WO2019109532A1 - 一种罗汉果果汁及其制备方法 - Google Patents

一种罗汉果果汁及其制备方法 Download PDF

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WO2019109532A1
WO2019109532A1 PCT/CN2018/078213 CN2018078213W WO2019109532A1 WO 2019109532 A1 WO2019109532 A1 WO 2019109532A1 CN 2018078213 W CN2018078213 W CN 2018078213W WO 2019109532 A1 WO2019109532 A1 WO 2019109532A1
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Prior art keywords
juice
han guo
luo han
guo juice
enzyme
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PCT/CN2018/078213
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English (en)
French (fr)
Inventor
杨文国
谢永富
宋云飞
李元元
蒋治舟
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桂林莱茵生物科技股份有限公司
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Priority to US17/059,996 priority Critical patent/US20210227857A1/en
Priority to AU2018379673A priority patent/AU2018379673B2/en
Publication of WO2019109532A1 publication Critical patent/WO2019109532A1/zh

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/80—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter by adsorption
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
    • A23L2/08—Concentrating or drying of juices
    • A23L2/082—Concentrating or drying of juices by membrane processes
    • A23L2/087—Concentrating or drying of juices by membrane processes by ultrafiltration, microfiltration
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes

Definitions

  • the invention relates to the field of food processing, in particular to a mangosteen fruit juice and a preparation method thereof.
  • Chinese patent document CN101167539A discloses a method for producing Luo Han Guo fresh fruit deodorizing concentrated juice, and the steps include: (1) Luo Han Guo crushing slurry; (2) adjusting pH to 3.5-7.5, 10 ° C -60 ° C immobilized pulp enzyme enzymatic hydrolysis 20-60 minutes to make Luo Han Guo juice; (3) ultrafiltration enzyme and ultrafiltration machine to clarify juice; (4) activated carbon deodorization; (5) concentration, pasteurization, ultraviolet or ultra-high temperature sterilization.
  • the comparative document 1 obtains Luo Han Guo juice by low temperature enzymatic hydrolysis of immobilized pulp enzyme, but the enzymatic hydrolysis process takes a long time, and after concentration, high temperature operation such as pasteurization and ultra-high temperature sterilization is adopted, and the heat-sensitive nutrition and flavor components are composed of This loss is more.
  • Chinese patent document CN101283831A discloses a preparation method of decolorized Luo Han Guo juice and a juice prepared by the method, and the steps include: (1) adding pectinase at 37 ° C for the first time for 2 h, adding the filter residue to the first time after initial filtration, etc.
  • the weight pectinase was extracted at the same temperature for 1 h, and the first and second filtrates were filtered again; (2) a supercationic column; (3) a decolorized resin; (4) a first acidification; (5) a concentration; ) The second acidification.
  • the comparison file 2 uses pectinase low temperature enzymatic hydrolysis to obtain mangosteen fruit juice, but it takes a long time, and the long time of enzymatic hydrolysis may cause excessive degradation of nutrients and flavor components by other biological enzymes.
  • the enzymatic hydrolysis of single enzymes of fruit pulp enzyme and pectinase is narrow, and the enzymatic hydrolysis efficiency is not as high as that of pectinase, cellulase, protease, amylase and xylanase.
  • the porridge enzyme is a complex enzyme obtained by solid state fermentation of Aspergillus niger. It is mainly composed of pectinase, cellulase and xylanase, and contains protease and amylase. In the processing of fruits and vegetables, the porridge enzyme can break the fruit, break the plant cells, and make the fruit and vegetable raw materials produce atherosclerosis, thereby increasing the juice yield, clarity and reducing the viscosity of the juice. At present, there is no report on the method for preparing Luo Han Guo juice by enzymatic hydrolysis of Luo Han Guo juice.
  • the efficiency and time of enzymatic hydrolysis are mainly affected by the activity of the enzyme.
  • the higher the activity of the enzyme the shorter the enzymatic hydrolysis time and the higher the efficiency.
  • the activity of the enzyme is affected by the temperature, pH and pressure of the environment. Adjusting the temperature, pH, pressure and other factors of the environment will increase or decrease the activity of the enzyme, thereby affecting the enzymatic hydrolysis time and the enzymatic hydrolysis efficiency.
  • Ultrahigh pressure also known as high hydrostatic press ⁇ re (HHP) refers to the use of water or other liquid as a pressure transmission medium, treated with pressure above 100MPa, at room temperature or even lower temperature
  • HP high hydrostatic press ⁇ re
  • the activity of the enzyme is the activity curve of the ordinate, and there is a peak-shaped region where the activity of the enzyme increases first and then decreases with the pressure increasing (hereinafter referred to as "activation". Area"). Therefore, treatment of enzymes under certain ultra-high pressure conditions can achieve a certain degree of activation of the enzymes.
  • enzymes are highly biospecific, and the activation regions of different enzymes may overlap or may not overlap, and some enzymes may not even exist in the activation region.
  • Temperature is one of the core factors affecting the activity of enzymes. In order to achieve the maximum enzymatic efficiency, it is the premise to achieve the maximum enzymatic efficiency in order to optimize the enzymatic hydrolysis environment.
  • the optimum temperature for enzyme activity is 45 ° C; but for Luo Han Guo, the temperature of 45 ° C causes a loss of some nutrients and flavor components, which ultimately affects the taste of the finished product.
  • the present invention provides a method for preparing Luo Han Guo juice.
  • the method not only adopts ultra-high pressure combined with porridge enzyme to cope with mangosteen, thereby improving enzymatic hydrolysis efficiency, shortening enzymatic hydrolysis time and increasing juice yield; and adopting low-temperature ultra-high pressure sterilization, the sterilization effect can satisfy food consumption, preservation, etc. At the same time as the quality requirements, it can also preserve the nutrients and flavor components in the juice.
  • the invention provides a preparation method of mangosteen fruit juice, characterized in that the fresh fruit of mangosteen fruit is added, and the porridge enzyme is added, and under the condition of ultra-high pressure of 120-220 MPa, the enzymatic hydrolysis of the mangosteen fruit juice is obtained.
  • the applicant of the present invention found in the study that the porridge enzyme was treated under the condition of ultra-high pressure of 120-220 MPa for 10-15 minutes at ⁇ 45 ° C (the recommended temperature of the porridge enzyme product specification), and the activity of each component in the porridge enzyme was observed. At the same time, it was further activated.
