CN113974099A - Preparation method and application of blood orange fermentation product with antioxidant activity - Google Patents
Preparation method and application of blood orange fermentation product with antioxidant activity Download PDFInfo
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- CN113974099A CN113974099A CN202111134635.7A CN202111134635A CN113974099A CN 113974099 A CN113974099 A CN 113974099A CN 202111134635 A CN202111134635 A CN 202111134635A CN 113974099 A CN113974099 A CN 113974099A
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- 235000005976 Citrus sinensis Nutrition 0.000 title claims abstract description 108
- 238000000855 fermentation Methods 0.000 title claims abstract description 48
- 230000004151 fermentation Effects 0.000 title claims abstract description 48
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- 238000002360 preparation method Methods 0.000 title claims abstract description 12
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- 241000186672 Lactobacillus delbrueckii subsp. bulgaricus Species 0.000 claims abstract description 22
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- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 229960003180 glutathione Drugs 0.000 description 1
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- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
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- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 description 1
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- 235000019154 vitamin C Nutrition 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
- A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
- A23L19/09—Mashed or comminuted products, e.g. pulp, purée, sauce, or products made therefrom, e.g. snacks
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/06—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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- 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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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/427—Pentosaceus
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Life Sciences & Earth Sciences (AREA)
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- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
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- Birds (AREA)
- Epidemiology (AREA)
- Gerontology & Geriatric Medicine (AREA)
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- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention discloses a preparation method and application of a blood orange fermentation product with antioxidant activity. The preparation method comprises the following steps: respectively activating and culturing lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus to make the strain number reach 107cfu/mL; mixing activated and cultured lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 0.5-1.5:1:0.5-1.5 to obtain mixed lactobacillus; inoculating the mixed lactobacillus into the blood orange stock solution according to the inoculation amount of 2-4% by volume fraction, and performing fermentation cultureAnd (5) obtaining the product. The blood orange fermentation product obtained by the method provided by the invention has obviously improved antioxidant activity and antioxidant enzyme activity relative to blood orange raw materials, can be widely applied to related fields such as food or cosmetics, and has the advantages of strong applicability, obvious economic benefit and the like.
Description
Technical Field
The invention relates to the field of biological utilization and function development of food. More particularly, relates to a preparation method and application of a blood orange fermentation product with antioxidant activity.
Background
Reactive Oxygen Species (ROS) are oxygen-containing chemically reactive chemical species, which in a biological context are natural byproducts of the normal metabolism of oxygen and play an important role in cell signaling and homeostasis. However, during environmental stress (e.g., ultraviolet light or heat exposure), ROS levels can increase dramatically, and excess ROS can attack cellular components, potentially causing severe damage to cellular structures, known as oxidative stress. Normally, the intracellular ROS are mainly balanced by the antioxidant protection system of the organism, which is divided into an enzyme system and a non-enzyme system. The enzyme system mainly comprises superoxide dismutase (SOD), Catalase (CAT), glutathione peroxidase (GSH-PX) and the like, and the non-enzyme system mainly comprises reduced Glutathione (GSH), vitamin C/E and the like. Although there is a set of effective endogenous antioxidant protection system in the body, its antioxidant regulation capacity is limited, especially when the oxidative stress is severe, it must rely on supplementing exogenous antioxidant to enhance the body's own antioxidant capacity. Therefore, the development of antioxidants has become a hot research point in the fields of food science, life science, and the like.
The blood orange is a cultivated variety plant of sweet orange of Citrus of Rutaceae, and is the only variety containing anthocyanidin in Citrus. The blood orange is rich in polyphenol such as anthocyanin, ascorbic acid, flavonoid and the like, and has various physiological effects of resisting oxidation, enhancing immunity, resisting aging, preventing cardiovascular diseases and the like, which are beneficial to body health. However, the conventional blood orange extract mainly contains an acidic solvent, and the acidic solvent can denature cell membranes of anthocyanin located near epidermal cells, so that the anthocyanin is degraded in the extraction and concentration process, and further the antioxidant activity of the blood orange is reduced. Lactic acid bacteria fermentation is a treatment mode for improving the nutritive value of fruits and vegetables, and through fermentation, not only can some active ingredients in original food be kept, but also partial original nutritive ingredients in food can be changed. The selection of excellent lactobacillus strains is the key influencing the fermentation of fruits and vegetables, and no special lactobacillus strains suitable for the fermentation of blood oranges exist at present.
Disclosure of Invention
The first purpose of the invention is to provide a preparation method of a blood orange fermentation product with antioxidant activity.
