CN110699402A - Method for preparing low-methoxyl pectin by using pomace - Google Patents
Method for preparing low-methoxyl pectin by using pomace Download PDFInfo
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- CN110699402A CN110699402A CN201911165097.0A CN201911165097A CN110699402A CN 110699402 A CN110699402 A CN 110699402A CN 201911165097 A CN201911165097 A CN 201911165097A CN 110699402 A CN110699402 A CN 110699402A
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- 241000228245 Aspergillus niger Species 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 25
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- PYMYPHUHKUWMLA-UHFFFAOYSA-N 2,3,4,5-tetrahydroxypentanal Chemical compound OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims abstract description 19
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- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
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- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
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- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical group [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 description 2
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- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 239000004366 Glucose oxidase Substances 0.000 description 1
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- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical group O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
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- 230000003247 decreasing effect Effects 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000005496 eutectics Effects 0.000 description 1
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- 239000002778 food additive Substances 0.000 description 1
- 235000021022 fresh fruits Nutrition 0.000 description 1
- 229940074391 gallic acid Drugs 0.000 description 1
- 235000004515 gallic acid Nutrition 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
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- 229940116332 glucose oxidase Drugs 0.000 description 1
- 235000019420 glucose oxidase Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
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- 229920000642 polymer Polymers 0.000 description 1
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- 241000894007 species Species 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0045—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid alpha-D-Galacturonans, e.g. methyl ester of (alpha-1,4)-linked D-galacturonic acid units, i.e. pectin, or hydrolysis product of methyl ester of alpha-1,4-linked D-galacturonic acid units, i.e. pectinic acid; Derivatives thereof
- C08B37/0048—Processes of extraction from organic materials
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
A method for preparing low methoxyl pectin by using pomace comprises the following steps: (1) preparing aspergillus niger liquid seeds, (2) performing aspergillus niger solid fermentation, (3) preparing a complex enzyme, (4) treating pomace by the complex enzyme, (5) performing acid hydrolysis and extraction, (6) performing ethanol precipitation and filtration, and (7) cleaning and drying. The method takes the squeezed pomace of yellow peaches, pears, apples, blueberries, oranges, grapefruits and the like as raw materials, prepares compound enzyme by solid fermentation of aspergillus niger, and treats the pomace with the compound enzyme to realize cell wall dissolution and pectin degreasing; then preparing the low methoxyl pectin through acidolysis, alcohol precipitation, cleaning, drying and the like.
Description
Technical Field
The invention belongs to the technical field of deep processing of agricultural products, and relates to a method for preparing low-methoxyl pectin by using pomace.
Background
Pectin is a non-toxic natural food additive and is an important component of plant cell walls. Pectin is a plant polysaccharide polymer consisting essentially of partially methyl-esterified galacturonic acid units, which are generally classified into two main groups according to their methoxy content: high Methoxyl Pectin (HMP), Low Methoxyl Pectin (LMP).
HMP generally refers to pectin having a degree of esterification or methyl esterification greater than 50%, i.e., a methoxyl content greater than 7%, LMP refers to pectin having a degree of methyl esterification between 25-50%, i.e., a methoxyl content less than 7%, HMP gels only when the soluble solids are above 55% and the pH is in the range of 2.0 ~ 3.5.5, and is primarily used in the production of high-sugar foods.
LMP is not completely limited by pH value and sugar degree (eutectic), and has low requirement on gel condition. The rate of gel formation by LMP increases with decreasing degree of methyl esterification of the pectin. The LMP has obvious significance in meeting the requirements of people on low-sugar, low-calorie and low-sweetness foods. In recent years, the demand for low-sugar foods has increased significantly due to the increasing number of obese and diabetic patients. LMP has resulted in a great market demand for LMP due to its unparalleled advantages as an edible gum for low-sugar foods.
Aspergillus niger, Deuteromycotina, class Hyphomycetes, order Hyphomycetales, family Moniliaceae, a common species of fungi of the genus Aspergillus. Widely distributed throughout the world in foodstuffs, vegetable products and soils. Is an important fermentation industrial strain, and can produce amylase, acid protease, cellulase, pectinase, glucose oxidase, citric acid, gluconic acid, gallic acid and the like.
Disclosure of Invention
The invention aims to provide a method for preparing low-methoxyl pectin by using pomace.
