises two chemical synthesis preparations and biological preparations, has the defects of high price, low absorption and utilization rate and the like, and the search for a natural iron supplement with high absorption rate becomes a hot spot in the field of dietary supplements. The viscera of crisped grass carp can generate small peptide and free amino acid after being hydrolyzed by protease, and can be chelated with trace elements to form small molecular complex, and the chelate can be directly absorbed by organism through small intestinal mucosa cells. Compared with the traditional iron supplement preparation, the iron peptide compound can promote the solubility of iron ions in intestinal tracts and improve the absorption rate and the utilization rate of the iron ions.

The invention relates to a method for comprehensively utilizing Peru squid viscera (publication No. CN1923052B), which comprises preparing refined squid oil by enzyme method, and preparing animal additive from lower liquid; pan scene, etc. utilizes alkaline protease to carry out enzymolysis on hemoglobin to prepare pig hemoglobin iron chelating peptide (publication number: CN105420323A) and invents an iron-containing chelating peptide iron supplement; hydrolyzing fish leftovers with pepsin and neutral protease, modifying by esterification reaction, and finally preparing fish oil with the content of EPA ethyl ester and DHA ethyl ester being more than 50%, and finally obtaining fish oil ethyl ester microcapsules by a spray drying method (publication No. CN 101940240B); li Bing Sheng, Lin Wan Ling, Yan Shao Ling, Yu Er Meng and Li Lin, etc. respectively invented the method for preparing fish fillet (publication No. CN103584181B, CN101965859B, CN110089695A) and detecting crispness of crisped grass carp (publication No. CN103994937B, CN109212173), which are not the researches on the utilization of viscera of crisped grass carp.

Disclosure of Invention

The invention aims to provide a production method for hydrolyzing crisped grass carp viscera in a biological enzymolysis way, and preparing an enzymolysis liquid with chelated ferrous ions while releasing fish oil with better quality by using proper protease types and enzymolysis conditions.

The fish oil is refined and then used for preparing feed grease, and a natural iron supplement is prepared by utilizing the chelation of the enzymolysis liquid and ferrous ions. The invention not only realizes the comprehensive utilization of leftovers, but also can obtain the bioactive zymolyte with chelated metal ions while preparing the fish oil.

In order to achieve the purpose, the invention adopts the following scheme:

a production method for preparing ferrous chelated peptide and high oleic acid fish oil by using crisped grass carp viscera is characterized by comprising the following steps:

A. preparing high oleic acid fish oil:

s1, cleaning the crisped grass carp viscera and draining off water, wherein 100 parts by weight of the crisped grass carp viscera are as follows: adding 100 parts of purified water in a ratio of 1:1, stirring, crushing, and uniformly mixing to obtain homogenate; heating the homogenate in an enzymolysis tank to 50-60 deg.C, adjusting pH, adding protease 1.5-3 parts, mixing, performing enzymolysis for 1-3 hr, boiling, and inactivating enzyme to obtain enzymolysis solution with amino acid content of 3.4-4.8 mg/ml;

s2, cooling the enzymolysis liquid to normal temperature, centrifuging at 6000-10000r/min and 1-6 ℃ for 10-20min, and collecting an upper oil phase to obtain crude fish oil;

s3, refining the crude fish oil obtained in the step S2 to obtain high oleic acid fish oil which is liquid at normal temperature and contains 44.52-45.92% of oleic acid;

