CN-121971482-A - Preparation method and application of rambutan polysaccharide and Acremonium composition

CN121971482ACN 121971482 ACN121971482 ACN 121971482ACN-121971482-A

Abstract

The invention relates to the technical field of probiotics, in particular to a preparation method of a trichur algae polysaccharide combined with Ackermansia composite and application thereof, wherein BFP extracting solution is degassed; pressurizing nitrogen to press the Acremonium suspension into the BFP extracting solution to supplement nitrogen, and stirring the nitrogen to obtain a mixed solution; the preparation method comprises the steps of mixing liquid, cooling and spraying to form powder embedded particles, pressing the powder embedded particles into a freeze dryer through pressurized nitrogen to remove water to obtain a porous rambutan polysaccharide and Ackermans combination, wherein the whole preparation process is under the protection of nitrogen, the rambutan polysaccharide and Ackermans combination can improve the structure and composition of intestinal flora, increase the field planting of the Ackermans, thereby achieving the effect of treating ulcerative colitis, ensuring that the dissolved gas of BFP extracting solution is nitrogen to form a strictly anaerobic condition, loosening the porous structure combination to conveniently and rapidly recover the activity, and facilitating BFP serving as the sole carbon source of the Ackermans to enhance the growth proliferation and the adhesion field planting of the Ackermans.

Inventors

ZHENG MINGJING
Zuo Jinhuai
ZHOU BINHUI
HONG TAO
JIANG ZEDONG
NI HUI
HU YANG

Assignees

集美大学

Dates

Publication Date: 20260505
Application Date: 20260305

Claims (10)