  • the residual enzyme activities of pectinase, cellulase, protease, amylase and xylanase were as high as (110.26 ⁇ 0.48)%, (125.72 ⁇ 0.39)%, (108.58 ⁇ 0.16)%. (110.52 ⁇ 0.46)%, (118.69 ⁇ 0.24)%.
  • the fresh fruit of Mangosteen is added, and the porridge enzyme is added, and under the condition of 140-180 MPa ultra-high pressure, the enzymatic hydrolysis of the Luo Han Guo juice is obtained by compressive enzymatic hydrolysis for 12 to 14 minutes.
  • the enzymatic hydrolysis is carried out under conditions of ultrahigh pressure at a temperature of ⁇ 30 °C.
  • the temperature is limited to avoid the destruction of nutrients and loss of flavor components caused by high temperature, effectively maintain the original flavor and nutrients of Luo Han Guo, and improve the taste and nutritional value of the obtained Luo Han Guo juice.
  • the poring enzyme is used in an amount of 0.5 to 1 times of the fresh fruit weight, and the composition ratio is: pectinase 3500-5000 ⁇ g/g, cellulase 100-200 ⁇ /g, amylase 2000-3000 ⁇ /g, Protease 4000-8000 ⁇ /g, xylanase 3500-5000 ⁇ /g.
  • the use of the defined amount and proportion of porridge enzymes effectively promotes the degradation of pectin, cellulose, starch, protein and other components in the Luo Han Guo in a short period of time, prompting the release of the Luo Han juice liquid from the cells.
  • the mogroside fruit juice after enzymatic hydrolysis is subjected to an autoclave of ⁇ 400 MPa for 5 to 10 minutes to obtain a Luo Han Guo juice after the enzyme is deactivated.
  • the applicant of the present invention found in the study that the pressure was further increased to 400 MPa or more, and the activity of each component in the poring enzyme gradually decreased with the increase of the dwell time. Therefore, treatment of the poring enzyme with the defined ultra-high pressure conditions can inactivate the enzyme passivation and achieve the effect of stopping enzymatic hydrolysis.
  • the mogroside fruit juice after the enzyme is removed is treated by a filter press or a high-speed centrifuge to obtain a clarified Luo Han Guo juice.
  • the Luo Han Guo juice obtained after the enzyme is passed through a sieve having a specification of 200 to 400 mesh at an operating pressure of ⁇ 0.2 MPa.
  • a sieve having a specification of 200 to 400 mesh at an operating pressure of ⁇ 0.2 MPa.
  • the rotation speed is 2000-4000 r/min, and the centrifugation time is 5-10 min.
  • the high-speed centrifuge defined, some large particles of impurities can be removed to improve the clarity of the Luo Han Guo juice.
  • the clarified Luo Han Guo juice is taken and added to the yeast for 30 to 480 minutes, and then the yeast is separated to obtain the yeast treated Luo Han Guo juice.
  • yeast treatment the yeast decomposes the disaccharides and monosaccharides in the juice to reduce the sugar and heat.
  • the yeast acts less on the mogroside, reduces the other sugar content in the juice, and can highlight the unique flavor of the mogroside, and improve the taste of the Luo Han Guo juice.
  • the yeast is used in an amount of 1.5 to 2.0 times the fresh fruit weight, and the yeast is fresh yeast or dry yeast.
  • the dried mangosteen juice after the yeast treatment is treated with activated carbon to obtain the Luo Han Guo juice after the activated carbon treatment.
  • the activated carbon is used in an amount of 2.5 times the weight of fresh fruit, has a particle size of 60-120 mesh, and is loaded on a chromatography column having a diameter of ⁇ 5 cm, and the yeast treated mangosteen fruit juice is passed through the chromatography column.
  • the activated carbon and the treatment method the acid, bitter and other odors can be better removed, the flavor of the product can be improved, and the heavy metals and pesticides of the residual materials in the raw materials can be removed to improve the food safety of the product.
  • the activated carbon treated Luo Han Guo juice is passed through an ultrafiltration membrane, filtered and concentrated through a nanofiltration membrane to obtain concentrated Luo Han Guo juice, and the pH of the concentrated Luo Han Guo juice is adjusted to 4.0-6.5, and the pH is weakly acidic. Mangosteen fruit juice.
  • the ultrafiltration membrane has a molecular weight cut off of 50,000 to 60,000 Daltons
  • the nanofiltration membrane has a molecular weight cutoff of 100 to 150 Daltons
  • the concentrated Luo Han Guo juice has a Brix of 5 to 65 degrees. Impurities having a molecular weight > 50,000 Daltons are further removed using the defined ultrafiltration membrane.
  • the defined nanofiltration membrane is used to remove excess liquid for further processing.
  • the Luo Han Guo juice with a pH of weakly acidic is subjected to pressure sterilization for 20 to 30 minutes under an ultra-high pressure condition of temperature ⁇ 30 ° C and 500 to 550 MPa, and the scented Luo Han Guo juice is obtained.
  • the applicant of the present invention found in the study that when the pressure is raised to 500 MPa or more in an environment of temperature ⁇ 30 ° C, the total number of colonies of bacterial microorganisms gradually decreases with the increase of the holding time, and can be the same as the sterilization of high temperature sterilization. Bacterial effect.
  • the use of the defined ultra-high pressure treatment of the slightly acidic Luo Han Guo juice not only the total number of bacterial microbial colonies of various types of bacteria to meet the food requirements, while low temperature operation is also conducive to maintaining the nutrients and flavor components of Luo Han Guo juice.
  • the present invention also provides a mangosteen fruit juice, characterized in that the mangosteen fruit juice is prepared by any of the above methods.
  • the method of the invention has the following advantages:
  • the invention adopts the porridge enzyme to treat the Luo Han Guo, the enzymatic hydrolysis efficiency is higher than that of the single enzyme, the juice time is fast, and the juice yield is high.
  • the present invention increases the activity of each component in the porridge enzyme by ultra-high pressure treatment, and further shortens the juice time and increases the juice yield.
  • the invention accelerates enzymatic hydrolysis, enzyme elimination, sterilization by ultra-high pressure instead of heating means, and operates at low temperature throughout the whole process to avoid damage of nutrients and loss of flavor components caused by high temperature.
