Phospholipid krill oil capable of promoting absorption of intestinal flora and preparation process of phospholipid krill oil

Abstract

The invention belongs to the technical field of food processing, and particularly relates to phospholipid krill oil capable of promoting intestinal flora absorption and a preparation technology of the phospholipid krill oil. Comprising the following steps: preparing a natural flora activity accelerant; performing four-phase extraction on the krill oil with high phospholipid content; constructing an oil phase system and a water phase system; and mixing and emulsifying. The fructus hippophae, the radix morindae officinalis powder, the honey, the herba erigerontis and the grapefruit are subjected to a multi-stage fermentation process, so that various natural components and probiotic metabolites in the formula are mutually coordinated, and the intestinal bacterial activity is jointly improved; in addition, the natural protein emulsifier generated through subsequent fermentation is compounded with the phospholipid krill oil to form a high-stability emulsion, and the prepared phospholipid krill oil not only has high stability, but also can promote the absorption and multiplication capacity of intestinal probiotics.

Description

Technical Field

[0001] The invention belongs to the technical field of food processing, and in particular relates to a phospholipid krill oil capable of promoting intestinal flora absorption and a preparation process thereof. Background Art

[0002] Antarctic krill oil is considered to be a valuable nutritional supplement because it is rich in active ingredients such as ω-3 polyunsaturated fatty acids (EPA / DHA), phospholipid-bound fatty acids and astaxanthin. Its unique phospholipid-bound form can significantly improve bioavailability. In recent years, with the development of Antarctic biological resources, Antarctic krill oil has become an emerging functional nutritional food raw material.

[0003] However, the active ingredients in Antarctic krill oil have poor stability and are easily oxidized and deteriorated. They have high viscosity and poor water solubility. Currently, liposomes, microcapsules and emulsion preparation technologies are mainly used to improve the stability of Antarctic krill oil and improve its bioavailability. However, due to the poor separation selectivity of existing extraction methods for components such as astaxanthin, phospholipids and carbohydrates, the nutritional components in phospholipid krill oil are unevenly distributed, which can easily reduce the stability of the prepared emulsion or microcapsule. In addition, existing krill oil products mostly focus on the supplementation of ω-3 fatty acids and the improvement of stability, and lack the ability to directional regulate intestinal flora.

[0004] Therefore, there is an urgent need to provide a phospholipid krill oil that can effectively promote the absorption capacity of intestinal flora while maintaining high stability. Summary of the invention

[0005] In order to solve the above technical defects, the present invention has developed a preparation process of phospholipid krill oil that can promote the absorption of intestinal flora. Through the reasonable compatibility fermentation of medicine and food and the extraction, separation and reorganization of composite solvents, phospholipid krill oil with high stability and the ability to effectively promote the activity and quantity of gastrointestinal flora is prepared.

[0006] A preparation process of phospholipid krill oil capable of promoting intestinal flora absorption comprises the following steps:

[0007] S1: Preparation of natural flora activity promoter

[0008] The sea buckthorn fruit, Morinda officinalis powder, honey, Erigeron brevis and grapefruit are crushed to prepare a fermentation substrate, and then Lactobacillus rhamnosus GG and Bifidobacterium breve B-3 are inoculated for primary fermentation, and then enzymatic hydrolysis is performed with laccase and then secondary fermentation is performed with thermophilic Streptococcus, and then sterilization, filtration and concentration are performed to obtain a natural flora activity promoter;

[0009] S2: Four-phase extraction of high-phospholipid krill oil

[0010] The Antarctic krill meat is enzymatically hydrolyzed to obtain an enzymatic hydrolyzate, and then sodium citrate, ethanol and a hydrophobic composite organic solvent are sequentially added and centrifuged to extract, and the caprylic capric triglyceride-astaxanthin mixed oil phase, a phospholipid krill oil phase, a sugar-containing ethanol phase and a protein flocculation phase are obtained by layering.

[0011] S3: Construction of oil phase and water phase system and mixed emulsification

[0012] The phospholipid krill oil phase, the caprylic capric triglyceride-astaxanthin mixed oil phase and rosemary oil are mixed to prepare an oil phase mixed system, the sugar-containing ethanol phase is desolventized and mixed with β-cyclodextrin and a natural flora activity promoter to obtain a water phase mixed system, the oil phase mixed system and the water phase mixed system are mixed, and then high-speed shearing, high-pressure homogenization and sterilization canning are performed to obtain the phospholipid krill oil that can promote intestinal flora absorption.

[0013] Furthermore, step S1 of preparing a natural flora activity promoter comprises the following steps:

[0014] S1.1: 40-50 parts by weight of seabuckthorn fruit, 15-25 parts by weight of Morinda officinalis powder, 20-40 parts by weight of honey, 5-10 parts by weight of Erigeron brevis and 10-30 parts by weight of grapefruit are mixed and crushed into jam, and then placed in a fermentation tank with 3-5 times the weight of sterile water and stirred evenly to obtain a fermentation substrate, inoculated with Lactobacillus rhamnosus GG seed liquid at an inoculation amount of 6-8% in the fermentation substrate, and inoculated with Bifidobacterium breve B-3 seed liquid at an inoculation amount of 2-4%, adjusted the initial pH of the fermentation to 6.8-7, and fermented at 37-40°C for 20-24 hours in a microaerobic manner to obtain a primary fermentation liquid;

