CN111296708A - Method for producing fructus momordicae extract by adopting immobilized enzyme technology - Google Patents
Method for producing fructus momordicae extract by adopting immobilized enzyme technology Download PDFInfo
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- CN111296708A CN111296708A CN202010112185.0A CN202010112185A CN111296708A CN 111296708 A CN111296708 A CN 111296708A CN 202010112185 A CN202010112185 A CN 202010112185A CN 111296708 A CN111296708 A CN 111296708A
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- exchange resin
- momordica grosvenori
- cation exchange
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- 229930189775 mogroside Natural products 0.000 abstract description 11
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- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical group OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 2
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- OHHNJQXIOPOJSC-UHFFFAOYSA-N stevioside Natural products CC1(CCCC2(C)C3(C)CCC4(CC3(CCC12C)CC4=C)OC5OC(CO)C(O)C(O)C5OC6OC(CO)C(O)C(O)C6O)C(=O)OC7OC(CO)C(O)C(O)C7O OHHNJQXIOPOJSC-UHFFFAOYSA-N 0.000 description 2
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
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Classifications
-
- 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
- A23L2/385—Concentrates of non-alcoholic beverages
- A23L2/39—Dry compositions
-
- 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
- A23L2/72—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter by filtration
- A23L2/74—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter by filtration using membranes, e.g. osmosis, ultrafiltration
-
- 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
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J17/00—Normal steroids containing carbon, hydrogen, halogen or oxygen, having an oxygen-containing hetero ring not condensed with the cyclopenta(a)hydrophenanthrene skeleton
- C07J17/005—Glycosides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Water Supply & Treatment (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a method for producing a fructus momordicae extract by adopting an immobilized enzyme technology, which comprises the following steps: extracting with hot water, filtering with ceramic membrane, desalting, decolorizing, performing enzymolysis with immobilized enzyme, and concentrating; if necessary, filtering a part of the enzymolysis liquid obtained in the step (4) by using a nanofiltration membrane, and respectively obtaining the momordica grosvenori condensed juice without mogroside and the momordica grosvenori extract dry powder by using the permeation liquid and the interception. The method of the invention simultaneously considers the production of the momordica grosvenori extract of liquid and solid products with various specifications, can meet various market demands, has high yield of momordica grosvenori glycoside V, and creates a precedent in the industry. Wherein the content of mogroside V in the concentrated momordica grosvenori juice is 3-8 wt%, and the mass content of the mogroside V in the momordica grosvenori extract dry powder is 40-60 wt%.
Description
Technical Field
The invention relates to a production method of a momordica grosvenori extract, in particular to a method for producing the momordica grosvenori extract by an immobilized enzyme technology.
Background
The momordica grosvenori is a famous and precious local specialty of Guilin, is mainly produced in Yongfu county and Longsheng county, and is also one of the first approved medicinal and edible materials in China. The natural high-power sweetener, namely mogroside, contained in the momordica grosvenori has the sweetness of 300 times that of cane sugar, has zero heat, has the effects of clearing heat, moistening lung, relieving cough, moistening intestines and relaxing bowels, and has the prevention and treatment effects on obesity, constipation, diabetes and the like. Mogroside as food is safe and nontoxic, and can be used for various foods in an unlimited way according to the national mandatory standard GB2760 food additive use standard. With the application of advanced extraction, separation and purification technologies, the production and processing of the momordica grosvenori extract and the momordica grosvenori glycoside in China are becoming mature day by day.
At present, the widely accepted momordica grosvenori extracts on the market mainly comprise two types, one is liquid momordica grosvenori condensed juice, and the other is solid momordica grosvenori extract dry powder. The two extracts have different specifications and different production purposes and modes. Producing the concentrated momordica grosvenori juice, aiming at obtaining the water-soluble components of the momordica grosvenori to the maximum extent, aiming at obtaining the finished product of the concentrated momordica grosvenori juice after impurity removal and concentration, wherein the higher the extraction rate of all the water-soluble components is, the better the extraction rate is; the production of the momordica grosvenori extract dry powder (particularly a product with a high-content mogroside V specification) aims to obtain the mogroside to the maximum extent, and aims to reduce the treatment amount and difficulty of subsequent separation and purification and obtain a high-content dry powder finished product, wherein the lower the extraction rate of water-soluble components except the mogroside is, the better the extraction rate is. Therefore, the traditional production method is difficult to meet the requirement of simultaneously obtaining the fructus momordicae extracts with two specifications. The invention provides a method for producing a momordica grosvenori extract product which simultaneously considers liquid and solid products with various specifications.