  • the invention has short operation time, and mainly adopts pressure regulation as an operating condition, has good process consistency, high production efficiency, and is suitable for industrial production.
  • the amount of activated carbon is 0.5kg, the particle size is 60 mesh, and it is loaded on a column with a diameter of 5 cm, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 50,000 Daltons, and the filtered portion is concentrated to 5 Brix through a nanofiltration membrane having a molecular weight cut off of 100 Daltons to obtain Luo Han Guo juice f;
  • the amount of activated carbon is 0.5kg, the particle size is 90 mesh, and it is loaded on a 6cm diameter chromatography column, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e was passed through an ultrafiltration membrane with a molecular weight cut off of 50,000 Daltons, and the filtered portion was concentrated to 20 Brix through a nanofiltration membrane having a molecular weight cut off of 150 Daltons to obtain Luo Han Guo juice f;
  • the amount of activated carbon is 1.0kg, the particle size is 120 mesh, and is loaded on a 7cm diameter chromatography column, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 60,000 Daltons, and the filtered portion is concentrated to 40 Brix through a nanofiltration membrane having a molecular weight cut off of 150 Daltons to obtain Luo Han Guo juice f;
  • the amount of activated carbon is 1.0kg, the particle size is 120 mesh, and is loaded on a column with a diameter of 8 cm, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is passed through an ultrafiltration membrane with a molecular weight cutoff of 60,000 Daltons, and the filtered portion is concentrated to 50 Brix through a nanofiltration membrane having a molecular weight cut off of 100 Daltons to obtain Luo Han Guo juice f;
  • the amount of activated carbon is 1.25kg, the particle size is 90 mesh, and it is loaded on a column with a diameter of 9cm, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 50,000 Daltons, and the filtered portion is concentrated to 65 Brix through a nanofiltration membrane having a molecular weight cut off of 100 Daltons to obtain Luo Han Guo juice f;
  • the amount of activated carbon is 1.25kg, the particle size is 60 mesh, and is loaded on a column with a diameter of 10 cm, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 60,000 Daltons, and the filtered portion is concentrated to 50 Brix through a nanofiltration membrane having a molecular weight cut off of 150 Daltons to obtain Luo Han Guo juice f;
  • the present comparative example is used to evaluate the technical effect of the technical solution obtained by enzymatic hydrolysis under normal pressure and sterilized by heating means and the technical scheme of the present invention, and the specific steps are as follows:
  • the amount of activated carbon is 1.0kg, the particle size is 120 mesh, and is loaded on a 7cm diameter chromatography column, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 60,000 Daltons, and the filtered portion is concentrated to 60 Brix through a nanofiltration membrane having a molecular weight cut off of 150 Daltons to obtain Luo Han Guo juice f;
  • the present comparative example is used to evaluate the technical effect of the technical solution obtained by enzymatic hydrolysis under normal pressure and sterilized by heating means and the technical scheme of the present invention, and the specific steps are as follows:
  • the amount of activated carbon is 1.0kg, the particle size is 120 mesh, and is loaded on a 7cm diameter chromatography column, so that the Luo Han Guo juice d passes through the chromatography column to obtain Luo Han Guo juice e;
  • Luo Han Guo juice e is subjected to an ultrafiltration membrane with a molecular weight cut off of 60,000 Daltons, and the filtered portion is concentrated to 60 Brix through a nanofiltration membrane having a molecular weight cut off of 150 Daltons to obtain Luo Han Guo juice f;
  • Comparative Examples 3 to 8 were used to evaluate the difference in technical effects between the technical solution for carrying out pressure-holding enzymatic hydrolysis using other high-pressure conditions and the technical solution of the present invention. Comparative examples 3 to 6 were subjected to pressure-holding enzymatic hydrolysis under pressure conditions of 40 MPa, 60 MPa, 80 MPa, 100 MPa, 240 MPa, and 260 MPa, respectively.
  • Comparative Example 9 was used to evaluate the technical effect difference between the Luo Han Guo juice prepared in Comparative Document 1 and the technical solution of the present invention. Specific steps are as follows:
  • Comparative Example 10 was used to evaluate the technical effect difference between the Luo Han Guo juice prepared in Comparative Document 2 and the technical solution of the present invention. Specific steps are as follows:
  • the decolorizing resin was introduced into D208 at a flow rate of 0.5 times (resin) per hour, washed with water after the completion of the feed, and the effluent was collected from the start of sweetness, and the sweetness of the effluent was collected until the sweetness was light.
  • the comparison results are shown in Table 2.
  • the comparison results are shown in Table 3.
  • the mangosteen fruit juice h was prepared according to the methods of Examples 1 to 6 and Comparative Examples 1 and 2, and the microbial content was measured by sampling (5 times in parallel). The comparison results are shown in Table 4.
  • Mangosteen fruit juice h was prepared according to the methods of Examples 1 to 6 and Comparative Examples 1 to 2, 9 to 10, and vitamin C, amino acid, and mogroside were sampled and measured (parallel measurement 5 times). The comparison results are shown in Table 5.
  • Vitamin C (mg/g) Total amino acid (mg/g) Mogroside% Absorbance (460nm, 0.1% w/v)
  • Example 1 0.45 ⁇ 0.18 2.46 ⁇ 0.37 0.51 ⁇ 0.11 0.025
  • Example 2 3.08 ⁇ 0.55 11.23 ⁇ 1.75 1.33 ⁇ 0.18 0.031
  • Example 3 2.79 ⁇ 0.21 15.61 ⁇ 0.97 2.72 ⁇ 0.23 0.107
  • Example 4 6.25 ⁇ 1.25 21.25 ⁇ 2.37 6.80 ⁇ 0.29 0.118
  • Example 5 6.27 ⁇ 1.16 20.58 ⁇ 2.28 8.91 ⁇ 1.76 0.129
  • Example 6 4.93 ⁇ 1.37 16.33 ⁇ 1.29 5.53 ⁇ 0.21 0.115
  • Comparative example 1 2.01 ⁇ 0.35 12.14 ⁇ 1.87 2.52 ⁇ 0.10 0.135
  • Comparative example 2 4.12 ⁇ 1.75 11.52 ⁇ 1.92 2.47 ⁇ 0.11 0.353
  • Comparative example 9 1.24 ⁇ 0.24 8.21 ⁇ 1.25 2.29 ⁇ 0.09 0.330
  • Comparative example 10 0.76 ⁇ 0.18 1.90 ⁇ 0.65 2.
