阅读说明：本技术 酶法辅助水磨同时提取糯米蛋白质和淀粉的方法 (Method for simultaneously extracting glutinous rice protein and starch by enzyme method-assisted water milling ) 是由 何述栋 廖新亚 孙汉巨 李兴江 王家良 于 2021-03-01 设计创作，主要内容包括：酶法辅助水磨同时提取糯米蛋白质和淀粉的方法,包括(1)微气泡气浮清除去杂、(2)第一次研磨、(3)超声波辅助果胶酶处理、(4)细化研磨、(5)微波辅助果胶酶处理、(6)离心、(7)制作糯米蛋白和(8)制作糯米淀粉。本发明采用酶法辅助破坏糯米结构,结合水磨法和离心处理,同时制备糯米蛋白质和淀粉,对后续指导产业化运行具有较好的作用。(A method for simultaneously extracting sticky rice protein and starch by using an enzymatic auxiliary water mill comprises (1) removing impurities by micro-bubble air flotation, (2) grinding for the first time, (3) ultrasonic-assisted pectinase treatment, (4) refining grinding, (5) microwave-assisted pectinase treatment, (6) centrifuging, (7) preparing sticky rice protein and (8) preparing sticky rice starch. The invention adopts an enzyme method to assist in destroying the structure of the glutinous rice, combines a water milling method and centrifugal treatment, simultaneously prepares glutinous rice protein and starch, and has better effect on subsequent guidance of industrialized operation.)

1. A method for simultaneously extracting glutinous rice protein and starch by an enzyme method assisted with water milling is characterized in that: comprises the following steps:

step 1: putting glutinous rice and purified water into a soaking and cleaning container with a micro-bubble generator, starting the micro-bubble generator, keeping the diameter of bubbles at 10-30 mu m, the pressure at 0.60-0.70MPa and the ventilation volume at 200 times of the volume of the purified water per hour, carrying out air floatation treatment for 5-15min, and continuously removing particle impurities floated by the bubbles until no particle impurities float; then taking out the cleaned glutinous rice;

step 2: arranging a spraying device in front of a feed inlet of a grinding device, spraying purified water onto the glutinous rice obtained in the step (1) at a constant speed in a spraying mode, and grinding the glutinous rice to the fineness of 60-80 meshes by the grinding device to obtain glutinous rice pulp;

and step 3: placing the glutinous rice pulp into an enzymolysis device, adding 0.5-1.0% hydrochloric acid solution to adjust pH of the glutinous rice pulp to 4.0-5.0, adding pectinase, keeping the temperature at 45-55 deg.C and the rotation speed at 15-45r/min, and performing enzymolysis for 50-70 min;

and 4, step 4: grinding the mixed material after enzyme treatment to the fineness of less than 50 mu m to obtain a refined mixed material;

and 5: placing the refined mixed material into a microwave oven, and starting the microwave oven once every 5-10 min; the microwave frequency of the microwave oven is 2450MHz, and the radiation intensity is 0.25-0.50w/cm²Each treatment time is 15-30 s;

step 6: treating the mixed material obtained in the step 5 for 5-10min at the rotating speed of 3000-;

and 7: placing the supernatant into vacuum reduced pressure concentration equipment, boiling and concentrating under vacuum gauge pressure of-0.09 to-0.10 MPa to 5-10% of the original volume to obtain a concentrated solution; carrying out vacuum freeze drying on the concentrated solution under the conditions of vacuum gauge pressure of-0.09 to-0.10 MPa and temperature of-60 to-30 ℃ until the water content is lower than 8 percent, thus obtaining the glutinous rice protein; drying the precipitate filter cake with 42-53 deg.C hot air until the water content is less than 12.5% to obtain glutinous rice starch.

2. The method for simultaneously extracting glutinous rice protein and starch by enzymatic auxiliary water milling according to claim 1, wherein: in the step 1, the mass ratio of the glutinous rice to the purified water is 1: 10-12.

