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CN-121975678-A - Strain Prescottella equi GXAS-2 for high-yield chitin deacetylase and fermentation medium and application thereof

CN121975678ACN 121975678 ACN121975678 ACN 121975678ACN-121975678-A

Abstract

The invention relates to a strain Prescottella equi GXAS-2 for high-yield chitin deacetylase, a fermentation medium and application thereof, wherein the strain Prescottella equi GXAS-2 has a preservation number of GDMCC NO:67488, is separated from mangrove soil, and can efficiently produce the enzyme in a medium taking N-acetylglucosamine (GlcNAc) as an inducer. By optimizing the medium components (colloidal chitin 3%, yeast powder 10%, glcNAc 0.3%) and the culture conditions (30 ℃, 180 rpm, pH 7, 48 h), the enzyme activity was significantly improved. The invention has the advantages of high induction efficiency, green process, stable enzyme activity and the like, and is suitable for the industry of chitin degradation and chitosan green preparation.

Inventors

  • PAN LIXIA
  • ZHANG ZIXUAN
  • YANG LIYAN
  • YANG DENGFENG
  • Jiang Siyin

Assignees

  • 广西科学院

Dates

Publication Date
20260505
Application Date
20251230

Claims (7)

  1. 1. A strain Prescottella equi GXAS-2 for high-yield chitin deacetylase is characterized in that the preservation number is GDMCC NO:67488, the strain is preserved in China center for type culture collection, and the preservation date is 2025, 12 months and 16 days.
  2. 2. A culture medium for fermentative production of chitin deacetylase according to claim 1, wherein the culture medium comprises colloidal chitin 0.1-5%, yeast powder 0.5-20%, glcNAc 0.05-0.5%, KH 2 PO 4 0.03%、MgSO 4 ·7H 2 O0.05%, naCl 0.1%, and water in balance, and has a natural pH.
  3. 3. The medium of claim 2, wherein the colloidal chitin is added in an amount of 3%, the yeast powder is added in an amount of 10%, and the GlcNAc is added in an amount of 0.3%.
  4. 4. A process for the fermentative production of chitin deacetylase as claimed in claim 1, The strain Prescottella equi GXAS-2 according to claim 1, which is obtained by fermentation culture in the medium according to claim 2 or 3 at 15-40℃and shaking table rotation speed of 100-250 rpm at an initial pH of 5-10 for 24-168 hours.
  5. 5. The method of claim 2, wherein the fermentation temperature is 30 ℃, the rotation speed of the shaker is 180 rpm, the initial pH is 7, and the fermentation time is 48 hours.
  6. 6. Use of GlcNAc for inducing Prescottella equi GXAS-2 chitin-producing deacetylase, wherein the GlcNAc is added in an amount of 0.05-0.5%.
  7. 7. Use of the strain Prescottella equi GXAS-2 according to claim 1 for chitin degradation, green chitosan production or industrial enzyme preparation production.

Description

Strain Prescottella equi GXAS-2 for high-yield chitin deacetylase and fermentation medium and application thereof Technical Field The invention relates to a strain Prescottella equi GXAS-2 for high-yield chitin deacetylase, a fermentation medium and application thereof, belonging to the technical field of microbial fermentation and enzyme engineering. Background The mangrove ecological system is widely distributed in tropical and subtropical coastal zones, is a transition zone of land and ocean juncture, has special ecological environment characteristics of high salt, low oxygen, tide fluctuation, rich organic matters and the like, also inoculates highly diversified animal and plant communities, and has a plurality of crustaceans, so that the mangrove becomes an important natural enrichment place of chitin degrading microorganisms, and is an important natural treasury for developing novel bioactive substances and industrial enzyme resources. Chitin (Chitin), a high molecular linear polysaccharide widely existing on the cell walls of crustaceans, insects, fungi and plants, has reserves in nature inferior to cellulose and annual output of about 100 hundred million tons. Chitin has a relatively large molecular weight, combines beta-1, 4-configuration and strong intermolecular or intramolecular hydrogen bonding force, so that the chitin has high crystallinity and indissolvable property, has limited application, and is a good alternative to chitosan through deacetylation treatment of chitin. Chitosan is a partially deacetylated form of chitin, has cationic property, can be combined with polyanion biomolecules such as cell membrane phospholipids, DNA, protein and the like, has solubility, biocompatibility, non-toxicity and various physiological activities (such as antibiosis, antioxidation and the like), and thus has wide application potential in the fields of medical treatment, agriculture, food, environment and the like. The chitosan is mainly prepared by a chemical method in industry, has the advantages of simple operation, high efficiency, large-scale production and the like, but has the limitations of high energy consumption, serious pollution, unstable product quality and the like. To overcome the above limitations, the development of green, efficient biological chitin deacetylation technology is a promising alternative strategy. The chitin deacetylase can be used for deacetylating under mild reaction conditions to retain the integrity and functional groups of chitin-sugar chain skeletons, can maintain the high molecular weight of products, has the advantages of environmental friendliness, low energy consumption and good uniformity of products, is easy to regulate and control the deacetylation degree and the acetylation mode, and is a key catalytic tool for realizing green preparation of chitosan. At present, it has been reported that chitin deacetylases are identified from various fungi and bacteria, and bacteria generally tend to produce chitin deacetylases with stronger activity than fungi, and have wider industrial application potential, so that the high-quality chitin deacetylases from bacterial sources are fully explored to have great significance for expanding enzyme resource libraries and industrial application. In addition, currently reported inducers for producing enzymes by chitin deacetylase are generally powder chitin or colloid chitin, have poor solubility and need to be pretreated by acid or alkali in advance, which may cause unstable induction effect and large fluctuation of enzyme production efficiency, and limit popularization in industrial application. Therefore, the research screens the high-efficiency chitin deacetylase-producing strain from soil samples of mangrove in urban harbor, creatively introduces a monomeric inducer GlcNAc with definite structure and easy water solubility as an inducer for starting the synthesis of target enzymes, optimizes enzyme production conditions through a single factor test, provides good enzyme resources for chitin bioconversion and provides reference for industrial application of chitin deacetylase. Disclosure of Invention In view of the above, the invention provides a Prescottella equi GXAS-2 strain with a preservation number of GDMCC NO:67488, which has strong enzyme production capability and stable enzyme activity, and an optimized culture medium containing colloidal chitin, yeast powder and GlcNAc, and a high-efficiency fermentation method for controlling temperature, rotating speed, pH and fermentation time, which can remarkably improve the yield and activity of chitin deacetylase and realize green degradation of chitin and sustainable preparation of chitosan. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: A strain Prescottella equi GXAS-2 for high-yield chitin deacetylase has the preservation number of GDMCC NO:67488, is preserved in China center for type culture collection, and has the preserva