The second purpose of the invention is to provide a blood orange ferment with antioxidant activity.
The third purpose of the invention is to provide the application of the blood orange fermentation product with antioxidant activity.
In order to achieve the purpose, the invention provides the following technical scheme:
in a first aspect, the present invention provides a method for preparing a fermented product of blood orange with antioxidant activity, comprising the following steps:
respectively activating and culturing lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus to make the strain number reach 107cfu/mL；
Mixing activated and cultured lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 0.5-1.5:1:0.5-1.5 to obtain mixed lactobacillus;
inoculating the mixed lactobacillus into the blood orange stock solution according to the inoculation amount of 2-4% of the volume fraction, and performing fermentation culture to obtain the product.
The invention discloses a method for extracting blood orange, which is characterized in that the traditional extraction method is easy to cause the loss of active ingredients of blood orange, and further the reduction of antioxidant activity. In addition, the invention also unexpectedly discovers that the blood orange stock solution is subjected to mixed fermentation by specific lactobacillus within the range of the mixture ratio of the invention, so that the synergistic effect can be achieved, and the oxidation resistance of the blood orange stock solution can be more effectively improved.
Preferably, the volume ratio of lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus in the mixed lactobacillus is 1-1.5:1: 0.5-1. The mixed lactic acid bacteria in the proportion can better improve the oxidation resistance of the blood orange stock solution.
More preferably, the volume ratio of lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus in the mixed lactic acid bacteria is 1:1: 1. The mixed lactic acid bacteria in the proportion can optimally improve the oxidation resistance of the blood orange stock solution.
Further, in the above method, the conditions of the activation culture are: the temperature of the activation culture is 30-37 ℃, and the time of the activation culture is 24-48 h.
The conditions of the fermentation culture are as follows: the temperature of fermentation culture is 30-37 deg.C, and the time of fermentation culture is 4-8 days.
The preparation method of the blood orange stock solution comprises the following steps:
peeling blood orange, pulping to obtain blood orange serous fluid, adjusting pH of the blood orange serous fluid to 4.0-6.0, adding mixed enzyme 0.16-0.26% of the blood orange serous fluid, and performing enzymolysis at 50 deg.C for 1.5-2 hr to obtain the final product; wherein the mixed enzyme consists of pectinase and cellulase with the mass ratio of 1: 0.5-2.
Further, the enzyme activity of the pectinase is 30-50U/mg, and the enzyme activity of the cellulase is 10-50U/mg.
According to the specific embodiment of the invention, the enzymolysis treatment further comprises sterilization at 115 ℃ for 20-30 min.
In a second aspect, the invention provides a blood orange ferment with antioxidant activity prepared by the preparation method.
The fermented product of blood orange may be in a solid state or a liquid state, which is not limited in the present invention.
In a third aspect, the invention provides an application of a blood orange fermentation product with antioxidant activity in preparing food or cosmetics for delaying senescence.
The invention has the following beneficial effects:
the invention provides a preparation method of a blood orange fermentation product with antioxidant activity, which has the advantages of cheap and easily-obtained raw materials, simple process and convenient operation, and is suitable for large-scale production and application.
The preparation method of the blood orange fermentation product provided by the invention not only can fully exert the potential functional activity of the blood orange, but also can avoid the loss of active substances of the blood orange. The blood orange fermentation product obtained by the method has obviously improved antioxidant activity and antioxidant enzyme activity relative to blood orange raw materials, can be widely applied to related fields such as food or cosmetics, and has the advantages of strong applicability, obvious economic benefit and the like.
Detailed Description
In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
Unless otherwise specified, the preparation methods of the present invention are conventional methods unless otherwise specified, and the raw materials used are commercially available from published sources or prepared according to the prior art, unless otherwise specified, and the percentages are mass percentages. Wherein, the lactobacillus casei used in the following examples is purchased from, and the lactobacillus bulgaricus and the pediococcus pentosaceus are purchased from the Guangdong province microorganism strain preservation center; both pectinase and cellulase were purchased from naning pompe bioengineering, ltd.
Example 1
Preparing a blood orange stock solution: peeling blood orange fruits, homogenizing to obtain blood orange serum, adjusting pH value of the blood orange serum to 4.0-6.0, adding mixed enzyme (the addition amount of the mixed enzyme is 0.2% of the mass of the blood orange serum), wherein the mixed enzyme comprises pectinase (50U/mg) and cellulase (10U/mg) in a mass ratio of 1:1, performing enzymolysis at 50 ℃ for 1.5h, and sterilizing at 115 ℃ for 20min to obtain blood orange stock solution.