[0015] S1.2: adding laccase to the primary fermentation broth at an enzyme-substrate ratio of 200-300 U / mg, then adding 0.1-0.2 wt% of a metal cofactor, fermenting under microaerobic conditions at 40-45° C. for 20-24 hours, sterilizing at 105-110° C., adding 0.3-0.5 wt% of xylo-oligosaccharide, and then inoculating a thermophilic Streptococcus seed solution at an inoculum amount of 4-6%, and aerobically fermenting at 40-42° C. and 160-180 rpm for 30-36 hours to obtain a crude fermentation broth;

[0016] S1.3: The crude fermentation liquid is sterilized at 105-110°C, passed through a ceramic membrane with a pore size of 8-10 μm, and then concentrated to 30-35% of the original volume to obtain a natural bacterial flora activity promoter.

[0017] Furthermore, step S2, four-phase extraction of high phospholipid content krill oil, comprises the following steps:

[0018] S2.1: The thawed Antarctic krill meat and deionized water are placed in a high-speed homogenizer at a material-liquid ratio of 1: (20-25), homogenized at a speed of 8000-10000 rpm for 0.5-1 minute, and then ultrasonically crushed at an ultrasonic frequency of 25-30 kHz for 10-15 minutes, the pH is adjusted to 4-4.5, and 0.25-0.3 wt% of enzyme preparation of fresh Antarctic krill meat is added respectively, and then placed in a constant temperature shaker at 50-55°C, enzymolyzed at 200-220 rpm for 4-5 hours, and then placed in a water bath at 95-100°C for inactivation for 15-20 minutes to obtain shrimp meat enzymolyzate;

[0019] S2.2: Add 15-16wt% sodium citrate to the shrimp meat hydrolysate, stir until the sodium citrate is completely dissolved, then add 18-22wt% ethanol, stir for 2-3 minutes, then add 18-20wt% hydrophobic composite organic solvent, stir evenly, centrifuge at 10000-12000rpm for 3-4 minutes to obtain a four-phase solution, separate the four-phase solution into layers, and obtain a caprylic / capric triglyceride-astaxanthin mixed oil phase, a phospholipid krill oil phase, a sugar-containing ethanol phase, and a protein flocculation phase.

[0020] Furthermore, step S3 of constructing the oil phase and water phase system and mixing and emulsifying comprises the following steps:

[0021] S3.1: 8-12 parts by weight of phospholipid krill oil phase, 1-1.2 parts by weight of caprylic capric triglyceride-astaxanthin mixed oil phase and 0.1-0.2 parts by weight of rosemary oil are mixed in a container, and stirred at 50-60° C. for 15-20 minutes to obtain an oil phase mixed system;

[0022] S3.2: 1.2-1.5 parts by weight of the sugar-containing ethanol phase is desolventized by vacuum distillation and then mixed with 0.8-1 parts by weight of β-cyclodextrin, and then added to 3-4 parts by weight of a natural flora activity promoter at 75-80°C and stirred for 10-12 minutes to obtain an aqueous phase mixing system, and the oil phase mixing system prepared in step S3.1 is added to the aqueous phase mixing system, and then transferred to a high-speed disperser, sheared at a speed of 8000-10000rpm for 3-4 minutes, and placed in a high-pressure homogenizer for homogenization at a pressure of 20-25MPa for 2-3 times, and sterilized by ultra-high temperature instantaneous sterilization technology, and then sealed and canned for storage to obtain phospholipid krill oil that can promote intestinal flora absorption.

[0023] Furthermore, the Lactobacillus rhamnosus GG seed solution and the Bifidobacterium breve B-3 seed solution in step S1 are obtained by selecting one ring of strains from the solid slant culture medium for preserving Lactobacillus rhamnosus GG and Bifidobacterium breve B-3, inoculating them in MRS liquid culture medium, and culturing them at 37°C and 150rpm for 12 hours. The thermophilic Streptococcus seed solution is obtained by selecting one ring of strains from the solid slant culture medium for preserving thermophilic Streptococcus, inoculating them in MRS liquid culture medium, and culturing them at 42°C and 180rpm for 12 hours.

[0024] Furthermore, the dissolved oxygen concentration of the microaerobic fermentation in step S1.1 is maintained at 0.03-0.6 mg / L, and the dissolved oxygen concentration of the microaerobic fermentation in step S1.2 is maintained at 1.2-2 mg / L.

[0025] Furthermore, the metal cofactor in step S1.2 is composed of zinc sulfate and manganese sulfate in a mass ratio of 1:(2.5-3), and both zinc sulfate and manganese sulfate are food grade.

[0026] Furthermore, the enzyme preparation in step S2.1 consists of pepsin and cellulase in a mass ratio of 1:(1-1.5).

[0027] Furthermore, the hydrophobic composite organic solvent of step S2.2 is composed of Antarctic krill oil and caprylic / capric triglyceride in a volume ratio of 1:(8-10).

[0028] A phospholipid krill oil capable of promoting intestinal flora absorption is prepared by the above-mentioned preparation process of the phospholipid krill oil capable of promoting intestinal flora absorption.

