CN100374046C - Method for preparing blueberry mixed juice by using enzymatic engineering tech. - Google Patents
Method for preparing blueberry mixed juice by using enzymatic engineering tech. Download PDFInfo
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Abstract
A method for preparing blueberry mixed juice by using the enzymatic engineering technology relates to the application of the enzymatic engineering technology in the food industry. In the present invention, the blueberry mixed juice is prepared by using the pectic enzyme liquifying technology and the enzymolysis end-point control technology; the pectic enzyme is added to blueberry fruit pulp for enzymolysis reaction by the pectic enzyme liquifying technology; the viscosity and the turbidity of the blueberry enzymolysis extract is controlled by the enzymolysis end-point control technology. The present invention has the advantages of scientific and reasonable process, and easy operation; thus, the blueberry has high juice yield and good quantity; the blueberry mixed juice keeps good colour stability and cloud stability.
Description
Technical Field
A method for preparing blueberry mixed juice by an enzyme engineering technology relates to the application of the enzyme engineering technology in the food industry, in particular to a method for preparing blueberry mixed juice by an enzyme method.
Background
The preparation method of fruit juice is various, and the fruit juice can be extracted by boiling extraction method, squeezing extraction method, enzyme extraction method and the like. With the continuous development of biotechnology, the application of enzymes in the fruit juice beverage industry is more and more extensive. The addition of pectinase preparation or cellulase preparation can help clarify and filter apple juice, grape juice, etc., prevent gel formation in concentrated clear juice, control turbidity of fruit and vegetable juice, etc., and improve quality and stability of fruit and vegetable juice. In the processing of fruit and vegetable juice, the juice extraction by enzyme method has started to gradually replace other juice extraction methods. The application of the enzyme can not only increase the economic benefits of fruit juice production, namely, improve the juice yield and reduce the energy consumption, but also is more specific and effective, can increase the color and flavor of the fruit juice, and has more flexibility.
Blueberries (blueberries) are plants of the genus Vaccinium (Vaccinium spp.) of the family Ericaceae, and are mainly distributed in the temperate regions of the northern hemisphere (cold). Blueberries mainly comprise four types, namely, dwarf (V.angustifolium), tall (V.corombosum), rabbit eye (V.ashei) and half-tall (V.angustifolium. Times.V.corombosum), and the economic life of the blueberries exceeds 50 years. At present, the cultivation area of the blueberries all over the world reaches more than 8 million hectares, wherein the north america accounts for about 90 percent, and the yield is increased from about 4.54 million tons in 1970 to over 13.5 million tons at present. Since the twentieth century, many countries have been successively developing the introduction and artificial cultivation of Blueberries, and China has started to study the introduction of Rabbiteye Blueberries (Rabbiteye Blueberries) and Southern Highbush Blueberries (Southern Highbush Blueberries) by the institute of plant, department of Chinese academy of Jiangsu province at the end of the eighties of the twentieth century. The observation and research for more than ten years shows that the rabbit-eye blueberries have strong adaptability to the acidity of acid soil and the change of soil moisture in the south Jiangsu hills. At present, there are large scale breeding bases in three provinces of Jiangxi, guizhou and Jiangsu. In addition, many companies and research units in China begin to research the planting and product development of blueberries.
The blueberry is the type of blueberry plant with the most abundant nutrient components, the content of natural pigments in fruits is also the highest, the content of anthocyanin and superoxide dismutase (SOD) is many times higher than that of other plants, and the blueberry is determined to be one of five kinds of human health food by grain and agriculture organization (FAO) of the United nations. The blueberry fruit is rich in Ve, va, vb, SOD, free radicals, arbutin, anthocyanin and other rare nutritional ingredients in other fruits, as well as rich potassium, iron, zinc, manganese and other trace elements, protein, edible fiber and fat, has a much higher nutritional value than apples, grapes, oranges and other fruits, can be eaten fresh, can be processed into fruit juice or fruit wine, is the most rapidly developed third-generation fruit tree variety in the world in recent years, and has minced fillet in Europe and America at present.
The blueberry has certain physiological functions and medicinal value. Compared with more than 40 fruits, researchers of human nutrition center (HNRCA) of the United states department of agriculture find that blueberry is the highest in the strong antioxidant capacity of eliminating free radicals of metabolic byproducts of human bodies, namely the blueberry has the effect of resisting human body aging. The anthocyanin in the blueberry fruit has the effects of improving brain memory and intelligence, reducing cholesterol accumulation in human body, improving cardiovascular function, preventing heart disease, preventing and treating urinary tract infection, enhancing collagen, regulating blood sugar, improving night vision, treating diarrhea and the like.
