CN115896209A - Extraction method of Feijoa pectin rich in RG-I high ester - Google Patents
Extraction method of Feijoa pectin rich in RG-I high ester Download PDFInfo
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
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- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses an extraction method of Fery fruit pectin rich in RG-I high ester, which comprises the following steps: pretreating the feijoa flesh into small pieces, inactivating pectinase, extracting pectin in a 50mM citric acid sodium salt solution in a microwave oven, cooling, adding enzyme for enzymolysis, centrifuging, purifying with ethanol, and drying to obtain the RG-I-rich high-ester pectin. According to the extraction method disclosed by the invention, pectin in feijoa pulp can be sufficiently extracted by microwave-assisted enzymolysis, the enzyme treatment time is shortened, and the extraction efficiency and the product yield are greatly improved; the pectin prepared by the method is high-esterification-degree pectin, the structure is more complete, the RG-I content is 41.38%, and the viscosity is higher.
Description
Technical Field
The invention belongs to the technical field of pectin extraction, and particularly relates to an extraction method of feijoa pectin rich in RG-I high ester.
Background
Feijoa, also known as guava, is a perennial subtropical evergreen shrub plant of the rhodomyrtus genus of myrtaceae family, and belongs to a new class of fruits in recent years in our country. Feijoa is rich in nutrients and functional active components, such as protein, vitamins, minerals and polyphenol. The pectin content in the feijoa fruits is reported to reach 20%, so the feijoa fruits are a resource treasure box for pectin, the feijoa fruits are to be popularized in a large-area planting scale in China, processed products of the feijoa fruits are to be developed, and related processed products, particularly pectin extraction reports and patents are few.
Pectin is an acidic heteropolysaccharide widely present in the intercellular layer of the cell wall of dicotyledonous plants. Pectin is widely used in the industries of food, medicine, cosmetics and the like due to its good emulsifying property, gelling property, stability and thickening property. Meanwhile, pectin is natural water-soluble dietary fiber, so that the risk of suffering from cancer, diabetes and obesity can be reduced, the intestinal microenvironment of a human body can be regulated, and blood fat can be reduced. The high-ester pectin is pectin with esterification degree (ratio of methyl-esterified galacturonic acid units to total galacturonic acid units in pectin) not less than 50%, and has good application prospect because the morphological structure is closest to natural pectin molecules. The esterification degree can influence the structure, functional property, the capability of forming a compound with other substances and the like of the pectin, the complete structure of the pectin is kept as much as possible, the performance of the pectin is improved, and the method has important significance for being better applied to industries such as food and the like.
The pectin major domains include Homogalacturonans (HG), rhamnogalacturonans I (RG-I), rhamnogalacturonans II (RG-II), and in addition minor amounts of substituted galacturonans. Commercial pectins, which are mainly composed of the HG region, are commonly used as thickeners and gelling agents in food-related fields. The use of high temperature and strong acid can remove most RG-I region to improve the stability and gel property of pectin. More and more researches show that the pectin rich in RG-I has better anticancer efficacy, chronic metabolic diseases regulation, immunological activity, prebiotic property and the like. Therefore, how to scientifically extract pectin having intact RG-I has received increasing attention from researchers and consumers.
The existing pectin extraction methods mainly comprise an ion exchange method, an acid/alkali extraction method, a supercritical extraction method, a biological enzyme method and the like, but the extraction methods have no ideal effect. In addition, strong acid or strong base is corrosive to equipment, and the extracted wastewater can cause environmental pollution. Therefore, how to prepare high-ester pectin with complete structure (RG-I), good performance and high yield is one of the problems to be solved at present.
Disclosure of Invention
The invention aims to provide an extraction method of Ferabout fruit pectin rich in RG-I high ester, which can obtain the Ferabout fruit high ester pectin rich in RG-I structure, good in quality and high in yield.
In order to achieve the purpose, the invention provides an extraction method of Fery fruit pectin rich in RG-I high ester, which comprises the following steps:
(1) Boiling the feijoa flesh, drying until the water content is lower than 6%, crushing, and uniformly mixing the powder with a buffer solution;
(2) Sequentially carrying out enzymolysis, centrifugation and purification on the mixed solution after microwave heating treatment, and drying the pectin;
the power of the microwave heating treatment is 350-400W, and the time is 5-15min.