3. The method for simultaneously extracting glutinous rice protein and starch by enzymatic auxiliary water milling according to claim 1, wherein: the enzyme activity of the pectinase is 5-20 ten thousand U, and the addition amount of the pectinase is 0.6-1.0 percent of the mass of the glutinous rice pulp.

Technical Field

The invention belongs to the technical field of deep processing of agricultural products, and relates to a method for simultaneously extracting glutinous rice protein and starch by using an enzyme method to assist a water mill.

Background

The method is characterized in that the topography of the abdominal land of Huaihe river basin in Huaiyuan county of Anhui province is mainly river plains, the climate is suitable, and glutinous rice is planted in more than 95% of the whole county in the paddy field, so that the method is the largest glutinous rice production base and transaction scattered land in China and is the leading industry for local farmers to lose poverty and become rich. Glutinous rice is a product obtained by hulling glutinous rice, is commonly called glutinous rice in northern China, and is divided into peduncle glutinous rice and indica glutinous rice according to shape, viscosity and planting area. The glutinous rice is rich in nutrition, and contains abundant starch and protein, as well as abundant metal elements such as calcium, phosphorus, iron, etc., and trace elements such as vitamin B1, vitamin B2, nicotinic acid, etc. In the traditional Chinese medicine, the glutinous rice has the effects of tonifying middle-jiao and Qi, strengthening spleen and nourishing stomach, and moistening and beautifying the skin. Therefore, the glutinous rice food is deeply loved by consumers in China, forms unique glutinous rice dietary culture after thousands of years, is an important raw material for various traditional foods in China, such as rice cakes, eight-treasure porridge, rice dumplings and the like, and is also a main source for brewing yellow rice wine. However, due to the lack of the deep processing capability of glutinous rice in the local area, the advantages of the traditional glutinous rice planting output type area are squeezed along with the enlargement of the glutinous rice planting area in the northeast, the southern Anhui and other areas, and the brand recognition of the 'Huaiyuan glutinous rice' is reduced. On the other hand, with the cognition of the health nutritive value of grain crops such as glutinous rice and the like, glutinous rice protein and starch are used as raw materials to develop health functional food, the formed industrial profit is attractive, and the prospect is huge. However, the related food crops in China are mainly researched by rice, and the research and development reports about the sticky rice are few. The traditional glutinous rice deep processing mainly uses a water milling method to prepare glutinous rice starch, and has the problems of low starch content, large wastewater discharge capacity, high energy consumption and the like.

Disclosure of Invention

The invention aims to provide a method for simultaneously extracting glutinous rice protein and starch by using an enzyme method to assist water milling.

In order to achieve the above objects and other related objects, the present invention provides the following technical solutions: a method for simultaneously extracting glutinous rice protein and starch by using an enzyme method to assist water milling comprises the following steps:

step 1: putting glutinous rice and purified water into a soaking and cleaning container with a micro-bubble generator, starting the micro-bubble generator, keeping the diameter of bubbles at 10-30 mu m, the pressure at 0.60-0.70MPa and the ventilation volume at 200 times of the volume of the purified water per hour, carrying out air floatation treatment for 5-15min, and continuously removing particle impurities floated by the bubbles until no particle impurities float; then taking out the cleaned glutinous rice;

step 2: arranging a spraying device in front of a feed inlet of a grinding device, spraying purified water onto the glutinous rice obtained in the step (1) at a constant speed in a spraying mode, and grinding the glutinous rice to the fineness of 60-80 meshes by the grinding device to obtain glutinous rice pulp;

and step 3: placing the glutinous rice pulp into an enzymolysis device, adding 0.5-1.0% hydrochloric acid solution to adjust pH of the glutinous rice pulp to 4.0-5.0, adding pectinase, keeping the temperature at 45-55 deg.C and the rotation speed at 15-45r/min, and performing enzymolysis for 50-70 min;

and 4, step 4: grinding the mixed material after enzyme treatment to the fineness of less than 50 mu m to obtain a refined mixed material;

and 5: placing the refined mixed material into a microwave oven, and starting the microwave oven once every 5-10 min; the microwave frequency of the microwave oven is 2450MHz, and the radiation intensity is 0.25-0.50w/cm²Each treatment time is 15-30 s;