Fermentation treatment: respectively inoculating Lactobacillus casei, Lactobacillus bulgaricus and Pediococcus pentosaceus to MRS culture medium, and respectively culturing at 30-37 deg.C for 24-48 hr to make the strain number reach 107cfu/mL, namely obtaining activated (expanded culture) strains, and mixing the activated lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 1:1:1 to obtainAnd (3) obtaining mixed lactic acid bacteria, adding the mixed lactic acid bacteria into the blood orange stock solution in an inoculation amount of which the volume fraction is 2%, standing at 30 ℃, performing fermentation culture for 4 days to obtain blood orange fermentation liquor, and performing vacuum freeze drying on the blood orange fermentation liquor to obtain a solid blood orange fermentation product.
Example 2
Preparing a blood orange stock solution: peeling blood orange fruits, homogenizing to obtain blood orange serum, adjusting pH value of the blood orange serum to 4.0-6.0, adding mixed enzyme (the addition amount of the mixed enzyme is 0.2% of the mass of the blood orange serum), wherein the mixed enzyme comprises pectinase (50U/mg) and cellulase (10U/mg) in a mass ratio of 1:1, performing enzymolysis at 50 ℃ for 1.5h, and sterilizing at 115 ℃ for 20min to obtain blood orange stock solution.
Fermentation treatment: respectively inoculating Lactobacillus casei, Lactobacillus bulgaricus and Pediococcus pentosaceus to MRS culture medium, and respectively culturing at 30-37 deg.C for 24-48 hr to make the strain number reach 107cfu/mL, namely obtaining activated (expanded culture) strains, mixing the activated lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 0.5:1:1.5 to obtain mixed lactobacillus, adding the mixed lactobacillus into the blood orange stock solution in an inoculation amount of 2%, standing at 30 ℃, performing fermentation culture for 4 days to obtain blood orange fermentation liquor, and performing vacuum freeze drying on the blood orange fermentation liquor to obtain a solid blood orange fermentation product.
Example 3
Preparing a blood orange stock solution: peeling blood orange fruits, homogenizing to obtain blood orange serum, adjusting pH value of the blood orange serum to 4.0-6.0, adding mixed enzyme (the addition amount of the mixed enzyme is 0.2% of the mass of the blood orange serum), wherein the mixed enzyme comprises pectinase (50U/mg) and cellulase (10U/mg) in a mass ratio of 1:1, performing enzymolysis at 50 ℃ for 1.5h, and sterilizing at 115 ℃ for 20min to obtain blood orange stock solution.
Fermentation treatment: respectively inoculating Lactobacillus casei, Lactobacillus bulgaricus and Pediococcus pentosaceus to MRS culture medium, and respectively culturing at 30-37 deg.C for 24-48 hr to make the strain number reach 107cfu/mL, namely obtaining each activated (expanded culture) strain, and obtaining activated cheese milk rodsMixing bacteria, lactobacillus bulgaricus and pediococcus pentosaceus according to a volume ratio of 1.5:1:0.5 to obtain mixed lactobacillus, adding the mixed lactobacillus into the blood orange stock solution in an inoculation amount of 2%, standing at 30 ℃, performing fermentation culture for 4d to obtain blood orange fermentation liquor, and performing vacuum freeze drying to obtain solid blood orange fermentation product.
Comparative example 1
Preparing blood orange fruit powder: peeling blood orange fruits, homogenizing to obtain blood orange serum, adjusting the pH value of the blood orange serum to 4.0-6.0, adding mixed enzyme (the addition amount of the mixed enzyme is 0.2% of the mass of the blood orange serum), wherein the mixed enzyme comprises pectinase (50U/mg) and cellulase (10U/mg) in a mass ratio of 1:1, performing enzymolysis at 50 ℃ for 1.5h, sterilizing at 115 ℃ for 20min to obtain blood orange stock solution, and performing vacuum freeze drying on the blood orange stock solution to obtain blood orange fruit powder.
Comparative example 2
Only the difference from example 1 is that: replacing the mixed lactobacillus with lactobacillus casei with equal bacterial amount.
Comparative example 3
Only the difference from example 1 is that: replacing the mixed lactobacillus with lactobacillus bulgaricus with equal bacterial amount.
Comparative example 4
Only the difference from example 1 is that: the mixed lactic acid bacteria are replaced by pediococcus pentosaceus with equal bacterial amount.