CN200810097509.7 discloses a method for preparing decolorized fructus momordicae juice and the juice prepared by the method, wherein fresh fructus momordicae is used as raw material, and the fructus momordicae juice is obtained by the steps of material selection, two times of pectinase enzymolysis, cation column passing, decolorization, first acidification, concentration, second acidification and the like. The method has excessive consumption of pectinase, and no targeted treatment on two kinds of macromolecular substances, namely protein and cellulose in the fructus momordicae extract liquid, can lead to the residue of the protein and the cellulose in the fructus momordicae juice, thereby causing the defects of the product in the aspects of taste, solubility, clarity and the like.
CN201610737621.7 discloses a method for preparing honey-flavored momordica grosvenori juice, which takes fresh momordica grosvenori as a raw material, and obtains momordica grosvenori glycoside V dry powder through the steps of raw material selection, extraction, pectinase enzymolysis, resin column adsorption, water washing, ethanol elution, concentration, drying and the like; and (3) carrying out membrane treatment, decoloring, pH value adjustment, concentration and other steps on the effluent of the adsorption resin and the water washing liquid to obtain the honey-flavored momordica grosvenori juice. The method depends on adsorption resin too much, a large amount of flammable ethanol solvent is needed, and the method is not suitable for safe civilized production; similarly, the consumption of pectinase is large, and two kinds of macromolecular substances, namely protein and cellulose in the momordica grosvenori extract are not subjected to targeted treatment, so that the protein and the cellulose are inevitably left in momordica grosvenori extract products (juice and dry powder); in addition, the content of mogroside in the fruit juice is very low.
CN201811397385.4 discloses a method for separating mogroside V, which is to take fresh momordica grosvenori fruits as raw materials, and obtain the mogroside V through the steps of adding water and pulping, carrying out plant enzyme enzymolysis, carrying out biological enzyme enzymolysis, carrying out yeast fermentation, purifying by composite anion and cation exchange resin, concentrating and drying and the like. The method uses too many kinds and dosage of enzyme, the enzymolysis process is complex, and the time is long; the subsequent yeast fermentation process is long in time, and the fermentation liquor is easy to infect mixed bacteria and rancidity, so that the yield is low due to degradation of the stevioside V; in addition, the method has small treatment capacity and is not suitable for industrial continuous batch production.
In the method disclosed above, all enzymes are disposable, cannot be reused, and have high production cost, and the concentrated juice and dry powder of the extract of momordica grosvenori cannot be obtained simultaneously.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects in the prior art and provide a method for producing a momordica grosvenori extract by using an immobilized enzyme which has the advantages of small enzyme dosage, cyclic and repeated use, good enzymolysis effect, low production cost, no use of macroporous adsorption resin and no use of organic solvent, and can simultaneously give consideration to three specifications of momordica grosvenori condensed juice, momordica grosvenori extract dry powder and momordica grosvenori condensed juice byproducts without momordica grosvenori sweet glycosides.
The technical scheme adopted by the invention for solving the technical problems is as follows: a method for producing a fructus momordicae extract by adopting an immobilized enzyme technology comprises the following steps:
(1) hot water extraction: crushing fresh fructus Siraitiae Grosvenorii, percolating with hot water, and cooling to obtain extractive solution;
(2) ceramic membrane filtration: filtering the fructus Siraitiae Grosvenorii extractive solution with ceramic membrane, and collecting the filtrate;
(3) desalting and decoloring: sequentially passing the ceramic membrane filtrate through a first anion exchange resin and a first cation exchange resin which are connected in series to obtain a desalting and decolorizing solution;
(4) carrying out enzymolysis on immobilized enzyme: passing the desalted and decolored liquid through an immobilized enzyme packed bed to obtain an enzymolysis liquid;
(5) concentration: and concentrating the enzymolysis liquid under reduced pressure to obtain the momordica grosvenori concentrated juice.
In a preferred technical scheme of the invention, in order to obtain the momordica grosvenori extract dry powder, the method further comprises the following steps of:
(6) and (4) nanofiltration: filtering a part or all of the enzymolysis liquid obtained in the step (4) by using a nanofiltration membrane, collecting nanofiltration membrane trapped fluid, and concentrating nanofiltration membrane permeate under reduced pressure until the sugar degree is 66brix to obtain a byproduct, namely momordica grosvenori concentrated juice without momordica grosvenori sweet glycosides;
(7) concentrating and drying: concentrating the nanofiltration membrane retentate under reduced pressure, and spray drying to obtain fructus Siraitiae Grosvenorii extract dry powder.
Preferably, in the step (1), the temperature of the hot water is 70-90 ℃, the consumption of the hot water is 1-2 times of the weight of the fresh momordica grosvenori, and the percolation flow rate is 0.2-1.0 BV/h. Cooling by plate heat exchanger to room temperature to obtain fructus Siraitiae Grosvenorii extractive solution. One purpose of the percolation extraction by hot water is to leach out the water-soluble components contained in the fresh fructus momordicae to the maximum extent and ensure the extraction yield of the water-soluble components including mogrosides, and the other purpose is to inactivate and denature the proteins (enzymes and the like) in the fresh fructus momordicae by utilizing the high-temperature condition so as to facilitate the subsequent filtration and removal. If the temperature of the hot water is too low, the dosage is too small or the percolation flow rate is too high, the purpose cannot be fully achieved; if the temperature of the hot water is too high, the dosage is too high or the percolation flow rate is too low, the waste of energy or materials is caused.
