Preparation method and application of small-molecule plant fermentation oil

Abstract

The present application relates to the technical field of vegetable oil processing, and particularly relates to a preparation method and application of small-molecule plant fermented oil. The preparation method of the small-molecule plant fermented oil comprises the following steps: step 1, raw material pretreatment; step 2, preparation of composite strains; step 3, segmented fermentation; and step 4, extraction and purification. The monkey bread tree seeds, black seed, jojoba seeds and white pool flower seeds are compounded and fermented according to a certain proportion for the first time, the active ingredients are complementary, and the fatty acid composition is balanced (the proportion of unsaturated fatty acids is greater than or equal to 85%), which lays a foundation for efficient fermentation degradation.

Description

Technical Field

[0001] This invention relates to the field of vegetable oil processing technology, specifically to a method for preparing small molecule fermented vegetable oil and its application. Background Technology

[0002] Plant oils are rich in unsaturated fatty acids, polyphenols, flavonoids, and other active ingredients, possessing physiological functions such as antioxidation, anti-inflammation, and lipid regulation, and are widely used in the food, cosmetics, and pharmaceutical industries. Traditional methods for preparing plant oils mainly include cold pressing, solvent extraction, and refining. These methods result in plant oils with larger molecular weights, making them less easily absorbed by the skin or intestines, and exhibiting poorer antioxidant properties and stability. For example, while cold pressing preserves natural components, the molecular structure of the oil remains unchanged, leading to low absorption rates; solvent extraction may leave harmful chemical residues, affecting safety.

[0003] To address these shortcomings, fermentation has been increasingly applied to the preparation of vegetable oils. However, existing technologies still have limitations: firstly, they often employ single-strain fermentation, resulting in limited metabolites that are difficult to effectively degrade large-molecule oils; secondly, the fermentation process lacks targeted regulation, leading to low retention rates of active ingredients (polyphenol loss rate > 40%); and thirdly, the preparation cycle is long (7-10 days), resulting in low production efficiency. Therefore, developing a method for preparing vegetable fermented oils with small molecular weight, high absorption efficiency, and good retention of active ingredients is of significant practical importance. Summary of the Invention

[0004] In view of the problems existing in the prior art, the purpose of this invention is to provide a method for preparing small molecule plant fermented oil and its application.

[0005] The objective of this invention is achieved through the following technical solution: In a first aspect, the present invention provides a method for preparing small molecule plant fermented oil, comprising the following steps: Step 1, Raw material pretreatment: Mix baobab seeds, black cumin seeds, jojoba seeds and meadowfoam seeds, dry and crush them, add deionized water, and ultrasonically break the cell walls to obtain raw material slurry; Step 2, preparation of compound bacterial strain: Lactobacillus plantarum, Saccharomyces boulardii, and Aspergillus oryzae are inoculated into the culture medium separately, and the activated bacterial solutions are mixed to prepare compound bacterial solutions; Step 3, Segmented fermentation: Add the compound bacterial liquid to the raw material slurry, first perform aerobic fermentation, then add 0.5-1.0% of the raw material mass of oligofructose, and ferment under anaerobic conditions to obtain the fermentation liquid; Step 4, extraction and purification: Centrifuge the fermentation broth to obtain the supernatant, extract it, purify it by molecular distillation, collect the light phase component, and obtain small molecule plant fermentation oil.

[0006] Preferably, in step 1, the mass ratio of baobab seeds, black cumin seeds, jojoba seeds, and meadowfoam seeds is 1-3:2-6:2-4:1-3.

[0007] Preferably, in step 1, the mass ratio of baobab seeds, black cumin seeds, jojoba seeds, and meadowfoam seeds is 2:4:3:2.

[0008] Preferably, in step 1, the ultrasonic power is 250W, the ultrasonic time is 18min, and the ultrasonic temperature is 38℃.

[0009] Preferably, in step 1, the amount of deionized water added is 5 times the mass of the raw materials.

[0010] Preferably, in step 2, the volume ratio of Lactobacillus plantarum solution, Saccharomyces boulardii solution, and Aspergillus oryzae solution is 1:2:1.

[0011] Preferably, in step 2, the Lactobacillus plantarum is numbered ATCC 14917, Saccharomyces boulardii is numbered CNCM I-1079, and Aspergillus oryzae is numbered ATCC 42149.