1. The preparation method of the rambutan polysaccharide combined with the Acremonium composition is characterized by comprising the following steps of: BFP extraction, namely extracting the cleaned and dried red Mao Zaotong by a water extraction and alcohol precipitation method to obtain BFP extract; akk culturing, weighing thioglycolate fluid culture medium, dissolving in distilled water, sterilizing, inoculating Achroman into culture medium, and standing =0.8( Centrifuging after CFU/mL/d), washing with sterile water, and then re-suspending to obtain Acremonium suspension; mixing and preparing, namely mixing BFP extracting solution and Ackermans suspension in a mixing system, and sterilizing the mixing system before mixing, wherein the mixing system comprises a BFP container, a Akk container, a nitrogen cylinder, a cooling spray tower, a freeze dryer and an air pump, wherein the nitrogen cylinder is internally provided with pressurized nitrogen; mixing comprises injecting BFP extract into BFP container, and injecting Acremonium suspension into Akk container; The preparation method comprises the steps of vacuumizing a BFP container through an air pump to degas BFP extracting solution, pressing the Acremonium suspension in the Akk container into the BFP container through pressurized nitrogen to supplement gas for the BFP extracting solution, stirring and mixing the Acremonium suspension and the BFP extracting solution through nitrogen flow to obtain mixed solution, vacuumizing a cooling spray tower through the air pump, simultaneously pressing the mixed solution into the cooling spray tower through the pressurized nitrogen to carry out cooling spraying to form powder embedded particles, and pressing the powder embedded particles into a freeze dryer through the pressurized nitrogen to remove water to obtain the porous triclopyr polysaccharide-containing Acremonium composition.
2. The preparation method of the mucopolysaccharide combined ackerman bacterium composition according to claim 1, wherein the mixing system further comprises a plurality of valves, the BFP container is provided with a first liquid inlet, a first liquid outlet, a mixing port and an air outlet, the Akk container is provided with a second liquid inlet, a second liquid outlet and an air inlet, the cooling spray tower comprises a sealing tower body and a cooler, the sealing tower body is provided with an air extraction port, a nozzle, an air charging port and a discharge port, the cooler is used for cooling the sealing tower body to a preset temperature, and the freeze dryer is provided with a feed port and an air discharge port; The nitrogen cylinder is respectively communicated with an air inlet of the Akk container and an air charging port of the sealing tower body through two valves, a second liquid outlet of the Akk container is communicated with a mixing port of the BFP container through a valve, a first liquid outlet of the BFP container is communicated with a nozzle of the sealing tower body through a valve, a feed inlet of the freeze dryer is communicated with a discharge port of the sealing tower body through a valve, and the air extracting pump is respectively communicated with the air outlet, the air extracting port and the air exhaust port through three valves; valves are respectively arranged on the first liquid inlet and the second liquid inlet; The mixing further comprises the steps of injecting BFP extracting solution into a BFP container through a first liquid inlet, closing a valve of the first liquid outlet, injecting Acremonium suspension into a Akk container through a second liquid inlet, closing a valve of the second liquid outlet, closing a valve of the first liquid inlet, a valve of the first liquid outlet and a valve of a mixing port, opening a valve of an air outlet and an air pump to vacuumize the BFP container, closing the valve of the air outlet and the air pump after the degassing is finished, opening a valve of an air inlet of the Akk container and a valve of a mixing port of the BFP container, pressing Acremonium suspension into the BFP container through pressurized nitrogen, stirring and mixing the Acremonium suspension and the BFP extracting solution to obtain mixed solution, closing the valve of the air inlet and the valve of the mixing port after the mixing is finished, opening a pump and the valve of the air pump for vacuumizing the sealing tower body, opening a cooler for cooling the sealing tower body to a preset temperature, opening the valve of the air inlet, opening the valve of the Akk container, the mixing port of the BFP container and the valve of the first liquid outlet, spraying the air inlet, the valve of the air pump for forming the air inlet, the air outlet of the air pump, the air outlet of the air pump, and the air pump, sealing the air outlet of the air pump, and the air pump.
3. The method for preparing the rambutan polysaccharide and ackermanni combination according to claim 2, wherein the preset temperature is lower than the coagulation temperature of the mixed solution.
4. The method of claim 2, wherein after sterilizing the mixing system, before mixing, the pump and all valves are opened and nitrogen is introduced until the oxygen concentration in the gas pumped by the pump is less than 0.5%.
5. The method for preparing a combination of trichlamydomonas reinhardtii and ackermanni composition according to claim 4, wherein the aspiration pump has a tail outlet, and an oxygen concentration sensor is disposed on the tail outlet; the oxygen concentration is obtained by an oxygen concentration sensor.
6. The method for preparing the trichomonas polysaccharide combined ackerman bacteria composition according to claim 2, wherein the first liquid inlet and the gas outlet are arranged at the top of a BFP container, and the first liquid outlet and the mixing port are arranged at the bottom of the BFP container; the nozzle and the air extraction opening are positioned at the top of the inner wall of the sealed tower body, and the discharge opening is positioned at the bottom of the inner wall of the sealed tower body; the second liquid inlet and the air inlet are arranged at the top of the Akk container, and the second liquid outlet is arranged at the bottom of the Akk container; the exhaust port is positioned at the top of the freeze dryer.
7. The method for preparing the combination of the polysaccharide from the red hair alga and the Acremonium acutum as claimed in claim 1, wherein the ratio of the combination of the polysaccharide from the red hair alga and the Acremonium acutum is 200-400 mg/kg/d, and the ratio of the polysaccharide from the red hair alga to the Acremonium acutum is - CFU/mL/d。
8. The method for preparing the trichuria polysaccharide combined ackermanni composition according to claim 1, wherein the BFP extraction further comprises: Step 11, crushing the cleaned and dried rambutan to obtain rambutan powder, adding ethanol solution with the volume of 10 times into the obtained rambutan powder, soaking, cleaning, filtering, adding distilled water with the volume of 30 times, extracting in a water bath at 90 ℃ for 2 hours, centrifuging, collecting supernatant, re-dissolving residues with the same amount of distilled water, extracting in a water bath at 90 ℃ for 2 hours again, combining the two supernatants, evaporating and concentrating the supernatant to 1/3 of the original volume to obtain concentrated solution; step 12, adding 3 times of ethanol solution with the mass concentration of 95% into the concentrated solution obtained in the step 1, precipitating with 4 ℃ ethanol overnight, centrifuging, collecting precipitate, and adding distilled water into the precipitate to form crude polysaccharide solution; And 13, mixing the crude polysaccharide solution obtained in the step 2 with a Sevage reagent according to the ratio of 3:1, oscillating, centrifuging, collecting supernatant, and dialyzing the collected supernatant to obtain BFP extract.
9. The method for preparing the mucopolysaccharide-ackermanni combination composition according to claim 1, wherein Akk culturing comprises: step 21, weighing thioglycolate fluid culture medium according to a proportion, dissolving in distilled water, and sterilizing for 15min at 121 ℃; Step 22, inoculating Acremonium to the culture medium in step 1 at an inoculum size of 10% until =0.8( CFU/mL/d), centrifugation, washing with sterile water for 2 times, and re-suspending to obtain ackermanni suspension.
10. Use of a composition obtained by combining a rhodophytan polysaccharide as claimed in any one of claims 1 to 9 with an akkermansia composition for the prevention and treatment of ulcerative colitis.