Preferably, in the step (3), the kind of the anion exchange resin is macroporous type strongly basic anion exchange resin, and the specific models are LSD762, D945 and LSA-700B, D941.
Preferably, in the step (3), the total volume consumption of the anion exchange resin is 0.1-0.2 times (L/Kg) of the weight of the fresh fructus momordicae, the height-diameter ratio of the anion exchange resin column is 5-8: 1, and the flow rate of the materials passing through the anion exchange resin column is 4-6 BV/h. The purpose of using an anion exchange resin column is decolorization. If the amount of the anion exchange resin is too small, the height-diameter ratio of the resin column is too small, or the material flow rate is too high, the above-mentioned object cannot be sufficiently achieved. If the dosage of the anion exchange resin is too much, the height-diameter ratio of the anion exchange resin column is too large or the flow rate is too slow, the waste of energy or materials is caused.
Preferably, in the step (3), the cation exchange resin is styrene type strongly acidic cation exchange resin, and the specific types are 001 × 16, 001 × 7, 001 × 12 and 001 × 8.
Preferably, in the step (3), the total volume consumption of the cation exchange resin is 0.1-0.2 times (L/Kg) of the weight of the fresh fructus momordicae, the height-diameter ratio of the cation exchange resin column is 5-8: 1, and the flow rate of the material passing through the cation exchange resin column is 4-6 BV/h. The purpose of using a cation exchange resin column is desalting. If the amount of the cation exchange resin used is too small, the aspect ratio of the cation exchange resin column is too small, or the flow rate of the material passing through the cation exchange resin column is too high, the above-mentioned object cannot be sufficiently achieved. If the dosage of the cation exchange resin is too much, the height-diameter ratio of the cation exchange resin column is too large or the flow rate of the material passing through the cation exchange resin column is too slow, the waste of energy or material is caused.
Preferably, in step (3), the feed liquid passes through the first anion exchange resin and the first cation exchange resin, and then passes through the second anion exchange resin and the second cation exchange resin.
In the preferred technical scheme of the invention, the ceramic membrane filtrate sequentially passes through four series-connected anion-cation-anion-cation ion exchange resin columns, and one of the purposes is that materials alternately and repeatedly pass through two groups of anion-cation ion exchange resin columns, so that the leakage of pigments and salts can be avoided when the materials pass through the resin columns at a high speed, the production speed is improved, and the desalting and decoloring effects can be guaranteed; the second purpose is that the second anion exchange resin and the second cation exchange resin do not reach the exchange saturation after being used in each batch, still have the decoloring and desalting capabilities, do not need to be regenerated, and can be used as the first anion exchange resin and the first cation exchange resin in the step (3) of the next batch of materials, so that the utilization rate of the resins can be improved, the regeneration frequency of the resins is reduced, and the purposes of reducing sewage discharge and reducing the production cost are achieved.
In the step (3), the order of the anion exchange resin column and the cation exchange resin column can not be changed, one reason is that only the last ion exchange resin is the cation exchange resin column, the pH values of the desalted and decolored liquid, the momordica grosvenori condensed juice and the momordica grosvenori extract dry powder can be ensured to be slightly acidic, and the requirements of the taste of consumers are met; the second reason is that the optimum pH value of the subsequent enzymolysis step of the immobilized enzyme is slightly acidic, so that the speed and the efficiency of enzymolysis can be improved.
Preferably, in step (4), the carrier of the immobilized enzyme is polysaccharide ion exchanger or synthetic polymer ion exchange resin, such as: diethylaminoethyl (DEAE) -cellulose, mixed amine (ECTEDLA) -cellulose, Tetraethylaminoethyl (TEAE) -cellulose, DEAE-sephadex, Amberlite IRA-93, 410, 900, Carboxymethyl (CM) -cellulose, cellulose-citrate, Amberlite CG-50, IRC-50, IR-200 and Dowex-50, wherein the dosage of the immobilized enzyme carrier is 0.05-0.1 time (L/Kg) of the weight of the fresh fructus momordicae. The purpose of using the immobilized enzyme carrier is to fix soluble enzyme on a water-insoluble carrier through ionic bonds, so that the local enzyme concentration of an enzymolysis reaction can be improved, the enzymolysis speed and efficiency are improved, the cyclic use and continuous production can be realized, enzyme deactivation and removal processes in the traditional enzymolysis method are not needed, and the operation is simplified.