B. preparing ferrous chelate peptide:

s-1, placing the lower layer enzymolysis liquid in the step S2 in the process of preparing the high oleic acid fish oil in an enzymolysis tank at 40-80 ℃ for continuous enzymolysis for 0.5-2h, and boiling to inactivate enzyme;

s-2, cooling the enzymatic hydrolysate in the step S-1 to normal temperature, centrifuging for 10-20min at 6000-10000r/min and 1-6 ℃, removing the upper oil phase, filtering the enzymatic hydrolysate to remove precipitates, adjusting the pH value of the enzymatic hydrolysate, taking 100 parts of the enzymatic hydrolysate by weight, adding 0.5-2 parts of activated carbon, standing for 10-30min to clarify the enzymatic hydrolysate from turbidity, using a freezing high-speed centrifuge, 10000-14000r/min and 2-6 ℃, centrifuging for 5-15 min, taking the centrifuged enzymatic hydrolysate, adding 1-3 parts of ferrous chloride, shaking for 20-90min at constant temperature of 20-60 ℃, placing in a rotary evaporator, concentrating in vacuum, and freeze-drying to obtain the ferrous chelate enzymatic hydrolysate.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by utilizing the crisped grass carp viscera, the crude fish oil is finely prepared in the step S3 in a degumming, neutralization, decoloration and deodorization mode. The degumming, neutralization, decolorization and deodorization processes in the invention are all existing preparation processes and are well known to the skilled person.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by using the crisped grass carp viscera, the pH value of the enzymolysis in the step S1 is 7.8-8.2.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by using the crisped grass carp viscera, the enzyme is boiled and inactivated for 10-30min in the step S1.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by utilizing the crisped grass carp viscera, the pH value of the enzymatic hydrolysate is adjusted to 6.9-7.1 in the step S-2.

In another improvement of the production method for preparing the ferrous chelated peptide and the high oleic acid fish oil by using the crisped grass carp viscera, in the step S1, the protease is a mixture of more than two of lcalse, papain, neutral protease, pepsin and trypsin.

As a further improvement of the protease, the protease is a mixture of alkaline protease Alcalse, papain, neutral protease, pepsin and trypsin in a mass ratio of 1:3:0.5:2.5: 1.5.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by utilizing the crisped grass carp viscera, the vacuum concentration temperature in the step S-2 is 20-40 ℃, and the concentration time is 2-4 h; freeze drying at-50 deg.C to-30 deg.C for 10-18 h.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by utilizing the crisped grass carp viscera, the molar concentration of the ferrous chloride in the step S-2 is 0.5-1.5 mol/L.

As another improvement of the production method for preparing the ferrous chelate peptide and the high oleic acid fish oil by utilizing the crisped grass carp viscera, the extraction rate of the crude fish oil in the step S2 is 56.50 to 67.27 percent, and the iodine value is 70.47 to 87.6gI₂100g of; the acid value is 7.11-13.62g/100 g; the peroxide value is 3.84-4.67milliequivalent/1000 g.

In summary, compared with the prior art, the invention has the beneficial effects that:

the crisped grass carp is huge in size, high in viscera occupation ratio and easy to collect and treat. The invention is based on controlling enzymolysis technology, prepares the oleic acid-rich crisped grass carp visceral oil and the hydrolysate with chelated ferrous ions by screening enzyme and controlling enzymolysis conditions, provides a new approach for feed oil, and generates a new iron-supplementing natural compound.

The grass carp high oleic acid fish oil of the invention conforms to the national edible animal oil sanitation standard (GB 10146-2005), the oil is liquid at normal temperature, the proportion of saturated fatty acid is 31.84%, the proportion of monounsaturated fatty acid is 47.23%, the proportion of polyunsaturated fatty acid is 19.93-20.45%, wherein the highest proportion of fatty acid is oleic acid (44.52-45.92%). The refined grease Atherosclerosis Index (AI) and coagulation promoting index (TI) are respectively 0.62-0.69 and 0.89-0.95.

Drawings

FIG. 1 is a schematic view of the process for preparing fish oil and ferrous chelate zymolyte simultaneously from the internal organs of crisped grass carp;

FIG. 2 is a schematic diagram of a crude fish oil product prepared by enzymatic hydrolysis of crisped grass carp viscera with different proteases of the present invention.

Detailed Description

The invention is further described below with reference to specific embodiments: