Search

CN-121992055-A - Deacetylated sophorolipid and preparation method and application thereof

CN121992055ACN 121992055 ACN121992055 ACN 121992055ACN-121992055-A

Abstract

The invention belongs to the technical field of microbial fermentation and daily chemicals, and particularly relates to deacetylated sophorolipid and a preparation method and application thereof. The method effectively solves the problem of overhigh production cost of the sophorolipid by using low-cost culture medium components and screening saccharomycetes for producing the lactone type sophorolipid, promotes the application of the sophorolipid in various fields, and solves the problems of long single fermentation period, low thallus utilization rate, high energy consumption and high cost of the traditional saccharomycetes by adopting the step of enzymolysis of fermentation substrates in advance. The multi-round fermentation circulating system is designed, bacterial liquid is settled and recycled for re-inoculation fermentation, so that bacterial body is efficiently and repeatedly utilized, unit yield is improved, fermentation period is shortened, and production cost is effectively controlled. The structure is regulated by a resin modification technology, and the final product is mainly acetylated sophorolipid, so that compared with the acetylated sophorolipid, the deacetylated sophorolipid has better foam stability and the efficacy of repairing sensitive muscles, and is suitable for the application fields of cosmetics, washing and the like.

Inventors

  • HE JUNJIE
  • CHENG JIANHUA

Assignees

  • 华南理工大学

Dates

Publication Date
20260508
Application Date
20260228

Claims (10)

  1. 1. A method for preparing deacetylated sophorolipid, which is characterized by comprising the following steps: s1, mixing a lysozyme solution and compound grease, and carrying out enzymolysis to obtain a grease substrate; S2, inoculating the bumblebee candida utilis seed solution into a fermentation substrate containing an oil substrate for fermentation culture, and standing after the culture is finished, wherein the bottom oily substance is crude sophorolipid; s3, performing modification treatment on the crude sophorolipid to obtain the deacetylated sophorolipid.
  2. 2. The preparation method according to claim 1, wherein in step S2, the bumblebee candida is named LSRH-007, and the bumblebee candida is preserved in the China center for type culture collection (cctccc) M20251461.
  3. 3. The preparation method according to claim 1, comprising at least one of (I) to (IV): (I) In the step S1, the enzyme solution is fermentation product enzyme of rhizopus oryzae; (II) in the step S1, the compound grease comprises soy sauce residue grease and rapeseed oil, wherein the mass ratio of the soy sauce residue grease to the rapeseed oil is (2-3); (III) in the step S1, the volume ratio of the lysozyme solution to the compound grease is (0.8-1.2): 1; In the step S1, the enzymolysis condition is that the temperature is 28-32 ℃ and the time is 20-60min, and the rotating speed of the stirring paddle is 80-150rpm.
  4. 4. A process according to claim 3, wherein, The preparation of the lysozyme solution comprises the following steps: a1, inoculating a rhizopus oryzae bacterial colony into a seed culture medium for amplification culture to obtain rhizopus oryzae seed liquid; A2, inoculating the rhizopus oryzae seed liquid into a liquid culture medium for fermentation to obtain rhizopus oryzae fermentation liquid; A3, centrifuging the rhizopus oryzae fermentation liquor to obtain a supernatant which is the lysozyme solution, In the step A1, the seed culture medium is a malt juice liquid culture medium; In the step A1, the conditions of the expansion culture are that the temperature is 25-30 ℃ and the time is 40-60 hours, and the rotating speed of a fermentation shake flask is 150-200rpm; In the step A2, the inoculation amount of the rhizopus oryzae seed liquid is 8-13%; In the step A2, the liquid culture medium comprises 30-50g/L glucose, 7-15g/L peptone, 1.5-3g/L KH 2 PO 4 、0.2-0.8g/L MgSO 4 ·7H 2 O and the balance of water; in the step A2, the fermentation condition is that the temperature is 25-30 ℃ and the time is 65-80 hours, and the dissolved oxygen in the tank is maintained at 30-50%; in the step A3, the condition of the centrifugal treatment is that the rotating speed is 7000-10000rpm, and the time is 12-20min.
  5. 5. The preparation method according to claim 1, comprising at least one of (V) to (VIII): (V) in the step S2, the inoculation amount of the bumblebee candida raw seed liquid is 3-7%; (VI) in step S2, the fermentation substrate for the lipid-containing substrate comprises the lipid substrate, a carbon source, a nitrogen source, an inorganic salt, a pH adjuster, and water; (VII) in the step S2, the conditions of the fermentation culture are that the temperature is 25-32 ℃, the time is 3-5 days, and the rotating speed of a fermentation shaking table is 180-220rpm; (VIII) in step S2, the standing time is 30-60min.
  6. 6. The method according to claim 5, wherein, The preparation method of the bumblebee candida raw seed liquid comprises the following steps: inoculating colony of the bumblebee candida into a seed culture medium, activating and culturing to obtain bumblebee candida seed liquid, wherein, The seed culture medium is YM liquid culture medium; the conditions of the activation culture are that the temperature is 25-32 ℃ and the time is 36-60 hours, and the rotating speed of a fermentation shaking table is 180-220rpm; the adding amount of the grease substrate is 60-100g/L in the fermentation substrate containing the grease substrate, wherein the carbon source comprises glucose, and the adding amount of the glucose is 50-80g/L; the nitrogen source comprises at least one of peptone, yeast extract powder and fish meal, and the adding amount of the nitrogen source is 4-8g/L; The inorganic salt comprises FeSO 4 ·7H 2 O and MgSO 4 ·7H 2 O, the adding amount of the inorganic salt is 1-3 g/L, and the mass ratio of the FeSO 4 ·7H 2 O to the MgSO 4 ·7H 2 O is (1.5-1); The pH regulator comprises KH 2 PO 4 and/or K 2 HPO 4 , and the pH of the fermentation substrate containing the grease substrate is regulated to 4.5-5.5; the balance of water.
  7. 7. The method for preparing deacetylated sophorolipid according to claim 1, wherein in the step S2, after the step of standing after the completion of the cultivation, the method further comprises the step of circularly fermenting by standing after the completion of the cultivation to obtain an upper bacterial liquid and the bottom oily substance, refluxing the upper bacterial liquid to a fermentation device, adding a fermentation substrate of the oily substrate, and performing fermentation cultivation; The volume ratio of the upper bacterial liquid to the fermentation substrate containing the grease substrate is 3-7%; the times of the cyclic fermentation are 1-3 times.
  8. 8. The deacetylated sophorolipid and the preparation method thereof according to claim 1, wherein in the step S3, the modification treatment comprises the steps of: s31, mixing the crude sophorolipid with ethanol, and centrifuging to obtain supernatant which is ethanol solution of sophorolipid; the ethanol is absolute ethanol; the volume ratio of the crude sophorolipid to the ethanol is 1 (1-3); The conditions of the centrifugal treatment are that the rotating speed is 7000-10000rpm, and the time is 20-40min; S32, adding an aqueous solution of NaOH into the ethanol solution of sophorolipid, and reacting to obtain a deacetylated sophorolipid mixed solution; The concentration of the NaOH aqueous solution is 0.05-0.2mol/L, and the addition amount of the NaOH aqueous solution is 0.3-0.8% of the volume ratio of the sophorolipid ethanol solution; the temperature of the reaction is 20-40 ℃, and the reaction time is 8-15min; s33, filtering the deacetylated sophorolipid mixed solution through a strong-alkaline anion exchange resin to obtain a purified deacetylated sophorolipid mixed solution; the strong-base anion exchange resin is D201 strong-base anion exchange resin; S34, concentrating the purified deacetylated sophorolipid mixed solution to obtain deacetylated sophorolipid; The concentration treatment is reduced pressure concentration under the conditions that the pressure is 100-150mbar, the temperature is 50-70 ℃ and the time is 2-4h.
  9. 9. A deacetylated sophorolipid prepared according to the method for preparing a deacetylated sophorolipid of claim 1.
  10. 10. Use of a deacetylated sophorolipid according to claim 9 for the preparation of a daily chemical.

