阅读说明：本技术 一种超声辅助提取鲍鱼内脏蛋白的方法 (Method for ultrasonic-assisted extraction of abalone visceral protein ) 是由 吴汶飞 汤宇轩 刘祉妤 毛臻彧 刘帅辰 任美霖 林松毅 李冬梅 姜鹏飞 于 2019-12-17 设计创作，主要内容包括：本发明公开了一种超声辅助提取鲍鱼内脏蛋白的方法,包括原料冻干粉碎、脱脂、蛋白提取、蛋白收集干燥步骤。本发明采用超声辅助提取这一食品加工新技术,有效解决了传统碱溶酸沉法提取时间长、提取率低的问题,具有工艺简单、提取率高、操作安全的优点。应用本发明所述方法,超声辅助提取时间为30～60min,鲍鱼内脏蛋白的提取率高于55％,最高可达到65.87％。本发明扩大了水产加工副产物鲍鱼内脏的消耗途径,促进了其高值化应用。(The invention discloses an ultrasonic-assisted method for extracting abalone viscera protein, which comprises the steps of raw material freeze-drying and crushing, degreasing, protein extraction, protein collection and drying. The invention adopts a new food processing technology of ultrasonic-assisted extraction, effectively solves the problems of long extraction time and low extraction rate of the traditional alkali-soluble acid precipitation method, and has the advantages of simple process, high extraction rate and safe operation. By applying the method disclosed by the invention, the ultrasonic-assisted extraction time is 30-60 min, and the extraction rate of the abalone visceral protein is higher than 55% and can reach 65.87% to the maximum extent. The invention expands the consumption way of the abalone viscera as the by-products in the aquatic product processing and promotes the high-valued application of the abalone viscera.)

1. A method for ultrasonic-assisted extraction of abalone visceral protein is characterized by comprising the following steps:

s1, freeze-drying and crushing raw materials: vacuum freeze-drying abalone viscera, pulverizing, and sieving with 50 mesh or 60 mesh sieve to obtain abalone viscera powder;

s2, degreasing: mixing the abalone viscera powder obtained in the step S1 with a degreasing solvent according to a material-to-liquid ratio of 1 (7-10) g/mL, stirring and leaching at 40-50 ℃ for 3-6 h, replacing the degreasing solvent every 1-2 h, performing suction filtration to collect abalone viscera powder, and volatilizing at room temperature to remove the residual degreasing solvent to obtain degreased abalone viscera powder; wherein the degreasing solvent is prepared by mixing n-hexane and ethanol according to the volume ratio of 3: 1;

s3, protein extraction: mixing the degreased abalone viscera powder obtained in the step S2 with 0.01-0.02 g/mL of NaOH solution according to a material-to-liquid ratio of 1 (15-20) g/mL, performing ultrasonic treatment and stirring for 30-60 min at the temperature of 30-50 ℃ and the ultrasonic power of 300-500W, and centrifuging to collect supernatant;

s4, collecting and drying protein: and (4) adjusting the pH value of the supernatant obtained in the step S3 to 3-4.5 by using a hydrochloric acid solution, standing for 1-2 h at 4-10 ℃, centrifuging, collecting precipitates, and performing vacuum freeze drying on the precipitates to obtain the abalone viscera protein.

2. The method for ultrasonically assisted extraction of abalone visceral protein according to claim 1, wherein the vacuum freeze-drying conditions of step S1 are specifically: the vacuum degree is below 10Pa, and the cold trap temperature is below-40 ℃.

3. The method for ultrasonically assisted extraction of abalone visceral protein according to claim 1, wherein the centrifugation conditions in step S3 are specifically: centrifuging at 8000-10000 g for 30-40 min at 4-10 ℃.

4. The method for ultrasonically assisted extraction of abalone visceral protein according to claim 1, wherein the centrifugation conditions in step S4 are specifically: centrifuging at 8000-10000 g for 10-20 min at 4-10 ℃.