Comparative example 5
Only the difference from example 1 is that: the mixed lactic acid bacteria are prepared from lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to a volume ratio of 0.3:1: 1.7.
Comparative example 6
Only the difference from example 1 is that: the mixed lactic acid bacteria are prepared from lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 1.7:1: 0.3.
Test examples
The blood orange fermentation product and the blood orange fruit powder prepared in the examples 1-3 and the comparative examples 1-6 are subjected to oxidation resistance index detection, and the specific detection method comprises the following steps:
1. DPPH radical scavenging Capacity determination
Diluting the blood orange fermentation samples prepared in examples 1-3 and comparative examples 1-6 and the blood orange powder samples to 1mg/mL by using deionized water respectively to obtain diluted solutions, taking 1mL of the diluted solutions, adding 1mL0.1mmol/L of DPPH-absolute ethyl alcohol solution into the diluted solutions respectively, mixing the solutions uniformly, reacting the mixed solutions for 30min at room temperature in a dark place to obtain sample solutions, and measuring the light absorption value (marked as A) of the sample solutions at the wavelength of 517nm1) And the blank group uses absolute ethyl alcohol to replace sample liquid to measure the light absorption value (marked as A)0) In the control group, absolute ethyl alcohol is used to replace DPPH-absolute ethyl alcohol solution, and the light absorption value (marked as A) of the solution obtained after the reaction is measured2). DPPH radical clearance was calculated according to the following formula:
DPPH radical scavenging ratio (%) - [1- (A)1–A2)/A0]×100
2. Determination of superoxide anion radical scavenging ability
Diluting the blood orange fermentation samples prepared in examples 1-3 and comparative examples 1-6 and the blood orange fruit powder samples to 1mg/mL by deionized water respectively to obtain diluted solutions, taking 1mL of the diluted solutions, adding 5mL of 50mmol/L Tris-HCl buffer solution (pH 8.2) into the diluted solutions respectively, mixing uniformly, adding 0.4mL of 25mmol/L pyrogallol solution which is also preheated to 25 ℃ after 20min of water bath in a constant-temperature water bath kettle at 25 ℃, reacting for 5min after mixing uniformly, adding 0.5mL of concentrated hydrochloric acid to stop the reaction to obtain sample solutions, and testing the absorbance value (marked as A) of the sample solutions at the wavelength of 325nm1). The blank group is measured for its absorbance value (marked as A) by using distilled water instead of sample liquid0) And in the control group, distilled water is used for replacing pyrogallol solution to measure the light absorption value (marked as A) of the solution obtained after the reaction2). Superoxide anion clearance was calculated according to the following equation:
superoxide anion scavenging rate (%) - [1- (A)1–A2)/A0]×100
3. Measurement of hydroxyl radical scavenging ability
The blood orange fermentation samples and the blood orange powder samples prepared in examples 1-3 and comparative examples 1-6 are respectively diluted to 1mg/mL by deionized water to obtain diluted solutions, 1mL of the diluted solutions are taken, and 1mL of 6mmol/L FeSO is respectively added into the diluted solutions4Solution and 1mL of 2.4mmol/L H2O2Mixing the solutions, reacting for 10min, adding 1mL of 6mmol/L salicylic acid solution, reacting at 37 deg.C for 30min to obtain sample solution, and measuring its absorbance (denoted as A) at 510nm1). The blank group is measured for its absorbance value (marked as A) by using distilled water instead of sample liquid0) The control group uses distilled water instead of H2O2Solution determination the absorbance value (denoted A) of the solution obtained after the measurement reaction2). Hydroxyl radical clearance was calculated according to the following formula:
hydroxyl radical clearance (%) - [1- (a)1–A2)/A0]×100
4. ABTS free radical scavenging Capacity assay
Preparing ABTS free radical (ABTS +) solution: mixing 2.45mmol/L potassium persulfate with 7mmol/L ABTS +, reacting at 25 deg.C in dark condition for 16h, and diluting ABTS + solution with methanol until the absorbance value at 734nm is 0.70 + -0.02.