step 6: treating the mixed material obtained in the step 5 for 5-10min at the rotating speed of 3000-;

and 7: placing the supernatant into vacuum reduced pressure concentration equipment, boiling and concentrating under vacuum gauge pressure of-0.09 to-0.10 MPa to 5-10% of the original volume to obtain a concentrated solution; carrying out vacuum freeze drying on the concentrated solution under the conditions of vacuum gauge pressure of-0.09 to-0.10 MPa and temperature of-60 to-30 ℃ until the water content is lower than 8 percent, thus obtaining the glutinous rice protein; drying the precipitate filter cake with 42-53 deg.C hot air until the water content is less than 12.5% to obtain glutinous rice starch.

The preferable technical scheme is as follows: in the step 1, the mass ratio of the glutinous rice to the purified water is 1: 10-12.

The preferable technical scheme is as follows: the enzyme activity of the pectinase is 5-20 ten thousand U, and the addition amount of the pectinase is 0.6-1.0 percent of the mass of the glutinous rice pulp.

Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:

the invention adopts an enzyme method to assist in destroying the structure of the glutinous rice, combines a water milling method and centrifugal treatment, simultaneously prepares glutinous rice protein and starch, and has better effect on subsequent guidance of industrialized operation.

Drawings

FIG. 1 shows the effect of enzyme species on water milling to simultaneously extract rice starch and protein.

FIG. 2 is a graph showing the effect of enzyme addition on water milling to simultaneously extract rice starch and protein.

FIG. 3 is a graph showing the effect of enzymatic hydrolysis time on water milling to simultaneously extract rice starch and protein.

FIG. 4 is a graph showing the effect of enzymatic pH on water milling to simultaneously extract rice starch and protein.

FIG. 5 is a graph showing the effect of enzymatic hydrolysis temperature on water milling to simultaneously extract rice starch and protein.

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Please refer to fig. 1-5. It should be understood that the structures, ratios, sizes, and the like shown in the drawings and described in the specification are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the present invention without affecting the efficacy and the achievable purpose of the present invention. In addition, the terms "upper", "lower", "left", "right", "middle" and "one" used in the present specification are for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the terms is not to be construed as a scope of the present invention.

Example 1: method for simultaneously extracting glutinous rice protein and starch by enzyme method-assisted water milling

1 materials and methods

1.1 materials and reagents

Glutinous rice raw materials: provided by Anhui Xiangyuan food science and technology Limited, the producing area is in Huaiyuan county of Anhui province, and the harvesting years are 2019. Cellulase (derived from Aspergillus niger, 10000u/g, Shanghai Aladdin Biotechnology Co., Ltd.), and pectinase (derived from Aspergillus niger, 10000u/g, Beijing carbofuran technology Co., Ltd.). All the chemical reagents used were analytical grade.

1.2 instruments and devices

UV-visible spectrophotometer (UV-1800, shanghai nyuan instrument limited), digital display constant temperature water bath (HH-2, guohua (chanzhou) instruments manufacturing limited), small high speed universal pulverizer (FW 100, shanghai chu dingding analytical instruments limited), colloid mill (JMF \ B-80, shanghai kolo mechanical equipment limited), desktop high speed refrigerated centrifuge (TGL 18M, salt city katt experimental instruments limited), electric heat constant temperature blast drying cabinet (DHG-9140A, shanghai essence macro experimental equipment limited), kakikni azotometer (K1160, ji nan hai instruments limited).

And (3) determining the protein content of the sticky rice:

the crude protein content of the raw material and the dried protein powder is measured by adopting a Kjeldahl method (the first method for measuring the protein in GB 5009.5-2016 food safety national standard food).

Protein content in the supernatant was determined by Coomassie blue staining, and standard curve was prepared with the absorbance values at 595 nm of bovine serum albumin stained with Coomassie blue G-250 at a series of concentrations (mg/mL).