Thus, the blueberry is a novel berry with rich nutrition, and has become a popular research agricultural product in the twenty-first century, japan, europe and America. In the international market, fresh blueberry fruits, blueberry pulp, blueberry health food and other commodities are popular food for consumers. In China, the research and production of blueberry juice still belong to the starting stage, and no commodity is on the market yet. Therefore, the development of high-quality blueberry juice beverage can not only meet the requirements of consumers on natural health-care juice beverage, but also create great economic benefit and social benefit.
For blueberry mixed juice, in the traditional juicing process, the cells cannot be well broken by pulping, and a series of problems of low juice yield, poor color and luster, poor turbidity stability, low anthocyanin retention rate and the like exist. In the literature reports, common enzymes are applied to the production of apple juice and other clear type fruit juice, but reports applied to the production of blueberry mixed juice are not found.
Disclosure of Invention
The invention aims to provide a method for preparing blueberry mixed juice by using an enzyme engineering technology, which applies a pectinase liquefaction technology to ensure that the juice yield of a product is high and better color stability and turbidity stability are maintained.
The technical scheme of the invention is as follows: the blueberry mixed juice prepared by the invention is the blueberry juice which is prepared from blueberry pulp, has higher soluble solid content and pigment dissolution rate, keeps proper viscosity and pectin content and maintains better turbidity stability.
1. The preparation process flow is as follows: the method comprises the following steps of taking frozen blueberries as raw materials, unfreezing at 4 ℃, blanching with steam, cooling and pulping to obtain blueberry pulp, then carrying out enzymolysis reaction, enzyme deactivation, cooling and juicing, centrifuging to obtain blueberry juice, and homogenizing, degassing, filling, sterilizing and cooling to obtain a blueberry mixed juice finished product; the enzymolysis reaction is carried out by applying a pectinase liquefaction technology and an enzymolysis end point control technology, pectinase is added into the blueberry pulp, the addition amount of the pectinase is 0.020 per mill to 0.050 per mill of the mass of the blueberry pulp, the enzymolysis temperature is 45 ℃ to 52 ℃, and the enzymolysis reaction time is 60 minutes to 100 minutes; the viscosity of the blueberry enzymolysis juice is controlled to be 1.70-2.00mPa.s, and the turbidity is controlled to be 250-350NTU.
2. Analytical method
2.1 protein: micro Kjeldahl method.
2.2 Total sugar: phenol-sulfuric acid colorimetry.
2.3 coarse fiber: an acid washing method.
2.4 pectin: m-hydroxybiphenyl colorimetric method.
2.5 soluble solids content (SS): refractometry, using a WZS-1 Abbe refractometer.
2.6 turbidity: measured with a model STZ-A24 turbidimeter.
2.7 juice yield: the net weight of the juice obtained by juicing after enzymolysis is the weight percentage of the added pulp.
2.8 determination of anthocyanin-pH differential method
Weighing appropriate amount of sample, adding 1% (v/v) HCl-methanol extractive solution (material-liquid ratio 1: 10), stirring in 35 deg.C water bath for 1 hr, centrifuging at 3000rpm for 15 min, collecting supernatant, and extracting the residue for 3 times. The supernatant was collected, concentrated in vacuo (40 ℃) to remove methanol, and 1% HCl-H 2 And O constant volume. Two 10mL volumetric flasks were each charged with 1mL of the pigment extract, and the mixture was washed with pH1.0 buffer [0.2 NKCl: 0.2NHCl = 25: 67 (V/V)]And pH4.5 buffer [1 NNaAc: 1 NHCl: H ] 2 O＝100∶60∶90(V/V)]The volume was determined, the mixture was left in a refrigerator for 2 hours, and the absorbance was measured at 510nm and 700nm, respectively. Anthocyanin content Totalantothoconinscontent (T) Acy ) Calculated according to the following formula (the result is calculated by cyanidin-3-glucoside):
A＝(O.D. 510nm -O.D. 700nm ) pH1.0 -(O.D. 510nm -O.D. 700nm ) pH4.5
V-Total volume of extract, mL
m-sample size, g
26900 molar extinction coefficient of cyanidin-3-glucoside
449.2 molar mass of cyanidin-3-glucoside
2.9 determination of Total phenols-Folin-Ciocalteu method (in terms of gallic acid)
A series of concentrations of gallic acid solution were used to plot a standard curve. Taking appropriate amount of sample, adding 1% HCl-methanol extract (ratio of feed to liquid 1: 10), stirring in 35 deg.C water bath for 1 hr, centrifuging at 3000rpm for 15 min, collecting supernatant, and extracting residue repeatedly for 3 times. The supernatant was collected, concentrated in vacuo (40 ℃) to remove methanol, and 1% HCl-H 2 And O constant volume. Diluting the extractive solution to appropriate concentration, placing 1mL into a 100mL brown volumetric flask, adding 60mL deionized water and 5mL LFolin-Ciocalteu reagent, shaking, and adding 20% Na in 0.5-8 min 2 CO 3 Shaking the solution evenly, fixing the volume to 100mL by deionized water, placing the solution in a dark place at the temperature of between 20 and 30 ℃ for 2 hours, and measuring the light absorption value at 765 nm.