Further, the boiling temperature of the step (1) is 85-100 ℃, and the boiling time is 3-8min.
Further, the drying temperature in the step (1) is 45-55 ℃.
Further, the powder crushed in the step (1) is sieved by a 60-mesh sieve.
Further, the proportion relation of the powder and the buffer solution in the step (1) is 10g:120-180mL, and the buffer is 50mM sodium citrate buffer with pH 6.8.
Further, the enzymolysis process specifically comprises: uniformly mixing beta-glucosidase and feijoa powder according to the amount of 250U/g, and carrying out enzymolysis for 90-100min at the temperature of 45-55 ℃ at the rotation speed of 120-180rpm.
Further, after the enzymolysis in the step (2), heating the extracted solution in a water bath environment at 90-100 ℃ for 3-5min.
Further, the rotation speed of the centrifugation in the step (2) is 4500-5500rpm, and the centrifugation time is 8-12min.
Further, the purification specifically comprises the following steps: collecting the centrifuged supernatant, adding anhydrous ethanol with a volume twice that of the supernatant, mixing uniformly, standing and precipitating for 2h, centrifuging at 4500-5500rpm after pectin is separated out, and cleaning the pectin in the test tube with anhydrous ethanol for 2-3 times.
Further, the temperature for drying pectin is 45-60 ℃.
In summary, the invention has the following advantages:
1. the extraction method disclosed by the invention can be used for sufficiently extracting pectin from feijoa flesh through microwave-assisted enzymolysis, so that the enzyme treatment time is shortened, and the extraction efficiency and the product yield are greatly improved; the pectin prepared by the method is high in esterification degree (68.43%), the structure is more complete, the RG I content is 41.38%, and the viscosity is higher.
2. The extraction method disclosed by the invention has the advantages of low energy consumption and low pollution, improves the utilization value of feijoa and provides theoretical guidance for further development and utilization of feijoa.
3. The method of the invention reserves RG-I area structure of pectin more, extracts high-ester pectin rich in RG-I, wherein pectin can be separated from raw materials by biological enzyme after microwave treatment of waste fruit pulp, and more protopectin is converted into soluble pectin to be extracted, so that the extraction method disclosed by the invention can obtain high-yield pectin.
Drawings
FIG. 1 is a graph showing the stable shear flow curves of pectin obtained by the microwave-assisted enzymatic Method (MEHE), the enzymatic method (EHE) and the conventional hot acid water method (CHE).
Detailed Description
The principles and features of this invention are described below in conjunction with embodiments, which are included to explain the invention and not to limit the scope of the invention. The examples, in which specific conditions are not specified, were carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The microwave heating method adopted in the invention is a microwave oven heating method, the model of the used microwave oven is MM823ESJ-PA American microwave oven, and the heating gear is medium-low fire. Therefore, the invention is also attached with a method for detecting the frequency of the medium-low fire gear so as to avoid the problem of unclear heating parameters. The detection method comprises the following steps: and (4) detecting the microwave power by adopting an IEC705 method. First, two 500mL plastic beakers were prepared and filled with 500mL water at a temperature of about 10 ℃, the beakers were placed in the center of the cooking area, the microwave oven was set to full power and the power was turned on, allowing the microwave oven to run precisely for 60s, and the magnetic control reached operating temperature of 2s. The beakers were then removed from the microwave oven and immediately stirred with a magnetic stirrer and rapidly temperature measured before measuring the temperature of each beaker with a thermometer, and the average was taken twice for each set.
Example 1
This example provides a method for extracting Fery fruit pectin rich in RG-I high ester, comprising the following steps:
(1) Washing picked feijoa, drying, peeling, cutting, adding into 90 deg.C deionized water, and boiling for 5min to inactivate pectinase. Properly controlling water, placing in a 50 ℃ forced air drying oven to dry until the water content is less than 6%, pulverizing with a pulverizer, sieving with a 60-mesh sieve to obtain waste fruit pulp powder (pulp and pulp), sealing in a self-sealing bag, and storing in a dryer for later use.