Diluting the blood orange fermentation samples and the blood orange powder samples prepared in examples 1-3 and comparative examples 1-6 to 1mg/mL by using deionized water, respectively taking 50uL in a test tube, adding 2mL of ABTS + solution, uniformly mixing, and reacting for 10min in the dark condition to obtain sample liquid. Measurement of the absorbance at a wavelength of 734nm (denoted A)1) The blank group is used for measuring the light absorption value (marked as A) of the blank group by replacing the sample liquid with methanol0) And the control group uses methanol to replace ABTS + solution to measure the light absorption value (marked as A) of the solution obtained after the reaction2). ABTS free radical clearance is calculated according to the following formula:
ABTS free radical scavenging rate (%) [1- (A)1–A2)/A0]×100
5. Determination of superoxide dismutase (SOD) Activity
SOD activity is measured by adopting a kit of Nanjing institute of bioengineering: respectively diluting the blood orange fermentation sample and the blood orange fruit powder sample prepared in examples 1-3 and comparative examples 1-6 to 1mg/mL by using deionized water to obtain sample solutions, and measuring the SOD content according to the requirements of a kit.
The results of the above tests are summarized in table 1.
Table 1:
and (4) conclusion: as can be seen from the data in Table 1, 1) the blood orange fermentation products of examples 1-3 and comparative examples 2-6 have higher antioxidant activity than the unfermented blood orange powder of comparative example 1, which indicates that the original antioxidant activity of the blood orange stock solution can be improved by fermentation; 2) the antioxidant activity of the blood orange fermentate obtained by fermenting the blood orange in the examples 1-3 and the comparative examples 5-6 with the mixed lactic acid bacteria is higher than that of the blood orange fermentate obtained by fermenting the blood orange with the single lactic acid bacteria in the comparative examples 2-4, which shows that the lactobacillus casei, the lactobacillus bulgaricus and the pediococcus pentosaceus can be matched with each other to synergistically improve the antioxidant activity of the blood orange; 3) the antioxidant activity of the blood orange fermentate obtained by fermenting the blood orange fermentate with the mixed lactic acid bacteria in the specific proportion is higher than that of the blood orange fermentate in the comparative examples 5-6, which shows that the antioxidant activity of the blood orange can be improved better and synergistically in the specific proportion range by lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus.
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention, and it will be obvious to those skilled in the art that other variations or modifications may be made on the basis of the above description, and all embodiments may not be exhaustive, and all obvious variations or modifications may be included within the scope of the present invention.
Claims (10)
1. A preparation method of a blood orange fermentation product with antioxidant activity is characterized by comprising the following steps:
respectively activating and culturing lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus to make the strain number reach 107cfu/mL；
Mixing activated and cultured lactobacillus casei, lactobacillus bulgaricus and pediococcus pentosaceus according to the volume ratio of 0.5-1.5:1:0.5-1.5 to obtain mixed lactobacillus;
inoculating the mixed lactobacillus into the blood orange stock solution according to the inoculation amount of 2-4% of the volume fraction, and performing fermentation culture to obtain the product.
2. The method according to claim 1, wherein the mixed lactic acid bacteria comprise Lactobacillus casei, Lactobacillus bulgaricus and Pediococcus pentosaceus in a volume ratio of 1-1.5:1: 0.5-1.
3. The method according to claim 1, wherein the mixed lactic acid bacteria comprise Lactobacillus casei, Lactobacillus bulgaricus and Pediococcus pentosaceus in a volume ratio of 1:1: 1.
4. The method according to claim 1, wherein the activation culture is carried out under the following conditions: the temperature of the activation culture is 30-37 ℃, and the time of the activation culture is 24-48 h.
5. The method according to claim 1, wherein the conditions of the fermentation culture are: the temperature of fermentation culture is 30-37 deg.C, and the time of fermentation culture is 4-8 days.
6. The method for preparing blood orange stock solution according to claim 1, comprising the following steps:
peeling blood orange, pulping to obtain blood orange serous fluid, adjusting pH of the blood orange serous fluid to 4.0-6.0, adding mixed enzyme with a mass of 0.16-0.26% of the blood orange serous fluid, and performing enzymolysis at 50 deg.C for 1.5-2 hr to obtain the final product; wherein the mixed enzyme consists of pectinase and cellulase with the mass ratio of 1: 0.5-2.
7. The method according to claim 6, wherein the enzyme activity of the pectinase is 30-50U/mg and the enzyme activity of the cellulase is 10-50U/mg.
8. The method of claim 6, wherein the enzymatic treatment is followed by sterilization at 115 ℃ for 20-30 min.
9. A fermented product of blood orange having antioxidant activity obtained by the method according to any one of claims 1 to 8.
10. Use of the fermented product of blood orange according to claim 9 with antioxidant activity for preparing food or cosmetic for delaying aging.
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| KR20200009723A (en) * | 2018-07-20 | 2020-01-30 | 뉴트리진 주식회사 | An anti-obesity composition comprising fruit extract and a weight-adjusting food comprising the same |
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