2.10 browning index: adding 5mL of blueberry mixed juice and 5mL of 95% ethanol into a centrifuge tube, fully shaking, centrifuging at 2000rpm for 20 minutes, filtering supernatant with filter paper, and measuring absorbance at 420 nm.
2.11 suspension stability: the juice was shaken well and centrifuged at 4200 Xg for 15 minutes, and the supernatant was taken to measure absorbance at 660 nm.
2.12 color index
Measured by a TC-PIIG type full-automatic colorimeter. Wherein: l is * Value representing luminance, L * The larger the value the larger the luminance; a is a * The value indicates the red and green bias of colored substances, the more positive the value is, the more red the bias is, the more negative the value isThe larger the value, the greater the degree of green bias; b * The value indicates the yellow-blue bias of a colored substance, and the more positive the value, the greater the degree of yellow bias, and the more negative the value, the greater the degree of blue bias.
2.13 relative viscosity: measured using an Ostwald viscometer.
2.14 pectinase activity: a spectrophotometer method is used. And drawing a galacturonic acid standard curve. An enzyme solution was prepared using a buffer solution of pH4.2, an appropriate amount of the enzyme solution was reacted at 40 ℃ for 30 minutes, and then developed with 3,5 dinitrosalicylic acid (DNS) and the O.D. value was measured at 520 nm. One unit of enzyme activity is the amount of enzyme required to produce 1ug galacturonic acid per minute (ug/min).
2.15 cellulase activity: a spectrophotometer method is used. And drawing a glucose standard curve. Preparing enzyme solution with phosphate buffer solution of pH4.8, reacting at 40 deg.C for 20 min, developing with 3,5 dinitrosalicylic acid (DNS), and measuring O.D. value at 520 nm. One unit of enzyme activity is the amount of enzyme required per minute to catalyze the production of 1umol of glucose from the substrate (umol/min).
3. Process conditions
3.1 thawing conditions
The frozen blueberries were thawed in a freezer at 4 ℃. Because part of the fruit peel of the blueberries is punctured by ice residues in the freezing process, if the blueberries are thawed at room temperature, the pigments are easily degraded and lost, and the blueberries can keep better color after being thawed at 4 ℃.
3.2 blanching Process
The thawed blueberries are blanched with steam for the following purposes: endogenous polyphenol oxidase in blueberry fruits is passivated, and the oxidative degradation of pigments in the processing process is reduced; the pigment in the blueberry peel and tissues is promoted to be dissolved out, and the pigment content in the juice is increased; the blueberry tissue is softened, the subsequent processing is facilitated, and the juice yield is improved; the microorganisms on the surfaces of the blueberries are killed, and the microbial pollution in the processing process is reduced. The blueberry contains a large amount of anthocyanin, and the hot condition is moderate, so that the purpose can be achieved. The preferred blanching conditions are: blanching in steam for 2-4 min to make the center temperature of blueberry fruit reach above 80 deg.C.
3.3 pectinase liquefaction technology
In the processing of fruit and vegetable juice, a compound enzyme system liquefaction technology of pectinase and cellulase is generally adopted to improve the juice yield, but different enzymes have different influences on the quality of the fruit and vegetable juice. The addition of cellulase to blueberry pulp can slightly improve the juice yield, but has great influence on the turbidity stability of the juice, so that the turbidity of the mixed juice is reduced, and the turbidity stability is reduced. The content of pectin in blueberries is high, and in order to improve the juice yield and the content of soluble solids, the soluble pectin in fruits is liquefied by pectinase generally so as to keep the juice at a certain viscosity, but excessive degradation of the pectin must be avoided, otherwise, the viscosity of the juice is too low, and the turbidity stability of turbid juice is influenced. Therefore, proper enzyme species must be screened, proper enzyme amount is selected, the enzymolysis reaction process is optimized, the juice yield is improved, and the color stability and turbidity stability of the blueberry mixed juice are kept.