5. The method for ultrasonically assisted extraction of abalone visceral protein according to claim 1, wherein the vacuum freeze-drying conditions of step S4 are specifically: the vacuum degree is below 10Pa, and the cold trap temperature is below-40 ℃.

6. A method for ultrasound-assisted extraction of abalone visceral proteins as claimed in claim 1, wherein the method for ultrasound-assisted extraction of abalone visceral proteins comprises the steps of:

s1, freeze-drying and crushing raw materials: vacuum freeze drying abalone viscera at vacuum degree of 10Pa and cold trap temperature of-40 deg.C, pulverizing with a pulverizer, and sieving with 60 mesh sieve to obtain abalone viscera powder;

s2, degreasing: mixing the abalone viscera powder obtained in the step S1 with a degreasing solvent according to a feed-liquid ratio of 1:10g/mL, stirring and degreasing for 6h at 50 ℃, replacing the degreasing solvent for 1 time every 2h, carrying out suction filtration to collect the abalone viscera powder, and volatilizing the residual degreasing solvent in a fume hood at room temperature to obtain degreased abalone viscera powder; wherein the degreasing solvent is prepared by mixing n-hexane and ethanol according to the volume ratio of 3: 1;

s3, protein extraction: mixing the degreased abalone viscera powder obtained in the step S2 with 0.015g/mL NaOH solution according to a material-to-liquid ratio of 1:20g/mL, carrying out ultrasonic treatment at 40 ℃ and an ultrasonic power of 400W, stirring for 45min, then centrifuging for 30min at 4 ℃ and 10000g, and collecting supernatant;

s4, collecting and drying protein: and (3) adjusting the pH value of the supernatant obtained in the step S3 to 4 by using 1mol/L hydrochloric acid solution, standing for 1h at 4 ℃, centrifuging for 15min at 10000g at 4 ℃, collecting precipitates, and carrying out vacuum freeze drying on the precipitates at the vacuum degree of 1Pa and the cold trap temperature of-50 ℃ to obtain the abalone visceral protein.

Technical Field

The invention belongs to the field of food processing, and particularly relates to a method for ultrasonic-assisted extraction of abalone visceral protein.

Background

Abalone is an important marine economic animal, and the abalone culture quantity of China is the first place in the world. In recent years, the culture quantity is rapidly increased, and the average annual growth rate reaches 39%. The processing and eating of the abalone are mainly carried out by the gastropod, and the viscera accounting for 15-20% of the total mass of the abalone are simply processed into low-value feed and fish meal or directly discarded, so that not only is serious economic loss caused, but also the environment is polluted. The abalone viscera has high nutritive value, contains rich nutritive substances such as protein, amino acid and the like, and has good research value. Therefore, there is a need to enhance the development and utilization of abalone viscera, a by-product of aquaculture processing.

The alkali-soluble acid precipitation method is a common method for extracting protein, but in specific application, the method has the problems of long treatment time and low extraction rate. Aiming at the nutritional characteristics of the abalone viscera, the invention combines ultrasonic wave which is a new food processing technology on the basis of the traditional alkali-dissolving acid-precipitating method, thereby effectively improving and overcoming the defects of the traditional method.

Disclosure of Invention

The invention provides a method for extracting protein from abalone viscera, wherein ultrasonic technology is used for auxiliary extraction in the treatment process, and the method has the advantages of high protein extraction rate and short extraction time, and is beneficial to high-valued utilization of the abalone viscera.

A method for ultrasonic-assisted extraction of abalone visceral protein, comprising the steps of:

s1, freeze-drying and crushing raw materials: vacuum freeze-drying abalone viscera, pulverizing, and sieving with 50 mesh or 60 mesh sieve to obtain abalone viscera powder;

s2, degreasing: mixing the abalone viscera powder obtained in the step S1 with a degreasing solvent according to a material-to-liquid ratio of 1 (7-10) g/mL, stirring and leaching at 40-50 ℃ for 3-6 h, replacing the degreasing solvent every 1-2 h, performing suction filtration to collect abalone viscera powder, and volatilizing to remove the residual degreasing solvent at room temperature to obtain degreased abalone viscera powder; wherein the degreasing solvent is prepared by mixing n-hexane and ethanol according to the volume ratio of 3: 1;