  • the vitamin C and amino acids of the mogroside fruit juice obtained in Examples 1 to 6 of the present invention were significantly improved.
  • the invention adopts ultra-high pressure treatment to activate the porridge enzyme activity, thereby improving the enzymatic hydrolysis effect, and significantly increasing the dissolution content of the target components such as vitamin C, amino acid and mogroside, and then using low temperature ultra-high pressure sterilization to avoid thermal sterilization. Vitamin C and amino acids are destroyed.
  • Mangosteen juice h was prepared according to Examples 1 to 6, 9 to 10, and the amount of juice was measured by sampling. The comparison results are shown in Table 6.
  • the mogroside fruits a to h obtained in Example 3 were each measured for texture.
  • Example 1 Each of the subjects was sequentially administered with the mogroside fruits a to h obtained in Example 1 as a test article, and the total amount was 10 ml, which was completed in 2 to 3 times in 10 minutes. Wait for 15 minutes after each test article is consumed, and continue to consume the next test after the waiting time. Before the start of the experiment and during the waiting period, the taste of the subject was cleaned with pure water and inorganic salt biscuits. The taste of each test article was evaluated from the aspects of characteristic odor, characteristic taste, sweet taste, bitterness, astringency, characteristic aftertaste and the like. For each indicator, 0 is divided into the minimum perceived value, and 6 is divided into the highest perceived value; each result is expressed as an average score.
  • the taste of the Luohan juice a to c is not much different, and there is a distinct sweetness, but there are still outstanding characteristic odors, tastes, aftertastes and bitterness, which affect the overall taste.
  • the Luohan juice d After the yeast treatment, the Luohan juice d, the taste scores except the sweetness are reduced, but the sweet taste is not affected, and the sweet taste can still be felt.
  • the above results show that the use of yeast to treat the Luo Han Guo juice has little effect on the pleasant taste such as sweetness, but it can obviously eliminate the aversive taste such as characteristic smell, taste, aftertaste and bitterness.
  • the technical scheme of the invention has better effects in improving the juice yield of the Luo Han Guo juice, shortening the juice discharge time, preserving the nutrient composition of the Luo Han Guo juice, and improving the overall taste of the Luo Han Guo juice, etc., which are superior to the prior art such as the comparison document 1 and the comparison document 2.

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Abstract

一种罗汉果果汁及其制备方法，所述方法为：取罗汉果鲜果，清洗，破碎，加入粥化酶，在120～220MPa超高压条件下，保压酶解10～15min。

Description

一种罗汉果果汁及其制备方法 技术领域
本发明涉及食品加工领域，特别是一种罗汉果果汁及其制备方法。
背景技术