(2) The feijoa powder of step (1) (10 g) was put in a 250mL triangular flask, and 150mL of 50mM sodium citrate buffer (stock-solution ratio: 1: 15) was added thereto, and the pH was adjusted to 6.8 with citric acid, followed by uniform mixing.
(3) And (3) putting the mixed solution in the step (2) into a microwave oven (MM 823ESJ-PA Mei microwave oven), setting the duration of the fire to be medium-low fire (380W IEC), treating for 9min, taking out, and cooling until the hands are not scalded.
(4) Weighing beta-glucosidase (250U/g, the quantitative ratio of enzyme activity to substrate) in a triangular flask, uniformly mixing, and carrying out enzymolysis treatment in a constant-temperature shaking incubator for 99min, wherein the enzymolysis temperature is set to be 50 ℃, and the rotating speed of a shaking table is 150rpm.
(5) After the extraction was completed, the enzyme was inactivated by heating in a water bath at 95 ℃ for 5 minutes.
(6) Centrifuging the extracted solution at 5000rpm for 10min, collecting supernatant, washing the residue with distilled water twice, mixing the supernatants, and recording the volume.
(7) Adding anhydrous ethanol with the volume twice that of the supernatant, uniformly mixing, standing and precipitating for two hours, centrifuging at 5000rpm after pectin is separated out, and cleaning the pectin in the test tube with the anhydrous ethanol for 2 times.
(8) Drying the cleaned pectin in an electrothermal blowing drying oven at a set temperature of 50 ℃, grinding the pectin by using a mortar, and sieving the pectin by using a 100-mesh sieve for later use.
Example 2
(1) Washing picked Fujiguo fruit, drying, peeling, cutting, adding into 95 deg.C deionized water, and boiling for 5min to inactivate pectinase. Properly controlling water, placing in a 55 ℃ forced air drying oven to dry until the water content is less than 6%, pulverizing with a pulverizer, sieving with a 60-mesh sieve to obtain waste fruit pulp powder (pulp and pulp), sealing in a self-sealing bag, and storing in a dryer for later use.
(2) The feijoa powder of step (1) (10 g) was put into a 250mL Erlenmeyer flask, and 160mL 50mM sodium citrate buffer (stock-solution ratio: 1: 16) was added thereto, and the pH was adjusted to 6.8 with citric acid, followed by mixing uniformly.
(3) And (3) putting the mixed solution in the step (2) into a microwave oven (MM 823ESJ-PA Mei microwave oven), setting the fire to be medium-low fire (350W IEC), treating for 10min, taking out, and cooling until the hands are not scalded.
(4) Weighing beta-glucosidase (250U/g, the amount ratio of enzyme activity to substrate) in a triangular flask, mixing uniformly, and carrying out enzymolysis treatment in a constant-temperature shaking incubator for 90min, wherein the enzymolysis temperature is set to be 50 ℃, and the rotation speed of a shaking table is 150rpm.
(5) After the extraction was completed, the enzyme was inactivated by heating in a water bath at 95 ℃ for 5 minutes.
(6) The extracted solution was centrifuged at 5500rpm for 8min, the supernatant was collected and the residue was washed twice with distilled water, and the supernatants were combined and the volume was recorded.
(7) Adding anhydrous ethanol with the volume twice that of the supernatant, uniformly mixing, standing and precipitating for two hours, centrifuging at 5000rpm after pectin is separated out, and cleaning the pectin in the test tube with the anhydrous ethanol for 3 times.
(8) Drying the cleaned pectin in an electrothermal blowing drying oven at a set temperature of 50 ℃, grinding the pectin by using a mortar, and sieving the pectin by using a 100-mesh sieve for later use.
Example 3
(1) Washing picked feijoa, drying, peeling, cutting, adding into 90 deg.C deionized water, and boiling for 5min to inactivate pectinase. Properly controlling water, placing the mixture in a 50 ℃ forced air drying oven to dry until the water content is less than 6%, pulverizing the mixture by a pulverizer, sieving the pulverized mixture by a 60-mesh sieve to prepare feijoa pulp powder (pulp and pulp slurry), sealing the feijoa pulp powder in a self-sealing bag, and storing the self-sealing bag in a dryer for later use.
