CN107853396B - Method for preparing goat milk active beverage by using lactobacillus rhamnosus protease - Google Patents
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/152—Milk preparations; Milk powder or milk powder preparations containing additives
- A23C9/156—Flavoured milk preparations ; Addition of fruits, vegetables, sugars, sugar alcohols or sweeteners
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- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/1203—Addition of, or treatment with, enzymes or microorganisms other than lactobacteriaceae
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- A—HUMAN NECESSITIES
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Abstract
A method for preparing goat milk active beverage by Lactobacillus rhamnosus protease comprises inoculating Lactobacillus rhamnosus inoculum size into Lactobacillus rhamnosus culture medium, culturing, centrifuging, collecting thallus precipitate, washing with sterile normal saline, and centrifuging to obtain Lactobacillus rhamnosus bacterial mud; adding a phosphate buffer solution into the lactobacillus rhamnosus bacterial mud, centrifuging to obtain an enzyme solution of muramidase, and freeze-drying to obtain lactobacillus rhamnosus muramidase bacterial powder; adding Lactobacillus rhamnosus muramidase powder and CaCl into defatted fresh goat milk2After enzymolysis, beta-cyclodextrin, hovenia dulcis thunb polysaccharide, erythritol, xylitol, L-malic acid, citric acid and sodium citrate are added, and after uniform mixing and sterilization, the goat milk active beverage containing antihypertensive peptide and antioxidant peptide is obtained, wherein the ACE inhibitory activity is 70-90%, the DPPH free radical scavenging rate is 60-80%, and the hydroxyl free radical scavenging rate is 75-95%. The goat milk active beverage prepared by the invention takes goat milk as a raw material, and can be used as a food for assisting in reducing blood pressure, resisting oxidation, reducing blood sugar and preventing dental caries.
Description
Technical Field
The invention relates to a preparation method of a functional goat milk beverage, in particular to a method for preparing a goat milk active beverage by using lactobacillus rhamnosus protease.
Background
Probiotics have proteolytic systems including CEP, aminopeptidases, endopeptidases, Pro-specific peptidases, etc., which hydrolyze proteins and release peptides of varying sizes and some free amino acids to meet the needs of bacterial growth, wherein some peptides produced by hydrolysis have biological activities such as antioxidant, hypotensive, immunomodulating, etc. Muramidase is the most important enzyme in the whole hydrolysis process, and several CEPs alone have been found to hydrolyze milk protein and produce ACE inhibitory peptides.
The cell surface of lactobacilli comprises a number of enzymes and proteins, mainly ribosomal proteinsLeukocytes, cell surface permeases, proteins of the surface layer (S-layer), cell membrane associated proteases (CEPs), and other protein-based molecules[10]. The mechanism of cell wall protease (CEP) hydrolysis of milk proteins is: firstly, milk protein is hydrolyzed into a series of short peptides by CEP, then the short peptides are transported into cells by oligopeptide, dipeptide and tripeptide transport systems, and finally the short peptides are further hydrolyzed into bioactive peptides such as ACE inhibitory peptide and antioxidant peptide by intracellular peptidase (endopeptidase, pro-specific peptidase, aminopeptidase and the like)
Hovenia dulcis, also called pawpaw, is named as Hovenia dulcis, and the polysaccharide of Hovenia dulcis has the function of reducing blood sugar, but the problem of low purity exists in the existing Hovenia dulcis polysaccharide preparation, and the main impurities are monosaccharide and disaccharide, so that the removal of monosaccharide and disaccharide and the retention of polysaccharide are the keys for preparing high-purity Hovenia dulcis polysaccharide and developing new products.
Shanxi province is the traditional main production area of milk goats in China, and the Guanzhong region is the largest production base of fine milk goat varieties in China. 180 million milk goats are stored in a whole province at the end of 2015, the goat milk yield is more than 50 million tons, the market share of the goat milk powder reaches more than 95%, the number of the milk goats, the goat milk yield and the goat milk powder yield are all the first nationwide, but the domestic goat milk products are few in variety and mainly comprise goat milk powder and liquid goat milk, most of the goat milk powder is sold abroad as industrial raw materials, and the problems of low technological content, low additional value, serious homogenization and the like exist, and the foreign goat milk products are various and mainly comprise fermented goat milk products (such as cheese, skim milk cheese or goat yogurt), frozen products (such as ice cream or frozen goat milk yogurt), liquid milk, butter, functional goat milk tablets and the like. Therefore, there is a need to develop new products of functional goat milk.
