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CN-122012536-A - S3G4685 gene and application thereof in synthesis of triterpene active product

CN122012536ACN 122012536 ACN122012536 ACN 122012536ACN-122012536-A

Abstract

The invention discloses an S3G4685 gene and application thereof in the synthesis of triterpene active products. The nucleotide sequence of the S3G4685 gene is shown as SEQ ID NO. 1, and the amino acid sequence of the protein is shown as SEQ ID NO. 2. The S3G4685 gene can obviously enhance the key metabolic flux of the terpene synthesis pathway in the host strain and improve the yield of squalene and downstream terpenoid thereof. The gene, the recombinant strain and the fermentation method provided by the invention provide a new biocatalysis strategy for efficiently and continuously producing terpenoid, and have important industrial application value.

Inventors

  • ZHANG LIWEN
  • XU YUQUAN
  • AI YUTONG
  • CAI MANJUN
  • YUE QUN
  • XIE LINAN

Assignees

  • 中国农业科学院生物技术研究所

Dates

Publication Date
20260512
Application Date
20260413

Claims (7)

  1. The S3G4685 gene is characterized in that the nucleotide sequence of the gene is shown as SEQ ID NO. 1.
  2. The S3G4685 protein is characterized in that the amino acid sequence of the protein is shown as SEQ ID NO. 2.
  3. 3. A recombinant vector comprising the S3G4685 gene of claim 1.
  4. 4. A recombinant strain comprising the recombinant vector of claim 3, wherein the strain is saccharomyces cerevisiae or a filamentous fungus.
  5. 5. Use of the S3G4685 gene of claim 1 in the synthesis of triterpene active products.
  6. 6. The use of the S3G4685 gene according to claim 5 for the synthesis of a triterpene active product, wherein said triterpene active product is squalene.
  7. 7. A method for increasing the production of terpenoid compounds in saccharomyces cerevisiae or filamentous fungi, wherein the S3G4685 gene of claim 1 is expressed in saccharomyces cerevisiae or filamentous fungi.

Description

S3G4685 gene and application thereof in synthesis of triterpene active product Technical Field The invention belongs to the field of biotechnology and metabolic engineering, and particularly relates to an S3G4685 gene and application thereof in synthesis of triterpene active products. Background Terpenoid is a natural product with various structures and wide biological activity, and is widely applied to the fields of medicines, cosmetics, fragrances, biofuels and the like. Terpenes can be classified into monoterpenes, sesquiterpenes, diterpenes, triterpenes, etc. according to the number of carbon atoms in the skeleton. Among them, triterpenes have been paid attention to for their remarkable pharmacological activities, for example, ganoderic acid (Ganoderic acids) as a main active triterpene component in ganoderma lucidum (Ganoderma lucidum) has been confirmed to have various biological activities such as antitumor, immunomodulation, liver protection, antiviral, etc., and have important development value in the fields of medicines and functional foods. Squalene is used as a key precursor of triterpenes, and is important in cholesterol synthesis, steroid hormone metabolism, and biosynthesis of higher terpenes such as ganoderic acid. At present, the production of terpenoids mainly depends on plant extraction or chemical synthesis. For example, the production of ganoderic acid still mainly depends on the cultivation and extraction of ganoderma lucidum fruiting bodies, and has the problems of long growth period, low extraction efficiency, unstable product content and the like, and the chemical synthesis has complex steps, low yield and large environmental pollution. Microbial cell factories provide a viable pathway for green sustainable production of terpenoids. Saccharomyces cerevisiae and filamentous fungi are common hosts for constructing terpene synthesis pathways because of their mature genetic manipulation, rapid growth, and easy large-scale cultivation. However, the flux of terpene synthesis pathways in natural strains is limited, metabolic regulation is complex, especially the metabolic flow upstream of squalene synthetase (SQS) often becomes a bottleneck, which results in low yield of target products such as ganoderic acid and the like, and limits the industrial application of the target products. In recent years, enhancement of expression of a key gene in a terpene synthesis pathway by genetic engineering means has become an effective strategy for improving yield. However, there is still a lack of efficient, versatile and stable endogenous or exogenous enhancement factors in the prior art, especially functional genes that efficiently promote squalene accumulation in filamentous fungi, and thus promote the synthesis of triterpene products. Therefore, a novel gene with the capability of broad-spectrum promotion of terpene synthesis is explored, and a high-efficiency stable recombinant strain is constructed, so that the method has important significance for promoting the industrialization process of high-value terpene compounds such as ganoderic acid and the like produced by a microbiological method. Disclosure of Invention The invention aims to provide a novel gene and a coded protein thereof which can obviously enhance the synthesis capability of a microbial terpenoid, solve the technical problems of low yield of the terpenoid and limited metabolic flux in the existing microbial cell factory, and provide a novel key element for efficiently and continuously producing high-value triterpenes such as squalene, ganoderic acid and the like. S3G4685 gene, the nucleotide sequence of the gene is shown as SEQ ID NO. 1. The amino acid sequence of the S3G4685 protein is shown as SEQ ID NO.2, or the amino acid sequence of the protein is substituted and/or deleted and/or added by one or more amino acid residues and still has the effect of producing squalene. Specifically, substitution and/or deletion and/or addition of one or several amino acid residues refers to substitution and/or deletion and/or addition of not more than 10, 5, 3 amino acid residues. In a more specific embodiment, a tag sequence, preferably Poly-Arg, poly-His, FLAG, strep-tag II, c-myc, more specifically RRRRR, HHHHHH, DYKDDDDK, WSHPQFEK, EQKLISEEDL, is attached at the amino-or carboxy-terminus. In order to facilitate purification of the protein of (a), a tag as shown in Table 1 may be attached to the amino-or carboxyl-terminus of the protein consisting of the amino acid sequence shown in SEQ ID NO. 1. TABLE 1 sequence of tags A recombinant vector containing the S3G4685 gene. Recombinant strains containing the recombinant vector, wherein the strains are Saccharomyces cerevisiae or filamentous fungi. The application of the S3G4685 gene in the synthesis of triterpene active products. The triterpene active product is squalene. The molecular structure of squalene is as follows: 。 a method for increasing the production of terpenoid compounds in sacchar