In order to achieve the above objects and other related objects, the present invention provides the following technical solutions: a method for preparing low methoxyl pectin by using pomace comprises the following steps:
step 1: adding sterile water into the aspergillus niger strain preservation inclined plane, scraping off spores in the inclined plane by using an inoculation shovel to prepare strain suspension; inoculating the strain suspension into the sterilized liquid seed culture medium according to the inoculum size of 2.5-5%, and culturing for 4-6d in a shaking table at the temperature of 30-35 ℃ and the rotation speed of 100-;
step 2: in a purification environment with the air cleaning level of 1 ten thousand, uniformly mixing Aspergillus niger liquid seeds with a solid culture medium subjected to steam sterilization according to the inoculation amount of 1-5% at the temperature of 25-30 ℃, then paving the mixture into a tray with the thickness of 1.5-2.5cm, keeping the relative humidity of the environment at 60-75%, and fermenting for 5-7d to obtain a solid fermentation product;
and step 3: adding distilled water with the mass 9-12 times of that of the solid culture medium into the solid fermentation product, soaking and filtering to obtain a filtrate, namely a crude enzyme solution; adding ammonium sulfate into the crude enzyme solution at the temperature of 25 +/-5 ℃, standing, filtering to obtain a precipitate, and cleaning the precipitate with purified water to obtain a complex enzyme obtained by fermenting aspergillus niger;
and 4, step 4: adding purified water 4-5 times the weight of the pomace, adding the complex enzyme prepared in the step 3 with the pomace weight of 0.5-1.0%, keeping the temperature at 40-45 ℃, and stirring at the rotating speed of 45-60r/min for 60-90min to obtain pomace mixed solution;
and 5: adding hydrochloric acid into the mixed liquid of the pomace, adjusting the pH value to 2.0-2.5, then heating and keeping the temperature at 90-92 ℃, and stirring at the rotating speed of 30-60r/min for 60-90 min; filtering the obtained solution to obtain filtrate as pectin extracting solution;
step 6: adding ethanol into the pectin extractive solution to make the ethanol content of the mixed solution 50-65%, and standing for 60-90 min; filtering the mixed solution by a 60-80 mesh filter screen, and collecting solid retentate;
and 7: repeatedly washing the solid retentate with purified water for 3-5 times, drying the solid retentate under the environment of-0.09 MPa to-0.10 MPa of vacuum gauge pressure and 60-65 ℃ until the water content is less than or equal to 5%, and crushing to obtain the low methoxyl pectin.
The preferable technical scheme is as follows: the fruit residue is at least one of fresh yellow peach, pear, apple, blueberry, orange and grapefruit, and is residue obtained after fruit juice is squeezed.
The preferable technical scheme is as follows: the raw material formula of the liquid seed culture medium comprises the following raw materials in percentage by mass: 25-30% of cane sugar, 0.25-0.50% of ammonium sulfate, 0.15-0.25% of dipotassium hydrogen phosphate, 0.15-0.25% of sodium chloride, 0.05-0.10% of ferrous sulfate and the balance of purified water, wherein the total content is 100%.
The preferable technical scheme is as follows: the raw material formula of the solid culture medium comprises the following raw materials in percentage by mass: 40-50% of pomace, 10-15% of bran, 10-15% of corn flour, 5% of soybean meal, 0.5% of ammonium sulfate and 0.25% of dipotassium hydrogen phosphate, wherein the total content is 100%; the preparation method of the solid culture medium comprises the following steps: weighing the raw materials according to the raw material formula of the solid culture medium, mixing uniformly, filling into a non-woven bag, sterilizing for 25-30min by using 120-grade 125-DEG saturated steam, and naturally cooling to room temperature in a purified environment with the air cleaning level of 1 ten thousand for later use.
Due to the application of the technical scheme, compared with the prior art, the invention has the advantages that:
the method takes the squeezed pomace of yellow peaches, pears, apples, blueberries, oranges, grapefruits and the like as raw materials, prepares compound enzyme by solid fermentation of aspergillus niger, and treats the pomace with the compound enzyme to realize cell wall dissolution and pectin degreasing; then preparing the low methoxyl pectin through acidolysis, alcohol precipitation, cleaning, drying and the like.
Detailed Description
The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.
Example 1: method for preparing low-methoxyl pectin by using pomace
A method for preparing low methoxyl pectin from fruit residue is provided, wherein the fruit residue is one or more of fresh fruits such as yellow peach, pear, apple, blueberry, orange, grapefruit, etc., and residue after juice is squeezed.
A method for preparing low methoxyl pectin by using pomace is characterized by comprising the following technical steps.