[0012] Preferably, in step 2, the concentration of the compound bacterial solution is 1×10⁻⁶. 8 -5×10 8 CFU / mL.

[0013] Preferably, in step 3, the compound bacterial solution is added to the raw material slurry at an inoculation rate of 8-12% of the raw material mass.

[0014] Preferably, in step 3, the aerobic fermentation conditions are 30-32℃, pH=5.5-6.0, and 24-36h.

[0015] Preferably, in step 3, the mass of fructooligosaccharide is 0.5-1.0% of the mass of the raw material.

[0016] Preferably, in step 3, the anaerobic fermentation conditions are 28-30℃ and 48-60h.

[0017] Preferably, in step 4, subcritical CO2 extraction is used, and the extraction conditions are: pressure 25-30 MPa, temperature 35-40℃, and time 2-3 h.

[0018] Preferably, in step 4, the molecular distillation temperature is 80-90℃ and the pressure is 1-5Pa.

[0019] Secondly, the present invention provides a small molecule plant fermented oil, which is prepared by the above-described preparation method.

[0020] Thirdly, this invention provides an application of small molecule plant fermented oil in functional foods, cosmetics, and pharmaceutical carriers.

[0021] Preferably, the functional food includes one of a nutrient fortifier and an oral emulsion, and the amount of fermented oil added is 8-20% of the total mass of the food.

[0022] Preferably, the cosmetic includes one of a repairing essence and an anti-aging cream, and the amount of fermented oil added is 5-10% of the total mass of the cosmetic.

[0023] Preferably, the pharmaceutical carrier is used in a sustained-release system for lipid-soluble drugs to increase the drug half-life by ≥2 times.

[0024] The beneficial effects of this invention are as follows: 1. This invention is the first to combine baobab seeds, black cumin seeds, jojoba seeds, and meadowfoam seeds in a specific ratio for compound fermentation. The active ingredients are complementary, and the fatty acid composition is balanced (unsaturated fatty acids account for ≥85%), laying the foundation for efficient fermentation and degradation. Baobab seeds are rich in polyphenols such as gallic acid, with antioxidant activity 2-3 times that of ordinary vegetable oils; black cumin seeds contain anti-inflammatory components such as nigerin, which can synergistically enhance the physiological activity of the fermented oil; jojoba seeds have oils in the form of liquid wax esters, exhibiting excellent oxidative stability (oxidation induction period >80h); and meadowfoam seeds contain long-chain fatty acids (C20-C24), which can regulate the fluidity and skin compatibility of the fermented oil.

[0025] 2. The preparation process of this invention first uses low-temperature ultrasonic cell disruption to precisely disrupt the cell wall structure of the four raw materials, increasing the release rate of oils and active ingredients by 40-50%, while avoiding high-temperature damage to heat-sensitive components; then, through synergistic fermentation of compound microorganisms, a "degradation-conversion-synthesis" process is achieved. First, aerobic fermentation promotes the proliferation of microorganisms, followed by anaerobic fermentation with the addition of galactooligosaccharides to enhance the degradation of macromolecules, with a triglyceride conversion rate of ≥90%; finally, efficient extraction and purification are performed, with subcritical CO2 extraction to specifically extract wax esters and fatty acids, and molecular distillation to separate macromolecular impurities, ultimately obtaining small molecule components with a molecular weight ≤300Da.

[0026] 3. The small molecule plant fermented oil prepared by this invention has the following advantages: (1) Small molecule and high absorption rate: The average molecular weight of the fermented oil is 260-290 Da, and the in vitro intestinal absorption rate is ≥68%, which is 2.2 times that of traditional cold-pressed oil; (2) High retention of active ingredients: Polyphenol retention rate is ≥90%, sterol retention rate is ≥88%, and anti-inflammatory activity (inhibition of NO generation rate) is ≥75%; (3) Excellent oxidative stability: After 30 days of storage at 60℃, the acid value is ≤1.2mgKOH / g, the peroxide value is ≤3.0mmol / kg, and the shelf life is extended to more than 18 months. Detailed Implementation

[0027] The technical solution of the present invention is illustrated below through specific examples. It should be understood that the one or more method steps mentioned in the present invention do not preclude the existence of other method steps before or after the combined steps, or the insertion of other method steps between these explicitly mentioned steps; it should also be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the present invention. Furthermore, unless otherwise stated, the numbering of each method step is merely a convenient tool for identifying each method step, and not for limiting the order of the method steps or defining the scope of the present invention. Changes or adjustments to their relative relationships, without substantially altering the technical content, should also be considered within the scope of the present invention.