Disclosure of Invention
The invention aims to provide a method for preparing a goat milk active beverage by using lactobacillus rhamnosus protease with the functions of reducing blood pressure, resisting oxidation, reducing blood sugar, preventing decayed teeth and the like.
In order to achieve the purpose, the invention adopts the technical scheme that:
1) preparation of lactobacillus rhamnosus culture medium:
adding 15-25g of lactose, 15-25g of beef extract, 10-20mg of magnesium sulfate, 800.5-1.0 mL of tween, 2-5g of galactooligosaccharide, 3-6g of peptone, 4-6g of disodium hydrogen phosphate and 5-10mg of alanine into purified water, fixing the volume to 1L, adjusting the pH value to 5.5-7.0, and sterilizing at 121 ℃ to obtain a rhamnose lactobacillus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 2-5%, culturing at constant temperature of 35-41 ℃ for 18-24h, centrifuging to collect thallus precipitate, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
3) preparation of lactobacillus rhamnosus muramidase:
adding 20-30mL of phosphate buffer solution into per g of lactobacillus rhamnosus bacterial mud, preserving heat at 35-45 ℃ for 1-3h, centrifuging to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 10-20mM, and freeze drying is carried out to obtain lactobacillus rhamnosus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 6.0-8.0 mass volume ratio, adding 1-3% Lactobacillus rhamnosus muramidase powder and 0.01-0.04% CaCl2Carrying out enzymolysis at 35-45 ℃ for 2-4h, then adding 0.5-1.0% of beta-cyclodextrin, 0.2-0.4% of hovenia dulcis thunb polysaccharide, 4-6% of erythritol, 3-5% of xylitol, 0.1-0.3% of L-malic acid, 0.10-0.25% of citric acid and 0.10-0.25% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing and sterilizing to obtain the goat milk active beverage containing antihypertensive peptide and antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 70-90%, the DPPH free radical scavenging rate is 60-80%, and the hydroxyl free radical scavenging rate is 75-95%.
The centrifugation in the step 2) is carried out for 15-25min at 4-10 ℃ and 6000-8000 r/min.
The pH value of the phosphate buffer solution of the step 3) is 7.0, and each liter of the phosphate buffer solution contains 15-30g of glycine and 0.5-1.0g of lysozyme.
The centrifugation in the step 3) is carried out for 20-30min at 4-10 ℃ and 6000-8000 r/min.
The homogenization in the step 4) is carried out at the temperature of 15-30 ℃.
The preparation process of the hovenia dulcis polysaccharide in the step 4) comprises the following steps: cleaning fresh hovenia dulcis, pulping, adding water with the weight being 20 times that of hovenia dulcis, uniformly stirring, preserving heat for 2 hours at 90 ℃, carrying out suction filtration to obtain hovenia dulcis polysaccharide extracting solution, cooling to 38 ℃, adding 2% of a bailadi yeast activating solution according to the mass-volume ratio, carrying out constant-temperature culture for 24 hours at 37 ℃, removing monosaccharide and disaccharide in the hovenia dulcis, centrifugally collecting the bailadi yeast thalli, concentrating the supernatant to 1/20 by using a rotary evaporator, adding absolute ethyl alcohol according to the volume ratio of 1:3, carrying out suction filtration by using a Buchner funnel to obtain hovenia dulcis crude polysaccharide, adding the hovenia crude polysaccharide into chloroform and n-butyl alcohol for deproteinization, adding chloroform and n-butyl alcohol into the hovenia dulcis crude polysaccharide again, taking water for deproteinization, and carrying out freeze drying to obtain the hovenia polysaccharide.