[0029] The beneficial effects are as follows: 1. The present invention uses Lactobacillus rhamnosus GG and Bifidobacterium breve B-3 to co-ferment seabuckthorn fruit, Morinda officinalis powder, honey, Erigeron breve and grapefruit, then adds laccase for enzymolysis and uses thermophilic streptococcus for secondary fermentation. Through multi-stage fermentation technology, multiple natural ingredients and probiotic metabolites in the formula cooperate with each other to jointly promote the proliferation of intestinal bacteria, regulate the balance of intestinal flora and improve the activity of intestinal probiotics. The bioavailability is high and the absorption of nutrients can be promoted. In the subsequent fermentation, the natural protein emulsifier produced is compounded with phospholipid krill oil into a high-stability emulsion, and the antioxidant capacity of the system is enhanced by substances such as polyphenols and organic acids. The prepared phospholipid krill oil not only has high stability, but also can improve the activity of intestinal flora, promote the absorption and proliferation capacity of intestinal probiotics, build a strong prebiotic network in the intestine, and improve the host immunity.

[0030] 2. The present invention uses a composite solvent system composed of sodium citrate, ethanol, Antarctic krill oil and caprylic capric triglyceride to simultaneously extract shrimp meat enzymatic hydrolysate, which can effectively separate and extract astaxanthin, phospholipids, sugars and proteins in the shrimp meat. The extraction process is simple and efficient, and the obtained phospholipid krill oil has high purity, avoiding excessive components from affecting the subsequent emulsion formation and the stability of the emulsion storage process, and can increase the phospholipid content and further improve the storage stability of the phospholipid krill oil.

[0031] 3. The present invention obtains an oil phase by mixing a phospholipid krill oil phase, caprylic capric triglyceride-astaxanthin and rosemary oil, desolventizes the sugar-containing ethanol phase, and mixes it with β-cyclodextrin and a natural flora activity promoter to obtain an aqueous phase, separates the nutrients in the shrimp meat enzymatic hydrolyzate and recombines them in a suitable proportion, and then adds rosemary oil, β-cyclodextrin and a natural flora activity promoter. The phospholipid krill oil can not only effectively mask the fishy smell, but also has good storage stability, can improve the activity of intestinal flora, and promote the absorption and utilization of nutrients. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is a flow chart of the preparation process of phospholipid krill oil that can promote intestinal flora absorption used in an embodiment of the present invention. DETAILED DESCRIPTION

[0033] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Example 1

[0035] A phospholipid krill oil capable of promoting intestinal flora absorption and a preparation process thereof, such as Figure 1 As shown, the following steps are included:

[0036] S1: Preparation of natural flora activity promoter

[0037] S1.1: 40 parts by weight of seabuckthorn fruit, 15 parts by weight of Morinda officinalis powder, 20 parts by weight of honey, 5 parts by weight of Erigeron brevis and 10 parts by weight of grapefruit are mixed and crushed into jam, and then placed in a fermentation tank with 3 times the weight of sterile water and stirred evenly to obtain a fermentation substrate, inoculated with 6% of Lactobacillus rhamnosus GG seed liquid in the fermentation substrate, and inoculated with 2% of Bifidobacterium breve B-3 seed liquid, and 1 ring of strains of Lactobacillus rhamnosus GG seed liquid and Bifidobacterium breve B-3 seed liquid are selected from the solid slant culture medium for preserving Lactobacillus rhamnosus GG and Bifidobacterium breve B-3, inoculated into MRS liquid culture medium, and cultured at 37°C and 150rpm for 12 hours to obtain, adjusting the initial pH of the fermentation to 6.8, and fermenting at 37°C and a dissolved oxygen concentration of 0.3 mg / L for 20 hours to obtain a primary fermentation liquid;

[0038] S1.2: Add laccase to the primary fermentation broth at an enzyme-substrate ratio of 200 U / mg, then add 0.1 wt% of a metal cofactor, the metal cofactor is composed of zinc sulfate and manganese sulfate in a mass ratio of 1:2.5, and both zinc sulfate and manganese sulfate are food grade, ferment at 40°C and a dissolved oxygen concentration of 1.2 mg / L for 20 hours, add 0.3 wt% of xylo-oligosaccharides after sterilization at 105°C, and then inoculate thermophilic Streptococcus seed liquid at an inoculation rate of 4%, the thermophilic Streptococcus seed liquid is selected from a solid slant culture medium for preserving thermophilic Streptococcus, inoculate in an MRS liquid culture medium, culture at 42°C and 180 rpm for 12 hours, and aerobically ferment at 40°C and 160 rpm for 30 hours to obtain a crude fermentation broth;

[0039] S1.3: The crude fermentation liquid is sterilized at 105°C, passed through a ceramic membrane with a pore size of 8 μm, and then concentrated to 30% of the original volume to obtain a natural bacterial flora activity promoter.

[0040] S2: Four-phase extraction of high-phospholipid krill oil

[0041] S2.1: The thawed Antarctic krill meat and deionized water were placed in a high-speed homogenizer at a material-liquid ratio of 1:20, homogenized at a speed of 8000 rpm for 0.5 minutes, and then ultrasonically crushed at an ultrasonic frequency of 25 kHz for 10 minutes, the pH was adjusted to 4, and 0.25 wt% of an enzyme preparation of fresh Antarctic krill meat was added, the enzyme preparation was composed of pepsin and cellulase in a mass ratio of 1:1, and then placed in a constant temperature shaker at 50°C, enzymolyzed at 200 rpm for 4 hours, and then placed in a water bath at 95°C for inactivation for 15 minutes to obtain a shrimp meat enzymolyzate;

[0042] S2.2: Add 15wt% sodium citrate to the shrimp meat hydrolysate, stir until the sodium citrate is completely dissolved, then add 18wt% ethanol, stir for 2 minutes, then add 18wt% hydrophobic composite organic solvent, the hydrophobic composite organic solvent consists of Antarctic krill oil and caprylic capric triglyceride in a volume ratio of 1:8, stir evenly, place in a centrifuge and centrifuge at 10000rpm for 3 minutes to obtain a four-phase solution, the four-phase solution is layered and taken out to obtain caprylic capric triglyceride-astaxanthin mixed oil phase, phospholipid krill oil phase, sugar-containing ethanol phase and protein flocculation phase.

