CN-121975827-A - Gastrodia elata acyl transferase GeBAHD gene and application thereof in preparation of barrison glycoside C

Abstract

The invention relates to a gastrodia elata acyl transferase GeBAHD gene and application thereof in preparation of barrison glycoside C, and belongs to the technical field of biology. The nucleotide sequence of the gastrodia elata acyl transferase GeBAHD gene is shown as SEQ ID NO.1, and the amino acid sequence of the coded protein is shown as SEQ ID NO. 2. The invention takes gastrodin and citrate-acyl coenzyme A as raw materials, and generates the balisonoside C by a one-pot method under the catalysis of the gastrodin-acyl transferase obtained by coding a citrate-acyl coenzyme A synthetase and the gastrodin-acyl transferase GeBAHD gene. The invention accurately analyzes the synthesis path and gene family evolution of the pareisen glycoside C in basic research, provides an accurate target point for directional synthesis of high-value components and innovative breeding in application development, and finally promotes the upgrading of the gastrodia elata from empirical medicine to molecular regulation and accurate utilization, which is a value difficult to cover in the prior art.

Inventors

• HAO BING
• CHEN LI
• LIU XIANGYU
• LI YUJIE
• SHEN YANRONG
• HE YUANFENG
• ZHU LIUYING
• OuYang Ruichen
• WANG LE

Assignees

• 云南农业大学

Dates

Publication Date

20260505

Application Date

20260206

Claims (9)

1. 1. The gastrodia elata acyl transferase GeBAHD gene is characterized in that the nucleotide sequence of the gastrodia elata acyl transferase GeBAHD925 gene is shown in SEQ ID NO. 1.
2. 2. The gastrodia elata acylase GeBAHD gene coded protein according to claim 1, which is characterized in that the amino acid sequence of the coded protein is shown as SEQ ID NO. 2.
3. 3. A recombinant plasmid comprising the gastrodia elata acylase GeBAHD925 gene according to claim 1.
4. 4. The recombinant plasmid containing the gastrodia elata acylase GeBAHD925 gene according to claim 3, wherein the gastrodia elata acylase GeBAHD925 gene is subjected to homologous recombination with the pET28a vector to obtain the pET28a-GeBAHD925 recombinant plasmid.
5. 5. A genetically engineered bacterium comprising the recombinant plasmid of claim 3, or having integrated into its genome an exogenous gene of gastrodia elata acyl transferase GeBAHD925 according to claim 1.
6. 6. The genetically engineered bacterium of claim 5, wherein the genetically engineered bacterium is a strain of e.coli BL21 (DE 3).
7. 7. The gastrodia elata acylase GeBAHD gene coding to obtain the gastrodia elata acylase according to claim 1.
8. 8. The use of the gastrodia elata acyl transferase GeBAHD gene according to claim 1 in the preparation of balisonoside C.
9. 9. The use of gastrodia elata acylase GeBAHD gene according to claim 8 in preparation of balison C, characterized in that the balison C is produced by using gastrodin and citrate-acyl-CoA as raw materials under the catalysis of the gastrodia elata acylase obtained by encoding the citrate-acyl-CoA synthetase and the gastrodia elata acylase GeBAHD925 gene.

Description

Gastrodia elata acyl transferase GeBAHD gene and application thereof in preparation of barrison glycoside C Technical Field The invention belongs to the technical field of biology, and particularly relates to a gastrodia elata acyl transferase GeBAHD gene and application thereof in preparation of barrison glycoside C. Background Orchid (Orchidaceae) is one of the largest families of the plant kingdom, covering about 750 genera, nearly 27000 species. In nature, most orchids (over 99%) live in a bacterial heterotrophic manner, with at least 200 chlorophyll-free orchids belonging to a complete bacterial heterotrophic population and distributed in more than 25 evolving branches. Gastrodia elata (Gastrodia elata Blume) is a typical representative of such plants, a root-free, leaf-free, chlorophyll-free whole-cell heterotrophic orchid plant, which is incapable of photosynthesis and must be symbiotic with mycorrhizal fungi to obtain the carbon source required for total growth. During its complex life cycle, nutrition needs to be obtained by establishing symbiotic relationship with fungus strains of the genus Pleurotus (Mycena) and Armillariella mellea (ARMILLARIA). With the advancement of isolation and analysis techniques, more than 100 new barrison glycoside compounds have been identified to date from gastrodia elata, many of which show significant biological activity. Its neuroprotective effect is widely accepted. Studies show that the antidepressant effect of gastrodia elata mainly depends on key active ingredients (such as gastrodin and PARISHIN C), and through targeting EGFR of sea Ma Ou nerve precursor cells (NSPCs), PI3K/Akt signal channels are activated, cell proliferation and differentiation are promoted, so that damage to hippocampal nerves is improved, and depression-like behaviors are relieved. Although the pharmacological action is of great concern, the plant extraction content is very small, and no research on the biosynthesis of the balison C is reported at present. The main method for early obtaining the barrison glycoside C is direct extraction or chemical processing from gastrodia tuber. However, the chemical method for synthesizing the barrison glycoside C is high in energy consumption and not friendly to the environment, the quality of the gastrodia elata cannot be guaranteed by an artificial cultivation and extraction method, the period of planting the gastrodia elata is long, and the problems of low extraction quantity, low extraction purity and the like exist. In recent years, with the rapid development of the field of synthetic biology, the above problems can be effectively solved by using synthetic biology techniques to produce natural drug monomers. However, to identify the biosynthetic pathways of these active ingredients, it is necessary to identify the key genes involved in these pathways, and to explore these catalytic enzyme genes as key links in the research of plant metabolite biosynthetic pathways. At present, the synthesis paths of gastrodin and citrate-CoA are clear, but the synthesis path of the balrison C is not resolved, which influences the promotion of the biosynthesis work of the balrison C. How to overcome the defects of the prior art is a problem to be solved in the prior art. Disclosure of Invention The invention aims to solve the defects in the prior art and provides a gastrodia elata acyl transferase GeBAHD gene and application thereof in preparation of barrison glycoside C. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: The first aspect of the invention provides a gastrodia elata acylase GeBAHD925 gene, the nucleotide sequence of the gastrodia elata acylase GeBAHD925 gene is shown as SEQ ID NO.1, and the total length of the sequence is 1323bp. The second aspect of the invention provides a gastrodia elata acyl transferase GeBAHD gene coding protein, the amino acid sequence of the coding protein is shown as SEQ ID NO.2, and 441 amino acid residues are coded. The third aspect of the invention provides a recombinant plasmid containing the gastrodia elata acylase GeBAHD925 gene. Further, the gastrodia elata acyl transferase GeBAHD gene and the pET28a vector are subjected to homologous recombination to obtain the pET28a-GeBAHD925 recombinant plasmid. The fourth aspect of the present invention provides a genetically engineered bacterium comprising the recombinant plasmid of claim 3, or wherein the genome of the genetically engineered bacterium has incorporated therein an exogenous gastrodia elata acylase GeBAHD925 gene according to claim 1. Further, the transgenic engineering bacteria are escherichia coli BL21 (DE 3) strains. The fifth aspect of the invention provides the gastrodia elata acylase GeBAHD gene coded by the gastrodia elata acylase GeBAHD gene. The sixth aspect of the invention provides an application of the gastrodia elata acyl transferase GeBAHD gene in preparation of the balisonoside C. Furthermore, taking