According to the invention, goat milk is taken as a raw material, cell wall protease of GRAS microorganism-lactobacillus rhamnosus is taken as a biocatalyst, protein in the goat milk is hydrolyzed into short peptide with ACE inhibitory activity and oxidation resistance, a consumer can take the short peptide to assist in reducing blood pressure and resisting oxidation, the added hovenia dulcis polysaccharide has the functions of a stabilizer and reducing blood sugar, and the functionality of the goat milk active beverage can be improved; the added erythritol and xylitol are functional sugar alcohols, and have the characteristics of low sweetness, low calorie, no decayed tooth, small influence on blood sugar and the like.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Example 1:
1) preparation of lactobacillus rhamnosus culture medium:
adding 20g of lactose, 15g of beef extract, 13mg of magnesium sulfate, 800.7 mL of tween, 3g of galacto-oligosaccharide, 3g of peptone, 6g of disodium hydrogen phosphate and 5mg of alanine into purified water, metering the volume to 1L, adjusting the pH value to 6.0, and sterilizing at 121 ℃ to obtain a lactobacillus rhamnosus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 3%, culturing at the constant temperature of 37 ℃ for 22h, centrifuging to collect thallus precipitates, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
the centrifugation is carried out for 20min at 8 ℃ and 7000 r/min;
3) preparation of lactobacillus rhamnosus muramidase:
adding 23mL of phosphate buffer solution with pH of 7.0 (20 g glycine and 1.0g lysozyme in each liter of phosphate buffer solution) into each g of lactobacillus rhamnosus bacterial sludge, keeping the temperature at 35 ℃ for 3h, centrifuging at 4 ℃ and 8000r/min for 20min to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl and CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 17mM, and freeze drying is carried out to obtain lactobacillus rhamnosus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 6.5 mass volume ratio, adding 1.5% Lactobacillus rhamnosus muramidase powder and 0.03% CaCl2Carrying out enzymolysis for 3h at 40 ℃, then adding 0.8% of beta-cyclodextrin, 0.2% of hovenia dulcis thunb polysaccharide, 5% of erythritol, 5% of xylitol, 0.3% of L-malic acid, 0.2% of citric acid and 0.15% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing at 25 ℃, and sterilizing to obtain the goat milk active beverage containing the antihypertensive peptide and the antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 80%, the DPPH free radical scavenging rate is 60%, and the hydroxyl free radical scavenging rate is 75%.
Example 2:
1) preparation of lactobacillus rhamnosus culture medium:
adding 15g of lactose, 20g of beef extract, 18mg of magnesium sulfate, 800.5 mL of tween, 4g of galacto-oligosaccharide, 5g of peptone, 4g of disodium hydrogen phosphate and 8mg of alanine into purified water, diluting to a constant volume of 1L, adjusting the pH value to 5.5, and sterilizing at 121 ℃ to obtain a lactobacillus rhamnosus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 5%, culturing at constant temperature of 35 ℃ for 24h, centrifuging to collect thallus precipitates, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
the centrifugation is carried out for 15min at the temperature of 4 ℃ and at the speed of 8000 r/min;
3) preparation of lactobacillus rhamnosus muramidase:
adding 30mL of phosphate buffer solution with pH of 7.0 (each liter of phosphate buffer solution contains 15g of glycine and 0.8g of lysozyme) into each g of lactobacillus rhamnosus bacterial sludge, preserving heat at 40 ℃ for 2h, centrifuging at 8 ℃ and 7000r/min for 5min to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl and CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 17mM, and freeze drying is carried out to obtain lactobacillus rhamnosus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 6.0 mass volume ratio, adding 3% Lactobacillus rhamnosus muramidase powder and 0.01% CaCl2Carrying out enzymolysis at 35 ℃ for 4h, then adding 0.5% of beta-cyclodextrin, 0.4% of hovenia dulcis thunb polysaccharide, 4% of erythritol, 3% of xylitol, 0.1% of L-malic acid, 0.15% of citric acid and 0.10% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing at 15 ℃, and sterilizing to obtain the goat milk active beverage containing antihypertensive peptide and antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 70%, the DPPH free radical scavenging rate is 80%, and the hydroxyl free radical scavenging rate is 85%.
Example 3:
1) preparation of lactobacillus rhamnosus culture medium:
adding 25g of lactose, 20g of beef extract, 20mg of magnesium sulfate, 801.0 mL of tween, 2g of galacto-oligosaccharide, 6g of peptone, 5g of disodium hydrogen phosphate and 10mg of alanine into purified water, metering the volume to 1L, adjusting the pH value to 6.5, and sterilizing at 121 ℃ to obtain a lactobacillus rhamnosus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 2%, culturing at the constant temperature of 39 ℃ for 20h, centrifuging to collect thallus precipitates, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
the centrifugation is carried out for 25min at 10 ℃ and 6000 r/min;
3) preparation of lactobacillus rhamnosus muramidase:
adding 28mL of phosphate buffer solution with pH of 7.0 (30 g glycine and 0.5g lysozyme in each liter of phosphate buffer solution) into per g of lactobacillus rhamnosus bacterial sludge, keeping the temperature at 45 ℃ for 1h, centrifuging at 6 ℃ and 6000r/min for 30min to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl and CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 20mM, and freeze drying is carried out to obtain lactobacillus rhamnosus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 8.0 mass volume ratio, adding 1% Lactobacillus rhamnosus muramidase powder and 0.04% CaCl2Carrying out enzymolysis for 2h at 45 ℃, then adding 1.0% of beta-cyclodextrin, 0.3% of hovenia dulcis thunb polysaccharide, 6% of erythritol, 5% of xylitol, 0.2% of L-malic acid, 0.10% of citric acid and 0.25% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing at 30 ℃, and sterilizing to obtain the goat milk active beverage containing antihypertensive peptide and antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 90%, the DPPH free radical scavenging rate is 75%, and the hydroxyl free radical scavenging rate is 95%.