传统的罗汉果果汁制备方法多采用水加热浸泡提取，近年来也出现了采用更为高效的酶解手段制备罗汉果鲜果汁的报道。如中国专利文件CN101167539A公开了一种罗汉果鲜果脱臭浓缩汁生产方法，步骤包括：(1)罗汉果破碎捣浆；(2)调节pH为3.5-7.5，10℃-60℃固定化果浆酶酶解20-60分钟使罗汉果出汁；(3)超滤酶及超滤机澄清果汁；(4)活性炭脱臭；(5)浓缩、巴氏消毒、紫外或超高温灭菌。所述对比文件1采用固定化果浆酶低温酶解获得罗汉果果汁，但酶解过程耗时较长，且浓缩之后采用了巴氏消毒、超高温灭菌等高温操作，热敏性营养及风味成分由此损失较多。中国专利文件CN101283831A公开了一种脱色罗汉果果汁制备方法及由所述方法制备的果汁，步骤包括：(1)第一次加入果胶酶37℃保温提取2h，初滤后滤渣加第一次等重量果胶酶相同温度保温提取1h，再次过滤合并第一、二次过滤液；(2)过阳离子柱；(3)过脱色树脂；(4)第一道酸化；(5)浓缩；(6)第二道酸化。所述对比文件2采用果胶酶低温酶解获得罗汉果果汁，但耗时更长，且酶解时间过长也会导致营养及风味成分被其他生物酶过度降解。
果浆酶、果胶酶单一酶类的可酶解对象范围较窄，酶解效率不如由果胶酶、纤维素酶、蛋白酶、淀粉酶、木聚糖酶等成分组成的粥化酶高。粥化酶是由黑曲霉经过固态发酵而获得的复合酶,它以果胶酶、纤维素酶、木聚糖酶为主,并含有蛋白酶、淀粉酶等。在果蔬加工中,粥化酶可以溃碎果实，破碎植物细胞，使果蔬原料产生粥样软化，从而提高果蔬汁的出 汁率、澄清度以及降低果汁粘度。目前尚未有以粥化酶酶解罗汉果出汁制备罗汉果果汁的方法的报道。
酶解的效率及时间主要受酶的活性影响，酶的活性越高，则所使用的酶解时间就更短，效率更高。此外，酶的活性受环境的温度、pH、压力所影响，调节环境的温度、pH、压力等因素将会使酶的活性提高或下降，从而影响酶解时间及酶解效率。
超高压处理(μltra high pressμre，ΜHP)又称为高静压处理(high hydrostatic pressμre，HHP)，是指以水或其他液体作为传压介质，经100MPa以上压力处理，在常温甚至更低的温度下达到杀菌、灭酶和改善食品功能特性等作用的新型技术。因此，常规的超高压处理不仅将抑制酶的活性，不利于酶解反应。文献《超高压处理对鲜橙汁中果胶酶及过氧化物酶活性的影响》(陈贺庆等，食品科学[J]，2011(15):54-57)公开了果胶酶在室温15℃，压力200MPa的超高压环境处理10min后，酶活性较常压提高1％～2％，而压力≥300MPa时，酶活性下降30％左右。因此，在一定处理时间下，若以压力为横坐标，酶的活性为纵坐标描绘活性曲线，存在着一个伴随压力不断增大而酶的活性先提高后下降的峰形区域(以下简称“活化区”)。因此，以一定超高压条件处理酶类，可使酶类达到一定程度的活化效果。然而，酶具有高度的生物特异性，不同酶类的活化区有可能重叠，也有可能不重叠，某些酶类甚至不存在所述的活化区。
目前尚未有超高压处理活化粥化酶的公开报道，也没有超高压处理联合粥化酶制备罗汉果果汁的公开报道。
温度作为影响酶类活性的核心因素之一，为了达到最大的酶解效率，使酶解环境处于最佳温度是实现最大酶解效率的前提。对于粥化酶而言，酶活性最高的适宜温度是45℃；但对于罗汉果而言，45℃的温度造成了一部分营养及风味成分的损失，最终影响成品的口感。
发明内容
为了克服现有技术存在的不足，本发明提供一种罗汉果果汁制备方法。所述方法不仅采用了超高压联合粥化酶共同处理罗汉果，从而提高酶解效率，缩短酶解时间，提高果汁得率；更采用低温超高压灭菌，灭菌效果除了能满足食用、保存等质量要求的同时，也能保全果汁中的营养、风味成分。
本发明的目的可以通过以下技术方案来实现：
本发明提供一种罗汉果果汁制备方法，其特征在于，取罗汉果鲜果，加入粥化酶，在120～220MPa超高压条件下，保压酶解10～15min，得酶解后的的罗汉果果汁。
本发明申请人在研究中发现，在＜45℃(粥化酶产品说明书推荐适宜温度)，120～220MPa的超高压条件下处理粥化酶10～15min，观察到粥化酶中各组分活性同时被进一步激活的现象，果胶酶、纤维素酶、蛋白酶、淀粉酶、木聚糖酶的残留酶活分别高达(110.26±0.48)％、(125.72±0.39)％、(108.58±0.16)％、(110.52±0.46)％、(118.69±0.24)％。与温度15℃，常压条件下的粥化酶处理比较，出汁率提高22％，达到相同出汁量所使用的时间减少52％；与温度45℃，常压条件下的粥化酶处理比较，出汁率提高23％，达到相同出汁量所使用的时间减少18％。
优选地，取罗汉果鲜果，加入粥化酶，在140～180MPa超高压条件下，保压酶解12～14min，得酶解后的的罗汉果果汁。
优选地，在温度≤30℃的超高压条件下进行酶解。采用所限定的温度，避免高温导致的营养成分破坏及风味成分损失，有效保持罗汉果原有风味及营养成分，提高所得罗汉果果汁的口感及营养价值。
优选地，所述粥化酶的用量为鲜果重的0.5～1‰倍，组成比例为：果胶酶3500-5000μ/g、纤维素酶100-200μ/g、淀粉酶2000-3000μ/g、蛋白酶4000-8000μ/g、木聚糖酶3500-5000μ/g。采用所限定的用量及比例的粥化酶，有效促使罗汉果中果胶、纤维素、淀粉、蛋白等成分在短时间 内降解，促使罗汉果汁液快速地从细胞中释放。
优选地，取酶解后的的罗汉果果汁，在≥400MPa的超高压条件下，保压灭酶5～10min，得灭酶后的罗汉果果汁。本发明申请人在研究中发现进一步将压力升高至400MPa或以上，粥化酶中各组分活性随保压时间的增加而逐渐下降。因此，采用所限定的超高压条件处理粥化酶，则可使酶钝化失活，达到停止酶解的作用。
优选地，取灭酶后的罗汉果果汁，采用压滤机或高速离心机处理，得澄清的罗汉果果汁。
优选地，采用压滤机处理时，以≥0.2MPa的操作压力使灭酶后得到的罗汉果果汁通过规格为200～400目的滤网。采用所限定的压滤机，能除去部分大颗粒杂质，提高罗汉果果汁的澄明度。
优选地，采用高速离心机处理时，转速2000～4000r/min，离心时间5～10min。采用所限定的高速离心机，能除去部分大颗粒杂质，提高罗汉果果汁的澄明度。
优选地，取澄清的罗汉果果汁，加入酵母处理30～480min，随后分离酵母，得酵母处理后的罗汉果果汁。加入酵母处理，使酵母分解果汁中的二糖、单糖，起到降低糖分及热量的作用。此外，在所限定的处理时间内，酵母较少作用于罗汉果甜苷，降低果汁中其他糖分含量的同时能起到凸显罗汉果甜苷的特有风味，改善罗汉果果汁风味口感的作用。
优选地，所述酵母的用量为鲜果重的1.5～2.0‰倍，所述酵母为鲜酵母或干酵母。
优选地，取酵母处理后的罗汉果果汁，以活性炭处理，得活性炭处理后的罗汉果果汁。
优选地，所述活性炭的用量为鲜果重的2.5‰倍，粒度为60-120目，并装载于直径≥5cm的层析柱上，使酵母处理后的罗汉果果汁通过所述层析柱。采用所限定的活性炭及处理方式，能更好地除去酸、苦等异味，提 高产品的风味口感；同时除去原料中残料的重金属、农药，提高产品的食用安全性。
优选地，使活性炭处理后的罗汉果果汁经过超滤膜，滤过部分再经纳滤膜浓缩，得浓缩的罗汉果果汁，将浓缩的罗汉果果汁的pH调节至4.0～6.5，得pH为弱酸性的罗汉果果汁。
优选地，所述超滤膜的截留分子量为50000～60000道尔顿，所述纳滤膜的截留分子量为100～150道尔顿，所述浓缩的罗汉果果汁Brix为5°～65°。采用所限定的超滤膜，进一步除去分子量＞5万道尔顿的杂质。采用所限定的纳滤膜，除去多余液体，便于进一步的制备加工。