Preferably, in the step (4), the enzyme is a compound of pectinase and cellulase, the dosage of the pectinase is 0.1-0.5 per mill of the weight of the fresh momordica grosvenori, and the dosage of the cellulase is 0.1-0.5 per mill of the weight of the fresh momordica grosvenori. The purpose of using pectinase and cellulase is to hydrolyze pectin and cellulase in the desalted and decolored solution into small molecular substances, so that the problem that precipitates are separated out when the momordica grosvenori condensed juice is stored for a long time can be solved, and the small molecular substances obtained after hydrolysis of the pectin and the cellulase can be removed through a subsequent nanofiltration step to obtain the momordica grosvenori extract dry powder with higher content of stevioside.
Preferably, in the step (4), the height-diameter ratio of the immobilized enzyme packed bed is 6-10: 1, and the flow speed of materials passing through the immobilized enzyme packed bed is 1-3 BV/h.
In the step (4), the temperature for enzymolysis is not particularly limited, and generally, the temperature at which the pectinase and cellulase exert activity is suitable is only required, and is generally 20-40 ℃.
In the step (4), the preparation of the enzyme immobilization is well known in the art, the enzyme is dissolved in warm water, the enzyme is activated for 20-30 minutes to obtain an enzyme solution, the carrier is added into the enzyme solution, the enzyme solution is stirred uniformly at a low speed, and the enzyme solution is kept stand for 2-4 hours. And finally, filling the mixture of the carrier and the enzyme liquid into a packed bed, and washing with deionized water to obtain the immobilized enzyme.
Preferably, in the step (5), the sugar degree of the momordica grosvenori condensed juice is 50-70 brix, and the content of momordica grosvenori glycoside V in the momordica grosvenori condensed juice is 3% -8%.
Preferably, in the step (6), the molecular weight cut-off of the nanofiltration membrane is 800-1000 Da. The nanofiltration membrane is used for filtering, and is used for removing pectin and micromolecular substances after cellulose hydrolysis in the enzymolysis liquid and micromolecular substances such as glucose, fructose and the like in water-soluble components of the momordica grosvenori so as to improve the content of the momordica grosvenori extract.
Preferably, in the step (7), the mogroside V content of the momordica grosvenori extract dry powder is 40-60% by mass.
In the method of the invention, 1BV is equal to 1 column volume.
The method has the following beneficial effects:
(1) the method of the invention simultaneously considers the production of the momordica grosvenori extract of liquid and solid products with various specifications, can meet various market demands, has high yield of momordica grosvenori glycoside V, and creates a precedent in the industry. Wherein the content of mogroside V in the concentrated momordica grosvenori juice is 3-8 wt%, and the mass content of the mogroside V in the momordica grosvenori extract dry powder is 40-60 wt%.
(2) The invention introduces the immobilized enzyme technology into the production field of the momordica grosvenori extract for the first time, the dosage of the enzyme is small, the enzyme can be recycled, meanwhile, the activity of the enzyme is not influenced, the enzymolysis effect is good, the enzymolysis speed and efficiency are improved, the production cost is reduced, and reference is provided for the popularization of the immobilized enzyme technology in the production field of natural plant products.
(3) The method does not need macroporous adsorption resin, has simple process, no use of any organic solvent, safety, greenness, environmental protection, low production cost, strong operability and suitability for industrial production, and can realize cyclic utilization and continuous operation.
(4) The nanofiltration membrane permeate liquid can be concentrated to obtain an important byproduct, namely momordica grosvenori concentrated juice without momordica grosvenori glycoside, the momordica grosvenori concentrated juice is composed of micromolecular saccharides (glucose, fructose and the like) from momordica grosvenori, main components of the momordica grosvenori concentrated juice are similar to honey, the momordica grosvenori concentrated juice has high nutritional and economic values, and a new channel is provided for comprehensive utilization of momordica grosvenori resources.
Detailed Description
The present invention will be further described with reference to the following examples.
The fresh momordica grosvenori fruits used in the embodiment of the invention are purchased from Guangxi Guilin, and the content of mogroside V in the fresh momordica grosvenori fruits is 0.52%; the anion and cation exchange resins used in the embodiment of the invention are purchased from Xian lan Xiao science and technology New materials Co., Ltd; the pectinase and cellulase used in the embodiment of the invention are purchased from Novoxin (China) Biotechnology limited; the auxiliary materials used in the examples of the present invention are obtained by conventional commercial methods unless otherwise specified.
In the embodiment of the invention, the content of mogroside V is detected by adopting a High Performance Liquid Chromatography (HPLC) external standard method.