[0028] To better understand the above technical solutions, exemplary embodiments of the present invention are described in more detail below. While exemplary embodiments of the present invention are shown, it should be understood that the present invention can be implemented in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided to enable a more thorough understanding of the present invention and to fully convey the scope of the invention to those skilled in the art.

[0029] The present invention will be further described below with reference to the following embodiments. Example

[0030] A method for preparing a small-molecule plant fermented oil includes the following steps: (1) Raw material pretreatment: Take 20g of baobab seeds, 40g of black cumin seeds, 30g of jojoba seeds and 20g of meadowfoam seeds (mass ratio 2:4:3:2), dry them at 60℃ until the moisture content is 5%, crush them through a 100-mesh sieve, add 550mL of deionized water, and ultrasonically break them at 300W and 40℃ for 22min to obtain raw material slurry.

[0031] (2) Preparation of compound strains: Lactobacillus plantarum (ATCC14917) was inoculated into MRS medium, Saccharomyces boulardii (CNCMI-1079) into YPD medium, and Aspergillus oryzae (ATCC42149) into PDA medium, respectively. After activation at 37℃ for 24 h, the mixtures were mixed at a volume ratio of 1:1:2 and diluted to 4×10⁻⁶. 8 CFU / mL was used to prepare a compound bacterial solution; The MRS medium composition (per liter of medium) is as follows: peptone (tryptone): 10.0 g; beef extract: 10.0 g; yeast extract: 5.0 g; glucose: 20.0 g; Tween-80 (polysorbate-80): 1.0 mL; dipotassium hydrogen phosphate (K2HPO4・3H2O): 2.0 g; sodium acetate (CH3COONa・3H2O): 5.0 g; triammonium citrate ((NH4)3C6H5O7): 2.0 g; magnesium sulfate (MgSO4・7H2O): 0.58 g; manganese sulfate (MnSO4・4H2O): 0.25 g; distilled water: 1000 mL.

[0032] YPD medium composition (per liter of medium): Yeast extract: 10.0g; Peptone: 20.0g; Dextrose: 20.0g; Distilled water: 1000mL.

[0033] PDA medium composition (per liter of medium): Potato (peeled): 200.0g; Glucose: 20.0g; Agar: 15-20g (for solid medium, not for liquid medium); Distilled water: 1000mL.

[0034] (3) Segmented fermentation: Add 13.2 mL of compound bacterial solution (12% inoculum) to the raw material slurry, and ferment aerobically at 33℃ and pH 6.0 for 36 h. Add 1.1 g of galactooligosaccharide and ferment anaerobically at 30℃ for 60 h to obtain fermentation liquid.

[0035] (4) Extraction and purification: The fermentation broth was centrifuged at 9000 r / min for 20 min and the supernatant was collected. Subcritical CO2 extraction was performed (pressure 30 MPa, temperature 40℃, time 3 h). The extract was purified by molecular distillation (temperature 90℃, pressure 4 Pa). The light phase components were collected to obtain 38.5 g of small molecule plant fermentation oil, with a yield of 35.0%. Example

[0036] A method for preparing a small-molecule plant fermented oil includes the following steps: (1) Raw material pretreatment: 10g of baobab seeds, 20g of black cumin seeds, 20g of jojoba seeds and 10g of meadowfoam seeds (mass ratio 1:2:2:1) were dried at 55℃ to a moisture content of 6%, pulverized through a 100-mesh sieve, and 400mL of deionized water were added. The mixture was ultrasonically broken at 250W and 38℃ for 25min to obtain raw material slurry.

[0037] (2) Preparation of compound strain: The strain was activated according to the method in Example 1, and then diluted to 2×10⁻⁶ after mixing. 8 CFU / mL was used to prepare a compound bacterial solution.

[0038] (3) Segmented fermentation: Add 6 mL of compound bacterial solution (10% inoculum), ferment aerobically at 32℃ and pH 5.8 for 40 h, add 0.48 g of galactooligosaccharide, and ferment anaerobically at 29℃ for 70 h to obtain fermentation broth.