(1) Preparation of Aspergillus niger liquid seed
The liquid seed culture medium formula comprises the following raw materials in percentage by mass: 27.5% of sucrose, 0.38% of ammonium sulfate, 0.2% of dipotassium hydrogen phosphate, 0.2% of sodium chloride, 0.07% of ferrous sulfate and the balance of purified water, wherein the total amount is 100%.
And (5) liquid seed culture. Adding sterile water into the aspergillus niger strain preservation inclined plane, scraping spores in the inclined plane by using an inoculation shovel to prepare strain suspension; inoculating the strain suspension into the sterilized liquid seed culture medium according to the inoculation amount of 4%, and culturing for 5d in a shaking table at the temperature of 32 ℃ and the rotating speed of 150r/min to obtain the Aspergillus niger liquid seed.
(2) Aspergillus niger solid fermentation
The solid fermentation culture medium formula comprises: 45% of pomace, 12% of bran, 12% of corn flour, 5% of soybean meal, 0.5% of ammonium sulfate and 0.25% of dipotassium hydrogen phosphate, wherein the total content is 100%. And (3) fully and uniformly mixing the solid culture medium according to the formula, filling the mixture into a non-woven fabric bag, sterilizing the mixture for 28min by using saturated steam at the temperature of 122 ℃, and naturally cooling the mixture to room temperature in a purified environment with the air cleaning level of 1 ten thousand for later use.
And (5) solid fermentation. In a clean environment with the air cleaning level of 1 ten thousand, the aspergillus niger liquid seeds and the solid culture medium after steam sterilization are fully mixed according to the inoculation amount of 3 percent at the temperature of 28 ℃, then the mixture is paved into a shallow tray with the thickness of 2cm, the relative humidity of the environment is kept at 70 percent, and the fermentation is carried out for 6 days.
(3) Preparation of Complex enzymes
And (4) leaching. After solid fermentation, distilled water is added according to 10 times of the weight of the solid culture medium, the mixture is soaked for 3 hours at room temperature, four layers of gauze are filtered, and the filtrate is the crude enzyme solution.
And (5) refining. Analytically pure grade ammonium sulfate was ground to a fine powder and screened through a 100 mesh screen for use. At the temperature of 25 ℃, ammonium sulfate after sieving is slowly and uniformly added into the filtrate, and the addition amount is 750g/L of the filtrate. Standing for 30min, filtering to obtain precipitate, and cleaning the precipitate with purified water. The precipitate is a complex enzyme obtained by fermenting Aspergillus niger, and comprises amylase, protease, pectinase and cellulase.
(4) Complex enzyme treatment of fruit residues
Adding food-grade purified water 4.5 times the weight of the pomace, adding 0.7% of the complex enzyme prepared in the third step, keeping the temperature at 42 ℃ and the rotating speed at 50r/min, and treating for 75 min.
(5) Acid hydrolysis and extraction
Adding a certain amount of hydrochloric acid into the mixed liquid of the pomace after the complex enzyme treatment is finished, heating and keeping the temperature at 91 ℃ under the condition that the pH value is 2.2, rotating at 45r/min, and treating for 75 min. Then filtering the solution by adopting a 80-mesh filter screen to obtain filtrate which is pectin extracting solution.
(6) Precipitating with ethanol and filtering
Adding 95% ethanol, or anhydrous ethanol or mixed solution thereof with ethanol content of 60% in other embodiments, and standing for 75 min; the solution was filtered through a 70 mesh screen and the solid retentate was collected.
(7) Cleaning and drying
Repeatedly extracting the solid retentate with food-grade purified water for 4 times, oven drying the solid retentate at vacuum gauge pressure of-0.095 MPa and 62 deg.C to 5%, and pulverizing to obtain low methoxyl pectin.