Description

Preparation method and application of rambutan polysaccharide and Acremonium composition Technical Field The invention relates to the technical field of probiotics, in particular to a preparation method and application of a trichur algae polysaccharide combined with Acremonium acetum composition. Background Inflammatory bowel Disease (Inflammatory Bowel Disease, IBD) is a type of chronic, recurrent intestinal inflammatory Disease, mainly including ulcerative colitis (Ulcerative Colitis, UC) and Crohn's Disease (CD). The global prevalence of UC is approximately 680 tens of thousands, whereas ulcerative colitis is as much as 1000 tens of thousands. At present, the means for clinically treating UC mainly comprises aminosalicylic acid medicines, glucocorticoids, immunosuppressants and the like, but the medicines have the common problems of large individual difference in curative effect, easy initiation of side effects such as liver and kidney function injury, intestinal flora imbalance and the like after long-term use, and difficult complete blocking of the recurrence progress of diseases. Therefore, there is an urgent need to find new intervention strategies that are safe, efficient and capable of modulating UC from the pathological mechanism level. Acremonium muciniphilum (AKKERMANSIA MUCINIPHILA, akk) (abbreviated as Acremonium muciniphilum) is a gram-negative bacterium colonized in intestinal mucus layers, accounting for about 1-3% of the total intestinal microbiota of healthy adults, and has been shown to be inversely related to diseases such as obesity, colitis and diabetes. Marine algae, which are important primary producers in marine ecosystems, are rich in a variety of bioactive substances such as polysaccharides, polyphenols, terpenes, etc. The polysaccharide (Bangia fusco-purpureaPolysaccharide, BFP) is a sulfated polysaccharide mainly composed of D-, L-galactose, 3, 6-diether-L-galactose and 6-O-methyl D-galactose, and is the main active ingredient of the rambutan, and has been proved to have the biological activities of anti-inflammatory, antioxidant, intestinal flora regulation and the like. Earlier studies found that BFP as a carbon source of Acremonium was able to enhance its growth proliferation and adhesion to colonize Caco-2 cells and to increase the expression of Caco-2 cell mucins and claudins. However, the rambutan polysaccharide solution is easy to mix with air in the extraction and preparation processes, when the BFP extracting solution and the Acremonium are mixed and cultured, the oxygen precipitation of the BFP extracting solution mixed with the air can inhibit the activity of the Acremonium, even irreversible damage occurs, so that the BFP and the Acremonium composition exert a slow or poor effect in intestinal tracts, and the expression of the Acremonium is insufficient, and the effect is not as expected. Therefore, a preparation method of a combination of the botrytis cinerea polysaccharide and the Achromyces avermitilis composition and application thereof are needed, wherein the preparation method can ensure better action and effect of the BFP and Akk composition. Disclosure of Invention The invention aims to solve the technical problem of providing a preparation method of a rambutan polysaccharide combined Acremonium composition capable of ensuring better action and effect of a BFP and Akk composition and application thereof. In order to solve the technical problems, the invention adopts the following technical scheme: a preparation method of a combination of rambutan polysaccharide and Ackermans comprises BFP extraction, akk culture and mixed preparation; BFP extraction, namely extracting the cleaned and dried red Mao Zaotong by a water extraction and alcohol precipitation method to obtain BFP extract; akk culturing, weighing thioglycolate fluid culture medium, dissolving in distilled water, sterilizing, inoculating Achroman into culture medium, and standing =0.8(CFU/mL), centrifuging, washing with sterile water, and re-suspending to obtain ackermanni suspension; Mixing and preparing, namely mixing BFP extracting solution and Ackermans suspension in a mixing system, and sterilizing the mixing system before mixing, wherein the mixing system comprises a BFP container, a Akk container, a nitrogen cylinder, a cooling spray tower, a freeze dryer and an air pump, wherein the nitrogen cylinder is internally provided with pressurized nitrogen; mixing comprises injecting BFP extract into BFP container, and injecting Acremonium suspension into Akk container; The preparation method comprises the steps of vacuumizing a BFP container through an air pump to degas BFP extracting solution, pressing the Acremonium suspension in the Akk container into the BFP container through pressurized nitrogen to supplement gas for the BFP extracting solution, stirring and mixing the Acremonium suspension and the BFP extracting solution through nitrogen flow to obtain mixed solution, vacuumizing a cooling spr