[0039] (4) Extraction and purification: After centrifugation, subcritical CO2 extraction (28MPa, 38℃, 3.5h) and molecular distillation (85℃, 2Pa) were performed to purify the fermented oil, yielding 26.2g of fermented oil with a yield of 32.8%. Example

[0040] A method for preparing a small-molecule plant fermented oil includes the following steps: (1) Raw material pretreatment: 30g of baobab seeds, 60g of black cumin seeds, 40g of jojoba seeds and 30g of meadowfoam seeds (mass ratio 3:6:4:3), dried at 65℃ to a moisture content of 7%, pulverized through a 100-mesh sieve, added to 900mL of deionized water, and ultrasonically broken at 350W and 42℃ for 20min to obtain raw material slurry.

[0041] (2) Preparation of compound strain: The strain was activated according to the method in Example 1, and then diluted to 6×10⁻⁶ after mixing. 8 CFU / mL was used to prepare a compound bacterial solution.

[0042] (3) Segmented fermentation: Add 24 mL of compound bacterial solution (15% inoculum), ferment aerobically at 34℃ and pH 6.2 for 30 h, add 1.6 g of galactooligosaccharide, and ferment anaerobically at 31℃ for 50 h to obtain fermentation broth.

[0043] (4) Extraction and purification: After centrifugation, subcritical CO2 extraction (32MPa, 42℃, 2.5h) and molecular distillation (95℃, 6Pa) were performed to purify the fermented oil, yielding 58.7g of fermented oil with a yield of 36.7%.

[0044] Compare with Example 1 Single-ingredient fermentation (black cumin seeds only) Except for replacing the raw material with 100g of black cumin seeds, the remaining steps were the same as in Example 1, yielding 22.3g of fermented oil with a yield of 22.3%.

[0045] Compare with Example 2 Single-strain fermentation (Saccharomyces boulardii only) Except for replacing the compound bacterial solution with a single strain of Saccharomyces boulardii (CGMCC2.382, concentration 4×10⁻⁶). 8 Except for the CFU / mL inoculum (12%), the remaining steps were the same as in Example 1, yielding 30.1g of fermented oil with a yield of 27.4%.

[0046] Compare with Example 3 Molecular distillation purification Except for omitting the molecular distillation step, the remaining steps were the same as in Example 1, yielding 41.2g of fermented oil with a yield of 37.5%.

[0047] Experimental testing and result analysis I. Detection Methods 1. Molecular weight distribution: High performance gel permeation chromatography (HPGPC), mobile phase: acetonitrile-water (80:20), flow rate: 0.8 mL / min; 2. In vitro intestinal absorption rate: The lipid permeation rate within 2 hours was calculated using a rat intestinal eversion sac model. 3. Content of active ingredients: polyphenols (Folin-phenol method), sterols (gas chromatography method), and black cumin (HPLC method). 4. Oxidative stability: The oxidation induction period was determined by the Rancimat method (100℃, air flow rate 20L / h), and the acid value and peroxide value were determined by the Schaal oven method (60℃, 30 days). 5. Anti-inflammatory activity: LPS-induced RAW264.7 cell model, NO production inhibition rate was detected.

[0048] II. Test Results

[0049] III. Results Analysis (1) Small molecular weight and absorption efficiency: The average molecular weight of Examples 1-3 is ≤290Da and the intestinal absorption rate is ≥68%, which is significantly better than the control example (molecular weight ≥360Da, absorption rate ≤45%), indicating that the composite raw materials and segmented fermentation can efficiently degrade macromolecular oils.

[0050] (2) Active ingredients and stability: The polyphenol and sterol retention rates of the examples were ≥88%, and the oxidation induction period was >80h, which was much higher than that of single raw materials (control example 1) and single strains (control example 2), indicating the complementarity of the active ingredients of the four raw materials and the metabolic advantages of the compound strains.

[0051] (3) Anti-inflammatory activity: The NO inhibition rate of the example is ≥75%, which is due to the retention of active ingredients in black cumin seeds and the transformation during the fermentation process, verifying its application potential in the field of anti-inflammatory.

[0052] Application Example 1 The fermented oil from Example 1 was used in an antioxidant oral emulsion, specifically: Formula: 15g fermented oil, 5g whey protein, 3g fructooligosaccharides, 0.2g vitamin E, 76.8g deionized water. Preparation: Dissolve whey protein in deionized water, stir at 60℃ until dissolved, add fermented oil, fructooligosaccharides, and vitamin E, homogenize under high pressure (30MPa, 3 times), fill and sterilize to obtain an oral emulsion. This emulsion has a DPPH free radical scavenging rate ≥90%, suitable for antioxidant health care for middle-aged and elderly people.