Example 2: method for preparing low-methoxyl pectin by using pomace
A method for preparing low methoxyl pectin by using pomace comprises the following steps:
step 1: adding sterile water into the aspergillus niger strain preservation inclined plane, scraping off spores in the inclined plane by using an inoculation shovel to prepare strain suspension; inoculating the strain suspension into the sterilized liquid seed culture medium according to the inoculum size of 2.5%, and culturing for 4d in a shaking table at the temperature of 30 ℃ and the rotating speed of 100r/min to obtain Aspergillus niger liquid seeds;
step 2: in a purification environment with the air cleaning level of 1 ten thousand, uniformly mixing Aspergillus niger liquid seeds with a solid culture medium subjected to steam sterilization according to the inoculation amount of 1% at 25 ℃, then paving the mixture into a tray with the thickness of 1.5cm, keeping the relative humidity of the environment at 60%, and fermenting for 5 days to obtain a solid fermentation product;
and step 3: adding distilled water with the mass 9 times of that of the solid culture medium into the solid fermentation product, soaking and filtering to obtain filtrate, namely crude enzyme liquid; adding ammonium sulfate into the crude enzyme solution at the temperature of 20 ℃, standing, filtering to obtain a precipitate, and cleaning the precipitate with purified water to obtain a complex enzyme obtained by fermenting aspergillus niger;
and 4, step 4: adding purified water 4 times the weight of the pomace, adding the complex enzyme prepared in the step 3 with the pomace weight of 0.5%, keeping the temperature at 40 ℃, and stirring at the rotating speed of 45r/min for 60min to obtain pomace mixed liquid;
and 5: adding hydrochloric acid into the pomace mixed solution, adjusting the pH value to 2.0, then heating and keeping the temperature at 90 ℃, and stirring at the rotating speed of 30r/min for 60 min; filtering the obtained solution to obtain filtrate as pectin extracting solution;
step 6: adding ethanol into the pectin extractive solution to make the ethanol content of the mixed solution 50%, and standing for 60 min; filtering the mixed solution by adopting a 60-mesh filter screen, and collecting solid retentate;
and 7: repeatedly cleaning the solid retentate with purified water for 3 times, oven drying the solid retentate under vacuum gauge pressure of-0.09 MPa and at 60 deg.C until the water content is 4.5%, and pulverizing to obtain low methoxyl pectin.
The preferred embodiment is: the fruit residue is residue obtained by squeezing juice from yellow peach.
The preferred embodiment is: the raw material formula of the liquid seed culture medium comprises the following raw materials in percentage by mass: 25% of sucrose, 0.25% of ammonium sulfate, 0.15% of dipotassium hydrogen phosphate, 0.15% of sodium chloride, 0.05% of ferrous sulfate and the balance of purified water, wherein the total amount is 100%.
The preferred embodiment is: the raw material formula of the solid culture medium comprises the following raw materials in percentage by mass: 40% of pomace, 10% of bran, 10% of corn flour, 5% of bean pulp, 0.5% of ammonium sulfate and 0.25% of dipotassium hydrogen phosphate, wherein the total content is 100%; the preparation method of the solid culture medium comprises the following steps: weighing the raw materials according to the raw material formula of the solid culture medium, mixing uniformly, filling into a non-woven fabric bag, sterilizing for 25min by using saturated steam at 120 ℃, and naturally cooling to room temperature in a purified environment with the air cleaning level of 1 ten thousand for later use.
Example 3: method for preparing low-methoxyl pectin by using pomace
A method for preparing low methoxyl pectin by using pomace comprises the following steps:
step 1: adding sterile water into the aspergillus niger strain preservation inclined plane, scraping off spores in the inclined plane by using an inoculation shovel to prepare strain suspension; inoculating the strain suspension into a sterilized liquid seed culture medium according to the inoculation amount of 5%, and culturing for 6d in a shaking table at the temperature of 35 ℃ and the rotating speed of 200r/min to obtain Aspergillus niger liquid seeds;
step 2: in a purification environment with the air cleaning level of 1 ten thousand, uniformly mixing Aspergillus niger liquid seeds with a solid culture medium subjected to steam sterilization according to the inoculation amount of 5% at the temperature of 30 ℃, then paving the mixture into a tray with the thickness of 2.5cm, keeping the relative humidity of the environment at 75%, and fermenting for 7d to obtain a solid fermentation product;
and step 3: adding distilled water with the mass 12 times of that of the solid culture medium into the solid fermentation product, soaking and filtering to obtain filtrate, namely crude enzyme liquid; adding ammonium sulfate into the crude enzyme solution at 30 ℃, standing, filtering to obtain a precipitate, and cleaning the precipitate with purified water to obtain a complex enzyme obtained by fermenting aspergillus niger;
and 4, step 4: adding purified water of which the mass is 5 times that of the pomace into the pomace, then adding the complex enzyme prepared in the step 3 of which the mass is 1.0 percent of the pomace, keeping the temperature at 45 ℃, and stirring at the rotating speed of 60r/min for 90min to obtain pomace mixed liquor;
and 5: adding hydrochloric acid into the pomace mixed solution, adjusting the pH value to 2.5, then heating and keeping the temperature at 92 ℃, and stirring at the rotating speed of 60r/min for 90 min; filtering the obtained solution to obtain filtrate as pectin extracting solution;
step 6: adding ethanol into the pectin extract to make the mass percent of the ethanol in the mixed solution be 65%, and standing for 90 min; filtering the mixed solution by adopting a 80-mesh filter screen, and collecting solid retentate;
and 7: repeatedly washing the solid retentate with purified water for 5 times, drying the solid retentate under vacuum gauge pressure of-0.10 MPa and at 65 deg.C until the water content is 4%, and pulverizing to obtain low methoxyl pectin.