[0043] S3: Construction of oil phase and water phase system and mixed emulsification

[0044] S3.1: 8 parts by weight of phospholipid krill oil phase, 1 part by weight of caprylic capric triglyceride-astaxanthin mixed oil phase and 0.1 parts by weight of rosemary oil are mixed in a container and stirred at 50° C. for 15 minutes to obtain an oil phase mixed system;

[0045] S3.2: 1.2 parts by weight of the sugar-containing ethanol phase is desolventized by vacuum distillation and then mixed with 0.8 parts by weight of β-cyclodextrin, added to 3 parts by weight of a natural flora activity promoter at 75°C and stirred for 10 minutes to obtain an aqueous phase mixing system, the oil phase mixing system prepared in step S3.1 is added to the aqueous phase mixing system, and then transferred to a high-speed disperser, sheared at a speed of 8000 rpm for 3 minutes, and placed in a high-pressure homogenizer for homogenization twice at a pressure of 20 MPa, sterilized by ultra-high temperature instantaneous sterilization technology, and then sealed and canned for storage to obtain phospholipid krill oil that can promote intestinal flora absorption.

[0046] Example 2

[0047] A phospholipid krill oil capable of promoting intestinal flora absorption and a preparation process thereof, such as Figure 1 As shown, the following steps are included:

[0048] S1: Preparation of natural flora activity promoter

[0049] S1.1: 50 parts by weight of seabuckthorn fruit, 25 parts by weight of Morinda officinalis powder, 40 parts by weight of honey, 10 parts by weight of Erigeron brevis and 30 parts by weight of grapefruit are mixed and crushed into jam, and then placed in a fermentation tank with 5 times the weight of sterile water and stirred evenly to obtain a fermentation substrate, inoculated with 6% of Lactobacillus rhamnosus GG seed liquid in the fermentation substrate, and inoculated with 2% of Bifidobacterium breve B-3 seed liquid, and 1 ring of strains of Lactobacillus rhamnosus GG seed liquid and Bifidobacterium breve B-3 seed liquid are selected from the solid slant culture medium for preserving Lactobacillus rhamnosus GG and Bifidobacterium breve B-3, inoculated into MRS liquid culture medium, and cultured at 37°C and 150rpm for 12 hours to obtain, adjusting the initial pH of the fermentation to 6.8, and fermenting at 37°C and a dissolved oxygen concentration of 0.3 mg / L for 20 hours to obtain a primary fermentation liquid;

[0050] S1.2: Add laccase to the primary fermentation broth at an enzyme-substrate ratio of 200 U / mg, then add 0.2 wt% of a metal cofactor, the metal cofactor is composed of zinc sulfate and manganese sulfate in a mass ratio of 1:2.5, and both zinc sulfate and manganese sulfate are food grade, ferment at 40°C and a dissolved oxygen concentration of 1.2 mg / L for 20 hours, sterilize at 105°C, add 0.5 wt% of xylo-oligosaccharide, and then inoculate thermophilic streptococcus seed liquid at an inoculation rate of 4%, the thermophilic streptococcus seed liquid is selected from a solid slant culture medium for preserving thermophilic streptococcus, inoculate in an MRS liquid culture medium, culture at 42°C and 180 rpm for 12 hours, and aerobically ferment at 40°C and 160 rpm for 30 hours to obtain a crude fermentation broth;

[0051] S1.3: The crude fermentation liquid is sterilized at 105°C, passed through a ceramic membrane with a pore size of 8 μm, and then concentrated to 30% of the original volume to obtain a natural bacterial flora activity promoter.

[0052] S2: Four-phase extraction of high-phospholipid krill oil

[0053] S2.1: The thawed Antarctic krill meat and deionized water were placed in a high-speed homogenizer at a material-liquid ratio of 1:25, homogenized at a speed of 8000 rpm for 0.5 minutes, and then ultrasonically crushed at an ultrasonic frequency of 25 kHz for 10 minutes, the pH was adjusted to 4, and 0.25 wt% of an enzyme preparation of fresh Antarctic krill meat was added, the enzyme preparation was composed of pepsin and cellulase in a mass ratio of 1:1, and then placed in a constant temperature shaker at 50°C, enzymolyzed at 200 rpm for 4 hours, and then placed in a water bath at 95°C for inactivation for 15 minutes to obtain a shrimp meat enzymolyzate;

[0054] S2.2: Add 16wt% sodium citrate to the shrimp meat hydrolysate, stir until the sodium citrate is completely dissolved, then add 22wt% ethanol, stir for 2 minutes, then add 20wt% hydrophobic composite organic solvent, the hydrophobic composite organic solvent consists of Antarctic krill oil and caprylic capric triglyceride in a volume ratio of 1:8, stir evenly, place in a centrifuge and centrifuge at 10000rpm for 3 minutes to obtain a four-phase solution, the four-phase solution is layered and taken out to obtain caprylic capric triglyceride-astaxanthin mixed oil phase, phospholipid krill oil phase, sugar-containing ethanol phase and protein flocculation phase.