Example 1
(1) Hot water extraction: taking 300kg of fresh fructus momordicae, crushing, putting into a percolator, percolating and extracting with 600kg of hot water at the temperature of 80 ℃, wherein the percolating flow rate is 0.3 BV/h, and cooling the percolate to room temperature by using a plate heat exchanger to obtain fructus momordicae extract;
(2) ceramic membrane filtration: filtering the fructus Siraitiae Grosvenorii extractive solution with ceramic membrane, and collecting the filtrate;
(3) desalting and decoloring: sequentially passing the ceramic membrane filtrate through four serially connected anion-cation-anion-cation ion exchange resin columns (a first anion exchange resin, a first cation exchange resin, a second anion exchange resin and a second cation exchange resin), wherein the type of the anion exchange resin is LSD762, the total dosage of the anion exchange resin is 45L, each column is 22.5L, the height-diameter ratio of each anion exchange resin column is 8:1, and the flow rate of materials passing through each anion exchange resin column is 5 BV/h; the model of the cation exchange resin is 001 multiplied by 16, the total dosage of the cation exchange resin is 45L, each column is 22.5L, the height-diameter ratio of each cation exchange resin column is 8:1, and the flow rate of materials passing through each cation exchange resin column is 5 BV/h, so as to obtain a desalting and decolorizing solution;
(4) carrying out enzymolysis on immobilized enzyme: passing the desalted and decolored liquid through a cylindrical immobilized enzyme packed bed with the height-diameter ratio of 8:1 (wherein the carrier of the immobilized enzyme is DEAE-cellulose, the using amount of the immobilized enzyme carrier is 30L, the packed bed contains pectinase and cellulase, the using amount of the pectinase is 0.06kg, and the using amount of the cellulase is 0.09kg) at the flow rate of 1.5 BV/h to obtain an enzymolysis liquid;
(5) concentration: taking half of the enzymolysis liquid in the step (4), and concentrating under reduced pressure until the sugar degree is 65brix to obtain 19.23kg of fructus momordicae concentrated juice with light yellow color;
(6) and (4) nanofiltration: taking the other half of the enzymolysis liquid in the step (4), filtering by using a nanofiltration membrane with the molecular weight cutoff of 800Da, and collecting nanofiltration membrane cutoff liquid for later use; concentrating the nanofiltration membrane permeate under reduced pressure to sugar degree of 66brix to obtain byproduct, namely fructus Siraitiae Grosvenorii concentrated juice containing no mogroside, 17.51 kg;
(7) concentrating and drying: concentrating the nanofiltration membrane retentate under reduced pressure, and spray drying to obtain fructus Siraitiae Grosvenorii extract dry powder 1.47kg, which is yellow-white.
Through detection of a High Performance Liquid Chromatography (HPLC) external standard method, the content of mogroside V in the obtained fructus momordicae concentrated juice is 4.03%, and the content of mogroside V in the fructus momordicae extract dry powder is 51.91%; the total yield of mogroside V was 98.6%.
Example 2
(1) Hot water extraction: taking 100kg of fresh fructus Siraitiae Grosvenorii, crushing, putting into a percolator, percolating with 200kg of 85 deg.C hot water at percolation flow rate of 0.5 BV/hr, and cooling the percolate to room temperature with a plate heat exchanger to obtain fructus Siraitiae Grosvenorii extract;
(2) ceramic membrane filtration: filtering the fructus Siraitiae Grosvenorii extractive solution with ceramic membrane, and collecting the filtrate;
(3) desalting and decoloring: sequentially passing the ceramic membrane filtrate through four serially connected anion-cation-anion-cation ion exchange resin columns (a first anion exchange resin, a first cation exchange resin, a second anion exchange resin and a second cation exchange resin), wherein the type of the anion exchange resin is D945, the total dosage of the anion exchange resin is 20L, the height-diameter ratio of each anion exchange resin column is 6:1, and the flow rate of the material passing through each anion exchange resin column is 4 BV/h; the model of the cation exchange resin is 001 multiplied by 7, the total dosage of the cation exchange resin is 20L, the height-diameter ratio of each cation exchange resin column is 6:1, and the flow rate of materials passing through each cation exchange resin column is 4 BV/h, so as to obtain a desalting and decolorizing solution;
(4) carrying out enzymolysis on immobilized enzyme: passing the desalted and decolored liquid through a cylindrical immobilized enzyme packed bed with the height-diameter ratio of 10:1 (wherein the carrier of the immobilized enzyme is Amberlite CG-50, the using amount of the immobilized enzyme carrier is 10L, the packed bed contains pectinase and cellulase, the using amount of the pectinase is 0.02kg, and the using amount of the cellulase is 0.03kg) at the flow rate of 2 BV/h to obtain an enzymolysis liquid;
(5) and (4) nanofiltration: filtering the enzymolysis liquid obtained in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 1000Da, and collecting nanofiltration membrane cutoff liquid;
(6) concentrating and drying: concentrating the nanofiltration membrane retentate under reduced pressure, and spray drying to obtain fructus Siraitiae Grosvenorii extract dry powder 0.95kg in yellow-white color.
The content of mogroside V in the obtained dry powder of the momordica grosvenori extract is 53.70% by detection of a High Performance Liquid Chromatography (HPLC) external standard method; the yield of mogroside V was 98.1%.