Protein extraction rate (%) = mass of protein in water-extracted supernatant (m 1)/mass of protein in raw material (m 2) × 100

And (3) determining the starch content of the glutinous rice:

according to the determination of starch in GB 5009.9-2016 national food safety standard, the content of crude glutinous rice starch in the raw materials is determined by an acid hydrolysis method (second method).

Starch yield (%) = glutinous rice flour cake starch mass (m 1) centrifugally precipitated by water mill/starch mass (m) in raw material₂）×100

And (3) determining the water content of the glutinous rice:

the content of crude water in glutinous rice is measured by a direct drying method (first method) in GB 5009.3-2016 national food safety standard for measuring water in food.

Selection of enzyme species:

washing a certain amount of glutinous rice sample twice with distilled water, coarsely crushing, adding distilled water in a mass ratio of 1:10 to the glutinous rice, adjusting the pH to 4.5, adding cellulase, pectinase, and a mixture of the cellulase and the pectinase (the activity ratio is 1: 1) in an amount which is 0.8% of the weight of the material, performing enzymolysis for 45 min at 50 ℃, grinding the mixture into glutinous rice pulp by using a colloid mill, performing low-temperature high-speed centrifugation treatment to obtain glutinous rice protein and starch products, and determining the content of the protein and the starch.

Determination of enzyme addition amount:

washing a certain amount of glutinous rice sample with distilled water twice, coarsely crushing, adding distilled water in a mass ratio of 1:10 to glutinous rice, adjusting pH to 4.5, adding optimal enzyme types of which the weight is 0.4, 0.6, 0.8, 1.0 and 1.2% of that of the materials respectively, performing enzymolysis for 45 min at 50 ℃, grinding the materials into glutinous rice pulp by using a colloid mill, performing low-temperature high-speed centrifugation treatment to obtain glutinous rice protein and starch products respectively, and determining the content of the protein and the starch.

Determination of enzymolysis time:

washing a certain amount of glutinous rice sample with distilled water twice, coarsely crushing, adding distilled water in a mass ratio of 1:10 to glutinous rice, adjusting pH to 4.5, adding an optimal enzyme type with an optimal material weight, performing enzymolysis at 50 ℃ for 30 min, 45 min, 60min, 75 min and 90 min respectively, milling into glutinous rice pulp by using a colloid mill, performing low-temperature high-speed centrifugation treatment to obtain glutinous rice protein and starch products respectively, and determining the content of protein and starch.

Selection of enzymolysis pH:

washing a certain amount of glutinous rice sample twice with distilled water, coarsely crushing, adding distilled water in a mass ratio of 1:10 to glutinous rice, adjusting the pH to 3.0, 3.5, 4.0, 4.5, 5.0, 5.5 and 6.0, adding the best enzyme type with the best material weight at 50 ℃, performing enzymolysis for the best time, grinding into glutinous rice pulp by using a colloid mill, performing low-temperature high-speed centrifugation treatment to obtain glutinous rice protein and starch products respectively, and determining the content of protein and starch.

Determination of enzymolysis temperature:

washing a certain amount of glutinous rice sample twice with distilled water, coarsely crushing, adding distilled water in a mass ratio of 1:10 to the glutinous rice, adjusting the optimal pH, adding the optimal enzyme type of the optimal material weight, performing enzymolysis at 40, 50 and 60 ℃ for the optimal time respectively, grinding the mixture into glutinous rice pulp by using a colloid mill, performing low-temperature high-speed centrifugation treatment to obtain glutinous rice protein and starch products respectively, and determining the content of the protein and the starch.

A method for simultaneously extracting glutinous rice protein and starch by an enzyme method assisted with water milling is characterized by comprising the following technical steps.

(1) Micro-bubble air floatation impurity removal

The glutinous rice raw material is fresh glutinous rice particles after shelling.

Putting glutinous rice raw materials and purified water into a food-grade soaking and cleaning container with a micro-bubble generator, starting the micro-bubble generator, keeping the diameter of bubbles at 10-30 mu m, the pressure at 0.65MPa and the ventilation volume at 150 times of the volume of the purified water per hour, carrying out air floatation treatment for 10min, and continuously removing particle impurities floated by the bubbles until no particle impurities float; and then taking out the cleaned glutinous rice for later use.