The preferred embodiment is: the fruit residues are residues obtained by squeezing juice from yellow peach, blueberry, orange and grapefruit. The mass ratio of the components is as follows: 2: 1: 1: 1
The preferred embodiment is: the raw material formula of the liquid seed culture medium comprises the following raw materials in percentage by mass: 30 percent of sucrose, 0.50 percent of ammonium sulfate, 0.25 percent of dipotassium hydrogen phosphate, 0.25 percent of sodium chloride, 0.10 percent of ferrous sulfate and the balance of purified water, and the total amount is 100 percent.
The preferred embodiment is: the raw material formula of the solid culture medium comprises the following raw materials in percentage by mass: 50% of pomace, 15% of bran, 15% of corn flour, 5% of soybean meal, 0.5% of ammonium sulfate and 0.25% of dipotassium hydrogen phosphate, wherein the total content is 100%; the preparation method of the solid culture medium comprises the following steps: weighing the raw materials according to the raw material formula of the solid culture medium, mixing uniformly, filling into a non-woven fabric bag, sterilizing for 30min by using saturated steam at 125 ℃, and naturally cooling to room temperature in a purified environment with the air cleaning level of 1 ten thousand for later use.
The foregoing is illustrative of the preferred embodiment of the present invention and is not to be construed as limiting thereof in any way, and any modifications or variations thereof that fall within the spirit of the invention are intended to be included within the scope thereof.
Claims (4)
1. A method for preparing low methoxyl pectin by using pomace is characterized by comprising the following steps: comprises the following steps:
step 1: adding sterile water into the aspergillus niger strain preservation inclined plane, scraping off spores in the inclined plane by using an inoculation shovel to prepare strain suspension; inoculating the strain suspension into the sterilized liquid seed culture medium according to the inoculum size of 2.5-5%, and culturing for 4-6d in a shaking table at the temperature of 30-35 ℃ and the rotation speed of 100-;
step 2: in a purification environment with the air cleaning level of 1 ten thousand, uniformly mixing Aspergillus niger liquid seeds with a solid culture medium subjected to steam sterilization according to the inoculation amount of 1-5% at the temperature of 25-30 ℃, then paving the mixture into a tray with the thickness of 1.5-2.5cm, keeping the relative humidity of the environment at 60-75%, and fermenting for 5-7d to obtain a solid fermentation product;
and step 3: adding distilled water with the mass 9-12 times of that of the solid culture medium into the solid fermentation product, soaking and filtering to obtain a filtrate, namely a crude enzyme solution; adding ammonium sulfate into the crude enzyme solution at the temperature of 25 +/-5 ℃, standing, filtering to obtain a precipitate, and cleaning the precipitate with purified water to obtain a complex enzyme obtained by fermenting aspergillus niger;
and 4, step 4: adding purified water 4-5 times the weight of the pomace, adding the complex enzyme prepared in the step 3 with the pomace weight of 0.5-1.0%, keeping the temperature at 40-45 ℃, and stirring at the rotating speed of 45-60r/min for 60-90min to obtain pomace mixed solution;
and 5: adding hydrochloric acid into the mixed liquid of the pomace, adjusting the pH value to 2.0-2.5, then heating and keeping the temperature at 90-92 ℃, and stirring at the rotating speed of 30-60r/min for 60-90 min; filtering the obtained solution to obtain filtrate as pectin extracting solution;
step 6: adding ethanol into the pectin extractive solution to make the ethanol content of the mixed solution 50-65%, and standing for 60-90 min; filtering the mixed solution by a 60-80 mesh filter screen, and collecting solid retentate;
and 7: repeatedly washing the solid retentate with purified water for 3-5 times, drying the solid retentate under the environment of-0.09 MPa to-0.10 MPa of vacuum gauge pressure and 60-65 ℃ until the water content is less than or equal to 5%, and crushing to obtain the low methoxyl pectin.