(2) The feijoa powder of step (1) (10 g) was put in a 250mL Erlenmeyer flask, 140mL of 50mM sodium citrate buffer (stock-solution ratio: 1: 14) was added, and the pH was adjusted to 6.8 with citric acid, and the mixture was mixed well.
(3) And (3) putting the mixed solution in the step (2) into a microwave oven (MM 823ESJ-PA Mei microwave oven), setting the duration of the fire to be medium-low fire (400W IEC), treating for 9min, taking out, and cooling until the hands are not scalded.
(4) Weighing beta-glucosidase (250U/g, the amount ratio of enzyme activity to substrate) in a triangular flask, mixing uniformly, and carrying out enzymolysis treatment in a constant-temperature shaking incubator for 100min, wherein the enzymolysis temperature is set to be 45 ℃, and the rotation speed of a shaking table is 150rpm.
(5) After the extraction was completed, the enzyme was inactivated by heating in a water bath at 90 ℃ for 5 minutes.
(6) Centrifuging the extracted solution at 5000rpm for 10min, collecting supernatant, washing the residue with distilled water twice, mixing the supernatants, and recording the volume.
(7) Adding anhydrous ethanol with the volume twice that of the supernatant, uniformly mixing, standing and precipitating for two hours, centrifuging at 5500rpm after pectin is separated out, and cleaning the pectin in the test tube with the anhydrous ethanol for 3 times.
(8) Drying the cleaned pectin in an electrothermal blowing drying oven at a set temperature of 50 ℃, grinding the pectin by using a mortar, and sieving the pectin by using a 100-mesh sieve for later use.
Comparative example 1
The comparative example provides a method for extracting feijoa pectin by a biological enzyme method, which comprises the following steps:
10g of feijoa pulp powder is accurately weighed into a conical flask, 150mL (1. The other processing was the same as in example 1.
Comparative example 2
The comparative example provides a method for extracting feijoa pectin by a conventional hot acid water method, which comprises the following steps:
(1) Weighing 10g of feijoa pulp powder in a beaker, adding 150mL of hydrochloric acid solution with pH of 1, and extracting for 130min in a heat collection type magnetic stirrer with set temperature of 95 ℃.
(2) Separating pulp residue from soluble compounds with centrifuge after extraction, and washing the separated pulp residue with distilled water.
(3) The filtrate was collected, centrifuged at 5000rpm for 10min to remove the fines, and 2 volumes of absolute ethanol were added to precipitate the pectin for 2 hours.
(4) Centrifuging the precipitated pectin at 5000rpm for 10min, washing pectin with anhydrous ethanol for 3 times, centrifuging again, and drying the obtained pectin at 50 deg.C with electrothermal blowing drying oven.
Test example 1 monosaccharide assay
Monosaccharide was measured by trichloroacetic acid (TFA) method, and a polysaccharide sample was accurately weighed at 10mg, and hydrolyzed at 110 ℃ for 4 hours by adding 1ml of a solution of LTFA (4M). Subsequently, the hydrolysate was co-distilled with ethanol several times under reduced pressure to completely remove TFA. The hydrolyzed sample and standard monosaccharide were acetylated by addition of a mixture of hydroxylamine hydrochloride, pyridine and acetic anhydride. The analysis was performed using a Japan Shimadzu GC/MS gas chromatography (QP 2010 Plus) system. The initial temperature was set at 150 ℃ and held for 17min, the temperature program was ramped from 150 ℃ to 165 ℃ at a rate of 1 ℃/min, then ramped to 200 ℃ at a rate of 10 ℃/min, and ramped to 380 ℃ at a ramp rate of 50 ℃/mi. The sample feeding amount is 1 mu L, the split ratio is 1; the ion source temperature is 230 ℃, the electron collision voltage is 70eV, the mass scanning range m/z is 20-650, the frequency is 0.2s/scan, and the solvent delay time is set to be 5.5min. Standard quantification was performed using β -phenyl-glucoside (0.05% w/v) as internal standard and the RG-I content was calculated according to the following formula:
RG-I(％)＝2Rha(％)+Ara(％)+Gal(％)
test example 2 determination of the Degree of Esterification (DE)
The method for measuring the degree of esterification is carried out by referring to the titration method in national standard for food safety GB25533 2010.