[0055] S3: Construction of oil phase and water phase system and mixed emulsification

[0056] S3.1: 12 parts by weight of phospholipid krill oil phase, 1.2 parts by weight of caprylic capric triglyceride-astaxanthin mixed oil phase and 0.2 parts by weight of rosemary oil are mixed in a container and stirred at 50° C. for 15 minutes to obtain an oil phase mixed system;

[0057] S3.2: 1.5 parts by weight of the sugar-containing ethanol phase is desolventized by vacuum distillation and mixed with 1 part by weight of β-cyclodextrin, and then added to 4 parts by weight of a natural flora activity promoter at 75°C and stirred for 10 minutes to obtain an aqueous phase mixing system. The oil phase mixing system prepared in step S3.1 is added to the aqueous phase mixing system, and then transferred to a high-speed disperser, sheared at a speed of 8000 rpm for 3 minutes, and placed in a high-pressure homogenizer for homogenization twice at a pressure of 20 MPa. After sterilization using ultra-high temperature instantaneous sterilization technology, the mixture is sealed and canned for storage to obtain phospholipid krill oil that can promote intestinal flora absorption.

[0058] Example 3

[0059] A phospholipid krill oil capable of promoting intestinal flora absorption and a preparation process thereof, such as Figure 1 As shown, the following steps are included:

[0060] S1: Preparation of natural flora activity promoter

[0061] S1.1: 40 parts by weight of seabuckthorn fruit, 15 parts by weight of Morinda officinalis powder, 20 parts by weight of honey, 5 parts by weight of Erigeron brevis and 10 parts by weight of grapefruit are mixed and crushed into jam, and then placed in a fermentation tank with 3 times the weight of sterile water and stirred evenly to obtain a fermentation substrate, inoculated with 8% of Lactobacillus rhamnosus GG seed liquid in the fermentation substrate, and inoculated with 4% of Bifidobacterium breve B-3 seed liquid, and one ring of strains of Lactobacillus rhamnosus GG seed liquid and Bifidobacterium breve B-3 seed liquid are selected from the solid slant culture medium for preserving Lactobacillus rhamnosus GG and Bifidobacterium breve B-3, inoculated into MRS liquid culture medium, and cultured at 37°C and 150rpm for 12 hours to obtain, adjusting the initial pH of the fermentation to 7, and fermenting at 40°C and a dissolved oxygen concentration of 0.6 mg / L for 24 hours to obtain a primary fermentation liquid;

[0062] S1.2: Add laccase to the primary fermentation broth at an enzyme-substrate ratio of 300 U / mg, then add 0.1 wt% of a metal cofactor, the metal cofactor is composed of zinc sulfate and manganese sulfate in a mass ratio of 1:3, and both zinc sulfate and manganese sulfate are food grade, ferment at 45°C and a dissolved oxygen concentration of 2 mg / L for 24 hours, sterilize at 110°C, add 0.5 wt% of xylo-oligosaccharide, and then inoculate thermophilic Streptococcus seed liquid at an inoculation rate of 6%, the thermophilic Streptococcus seed liquid is selected from a solid slant culture medium for preserving thermophilic Streptococcus, inoculate in an MRS liquid culture medium, culture at 42°C and 180 rpm for 12 hours, and aerobically ferment at 42°C and 180 rpm for 36 hours to obtain a crude fermentation broth;

[0063] S1.3: The crude fermentation broth is sterilized at 110°C, passed through a ceramic membrane with a pore size of 10 μm, and then concentrated to 35% of the original volume to obtain a natural bacterial flora activity promoter.

[0064] S2: Four-phase extraction of high-phospholipid krill oil

[0065] S2.1: The thawed Antarctic krill meat and deionized water were placed in a high-speed homogenizer at a material-liquid ratio of 1:20, homogenized at a speed of 10000 rpm for 1 minute, and then ultrasonically broken at an ultrasonic frequency of 30 kHz for 15 minutes, the pH was adjusted to 4.5, and 0.25 wt% of an enzyme preparation of fresh Antarctic krill meat was added, the enzyme preparation consisting of pepsin and cellulase in a mass ratio of 1:1.5, and then placed in a constant temperature shaker at 55°C, enzymolyzed at 220 rpm for 5 hours, and then placed in a water bath at 100°C for inactivation for 15 minutes to obtain a shrimp meat enzymolyzate;

[0066] S2.2: Add 15wt% sodium citrate to the shrimp meat hydrolysate, stir until the sodium citrate is completely dissolved, then add 18wt% ethanol, stir for 3 minutes, then add 18wt% hydrophobic composite organic solvent, the hydrophobic composite organic solvent consists of Antarctic krill oil and caprylic capric triglyceride in a volume ratio of 1:10, stir evenly, place in a centrifuge and centrifuge at 12000rpm for 4 minutes to obtain a four-phase solution, the four-phase solution is layered and taken out to obtain caprylic capric triglyceride-astaxanthin mixed oil phase, phospholipid krill oil phase, sugar-containing ethanol phase and protein flocculation phase.