2. The method for preparing low methoxyl pectin by using pomace according to claim 1, wherein: the fruit residue is at least one of fresh yellow peach, pear, apple, blueberry, orange and grapefruit, and is residue obtained after fruit juice is squeezed.
3. The method for preparing low methoxyl pectin by using pomace according to claim 1, wherein: the raw material formula of the liquid seed culture medium comprises the following raw materials in percentage by mass: 25-30% of cane sugar, 0.25-0.50% of ammonium sulfate, 0.15-0.25% of dipotassium hydrogen phosphate, 0.15-0.25% of sodium chloride, 0.05-0.10% of ferrous sulfate and the balance of purified water, wherein the total content is 100%.
4. The method for preparing low methoxyl pectin by using pomace according to claim 1, wherein: the raw material formula of the solid culture medium comprises the following raw materials in percentage by mass: 40-50% of pomace, 10-15% of bran, 10-15% of corn flour, 5% of soybean meal, 0.5% of ammonium sulfate and 0.25% of dipotassium hydrogen phosphate, wherein the total content is 100%; the preparation method of the solid culture medium comprises the following steps: weighing the raw materials according to the raw material formula of the solid culture medium, mixing uniformly, filling into a non-woven bag, sterilizing for 25-30min by using 120-grade 125-DEG saturated steam, and naturally cooling to room temperature in a purified environment with the air cleaning level of 1 ten thousand for later use.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111543629A (en) * | 2020-04-07 | 2020-08-18 | 杭州莱恩纳尔生物科技有限公司 | Method for simultaneously preparing pectin, fruit vinegar, ferment, probiotics and carbon black from peel residues |
| CN112006202A (en) * | 2020-09-17 | 2020-12-01 | 巢湖学院 | Apple-flavored beverage and preparation method thereof |
| CN113731307A (en) * | 2021-08-30 | 2021-12-03 | 华南理工大学 | Ammonium sulfate-containing high-ester pectin gel and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4200694A (en) * | 1977-10-08 | 1980-04-29 | Kikkoman Shoyu Co., Ltd. | Novel pectin esterase, process for its production, and process for producing demethoxylated pectin by the use of said pectin esterase |
| CN101073384A (en) * | 2006-05-17 | 2007-11-21 | 烟台安德利果胶有限公司 | Production of low-methoxy apple pectin |
| CN101153306A (en) * | 2007-09-30 | 2008-04-02 | 湖南省农产品加工研究所 | Method for producing tangerine peel low methoxy group pectin by using pectinase |
| CN101665769A (en) * | 2009-04-03 | 2010-03-10 | 中南大学 | Bacterial strain with high protopectinase yield and method for extracting pectin by fermentation |
-
2019
- 2019-11-25 CN CN201911165097.0A patent/CN110699402A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4200694A (en) * | 1977-10-08 | 1980-04-29 | Kikkoman Shoyu Co., Ltd. | Novel pectin esterase, process for its production, and process for producing demethoxylated pectin by the use of said pectin esterase |
| CN101073384A (en) * | 2006-05-17 | 2007-11-21 | 烟台安德利果胶有限公司 | Production of low-methoxy apple pectin |
| CN101153306A (en) * | 2007-09-30 | 2008-04-02 | 湖南省农产品加工研究所 | Method for producing tangerine peel low methoxy group pectin by using pectinase |
| CN101665769A (en) * | 2009-04-03 | 2010-03-10 | 中南大学 | Bacterial strain with high protopectinase yield and method for extracting pectin by fermentation |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111543629A (en) * | 2020-04-07 | 2020-08-18 | 杭州莱恩纳尔生物科技有限公司 | Method for simultaneously preparing pectin, fruit vinegar, ferment, probiotics and carbon black from peel residues |
| CN112006202A (en) * | 2020-09-17 | 2020-12-01 | 巢湖学院 | Apple-flavored beverage and preparation method thereof |
| CN112006202B (en) * | 2020-09-17 | 2023-06-09 | 巢湖学院 | Apple-flavored beverage and preparation method thereof |
| CN113731307A (en) * | 2021-08-30 | 2021-12-03 | 华南理工大学 | Ammonium sulfate-containing high-ester pectin gel and preparation method and application thereof |
| CN113731307B (en) * | 2021-08-30 | 2022-05-24 | 华南理工大学 | Ammonium sulfate-containing high-ester pectin gel and preparation method and application thereof |
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