Example 4:
1) preparation of lactobacillus rhamnosus culture medium:
adding 18g of lactose, 22g of beef extract, 10mg of magnesium sulfate, 800.8 mL of tween, 5g of galacto-oligosaccharide, 4g of peptone, 4g of disodium hydrogen phosphate and 6mg of alanine into purified water, diluting to 1L, adjusting the pH value to 7.0, and sterilizing at 121 ℃ to obtain a lactobacillus rhamnosus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 4%, culturing at constant temperature of 41 ℃ for 18h, centrifuging to collect thallus precipitates, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
the centrifugation is carried out for 18min at the temperature of 6 ℃ and at the speed of 8000 r/min;
3) preparation of lactobacillus rhamnosus muramidase:
adding 20mL of phosphate buffer solution with pH of 7.0 (25 g glycine and 0.6g lysozyme in each liter of phosphate buffer solution) into per g of lactobacillus rhamnosus bacterial sludge, keeping the temperature at 38 ℃ for 3h, centrifuging at 10 ℃ and 6000r/min for 28min to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl and CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 10mM, and freeze drying is carried out to obtain rhamnose lactobacillus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 7.0 mass volume ratio, adding 2% Lactobacillus rhamnosus muramidase powder and 0.02% CaCl2Carrying out enzymolysis at 42 ℃ for 3h, then adding 0.6% of beta-cyclodextrin, 0.4% of hovenia dulcis thunb polysaccharide, 5% of erythritol, 4% of xylitol, 0.1% of L-malic acid, 0.25% of citric acid and 0.20% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing at 20 ℃, and sterilizing to obtain the goat milk active beverage containing antihypertensive peptide and antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 85%, the DPPH free radical scavenging rate is 70%, and the hydroxyl free radical scavenging rate is 90%.
The hovenia dulcis thunb polysaccharide in the embodiment is prepared by the following method: cleaning fresh hovenia dulcis, pulping, adding water with the weight being 20 times that of hovenia dulcis, uniformly stirring, preserving heat for 2 hours at 90 ℃, carrying out suction filtration to obtain hovenia dulcis polysaccharide extracting solution, cooling to 38 ℃, adding 2% of a bailadi yeast activating solution according to the mass-volume ratio, carrying out constant-temperature culture for 24 hours at 37 ℃, removing monosaccharide and disaccharide in the hovenia dulcis, centrifugally collecting the bailadi yeast thalli, concentrating the supernatant to 1/20 by using a rotary evaporator, adding absolute ethyl alcohol according to the volume ratio of 1:3, carrying out suction filtration by using a Buchner funnel to obtain hovenia dulcis crude polysaccharide, adding the hovenia crude polysaccharide into chloroform and n-butyl alcohol for deproteinization, adding chloroform and n-butyl alcohol into the hovenia dulcis crude polysaccharide again, taking water for deproteinization, and carrying out freeze drying to obtain the hovenia polysaccharide.