Example 3
(1) Hot water extraction: taking 200kg of fresh fructus Siraitiae Grosvenorii, crushing, putting into a percolator, percolating and extracting with 300kg of 90 deg.C hot water at a percolation flow rate of 0.2 BV/hr, and cooling the percolate to room temperature with a plate heat exchanger to obtain fructus Siraitiae Grosvenorii extract;
(2) ceramic membrane filtration: filtering the fructus Siraitiae Grosvenorii extractive solution with ceramic membrane, and collecting the filtrate;
(3) desalting and decoloring: sequentially passing the ceramic membrane filtrate through four serially connected 'anion-cation-anion-cation' ion exchange resin columns (wherein the type of anion exchange resin is LSA-700B, the total dosage of the anion exchange resin is 40L, the height-diameter ratio of each anion exchange resin column is 7:1, the flow rate of the material passing through each anion exchange resin column is 5.5 BV/h, the type of cation exchange resin is 001 x 12, the total dosage of the cation exchange resin is 40L, the height-diameter ratio of each cation exchange resin column is 7:1, and the flow rate of the material passing through each cation exchange resin column is 5.5 BV/h to obtain a desalting and decolorizing solution;
(4) carrying out enzymolysis on immobilized enzyme: passing the desalted and decolored liquid through a cylindrical immobilized enzyme packed bed with a height-diameter ratio of 9:1 (wherein, the carrier of the immobilized enzyme is Dowex-50, the dosage of the immobilized enzyme carrier is 20L, the packed bed contains pectinase and cellulase, the dosage of the pectinase is 0.04kg, and the dosage of the cellulase is 0.04kg) at the flow rate of 1 BV/h to obtain an enzymolysis liquid;
(5) concentration: concentrating the enzymolysis solution under reduced pressure to sugar degree of 67brix to obtain fructus Siraitiae Grosvenorii concentrated juice 24.66kg, which is light yellow.
The content of mogroside V in the obtained fructus momordicae concentrated juice is 4.20% by detecting with a High Performance Liquid Chromatography (HPLC) external standard method; the yield of mogroside V was 99.6%.
Example 4
The other conditions, procedure and example 1 were the same except that the first anion exchange resin and the first cation exchange resin used in step (3) were derived from the second anion exchange resin and the second cation exchange resin after use in example 1. 19.18kg of fructus momordicae concentrated juice, 17.51kg of fructus momordicae concentrated juice without mogroside and 1.45kg of fructus momordicae extract dry powder are finally obtained, and the content of mogroside V in the fructus momordicae concentrated juice obtained in the embodiment is 3.97 percent and the content of mogroside V in the fructus momordicae extract dry powder is 51.75 percent through the detection of a High Performance Liquid Chromatography (HPLC) external standard method; the total yield of mogroside V was 96.9%.
In the desalting and decoloring step of this example, the first group of anion exchange resin and cation exchange resin in four "anion-cation-anion-cation" ion exchange resin columns connected in series was used as the second group of anion exchange resin and cation exchange resin, and it can be seen that, although the used ion exchange resin without regeneration was used, the desalting and decoloring ability was not decreased, and satisfactory exchange efficiency was maintained. Has no obvious adverse effect on the content and yield of mogroside V in the final product. The invention is illustrated by the advancement of using four series-connected anion-cation-anion-cation ion exchange resin columns.
Example 5
The other conditions and steps are the same as those in example 1, except that in the step (4), the dosage of the immobilized enzyme carrier DEAE-cellulose is 15L, the dosage of the pectinase is 0.03kg, and the dosage of the cellulase is 0.05kg, so that 19.25kg of the fructus momordicae concentrated juice and 1.46kg of the dry powder of the fructus momordicae extract are finally obtained, and the content of mogroside V in the fructus momordicae concentrated juice obtained in the embodiment is 3.97% and the content of the mogroside V in the dry powder of the fructus momordicae extract is 50.69% through detection of a High Performance Liquid Chromatography (HPLC) external standard method; the total yield of mogroside V was 96.4%.