[0067] S3: Construction of oil phase and water phase system and mixed emulsification

[0068] S3.1: 8 parts by weight of phospholipid krill oil phase, 1 part by weight of caprylic capric triglyceride-astaxanthin mixed oil phase and 0.1 parts by weight of rosemary oil are mixed in a container and stirred at 60° C. for 20 minutes to obtain an oil phase mixed system;

[0069] S3.2: 1.2 parts by weight of the sugar-containing ethanol phase is desolventized by vacuum distillation and then mixed with 0.8 parts by weight of β-cyclodextrin, and then added to 3 parts by weight of a natural flora activity promoter at 80°C and stirred for 12 minutes to obtain an aqueous phase mixing system. The oil phase mixing system prepared in step S3.1 is added to the aqueous phase mixing system, and then transferred to a high-speed disperser, sheared at a speed of 10,000 rpm for 4 minutes, and placed in a high-pressure homogenizer for homogenization at a pressure of 25 MPa for 3 times. After sterilization using ultra-high temperature instantaneous sterilization technology, the mixture is sealed and canned for storage to obtain phospholipid krill oil that can promote intestinal flora absorption.

[0070] Comparative Example 1

[0071] A preparation process of phospholipid krill oil capable of promoting intestinal flora absorption, which is different from Example 1 in that, in Comparative Example 1, step S1 is removed, 40 parts by weight of seabuckthorn fruit, 15 parts by weight of Morinda officinalis powder, 20 parts by weight of honey, 5 parts by weight of Erigeron breviscapus and 10 parts by weight of grapefruit are mixed and crushed into jam and then freeze-dried, and then the obtained freeze-dried powder is evenly mixed with 3 times the weight of sterile water to obtain a fruit powder solution, and the crude fermentation liquid of step S1.3 is replaced with an equal weight of fruit powder solution to obtain a natural flora activity promoter, and the remaining steps are the same as in Example 1 to obtain phospholipid krill oil, which is recorded as Comparative Example 1.

[0072] Comparative Example 2

[0073] A process for preparing phospholipid krill oil capable of promoting intestinal flora absorption is disclosed, which is different from Example 1 in that, in Comparative Example 2, the natural flora activity promoter in step S3.2 is replaced with an equal mass of physiological saline, and the remaining steps are the same as in Example 1 to prepare phospholipid krill oil, which is recorded as Comparative Example 2.

[0074] Comparative Example 3

[0075] A preparation process of phospholipid krill oil capable of promoting intestinal flora absorption, which is different from Example 1 in that, in Comparative Example 3, the shrimp meat enzymatic hydrolyzate obtained in step S2.1 is extracted by supercritical CO2 extraction technology to obtain phospholipid krill oil, and the subsequent phospholipid krill oil phase is replaced with an equal mass of phospholipid krill oil, and no caprylic capric triglyceride-astaxanthin mixed oil phase and sugar-containing ethanol phase are added in step S3. The remaining steps are the same as in Example 1 to obtain phospholipid krill oil, which is recorded as Comparative Example 3.

[0076] Comparative Example 4

[0077] A process for preparing phospholipid krill oil capable of promoting intestinal flora absorption is different from Example 1 in that rosemary oil is not added in step S3.2 of Comparative Example 4, and the remaining steps are the same as those of Example 1 to prepare phospholipid krill oil, which is recorded as Comparative Example 4.

[0078] Experiment 1: 40 mice were selected, half of them were male and half were female, and then randomly divided into four groups, 10 mice in each group, half of them were male and half were female. The normal group used feed to feed the mice normally, and the comparative example 2 group was given 10 mL / kg of comparative example 2 twice a day by gavage on the basis of normal feeding, and the comparative example 1 group was given 10 mL / kg of comparative example 1 twice a day by gavage on the basis of normal feeding. The example 1 group was given 10 mL / kg of phospholipid krill oil prepared in example 1 that can promote intestinal flora absorption twice a day by gavage on the basis of normal feeding. After feeding for 14 days, 3 mice were randomly selected from each group and killed, and then the cecal feces of each group were aseptically collected and placed in 2 mL sterile tubes, and sent to the biological company for high-throughput sequencing of intestinal flora. The test results are shown in Table 1.

[0079] Table 1: Significance of abundance of various intestinal bacterial species after 14 days of feeding

[0080] Abundance significance Normal group Comparative Example 1 Comparative Example 2 Example 1 Group Firmicutes 48.5％±3.0a 50.1％±2.8a 56.3％±3.5b 63.8％±4.2c Lactobacillus 4.2％±0.5a 7.6％±0.8b 12.3％±1.2c 21.5％±2.0d Bacteroidetes 35.2％±2.1a 32.8％±1.9a 28.5％±2.3b 22.4％±1.7c Verrucomicrobia 0.5％±0.1a 0.8％±0.2a 1.2％±0.3b 2.7％±0.4c Enterococcus 8.3％±0.9a 6.7％±0.7b 4.2％±0.5c 1.5％±0.3d Staphylococcus 5.1％±0.6a 3.9％±0.5b 2.4％±0.4c 0.8％±0.2d Escherichia group_Shigella 2.3％±0.3a 1.8％±0.2b 0.9％±0.1c 0.3％±0.1d Proteobacteria 7.6％±0.8a 5.5％±0.6b 3.1％±0.4c 1.2％±0.3d