s3, protein extraction: mixing the degreased abalone viscera powder obtained in the step S2 with 0.01-0.02 g/mL of NaOH solution according to a material-to-liquid ratio of 1 (15-20) g/mL, performing ultrasonic extraction and stirring extraction at an extraction temperature of 30-50 ℃, under the condition of an ultrasonic power of 300-500W and a time of 30-60 min, and centrifuging to collect supernatant after extraction is finished; the stirring is carried out in the ultrasonic process in order to uniformly disperse the abalone viscera powder in the solution and prevent the abalone viscera powder from sinking so as to improve the extraction effect;

s4, collecting and drying protein: and (3) adjusting the pH value of the supernatant obtained in the step S3 to 3-4.5 by using 1mol/L hydrochloric acid solution, standing for 1-2 h at 4-10 ℃, centrifuging, collecting precipitate, and performing vacuum freeze drying on the precipitate to obtain the abalone viscera protein.

Preferably, the vacuum freeze-drying conditions in step S1 are specifically: the vacuum degree is not higher than 10Pa, and the temperature of the cold trap is not higher than-40 ℃.

Preferably, the centrifugation conditions in step S3 are specifically: centrifuging at 8000-10000 g for 30-40 min at 4-10 ℃.

Preferably, the centrifugation conditions in step S4 are specifically: centrifuging at 8000-10000 g for 10-20 min at 4-10 ℃; the vacuum freeze-drying conditions are as follows: the vacuum degree is not higher than 10Pa, and the temperature of the cold trap is not higher than-40 ℃.

In a preferred mode, the method for ultrasonically assisting in extracting the abalone visceral protein comprises the following steps:

s1, freeze-drying and crushing raw materials: vacuum freeze drying abalone viscera (vacuum degree 10Pa, cold trap temperature-40 deg.C), pulverizing with pulverizer, and sieving with 60 mesh sieve to obtain abalone viscera powder;

s2, degreasing: mixing the abalone viscera powder obtained in the step S1 with a degreasing solvent according to a feed-liquid ratio of 1:10g/mL, stirring and degreasing for 6h at 50 ℃, replacing the degreasing solvent every 2h for 1 time, performing suction filtration to collect the abalone viscera powder, and volatilizing the residual degreasing solvent in a fume hood at room temperature to obtain degreased abalone viscera powder; wherein the degreasing solvent is prepared by mixing n-hexane and ethanol according to the volume ratio of 3: 1;

s3, protein extraction: mixing the degreased abalone viscera powder obtained in the step S2 with 0.015g/mL NaOH solution according to a material-liquid ratio of 1:20g/mL, performing ultrasonic extraction and stirring under the conditions of 40 ℃, ultrasonic power of 400W and ultrasonic time of 45min, centrifuging for 30min at 4 ℃ at 10000g, and collecting supernatant;

s4, collecting and drying protein: and (4) performing acid precipitation treatment, adjusting the pH of the supernatant obtained in the step S3 to 4 by using 1mol/L hydrochloric acid solution, standing for 1h at 4 ℃, centrifuging for 15min at 10000g at 4 ℃, collecting precipitates, and performing vacuum freeze drying on the precipitates (the vacuum degree is 1Pa, and the cold trap temperature is-50 ℃) to obtain the abalone viscera protein.

The invention has the beneficial effects that:

(1) the invention adopts a new food processing technology of ultrasonic-assisted extraction, and effectively solves the problems of long extraction time and low extraction rate of the traditional alkali-soluble acid precipitation method. The method for extracting the abalone viscera protein has the advantages that the ultrasonic-assisted extraction time is 30-60 min, and the highest protein extraction rate can reach 65.87%.

(2) The invention enriches the development and utilization methods of the abalone viscera which are byproducts of aquatic product processing, and is beneficial to high-valued application.

Detailed Description

The invention is further illustrated by the examples.