Claims (6)
1. A method for preparing goat milk active beverage by lactobacillus rhamnosus protease is characterized in that:
1) preparation of lactobacillus rhamnosus culture medium:
adding 15-25g of lactose, 15-25g of beef extract, 10-20mg of magnesium sulfate, 800.5-1.0 mL of tween, 2-5g of galactooligosaccharide, 3-6g of peptone, 4-6g of disodium hydrogen phosphate and 5-10mg of alanine into purified water, fixing the volume to 1L, adjusting the pH value to 5.5-7.0, and sterilizing at 121 ℃ to obtain a rhamnose lactobacillus culture medium;
2) culturing lactobacillus rhamnosus and collecting thalli:
inoculating lactobacillus rhamnosus into a lactobacillus rhamnosus culture medium according to the inoculation amount of 2-5%, culturing at constant temperature of 35-41 ℃ for 18-24h, centrifuging to collect thallus precipitate, washing with sterile normal saline, and centrifuging to obtain lactobacillus rhamnosus bacterial sludge;
3) preparation of lactobacillus rhamnosus muramidase:
adding 20-30mL of phosphate buffer solution into per g of lactobacillus rhamnosus bacterial mud, preserving heat at 35-45 ℃ for 1-3h, centrifuging to obtain supernatant, namely enzyme solution of muramidase, removing small molecular substances by using a 1-3KDa ultrafiltration membrane, and adding CaCl into the retention solution2Controlling CaCl2The concentration of the lactobacillus rhamnosus is 10-20mM, and freeze drying is carried out to obtain lactobacillus rhamnosus muramidase powder;
4) preparing the goat milk active beverage:
defatting fresh goat milk, adjusting pH to 6.0-8.0 mass volume ratio, adding 1-3% Lactobacillus rhamnosus muramidase powder and 0.01-0.04% CaCl2Carrying out enzymolysis at 35-45 ℃ for 2-4h, then adding 0.5-1.0% of beta-cyclodextrin, 0.2-0.4% of hovenia dulcis thunb polysaccharide, 4-6% of erythritol, 3-5% of xylitol, 0.1-0.3% of L-malic acid, 0.10-0.25% of citric acid and 0.10-0.25% of sodium citrate according to the mass-volume ratio, uniformly mixing, homogenizing and sterilizing to obtain the goat milk active beverage containing the antihypertensive peptide and the antioxidant peptide, wherein the ACE inhibitory activity of the goat milk active beverage is 70-90%, the DPPH free radical scavenging rate is 60-80%,the hydroxyl radical clearance rate is 75-95%.
2. Method for the preparation of a goat milk active drink with lactobacillus rhamnosus protease according to claim 1 characterized by: the centrifugation in the step 2) is carried out for 15-25min at 4-10 ℃ and 6000-8000 r/min.
3. Method for the preparation of a goat milk active drink with lactobacillus rhamnosus protease according to claim 1 characterized by: the pH value of the phosphate buffer solution of the step 3) is 7.0, and each liter of the phosphate buffer solution contains 15-30g of glycine and 0.5-1.0g of lysozyme.
4. Method for the preparation of a goat milk active drink with lactobacillus rhamnosus protease according to claim 1 characterized by: the centrifugation in the step 3) is carried out for 20-30min at 4-10 ℃ and 6000-8000 r/min.
5. Method for the preparation of a goat milk active drink with lactobacillus rhamnosus protease according to claim 1 characterized by: the homogenization in the step 4) is carried out at the temperature of 15-30 ℃.
6. Method for the preparation of a goat milk active drink with lactobacillus rhamnosus protease according to claim 1 characterized by: the preparation process of the hovenia dulcis polysaccharide in the step 4) comprises the following steps: cleaning fresh hovenia dulcis, pulping, adding water with the weight being 20 times that of hovenia dulcis, uniformly stirring, preserving heat for 2 hours at 90 ℃, carrying out suction filtration to obtain hovenia dulcis polysaccharide extracting solution, cooling to 38 ℃, adding 2% of a bailadi yeast activating solution according to the mass-volume ratio, carrying out constant-temperature culture for 24 hours at 37 ℃, removing monosaccharide and disaccharide in the hovenia dulcis, centrifugally collecting the bailadi yeast thalli, concentrating the supernatant to 1/20 by using a rotary evaporator, adding absolute ethyl alcohol according to the volume ratio of 1:3, carrying out suction filtration by using a Buchner funnel to obtain hovenia dulcis crude polysaccharide, adding the hovenia crude polysaccharide into chloroform and n-butyl alcohol for deproteinization, adding chloroform and n-butyl alcohol into the hovenia dulcis crude polysaccharide again, taking water for deproteinization, and carrying out freeze drying to obtain the hovenia polysaccharide.
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| CN113186176B (en) * | 2021-04-22 | 2023-04-04 | 华东理工大学 | Method for producing rhizomucor miehei lipase by total synthesis culture medium and fermentation method |
| CN113755365B (en) * | 2021-08-11 | 2024-02-27 | 天津科技大学 | Preparation method and application of lactobacillus extract |
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