优选地，取pH为弱酸性的罗汉果果汁，在温度≤30℃，500～550MPa的超高压条件下，保压灭菌20～30min，得灭菌后的罗汉果果汁。本发明申请人在研究中发现，在温度≤30℃的环境中将压力升高至500MPa或以上，细菌微生物的菌落总数随保压时间的增加而逐渐减少，并可达到与高温杀菌相同的灭菌效果。因此，采用所限定的超高压处理pH为弱酸性的罗汉果果汁，不仅将各类细菌微生物菌落总数杀灭至符合食品要求，同时低温操作也有利于保持罗汉果果汁的营养物质及风味成分。
本发明还提供一种罗汉果果汁，其特征在于，所述罗汉果果汁由上述任一方法制备获得。
与现有技术比较，本发明的方法具有以下优点：
1.本发明采用粥化酶处理罗汉果，酶解效率较单一酶类高，出汁时间快，出汁率高。
2.本发明以超高压处理提高粥化酶中各成分的活性，继而进一步缩短出汁时间，提高出汁率。
3.本发明以超高压替代加热手段加速酶解、灭酶、灭菌，全程低温操作，避免高温可能导致的营养成分破坏及风味成分损失。
4.本发明操作时间短，并主要以压力调控作为操作条件，工艺连贯性 好，生产效率高，适合工业化生产。
具体实施方式
以下实施例用于说明本发明，但不用来限制本发明的范围。
实施例1
(1)取罗汉果鲜果200kg，清洗，破碎，加入粥化酶0.1kg,在220MPa的超高压条件下，保压酶解10min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶5min，得罗汉果果汁b；
(3)采用压滤机处理罗汉果果汁b，以0.2MPa的操作压力使罗汉果果汁b通过规格为200目的滤网，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.3kg干酵母处理30min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为0.5kg，粒度为60目，装载于直径为5cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为50000道尔顿的超滤膜，滤过部分再经截留分子量为100道尔顿的纳滤膜浓缩至5Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至4.0，得罗汉果果汁g；
(8)取罗汉果果汁g，在500MPa的超高压条件下，保压灭菌30min，得罗汉果果汁h。
上述步骤均在温度15℃的环境下进行。
实施例2
(1)取罗汉果鲜果200kg，清洗，破碎，加入粥化酶0.2kg,在200MPa的超高压条件下，保压酶解11min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶6min，得罗汉果果汁b；
(3)采用压滤机处理罗汉果果汁b，以0.6MPa的操作压力使罗汉果果汁b通过规格为300目的滤网，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.3kg鲜酵母处理120min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为0.5kg，粒度为90目，装载于直径为6cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为50000道尔顿的超滤膜，滤过部分再经截留分子量为150道尔顿的纳滤膜浓缩至20Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至4.5，得罗汉果果汁g；
(8)取罗汉果果汁g，在500MPa的超高压条件下，保压灭菌30min，得罗汉果果汁h。
上述步骤均在温度20℃的环境下进行。
实施例3
(1)取罗汉果鲜果400kg，清洗，破碎，加入粥化酶0.2kg,在180MPa的超高压条件下，保压酶解12min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶7min，得罗汉果果汁b；
(3)采用压滤机处理罗汉果果汁b，以1MPa的操作压力使罗汉果果汁b通过规格为400目的滤网，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.6kg干酵母处理90min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.0kg，粒度为120目，装载于直径为7cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为60000道尔顿的超滤膜，滤过 部分再经截留分子量为150道尔顿的纳滤膜浓缩至40Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至6.5，得罗汉果果汁g；
(8)取罗汉果果汁g，在530MPa的超高压条件下，保压灭菌25min，得罗汉果果汁h。
上述步骤均在温度30℃的环境下进行。
实施例4
(1)取罗汉果鲜果400kg，清洗，破碎，加入粥化酶0.4kg,在160MPa的超高压条件下，保压酶解13min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶8min，得罗汉果果汁b；
(3)采用高速离心机处理罗汉果果汁b，转速2000r/min，离心时间10min，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.8kg鲜酵母处理240min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.0kg，粒度为120目，装载于直径为8cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为60000道尔顿的超滤膜，滤过部分再经截留分子量为100道尔顿的纳滤膜浓缩至50Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至5.5，得罗汉果果汁g；
(8)取罗汉果果汁g，在530MPa的超高压条件下，保压灭菌25min，得罗汉果果汁h。
上述步骤均在温度15℃的环境下进行。
实施例5
(1)取罗汉果鲜果500kg，清洗，破碎，加入粥化酶0.25kg,在140MPa的超高压条件下，保压酶解14min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶9min，得罗汉果果汁b；
(3)采用高速离心机处理罗汉果果汁b，转速3000r/min，离心时间7min，得罗汉果果汁c；
(4)取罗汉果果汁c,加入1.0kg干酵母处理480min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.25kg，粒度为90目，装载于直径为9cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为50000道尔顿的超滤膜，滤过部分再经截留分子量为100道尔顿的纳滤膜浓缩至65Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至6.0，得罗汉果果汁g；
(8)取罗汉果果汁g，在550MPa的超高压条件下，保压灭菌10min，得罗汉果果汁h。
上述步骤均在温度25℃的环境下进行。