Example 6
The other conditions and steps are the same as those in example 1, except that in step (4), the used immobilized enzyme packed bed is repeatedly recycled for 5 times, and the content and yield of the concentrated juice and the dry powder of momordica grosvenori are recorded each time. The results are shown in table 1 below:
TABLE 1
| Product(s) | For the first time | For the second time | The third time | Fourth time | Fifth time |
| Sweet glycoside V content (wt%) in momordica grosvenori concentrated juice | 4.03 | 4.03 | 3.97 | 3.94 | 3.85 |
| Content (wt%) of mogroside V in dry momordica grosvenori powder juice | 51.91 | 51.87 | 51.83 | 51.75 | 51.63 |
| Total yield of dulcoside V (%) | 98.6 | 98.6 | 98.4 | 98.3 | 98.1 |
Example 7
The other conditions and procedures were the same as in example 1 except that in step (3), the ceramic membrane filtrate was passed through two anion exchange resins and a cation exchange resin in series, the total amount of anion exchange resin was 45L and the total amount of cation exchange resin was 45L. 19.42kg of fructus momordicae concentrated juice, 18.53kg of fructus momordicae concentrated juice without mogroside and 1.51kg of fructus momordicae extract dry powder are finally obtained, and the content of mogroside V in the fructus momordicae concentrated juice obtained in the embodiment is 3.76 percent, the color is yellow brown, the color of the fructus momordicae extract dry powder is yellow brown, and the content of the mogroside V is 48.36 percent through the detection of a High Performance Liquid Chromatography (HPLC) external standard method; the total yield of mogroside V was 91.5%.
Comparative example 1
The other conditions and steps are the same as the example 1, except that in the step (4), the immobilized enzyme is not adopted, but 0.15kg of pectinase and 0.08kg of cellulase are directly added, the temperature is kept at 40 ℃ for enzymolysis for 4 hours, after the enzymolysis is finished, the high-temperature boiling is carried out to inactivate the enzyme, and the temperature is reduced to obtain the enzymolysis liquid. 18.67kg of momordica grosvenori condensed juice and 1.48kg of momordica grosvenori dry powder are finally obtained, and the momordica grosvenori condensed juice obtained in the embodiment is brown in color through detection of a High Performance Liquid Chromatography (HPLC) external standard method, wherein the content of mogroside V is 3.42%, the color of momordica grosvenori extract dry powder is brown, and the content of mogroside V is 42.70%; the total yield of mogroside V was 81.4%.
By adopting a common enzymolysis method, the yield is obviously reduced mainly because the enzymolysis time is too long, microorganisms are most easily bred at the temperature and the pH value under the enzymolysis condition, so that the microbial fermentation and the material rancidity are caused, and the mogrosides are degraded by the microorganisms.
The preferred embodiments of the present invention have been described above in detail, but the present invention is not limited thereto. Within the scope of the technical idea of the invention, many simple modifications can be made to the technical solution of the invention, including combinations of various technical features in any other suitable way, and these simple modifications and combinations should also be regarded as the disclosure of the invention, and all fall within the scope of the invention.
Claims (10)
1. A method for producing a fructus momordicae extract by adopting an immobilized enzyme technology comprises the following steps:
(1) hot water extraction: crushing fresh fructus Siraitiae Grosvenorii, percolating with hot water, and cooling to obtain extractive solution;
(2) ceramic membrane filtration: filtering the fructus Siraitiae Grosvenorii extractive solution with ceramic membrane, and collecting the filtrate;
(3) desalting and decoloring: sequentially passing the ceramic membrane filtrate through a first anion exchange resin and a first cation exchange resin which are connected in series to obtain a desalting and decolorizing solution;
(4) carrying out enzymolysis on immobilized enzyme: passing the desalted and decolored liquid through an immobilized enzyme packed bed to obtain an enzymolysis liquid;
(5) concentration: and concentrating the enzymolysis liquid under reduced pressure to obtain the momordica grosvenori concentrated juice.
2. The method of claim 1, further comprising the steps of:
(6) and (4) nanofiltration: filtering the enzymolysis liquid obtained in the step (4) by using a nanofiltration membrane, collecting nanofiltration membrane trapped fluid, and concentrating nanofiltration membrane permeated fluid under reduced pressure until the sugar degree is 66brix to obtain a byproduct, namely the momordica grosvenori concentrated juice without momordica grosvenori sweet glycosides;
(7) concentrating and drying: concentrating the nanofiltration membrane retentate under reduced pressure, and spray drying to obtain fructus Siraitiae Grosvenorii extract dry powder.
3. The method according to claim 1 or 2, wherein in the step (1), the temperature of the hot water is 70-90 ℃, the amount of the hot water is 1-2 times of the weight of the fresh momordica grosvenori, and the percolation flow rate is 0.2-1.0 BV/h.
4. The process according to claim 1 or 2, wherein in step (3), the kind of anion exchange resin is a macroporous strongly basic anion exchange resin, preferably LSD762, D945, LSA-700B, D941; and/or
The cation exchange resin is a styrene type strongly acidic cation exchange resin, preferably 001 × 16, 001 × 7, 001 × 12, 001 × 8.
5. The method according to claim 1 or 2, wherein in the step (3), the total volume consumption of the anion exchange resin is 0.1-0.2 times (L/Kg) of the weight of the fresh fructus momordicae, the height-diameter ratio of the anion exchange resin column is 5-8: 1, and the flow rate of the material passing through the cation exchange resin column is 4-6 BV/h; and/or
The total volume consumption of the cation exchange resin is 0.1-0.2 times (L/Kg) of the weight of fresh fructus momordicae, the height-diameter ratio of the cation exchange resin column is 5-8: 1, and the flow rate of materials passing through the cation exchange resin column is 4-6 BV/h.