1) Influence of different pectinases on the quality of the blueberry mixed juice: most of the pectinase for treating the fruit pulp provided in the market is complex enzyme consisting of pectinase and cellulase, the variety and the dosage of the pectinase have great influence on the quality of the blueberry mixed juice, and the pectinase can be selected for a pectinase formula.
2) Influence of enzyme dosage on blueberry mixed juice quality: generally, the pectinase is a complex enzyme and has pectinase activity and cellulase activity, so the addition amount of the pectinase should be proper.
3) Influence of enzymolysis reaction temperature on blueberry mixed juice quality: an enzymolysis reaction experiment is carried out at the temperature of 35-55 ℃ with relatively stable pectinase activity, and the result shows that the blueberry mixed juice has better quality by adopting the enzymolysis reaction at the temperature of 45-52 ℃.
4) Influence of enzymolysis reaction time on blueberry mixed juice quality: the enzymolysis reaction time is preferably 60-100 minutes.
3.4 enzymolysis end-point control technique
The higher the macromolecular pectin content in the blueberry mixed juice is, the better the turbidity stability of the product is. Therefore, the existence of the macromolecular pectin can not only play a role in stabilizing the blueberry mixed juice, but also endow the blueberry mixed juice with rich nutritional value, and improve other characteristics of the blueberry mixed juice such as turbidity and the like.
The method is to keep the soluble solid content of the blueberry mixed juice, avoid flocculation and have good turbidity stability, and the essence is to control the relative molecular mass distribution of pectin in the blueberry mixed juice and keep the percentage content of macromolecular pectin at a higher level, so that the enzymolysis end point must be controlled. However, in industrial production, the viscosity and the turbidity of the blueberry enzymolysis juice can be controlled only by indirectly controlling the relative molecular mass distribution of pectin so as to effectively control the enzymolysis end point, wherein the viscosity of the blueberry enzymolysis juice is controlled to be 1.70-2.00mPa.s, and the turbidity is controlled to be 250-350NTU.
The quality of the blueberry mixed juice has close relation with the variety of the used enzyme, the enzyme dosage, the enzymolysis reaction time and the temperature, so the comprehensive control should be carried out within the range of the invention.
3.5, enzyme deactivation: heated and stirred at 80-90 ℃ for 5 minutes.
3.6, a sterilization process: the filled blueberry mixed juice is heated in a boiling water bath for 10 minutes, and the sterilized blueberry mixed juice can keep better color stability and turbidity stability within 6 months or even longer.
The invention has the beneficial effects that: the pectinase liquefaction technology decomposes pectin in blueberries through the action of biological enzyme, reduces the viscosity of juice, improves the squeezing performance of blueberry pulp, and improves the juice yield, the content of soluble solid matters and the dissolution rate of pigments. The enzymolysis control technology is to avoid the excessive action of enzyme by controlling the enzymolysis end point so as to keep the proper viscosity and turbidity of the blueberry mixed juice and maintain better turbidity stability. By adopting the two technologies, the tissue cells of the blueberries are broken, the degradation of pectin in the blueberries is controlled, and the effective components of the blueberries are transferred into the fruit juice to keep the color, the fragrance, the taste and the nutritional value of the fruit juice. The dark berry fruits mostly contain nutritional ingredients such as anthocyanin, vitamin, water-soluble mineral substances, cellulose and the like, have good organism balance function, particularly the turbid juice is used as an acidic juice, has important function on maintaining the acid-base balance of human body, and has physiological function irreplaceable for other foods. In addition, berry fruits, especially blueberries, also contain certain special nutrient elements such as malvidin, cyanidin and the like, and have certain prevention and adjuvant treatment effects on certain diseases, for example, the blueberries can prevent myopia, relieve asthenopia, improve the function of the urinary system, prevent and treat urinary tract infection and the like.
After pectinase is added for enzymolysis, the juice yield and the content of soluble solid matters of blueberry mixed juice are improved, but excessive enzymolysis can lead to excessive hydrolysis of macromolecular pectin to generate pectic acid, the content of the macromolecular pectin in the juice is reduced, the viscosity and the turbidity of the juice are reduced, and the stability of suspension of other macromolecular substances in the juice, such as protein, cellulose and partial cell fragments, is not facilitated. Therefore, the end point of the enzyme reaction should be controlled.
Therefore, the pectinase liquefaction technology and the enzyme reaction end point control technology are adopted to develop the color with good color
The blueberry mixed juice with turbidity stability has great economic and social values.