实施例6
(1)取罗汉果鲜果500kg，清洗，破碎，加入粥化酶0.5kg,在120MPa的超高压条件下，保压酶解15min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶10min，得罗汉果果汁b；
(3)采用高速离心机处理罗汉果果汁b，转速4000r/min，离心时间5min，得罗汉果果汁c；
(4)取罗汉果果汁c,加入1.0kg鲜酵母处理360min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.25kg，粒度为60目，装载于直径为10cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉 果果汁e；
(6)使罗汉果果汁e经过截留分子量为60000道尔顿的超滤膜，滤过部分再经截留分子量为150道尔顿的纳滤膜浓缩至50Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至5.0，得罗汉果果汁g；
(8)取罗汉果果汁g，在550MPa的超高压条件下，保压灭菌10min，得罗汉果果汁h。
上述步骤均在温度30℃的环境下进行。
对比例1
本对比例用于评价温度为15℃，常压条件下酶解且以加热手段灭菌的技术方案与本发明技术方案取得的技术效果差异，具体步骤如下：
(1)取罗汉果鲜果400kg，清洗，破碎，加入粥化酶0.2kg,在温度为15℃、常压条件下，酶解12min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶10min，得罗汉果果汁b；
(3)采用压滤机处理罗汉果果汁b，以1MPa的操作压力使罗汉果果汁b通过规格为400目的滤网，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.6kg干酵母处理640min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.0kg，粒度为120目，装载于直径为7cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为60000道尔顿的超滤膜，滤过部分再经截留分子量为150道尔顿的纳滤膜浓缩至60Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至5.0，得罗汉果果汁g；
(8)取罗汉果果汁g，以温度120℃处理10s，得罗汉果果汁h。
对比例2
本对比例用于评价温度为45℃，常压条件下酶解且以加热手段灭菌的技术方案与本发明技术方案取得的技术效果差异，具体步骤如下：
(1)取罗汉果鲜果400kg，清洗，破碎，加入粥化酶0.2kg,在温度为45℃，常压条件下，酶解12min，得罗汉果果汁a；
(2)取罗汉果果汁a，在400MPa的超高压条件下，保压灭酶10min，得罗汉果果汁b；
(3)采用压滤机处理罗汉果果汁b，以1MPa的操作压力使罗汉果果汁b通过规格为400目的滤网，得罗汉果果汁c；
(4)取罗汉果果汁c,加入0.6kg干酵母处理60min,随后分离酵母，得罗汉果果汁d；
(5)以活性炭处理罗汉果果汁d，活性炭的用量为1.0kg，粒度为120目，装载于直径为7cm的层析柱上，使罗汉果果汁d通过层析柱，得罗汉果果汁e；
(6)使罗汉果果汁e经过截留分子量为60000道尔顿的超滤膜，滤过部分再经截留分子量为150道尔顿的纳滤膜浓缩至60Brix，得罗汉果果汁f；
(7)加入柠檬酸，将罗汉果果汁f的pH调节至5.0，得罗汉果果汁g；
(8)取罗汉果果汁g，以温度65℃处理30min，得罗汉果果汁h。
对比例3～8
对比例3～8用于评价采用其他高压条件进行保压酶解的技术方案与本发明技术方案取得的技术效果差异。对比例3～6分别以40MPa、60MPa、80MPa、100MPa、240MPa、260MPa的压力条件进行保压酶解，其他步骤同
实施例3。
对比例9
对比例9用于评价对比文件1制备的罗汉果果汁与本发明技术方案取得的技术效果差异。具体步骤如下：
(1)取1吨罗汉果鲜果，制成果浆；
(2)加入柠檬酸调节至pH＝5.5，控制温度为50℃，加入4kg固定化果浆酶；
(3)酶解30分钟后，用板框过滤机滤出果汁，在果汁中加入5g超滤酶，60分钟后，进行超滤；
(4)得到澄清的果汁中加入1kg产活性炭，搅拌5分钟后离心分离活性炭；
(5)进行真空浓缩至50Brix，经超高温瞬间灭菌后包装，制得罗汉果果汁h。
对比例10
对比例10用于评价对比文件2制备的罗汉果果汁与本发明技术方案取得的技术效果差异。具体步骤如下：
(1)取鲜罗汉果400kg，用水清洗、灭活、破碎后，加入0.4kg的果胶酶37℃保温提取2h，初滤、滤渣加4000L去离子水，加入第一次等重量果胶酶、相同温度保温提取1h，再次过滤，合并第一、二次过滤液，再过微滤，收集微滤液，浓缩；
(2)用3倍量去离子水将浓缩后的罗汉果果汁搅拌溶解,以流速0.3倍量(树脂)/每小时进型号为SPC-1阳离子交换树脂，进完料后水洗，从有甜味开始收集流出液，收集到流出液甜味很淡为止；
(3)以流速在0.5倍量(树脂)/每小时进D208脱色树脂，进完料后水洗，从有甜味开始收集流出液，收集到流出液甜味很淡为止。
(4)调节脱色后的果汁PH为6.2；
(5)在真空度0.07MPa，温度50℃，压力为0.04MPa条件下，浓缩脱色罗汉果果汁，将果汁浓缩到60Brix，制得罗汉果果汁h。
理化指标
1.出汁率、出汁时间比较
按实施例1～6及对比例1～2方法制备罗汉果果汁a，计算出汁率(出 汁率＝(罗汉果果汁a质量/罗汉果鲜果质量)×100％)；计算达到20％出汁率时所使用的时间。比较结果见表2。
表2 出汁率、出汁时间比较
  出汁率％ 出汁时间min
实施例1 74.5 20
实施例2 72.8 15
实施例3 71.6 14
实施例4 73.2 18
实施例5 74.1 15
实施例6 77.9 16
对比例1 59.7 42
对比例2 60.5 26
由表2所示可知，与未采用超高压处理的对比例1～2比较，采用超高压处理粥化酶的本发明技术方案实施例1～6出汁率均显著高于对比例1～2，达到20％出汁率时所使用的时间显著少于对比例1～2。说明本发明技术方案采用超高压处理粥化酶的酶解出汁效果优于普通条件下的粥化酶的酶解出汁效果。
2.残留酶活比较
分别按实施例1～6及对比例1～6中的温度、压力条件处理粥化酶，分别测定保压酶解开始前及保压酶解结束后的酶活力(平行测定5次)，根据酶活力计算残留酶活(RA％，RA％＝-A t/A o×100％,A t为经时长tmin高压处理后的酶活Μ/ml,A o为保压时间0min的酶活Μ/ml)。比较结果见表3。
表3 残留酶活比较
Figure PCTCN2018078213-appb-000001
Figure PCTCN2018078213-appb-000002
由表3所示可知，与对比例1～8比较，实施例1～6的粥化酶中各组的残留酶活均显著提高。以上结果表明，在120～220MPa的超高压条件下处理粥化酶10～15min，粥化酶中各组分激活程度分别较低温常压条件、适温常压条件处理显著提高；而无论是低于本发明所限定的压力范围或高于本发明所限定的压力范围，不但不能进一步粥化酶各组分活性，反而随着时间的进行持续抑制粥化酶各组分活化。