[0081] (Note: mean ± standard deviation, different letters in the same column indicate significant differences between groups (p < 0.05), and the same letters indicate no significant differences)

[0082] It can be seen from Table 1 that the abundance of intestinal flora in mice fed with the phospholipid krill oil prepared in Examples 1-3 that can promote intestinal flora absorption is higher than that in mice fed with the phospholipid krill oil prepared in Comparative Example 1 and Comparative Example 2, and is greatly improved compared with normally fed mice. This proves that the phospholipid krill oil prepared in the present application by preparing a natural flora activity promoter and compounding it with krill oil into an emulsion can improve the activity of intestinal flora, promote the absorption and proliferation ability of intestinal probiotic flora, inhibit the reproduction of harmful bacteria, and build a strong prebiotic network in the intestine.

[0083] Experiment 2: Take the phospholipid krill oil capable of promoting intestinal flora absorption prepared in Examples 1-3 and Comparative Examples 1-4, take 20 mL of each, use Turbiscan Lab Expert (TSI) stability analyzer to test the stability, and use TurbiSoft 2.1.0.52 version software to analyze the obtained data to obtain data. The lower the value, the better the stability. At the same time, the smell, taste and taste are comprehensively scored. The score adopts a 0-10 point system, with 0 points as the lowest and 10 points as the highest. A sensory evaluation team composed of 10 professionally trained personnel conducts sensory evaluation and testing. The scoring criteria are shown in Table 2. The comprehensive sensory score data is recorded and the average value is taken, as shown in Table 3.

[0084] Table 2: Sensory evaluation score table

[0085]

[0086] Table 3: TSI stability and comprehensive sensory scores

[0087] TSI Stability Comprehensive sensory score Example 1 1.18 34.6 Example 2 1.21 33.8 Example 3 1.15 34.8 Comparative Example 1 3.21 24.2 Comparative Example 2 5.13 22.3 Comparative Example 3 3.68 28.4 Comparative Example 4 1.12 27.6

[0088] It can be seen from the data of Example 1 and Comparative Example 1 in Table 2 that the phospholipid krill oil capable of promoting intestinal flora absorption obtained by compounding the natural flora activity promoter with krill oil into an emulsion has high stability and can effectively cover the fishy smell and improve the overall sensory experience.

[0089] It can be seen from the data of Comparative Example 3 in Table 2 that the simultaneous extraction of shrimp meat enzymatic hydrolysate by a composite solvent system consisting of sodium citrate, ethanol, Antarctic krill oil and caprylic capric triglyceride can avoid excessive components affecting the subsequent formation of the emulsion and the stability of the emulsion storage process. At the same time, the nutrients can be reorganized in a suitable proportion in the subsequent process to further improve the stability of the emulsion.

[0090] It can be seen from the data of Comparative Example 4 in Table 2 that adding rosemary oil in the process of preparing the oil phase mixed system can further mask the fishy smell, effectively improve the acceptance of phospholipid krill oil among the public, and improve the overall sensory experience.

[0091] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A process for preparing phospholipid krill oil capable of promoting intestinal flora absorption, characterized in that: The following steps are involved: S1: Preparation of natural flora activity promoter The sea buckthorn fruit, Morinda officinalis powder, honey, Erigeron brevis and grapefruit are crushed to prepare a fermentation substrate, and then Lactobacillus rhamnosus GG and Bifidobacterium breve B-3 are inoculated for primary fermentation, and then enzymatic hydrolysis is performed with laccase and then secondary fermentation is performed with thermophilic Streptococcus, and then sterilization, filtration and concentration are performed to obtain a natural flora activity promoter; S2: Four-phase extraction of high-phospholipid krill oil The Antarctic krill meat is enzymatically hydrolyzed to obtain an enzymatic hydrolyzate, and then sodium citrate, ethanol and a hydrophobic composite organic solvent are sequentially added and centrifuged to extract, and the caprylic capric triglyceride-astaxanthin mixed oil phase, a phospholipid krill oil phase, a sugar-containing ethanol phase and a protein flocculation phase are obtained by layering. S3: Construction of oil phase and water phase system and mixed emulsification The phospholipid krill oil phase, the caprylic capric triglyceride-astaxanthin mixed oil phase and rosemary oil are mixed to prepare an oil phase mixed system, the sugar-containing ethanol phase is desolventized and mixed with β-cyclodextrin and a natural flora activity promoter to obtain a water phase mixed system, the oil phase mixed system and the water phase mixed system are mixed, and then high-speed shearing, high-pressure homogenization and sterilization canning are performed to obtain the phospholipid krill oil that can promote intestinal flora absorption.

2. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 1, characterized in that: Step S1: preparing a natural flora activity promoter, comprising the following steps: S1.1: 40-50 parts by weight of seabuckthorn fruit, 15-25 parts by weight of Morinda officinalis powder, 20-40 parts by weight of honey, 5-10 parts by weight of Erigeron brevis and 10-30 parts by weight of grapefruit are mixed and crushed into jam, and then placed in a fermentation tank with 3-5 times the weight of sterile water and stirred evenly to obtain a fermentation substrate, inoculated with Lactobacillus rhamnosus GG seed liquid at an inoculation amount of 6-8% in the fermentation substrate, and inoculated with Bifidobacterium breve B-3 seed liquid at an inoculation amount of 2-4%, adjusted the initial pH of the fermentation to 6.8-7, and fermented at 37-40°C for 20-24 hours in a microaerobic manner to obtain a primary fermentation liquid; S1.2: adding laccase to the primary fermentation broth at an enzyme-substrate ratio of 200-300 U / mg, then adding 0.01-0.02 wt% of a metal cofactor, fermenting at 40-45° C. under microaerobic conditions for 20-24 hours, sterilizing at 105-110° C., adding 0.3-0.5 wt% of xylo-oligosaccharide, and then inoculating a thermophilic Streptococcus seed solution at an inoculum amount of 4-6%, and aerobically fermenting at 40-42° C. and 160-180 rpm for 30-36 hours to obtain a crude fermentation broth; S1.3: The crude fermentation liquid is sterilized at 105-110°C, passed through a ceramic membrane with a pore size of 8-10 μm, and then concentrated to 30-35% of the original volume to obtain a natural bacterial flora activity promoter.

3. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 2, characterized in that: Step S2: Four-phase extraction of high phospholipid content krill oil, comprising the following steps: S2.1: The thawed Antarctic krill meat and deionized water are placed in a high-speed homogenizer at a material-liquid ratio of 1: (20-25), homogenized at a speed of 8000-10000 rpm for 0.5-1 minute, and then ultrasonically crushed at an ultrasonic frequency of 25-30 kHz for 10-15 minutes, the pH is adjusted to 4-4.5, and 0.25-0.3 wt% of enzyme preparation of fresh Antarctic krill meat is added respectively, and then placed in a constant temperature shaker at 50-55°C, enzymolyzed at 200-220 rpm for 4-5 hours, and then placed in a water bath at 95-100°C for inactivation for 15-20 minutes to obtain shrimp meat enzymolyzate; S2.2: Add 15-16wt% sodium citrate to the shrimp meat hydrolysate, stir until the sodium citrate is completely dissolved, then add 18-22wt% ethanol, stir for 2-3 minutes, then add 18-20wt% hydrophobic composite organic solvent, stir evenly, centrifuge at 10000-12000rpm for 3-4 minutes to obtain a four-phase solution, separate the four-phase solution into layers, and obtain a caprylic / capric triglyceride-astaxanthin mixed oil phase, a phospholipid krill oil phase, a sugar-containing ethanol phase, and a protein flocculation phase.

4. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 3, characterized in that: Step S3: Construction of the oil phase and water phase system and mixed emulsification, including the following steps: S3.1: 8-12 parts by weight of phospholipid krill oil phase, 1-1.2 parts by weight of caprylic capric triglyceride-astaxanthin mixed oil phase and 0.1-0.2 parts by weight of rosemary oil are mixed in a container, and stirred at 50-60° C. for 15-20 minutes to obtain an oil phase mixed system; S3.2: 1.2-1.5 parts by weight of the sugar-containing ethanol phase is desolventized by vacuum distillation and then mixed with 0.8-1 parts by weight of β-cyclodextrin, and then added to 3-4 parts by weight of a natural flora activity promoter at 75-80°C and stirred for 10-12 minutes to obtain an aqueous phase mixing system, and the oil phase mixing system prepared in step S3.1 is added to the aqueous phase mixing system, and then transferred to a high-speed disperser, sheared at a speed of 8000-10000rpm for 3-4 minutes, and placed in a high-pressure homogenizer for homogenization at a pressure of 20-25MPa for 2-3 times, and sterilized by ultra-high temperature instantaneous sterilization technology, and then sealed and canned for storage to obtain phospholipid krill oil that can promote intestinal flora absorption.

5. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 2, characterized in that: The Lactobacillus rhamnosus GG seed solution and the Bifidobacterium breve B-3 seed solution in step S1 are obtained by selecting one ring of strains from the solid slant culture medium for preserving Lactobacillus rhamnosus GG and Bifidobacterium breve B-3, inoculating them in MRS liquid culture medium, and culturing them at 37°C and 150rpm for 12 hours. The thermophilic Streptococcus seed solution is obtained by selecting one ring of strains from the solid slant culture medium for preserving thermophilic Streptococcus, inoculating them in MRS liquid culture medium, and culturing them at 42°C and 180rpm for 12 hours.

6. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 2, characterized in that: The dissolved oxygen concentration of the microaerobic fermentation in step S1.1 is maintained at 0.3-0.6 mg / L, and the dissolved oxygen concentration of the microaerobic fermentation in step S1.2 is maintained at 1.2-2 mg / L.

7. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 2, characterized in that: The metal cofactor in step S1.2 is composed of zinc sulfate and manganese sulfate in a mass ratio of 1:(2.5-3), and both zinc sulfate and manganese sulfate are food grade.

8. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 3, characterized in that: The enzyme preparation in step S2.1 consists of pepsin and cellulase in a mass ratio of 1:(1-1.5).

9. The process for preparing phospholipid krill oil capable of promoting intestinal flora absorption according to claim 3, characterized in that: The hydrophobic composite organic solvent of step S2.2 is composed of Antarctic krill oil and caprylic / capric triglyceride in a volume ratio of 1:(8-10).

10. A phospholipid krill oil capable of promoting intestinal flora absorption, characterized in that: The phospholipid krill oil capable of promoting intestinal flora absorption is prepared by the preparation process of any one of claims 1 to 9.

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