[0053] Application Example 2 The fermented oil from Example 1 was used in a repair and anti-aging face cream, specifically: Formula: Example 1: 8g fermented oil, 0.5g ceramide, 3g squalane, 5g glycerin, 0.2g xanthan gum, 83.3g deionized water. Preparation: The oil phase (fermented oil, ceramide, squalane) and the aqueous phase (glycerin, xanthan gum, deionized water) were heated to 75°C respectively, mixed, homogenized (5000r / min, 10min), and cooled to 35°C to obtain a face cream. Human patch experiments showed no irritation and a 32% reduction in transdermal water loss, making it suitable for anti-aging and repair of sensitive skin.

[0054] Application Example 3 The fermented oil from Example 1 was used as a sustained-release carrier for lipid-soluble drugs, specifically: 10g of fermented oil from Example 1 was mixed with 0.1g of paclitaxel and stirred until dissolved to form a nanoemulsion (particle size 150-200nm). A mouse tail vein injection experiment showed that the drug half-life increased from 4.2h to 9.8h, and bioavailability increased by 2.3 times, verifying its effectiveness as a sustained-release carrier.

[0055] In the description of this specification, the references to terms such as "one embodiment," "some embodiments," "example," "specific example," or "some examples," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. The illustrative expressions of the above terms in this specification should not be construed as necessarily referring to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples. In addition, those skilled in the art can combine and integrate the different embodiments or examples described in this specification.

[0056] Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention. Those skilled in the art can make changes, modifications, substitutions and variations to the above embodiments within the scope of the present invention.

Claims

1. A method for preparing small molecule plant fermented oil, characterized in that, Includes the following steps: Step 1, Raw material pretreatment: Mix baobab seeds, black cumin seeds, jojoba seeds and meadowfoam seeds, dry and crush them, add deionized water, and ultrasonically break the cell walls to obtain raw material slurry; Step 2, preparation of compound bacterial strain: Lactobacillus plantarum, Saccharomyces boulardii, and Aspergillus oryzae are inoculated into the culture medium separately, and the activated bacterial solutions are mixed to prepare compound bacterial solutions; Step 3, Segmented fermentation: Add the compound bacterial liquid to the raw material slurry, first perform aerobic fermentation, then add 0.5-1.0% of the raw material mass of oligofructose, and ferment under anaerobic conditions to obtain the fermentation liquid; Step 4, extraction and purification: Centrifuge the fermentation broth to obtain the supernatant, extract it, purify it by molecular distillation, collect the light phase component, and obtain small molecule plant fermentation oil.

2. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 1, the mass ratio of baobab seeds, black cumin seeds, jojoba seeds, and meadowfoam seeds is 1-3:2-6:2-4:1-3.

3. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 2, the volume ratio of Lactobacillus plantarum solution, Saccharomyces boulardii solution, and Aspergillus oryzae solution is 1:2:

1.

4. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 2, the Lactobacillus plantarum is ATCC 14917, Saccharomyces boulardii is CNCM I-1079, and Aspergillus oryzae is ATCC 42149.

5. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 2, the concentration of the compound bacterial solution is 1×10⁻⁶. 8 -5×10 8 CFU / mL.

6. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 3, the compound bacterial solution is added to the raw material slurry at an inoculation rate of 8-12% of the raw material mass.

7. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 3, the conditions for aerobic fermentation are 30-32℃, pH=5.5-6.0, and 24-36h; the conditions for anaerobic fermentation are 28-30℃ and 48-60h.

8. The method for preparing a small molecule plant fermented oil according to claim 1, characterized in that, In step 4, subcritical CO2 extraction is used, and the extraction conditions are: pressure 25-30 MPa, temperature 35-40℃, and time 2-3 h. Preferably, in step 4, the molecular distillation temperature is 80-90℃ and the pressure is 1-5Pa.

9. A small-molecule plant fermented oil, characterized in that, It was prepared using the preparation method described in claim 1.

10. The application of the small molecule plant fermented oil according to claim 9 in functional foods, cosmetics, and pharmaceutical carriers.

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