6. The method of claim 1 or 2, wherein in step (3), the feed liquid passes through the first anion exchange resin and the first cation exchange resin, and then passes through the second anion exchange resin and the second cation exchange resin, and the second anion exchange resin and the second cation exchange resin can be used as the first anion exchange resin and the first cation exchange resin in step (3) of the next batch.
7. The method of claim 1 or 2, wherein in step (4), the carrier of the immobilized enzyme is polysaccharide ion exchanger or synthetic polymer ion exchange resin, preferably Diethylaminoethyl (DEAE) -cellulose, mixed amine (ECTEDLA) -cellulose, Tetraethylaminoethyl (TEAE) -cellulose, DEAE-sephadex, Amberlite ira-93, 410, 900, Carboxymethyl (CM) -cellulose, cellulose-citrate, Amberlite CG-50, IRC-50, IR-200 or Dowex-50.
8. The method of claim 1 or 2, wherein in the step (4), the amount of the immobilized enzyme carrier is 0.05 to 0.1 times (L/Kg) of the weight of the fresh fructus momordicae.
9. The method according to claim 1 or 2, wherein in the step (4), the enzyme is a compound of pectinase and cellulase, the amount of the pectinase is 0.1-0.5 per mill of the weight of the fresh momordica grosvenori fruit, and the amount of the cellulase is 0.1-0.5 per mill of the weight of the fresh momordica grosvenori fruit.
10. The method of claim 1 or 2, wherein the aspect ratio of the immobilized enzyme packed bed is 6-10: 1, and the flow rate of the materials passing through the immobilized enzyme packed bed is 1-3 BV/h.
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| WO2021169133A1 (en) * | 2020-02-24 | 2021-09-02 | 湖南华诚生物资源股份有限公司 | Method for producing siraitia grosvenorii extract using immobilized enzyme technology |
| CN113455550A (en) * | 2021-06-30 | 2021-10-01 | 湖南绿蔓生物科技股份有限公司 | Preparation method of momordica grosvenori bean curd |
| CN115581266A (en) * | 2022-09-02 | 2023-01-10 | 湖南华诚生物资源股份有限公司 | Clean production method of momordica grosvenori dry powder and momordica grosvenori condensed juice |
| CN116098249A (en) * | 2021-11-10 | 2023-05-12 | 桂林莱茵生物科技股份有限公司 | Preparation method of multi-fold sweet momordica grosvenori pure juice with sugar replacing function |
| WO2023173705A1 (en) * | 2022-03-16 | 2023-09-21 | 桂林莱茵生物科技股份有限公司 | Sweetening agent composition, preparation method therefor and use thereof |
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| CN107156563B (en) * | 2017-04-25 | 2020-08-28 | 湖南华诚生物资源股份有限公司 | Preparation process of decolorized concentrated juice of fresh momordica grosvenori |
| CN111296708A (en) * | 2020-02-24 | 2020-06-19 | 湖南华诚生物资源股份有限公司 | Method for producing fructus momordicae extract by adopting immobilized enzyme technology |
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| CN101167539A (en) * | 2006-10-26 | 2008-04-30 | 湖南金农生物资源股份有限公司 | Method for producing grosvenor momordca fresh fruit deodorant concentrated juice |
| CN107397103A (en) * | 2017-08-25 | 2017-11-28 | 湖南华诚生物资源股份有限公司 | A kind of preparation method of Momordica grosvenori decolouring flavor inspissated juice |
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| WO2021169133A1 (en) * | 2020-02-24 | 2021-09-02 | 湖南华诚生物资源股份有限公司 | Method for producing siraitia grosvenorii extract using immobilized enzyme technology |
| CN113455550A (en) * | 2021-06-30 | 2021-10-01 | 湖南绿蔓生物科技股份有限公司 | Preparation method of momordica grosvenori bean curd |
| CN116098249A (en) * | 2021-11-10 | 2023-05-12 | 桂林莱茵生物科技股份有限公司 | Preparation method of multi-fold sweet momordica grosvenori pure juice with sugar replacing function |
| WO2023173705A1 (en) * | 2022-03-16 | 2023-09-21 | 桂林莱茵生物科技股份有限公司 | Sweetening agent composition, preparation method therefor and use thereof |
| CN115581266A (en) * | 2022-09-02 | 2023-01-10 | 湖南华诚生物资源股份有限公司 | Clean production method of momordica grosvenori dry powder and momordica grosvenori condensed juice |
| CN115581266B (en) * | 2022-09-02 | 2024-03-12 | 湖南华诚生物资源股份有限公司 | Clean production method of momordica grosvenori dry powder and momordica grosvenori concentrated juice |
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