Detailed Description
Example 1
Preparing blueberry mixed juice according to the process route in the specification, wherein the enzymolysis reaction process comprises the following steps: the reaction temperature is 52 ℃, the reaction pH is naturally, the used pectinase is the pectinase RAPIDASE C80MAX, the dosage is 0.050 per thousand of the weight of the blueberries, the enzyme reaction time is 60 minutes, the juice yield is 80.0 percent, the content of soluble solids is 10.20 degrees Brix, the viscosity is 1.80mPa.s, and the turbidity is 320NTU. The fruit juice obtained by enzymolysis under the condition has good color stability and mixed juice stability. The pectinase RAPIDASE C80MAX is supplied by DSM.
Example 2
The operation is the same as example 1, and the enzymolysis reaction process: the reaction temperature is 48 ℃, the reaction time is 100 minutes, the juice yield is 81.7%, the content of soluble solids is 10.10 DEG Brix, the viscosity is 1.76mPa.s, and the turbidity is 255NTU, wherein the used pectinase is pectinase RAPIDASE C80MAX, the dosage is 0.041 per thousand of the mass of the blueberries. The juice obtained by enzymolysis under the condition has good color stability and turbidity stability. The pectinase RAPIDASE C80MAX is supplied by DSM.
Example 3
The operation is the same as example 1, and the enzymolysis reaction process: the reaction temperature is 45 ℃, the natural pH value is reached, the used pectinase is the pectinase RAPIDASE PRESS, the dosage is 0.022 per thousand of the mass of the blueberries, the enzyme reaction time is 80 minutes, the juice yield is 80.5 percent, the content of soluble solids is 10.40 degrees Brix, the viscosity is 1.92mPa & s, and the turbidity is 325NTU. The juice obtained by enzymolysis under the condition has good color stability and turbidity stability. The pectinase RAPIDASE PRESS is provided by DSM.
Claims (3)
1. A method for preparing blueberry mixed juice by using an enzyme engineering technology is characterized in that frozen blueberries are used as raw materials, and blueberry pulp is obtained by unfreezing at 4 ℃, blanching with steam, cooling and pulping, then enzymolysis reaction, enzyme deactivation, cooling and juicing are carried out, and the blueberry juice is obtained by centrifugation, and then a finished product of the blueberry mixed juice is obtained by homogenizing, degassing, filling, sterilizing and cooling; the enzymolysis reaction is carried out by applying a pectinase liquefaction technology and an enzymolysis end point control technology, pectinase is added into the blueberry pulp, the enzyme addition amount is 0.020 per mill to 0.050 per mill of the mass of the blueberry pulp, the enzymolysis temperature is 45 ℃ to 52 ℃, and the enzymolysis reaction time is 60 minutes to 100 minutes; the viscosity of the blueberry enzymolysis juice is controlled to be 1.70-2.00mPa.s, and the turbidity is controlled to be 250-350NTU.
2. The method as claimed in claim 1, wherein the blanching is a blanching in steam for 2-4 minutes to achieve a blueberry core temperature above 80 ℃.
3. The method as claimed in claim 1, wherein the sterilizing is carried out by heating the filled blueberry juice in a boiling water bath for 10 minutes.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102613624A (en) * | 2012-03-02 | 2012-08-01 | 奉化市以勒食品有限公司 | Blueberry fruit juice beverage and preparation method thereof |
| CN107637735A (en) * | 2017-10-31 | 2018-01-30 | 格瑞果汁工业（天津）有限公司 | Composite fruit juice and preparation method thereof made of a kind of blueberry banana |
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| CN102715605B (en) * | 2012-06-28 | 2014-01-15 | 安徽农业大学 | Preparation process of blueberry concentrated juice with high content of anthocyanin |
| CN102793239B (en) * | 2012-08-30 | 2014-04-16 | 山东省农业科学院农产品研究所 | Blueberry/purple sweet potato compound beverage and preparation method thereof |
| CN103230067A (en) * | 2013-05-28 | 2013-08-07 | 贵州苗都酒业有限公司 | Blueberry juice processing method |
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| 蓝莓饮料生产工艺. 陆新龙.食品与机械，第2001年第6期. 2001 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102613624A (en) * | 2012-03-02 | 2012-08-01 | 奉化市以勒食品有限公司 | Blueberry fruit juice beverage and preparation method thereof |
| CN107637735A (en) * | 2017-10-31 | 2018-01-30 | 格瑞果汁工业（天津）有限公司 | Composite fruit juice and preparation method thereof made of a kind of blueberry banana |
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