(2) First grinding

Feeding the glutinous rice in the step (1) into food-grade grinding equipment at a constant speed, arranging a spraying device at a feed inlet of the food-grade grinding equipment, spraying food-grade purified water onto the fed glutinous rice at a constant speed in a spraying mode, feeding the glutinous rice and the purified water into the grinding equipment, wherein the water spraying amount is 10 times of the weight of the soaked glutinous rice; fully grinding the materials to the fineness of 60-80 meshes to obtain glutinous rice pulp for later use.

(3) Ultrasonic-assisted pectinase treatment

Putting the glutinous rice pulp obtained in the step (2) into a heat-preservation enzymolysis device with a stirring device, adding a hydrochloric acid solution with the mass fraction of 0.8% to adjust the pH value of the glutinous rice pulp to 4.5, then adding pectinase, keeping the temperature at 50 ℃, rotating speed at 30r/min, and carrying out enzymolysis for 60 min; the enzyme activity of the pectinase is 10 ten thousand U, and the addition amount of the pectinase is 0.8 percent of the mass of the glutinous rice pulp.

(4) Refining grinding

And (4) grinding the mixed material treated by the pectinase in the step (3) by adopting a food-grade colloid mill until the fineness is lower than 50 mu m to obtain a refined mixed material for later use.

(5) Microwave assisted pectinase treatment

Placing the refined mixed material in the step (4) into microwave treatment equipment at the normal temperature, wherein the microwave treatment is started once every 7min, the frequency is 2450MHz, and the radiation intensity is as follows: 0.35w/cm²And each treatment time is 20 s.

(6) Centrifugation

Treating the mixture treated by the pectinase in the step (5) for 6min at the rotating speed of 4000 r/min, and respectively collecting supernatant and a precipitate filter cake; the obtained supernatant is glutinous rice protein solution; the obtained filter cake is rich in glutinous rice starch.

(7) Preparation of glutinous rice protein

And (4) concentrating in vacuum. And (4) placing the supernatant in the step (6) into vacuum reduced pressure concentration equipment, and boiling and concentrating the supernatant to 6% of the original volume under the condition of vacuum gauge pressure of-0.09 to-0.10 MPa to obtain a concentrated solution for later use.

And (5) freeze drying. And (3) carrying out vacuum freeze drying on the concentrated solution under the conditions of vacuum gauge pressure of-0.09 to-0.10 MPa and temperature of-45 ℃ until the water content is lower than 8%, thus obtaining the glutinous rice protein.

(8) Glutinous rice starch

And (4) drying the filter cake precipitated in the step (6) by adopting hot air at 48 ℃ until the water content is lower than 12.5 percent to obtain the glutinous rice starch.

Example 2: method for simultaneously extracting glutinous rice protein and starch by enzyme method-assisted water milling

A method for simultaneously extracting glutinous rice protein and starch by using an enzyme method to assist water milling comprises the following steps:

step 1: putting glutinous rice and purified water into a soaking and cleaning container with a micro-bubble generator, starting the micro-bubble generator, keeping the diameter of bubbles at 10 μm and the pressure at 0.60MPa, wherein the ventilation volume is 100 times of the volume of the purified water per hour, carrying out air floatation treatment for 5min, and continuously removing particle impurities floated by the bubbles until no particle impurities float; then taking out the cleaned glutinous rice;

step 2: arranging a spraying device in front of a feed inlet of a grinding device, spraying purified water onto the glutinous rice obtained in the step (1) at a constant speed in a spraying mode, and grinding the glutinous rice to the fineness of 60 meshes by the grinding device to obtain glutinous rice pulp;

and step 3: putting the glutinous rice pulp into an enzymolysis device, adding a hydrochloric acid solution with the mass concentration of 0.5% to adjust the pH value of the glutinous rice pulp to 4.0, then adding pectinase, keeping the temperature at 45 ℃ and the rotating speed at 15r/min, and carrying out enzymolysis for 50 min;