3.灭菌效果比较
按实施例1～6及对比例1～2方法制备罗汉果果汁h，取样测定微生物含量(平行测定5次)。比较结果见表4。
表4 灭菌效果比较
Figure PCTCN2018078213-appb-000003
Figure PCTCN2018078213-appb-000004
由表4所示可知，与对比例1～2比较，本发明技术方案实施例1～6的细菌总数等各项指标均无明显差异，说明采用的低温超高压灭菌所取得的灭菌效果与热力灭菌的效果相当，并达到食用要求标准。
4.成分比较
按实施例1～6及对比例1～2、9～10方法制备罗汉果果汁h，取样测定维生素C、氨基酸、罗汉果甜苷(平行测定5次)。比较结果见表5。
表5 成分比较
  维生素C(mg/g) 总氨基酸(mg/g) 罗汉果甜苷％ 吸光度(460nm，0.1％w/v)
实施例1 0.45±0.18 2.46±0.37 0.51±0.11 0.025
实施例2 3.08±0.55 11.23±1.75 1.33±0.18 0.031
实施例3 2.79±0.21 15.61±0.97 2.72±0.23 0.107
实施例4 6.25±1.25 21.25±2.37 6.80±0.29 0.118
实施例5 6.27±1.16 20.58±2.28 8.91±1.76 0.129
实施例6 4.93±1.37 16.33±1.29 5.53±0.21 0.115
对比例1 2.01±0.35 12.14±1.87 2.52±0.10 0.135
对比例2 4.12±1.75 11.52±1.92 2.47±0.11 0.353
对比例9 1.24±0.24 8.21±1.25 2.29±0.09 0.330
对比例10 0.76±0.18 1.90±0.65 2.50±0.11 0.175
由表5所示可知，与对比例1、2、9、10比较，本发明实施例1～6所得罗汉果果汁的维生素C、氨基酸等成分均明显提高。说明本发明采用超高压处理激活粥化酶活性，从而提高酶解效果，使维生素C、氨基酸、罗汉果甜苷等目标成分溶出含量显著增加；随后，采用低温超高压灭菌，避免热力灭菌造成的维生素C、氨基酸被破坏。
5.果汁糖量比较
按实施例1～6、9～10制备罗汉果果汁h，取样测定果汁糖量。比较结 果见表6。
表6 果汁糖量比较
Figure PCTCN2018078213-appb-000005
由表6所示可知，与对比例9、10比较，本发明实施例1～6所得罗汉果果汁的蔗糖、果糖、葡萄糖等含量明显降低。说明本发明采用酵母处理果汁，能降低果汁中糖分及热量。
6.产品口感比较
取实施例3得到的罗汉果果汁a～h,分别测定口感。
每名受试者依次给予实施例1得到的罗汉果果汁a～h作为受试品食用，总量10ml，在10min内分2～3次食用完毕。每样受试品食用完毕后需等待15min，等待时间结束后继续食用下一受试品。实验开始前及等待期间，以纯净水及无机盐饼干清理受试者的味觉。从特征气味、特征口味、甜味、苦味、涩味、特征余味等方面评价各受试品的口感。各项指标，0分为最小感受值，6分为最高感受值；各项结果以平均分表示。
表7 产品口感比较
Figure PCTCN2018078213-appb-000006
由表7所示可知，罗汉果汁a～c的口感差异不大，有较明显的甜味，但仍然有突出的特征气味、口味、余味以及苦涩味，从而影响整体口感。酵母处理后的罗汉果汁d，除甜度外的各项口感评分都降低，但对甜味影响不大，仍然能感受到明显的甜味。以上结果表明，采用酵母处理罗汉果果汁后，对甜味等愉悦型口感影响不大，但能明显祛除特征气味、口味、余味以及苦涩味等厌恶型口感。
结论
本发明技术方案在提高罗汉果果汁出汁率、缩短出汁时间、保全罗汉果果汁营养成分、提高罗汉果果汁整体口感等方面的作用效果均优于对比文件1、对比文件2等现有技术。
虽然，上文中已经用一般性说明、具体实施方式及试验，对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的这些修改或改进，均属于本发明要求保护的范围。

Claims (16)

  1. 一种罗汉果果汁制备方法，其特征在于，取罗汉果鲜果，加入粥化酶，在120～220MPa超高压条件下，保压酶解10～15min，得酶解后的的罗汉果果汁。
  2. 根据权利要求1所述的方法，其特征在于，取罗汉果鲜果，加入粥化酶，在140～180MPa超高压条件下，保压酶解12～14min，得酶解后的的罗汉果果汁。
  3. 根据权利要求1所述的方法，其特征在于，在温度≤30℃的超高压条件下进行酶解。
  4. 根据权利要求1所述的方法，其特征在于，所述粥化酶的用量为鲜果重的0.5～1‰倍，组成比例为：果胶酶3500-5000μ/g、纤维素酶100-200μ/g、淀粉酶2000-3000μ/g、蛋白酶4000-8000μ/g、木聚糖酶3500-5000μ/g。
  5. 根据权利要求1所述的方法，其特征在于，取所述酶解后的的罗汉果果汁，在≥400MPa的超高压条件下，保压灭酶5～10min，得灭酶后的罗汉果果汁。
  6. 根据权利要求5所述的方法，其特征在于，取所述灭酶后的罗汉果果汁，采用压滤机或高速离心机处理，得澄清的罗汉果果汁。
  7. 根据权利要求6所述的方法，其特征在于，采用压滤机处理时，以≥0.2MPa的操作压力使灭酶后得到的罗汉果果汁通过规格为200～400目的滤网。
  8. 根据权利要求6所述的方法，其特征在于，采用高速离心机处理时，转速2000～4000r/min，离心时间5～10min。
  9. 根据权利要求6所述的方法，其特征在于，取所述澄清的罗汉果果汁，加入酵母处理30～480min，随后分离酵母，得酵母处理后的罗汉果果 汁。
  10. 根据权利要求9所述的方法，其特征在于，所述酵母的用量为鲜果重的1.5～2.0‰倍，所述酵母为干酵母或鲜酵母。
  11. 根据权利要求9所述的方法，其特征在于，取所述酵母处理后的罗汉果果汁，以活性炭处理，得活性炭处理后的罗汉果果汁。
  12. 根据权利要求11所述的方法，其特征在于，所述活性炭的用量为鲜果重的2.5‰倍，粒度为60-120目，并装载于直径≥5cm的层析柱上，使酵母处理后的罗汉果果汁通过所述层析柱。
  13. 根据权利要求11所述的方法，其特征在于，使活性炭处理后的罗汉果果汁经过超滤膜，滤过部分再经纳滤膜浓缩，得浓缩的罗汉果果汁，将浓缩的罗汉果果汁的pH调节至4.0～6.5，得pH为弱酸性的罗汉果果汁。
  14. 根据权利要求13所述的方法，其特征在于，所述超滤膜的截留分子量为50000～60000道尔顿，所述纳滤膜的截留分子量为100～150道尔顿，所述浓缩的罗汉果果汁的Brix为5°～65°。
  15. 根据权利要求13所述的方法，其特征在于，取pH为弱酸性的罗汉果果汁，在温度≤30℃，500～550MPa的超高压条件下，保压灭菌20～30min，得灭菌后的罗汉果果汁。
  16. 一种罗汉果果汁，其特征在于，所述罗汉果果汁由权利要求1～15任一所述的方法制备获得。
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