Description

Deacetylated sophorolipid and preparation method and application thereof Technical Field The invention belongs to the technical field of microbial fermentation and daily chemicals, and particularly relates to deacetylated sophorolipid and a preparation method and application thereof. Background Sophorolipid is used as a natural biosurfactant, and has wide application in the fields of daily chemicals, agriculture, petroleum and the like due to good emulsifying, foaming, antibacterial, anti-inflammatory performances and biodegradability. Microbial fermentation is the main method for obtaining sophorolipids. The sophorolipid molecule consists of sophorose and hydrophobic group, the sophorose is disaccharide formed by connecting 2 glucose beta-1, 2, and can be acetylated at the 6' or 6' ' position of glucose. In natural fermentation products, the hydroxyl groups on the sophorose hydrophilic groups of sophorolipids are usually modified by acetyl groups to produce acetylated sophorolipids. The degree of acetylation has a critical effect on its surface-to-surface activity and physicochemical properties. Traditional sophorolipids generally have excellent surface activity, anticancer properties, antibacterial properties, and lipophilicity. However, because the hydroxyl group of the hydrophilic group is substituted by the acetyl group, on one hand, the acetylated sophorolipid has poor water solubility and limits the application of the sophorolipid in an aqueous medium, and on the other hand, the acetyl group has strong toxicity and cell stimulation, so that the sophorolipid can be limited in the aspects of food, daily chemicals, cosmetics, pharmacy and the like. The deacetylated sophorolipid has higher water solubility because the acetyl group of the sophorolipid is hydrolyzed, and has the advantages of being used as a detergent, a wetting agent and the like in an aqueous medium environment, such as being used as an emulsifying dispersant for nanoparticle synthesis, and being used as a high-end essence, a special cleansing agent for sensitive muscles, a mild cleansing agent, a surfactant for washing and protecting infants and the like. However, few studies have been made on the preparation method of deacetylated sophorolipids at present, and there is still a lack of in-depth study on the efficacy of deacetylated sophorolipids. Disclosure of Invention The invention aims at solving the existing problems and provides deacetylated sophorolipid and a preparation method and application thereof. The invention is realized by the following technical scheme: the first aspect of the present invention provides a method for producing deacetylated sophorolipids, comprising: s1, mixing a lysozyme solution and compound grease, and carrying out enzymolysis to obtain a grease substrate; S2, inoculating the bumblebee candida utilis seed solution into a fermentation substrate containing an oil substrate for fermentation culture, and standing after the culture is finished, wherein the bottom oily substance is crude sophorolipid; s3, performing modification treatment on the crude sophorolipid to obtain the deacetylated sophorolipid. Alternatively, in some embodiments of the invention, in step S1, the lysate is a fermentation product enzyme of rhizopus oryzae. Optionally, in some embodiments of the present invention, in step S1, the preparation of the lysozyme solution includes the following steps: a1, inoculating a rhizopus oryzae bacterial colony into a seed culture medium for amplification culture to obtain rhizopus oryzae seed liquid; A2, inoculating rhizopus oryzae seed liquid into a liquid culture medium for fermentation to obtain rhizopus oryzae fermentation liquid; And A3, performing centrifugal treatment on the rhizopus oryzae fermentation liquor, wherein the obtained supernatant is the lysozyme solution. Alternatively, in some embodiments of the invention, in step A1, the seed medium comprises a malt juice liquid medium. Alternatively, in some embodiments of the present invention, in step A1, the conditions of the expansion culture are a temperature of 25-30 ℃ for 40-60 hours, and a rotational speed of 150-200rpm of the fermentation flask. Alternatively, in some embodiments of the invention, in step A2, the rhizopus oryzae seed fluid is inoculated in an amount of 8-13% (v/v). Alternatively, in some embodiments of the invention, in step A2, the liquid medium comprises 30-50g/L glucose, 7-15g/L peptone, 1.5-3g/L KH 2PO4、0.2-0.8g/L MgSO4·7H2 O, and the balance water. Alternatively, in some embodiments of the invention, the fermentation is carried out at a temperature of 25-30 ℃ for a period of 65-80 hours, with dissolved oxygen in the tank maintained at 30-50%. Alternatively, in some embodiments of the present invention, in step A3, the centrifugation is performed at 7000-10000rpm for 12-20min. Alternatively, in some embodiments of the present invention, in step S1, the compound grease includes soy sauce residue grease and rapeseed