and 4, step 4: grinding the mixed material after enzyme treatment to the fineness of less than 50 mu m to obtain a refined mixed material;

and 5: placing the refined mixed material into a microwave oven, and starting the microwave oven once every 5 min; the microwave frequency of the microwave oven is 2450MHz, and the radiation intensity is 0.25w/cm²Each processing time is 15 s;

step 6: treating the mixed material obtained in the step 5 for 5min at a rotating speed of 3000r/min, and respectively collecting supernatant and a precipitate filter cake;

and 7: placing the supernatant into vacuum decompression concentration equipment, boiling and concentrating to 5% of the original volume under the vacuum gauge pressure of-0.09 MPa to obtain a concentrated solution; vacuum freeze drying the concentrated solution at-60 deg.C under-0.09 MPa to water content below 8% to obtain glutinous rice protein; drying the precipitate filter cake with 42 hot air until the water content is lower than 12.5% to obtain glutinous rice starch.

The preferred embodiment is: in the step 1, the mass ratio of the glutinous rice to the purified water is 1: 10.

the preferred embodiment is: the enzyme activity of the pectinase is 5 ten thousand U, and the addition amount of the pectinase is-1.0 percent of the mass of the glutinous rice pulp.

Example 3: method for simultaneously extracting glutinous rice protein and starch by enzyme method-assisted water milling

A method for simultaneously extracting glutinous rice protein and starch by using an enzyme method to assist water milling comprises the following steps:

step 1: putting glutinous rice and purified water into a soaking and cleaning container with a micro-bubble generator, starting the micro-bubble generator, keeping the diameter of bubbles at 30 mu m, the pressure at 0.70MPa and the ventilation volume at 200 times of the volume of the purified water per hour, carrying out air floatation treatment for 15min, and continuously removing particle impurities floated by the bubbles until no particle impurities float; then taking out the cleaned glutinous rice;

step 2: arranging a spraying device in front of a feed inlet of a grinding device, spraying purified water onto the glutinous rice obtained in the step (1) at a constant speed in a spraying mode, and grinding the glutinous rice to the fineness of 80 meshes by the grinding device to obtain glutinous rice pulp;

and step 3: putting the glutinous rice pulp into an enzymolysis device, adding 1.0% hydrochloric acid solution by mass concentration to adjust the pH value of the glutinous rice pulp to 5.0, then adding pectinase, keeping the temperature at 55 ℃ and the rotating speed at 45r/min, and carrying out enzymolysis for 70 min;

and 4, step 4: grinding the mixed material after enzyme treatment to the fineness of less than 50 mu m to obtain a refined mixed material;

and 5: placing the refined mixed material into a microwave oven, and starting the microwave oven once every 10 min; the microwave frequency of the microwave oven is 2450MHz, and the radiation intensity is 0.50w/cm²30s of each treatment time;

step 6: treating the mixed material obtained in the step 5 for 10min at the rotating speed of 5000 r/min, and respectively collecting supernatant and a precipitate filter cake;

and 7: placing the supernatant into vacuum decompression concentration equipment, boiling and concentrating to 10% of the original volume under the vacuum gauge pressure of-0.10 MPa to obtain a concentrated solution; vacuum freeze drying the concentrated solution under vacuum gauge pressure of-0.10 MPa and at-30 deg.C until the water content is less than 8% to obtain glutinous rice protein; drying the precipitate filter cake with 53 deg.C hot air until the water content is less than 12.5% to obtain glutinous rice starch.

The preferred embodiment is: in the step 1, the mass ratio of the glutinous rice to the purified water is 1: 12.

the preferred embodiment is: the enzyme activity of the pectinase is 20 ten thousand U, and the addition amount of the pectinase is 0.6 percent of the mass of the glutinous rice pulp.

The foregoing is illustrative of the preferred embodiment of the present invention and is not to be construed as limiting thereof in any way, and any modifications or variations thereof that fall within the spirit of the invention are intended to be included within the scope thereof.