Test example 3 determination of rheological Properties
Pectin samples were dissolved in deionized water to make a concentration of 1.0% (w/v). The rotational rheometer was HaakeRS6000 under the U.S. Thermo Corp.A.A.flat circular steel plate (diameter 60 mm) was used as the probe, and the apparent viscosity of the sample was measured with a gap set at 1.0 mm. Measured at 25 ℃ with shear rate from 0.1s -1 Gradually increase to 100s -1 Apparent viscosity curve of (b).
The results of the experiment are shown in table 1 and fig. 1.
TABLE 1 pectin physico-chemical parameters
As can be seen from Table 1 and FIG. 1, the extraction method disclosed by the invention can greatly improve the yield of pectin, and keep higher levels of both the esterification degree and the RG-I content. The extraction method disclosed by the invention can be used for reserving more RG-I region structures of the pectin and extracting the high-ester pectin rich in RG-I. Meanwhile, the extracted pectin has the esterification degree of 68.43 percent, belongs to high-ester pectin, has the viscosity equivalent to that of the pectin extracted by an enzyme method, and is superior to the pectin extracted by a conventional hot acid-water method.
While the present invention has been described in detail with reference to the specific embodiments thereof, it should not be construed as limited by the scope of the present patent. Various modifications and changes may be made by those skilled in the art without inventive step within the scope of the appended claims.
Claims (10)
1. An extraction method of Ferula pectin rich in RG-I high ester is characterized by comprising the following steps:
(1) Boiling the feijoa flesh, drying until the water content is lower than 6%, crushing, and uniformly mixing the powder with a buffer solution;
(2) Sequentially carrying out enzymolysis, centrifugation and purification on the mixed solution after microwave heating treatment, and drying the pectin;
the power of the microwave heating treatment is 350-400W, and the time is 5-15min.
2. The extraction process of feijoa pectin enriched in RG-I high esters according to claim 1, wherein the boiling temperature in step (1) is 85-100 ℃ and the boiling time is 3-8min.
3. The extraction process of feijoa pectin enriched in RG-I high esters according to claim 1, wherein the drying temperature in step (1) is 45-55 ℃.
4. The extraction method of feijoa pectin enriched in RG-I high esters according to claim 1, wherein the powder obtained after pulverization in step (1) is sieved through a 60 mesh sieve.
5. The extraction method of feijoa pectin enriched in RG-I high esters according to claim 1 or 4, wherein the ratio of powder to buffer in step (1) is 10g:120-180mL, and the buffer is 50mM sodium citrate buffer with the pH value of 6.8.
6. The extraction method of the RG-I high-ester enriched Feijoa pectin according to claim 1, characterized in that the enzymatic hydrolysis process comprises: uniformly mixing beta-glucosidase and feijoa powder according to the amount of 250U/g, and performing enzymolysis for 90-100min at 45-55 ℃ at the rotation speed of 120-180rpm.
7. The method for extracting feijoa pectin rich in RG-I high ester according to claim 1, wherein after the enzymatic hydrolysis in step (2), the extracted solution is heated in a water bath environment at 90-100 ℃ for 3-5min.
8. The extraction method of Fery pectin enriched in RG-I high esters according to claim 1, characterized in that the centrifugation speed of step (2) is 4500-5500rpm, and the centrifugation time is 8-12min.
9. The extraction method of feijoa pectin enriched in RG-I high esters according to claim 1, wherein the purification specifically comprises the following steps: collecting the centrifuged supernatant, adding anhydrous ethanol with a volume twice that of the supernatant, mixing uniformly, standing and precipitating for 2h, centrifuging at 4500-5500rpm after pectin is separated out, and cleaning the pectin in the test tube with anhydrous ethanol for 2-3 times.
10. A method of extracting feijoa pectin enriched in RG-I high esters according to claim 1, wherein the pectin is dried at a temperature of 45-60 ℃.
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