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Protein from centella asiatica and application of protein in the preparation of maslinic acid and corosolic acid

A technology of corosolic acid and maslinic acid is applied in the application field of preparing maslinic acid and corosolic acid, and can solve the problems of difficult product purification, low content, destruction of wild resources of biological resources and the like

Inactive Publication Date: 2019-01-01
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, such compounds are mainly isolated and extracted from hawthorn, crape myrtle, loquat and other plants, but this method has many disadvantages, including low content and large differences, difficult product purification, long plant growth cycle, and harmful to biological resources. In particular, wild resources have caused serious damage, etc.

Method used

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  • Protein from centella asiatica and application of protein in the preparation of maslinic acid and corosolic acid
  • Protein from centella asiatica and application of protein in the preparation of maslinic acid and corosolic acid
  • Protein from centella asiatica and application of protein in the preparation of maslinic acid and corosolic acid

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Embodiment 1

[0057] Embodiment 1, SynCaMAA45 can be used to prepare maslinic acid and corosolic acid

[0058] The present invention provides a protein from Centella asiatica, its name is maslinic acid / corosolic acid-related protein (CaMAA45), CaMAA45 can catalyze the α-hydroxylation of the 2-position of triterpene, and belongs to the 2-position of triterpene The amino acid sequence of α-hydroxylase, CaMAA45 is shown in Sequence 1 in the Sequence Listing, which can be encoded by the SynCaMAA45 gene shown in the 444-1871 position of Sequence 2.

[0059] 1. Preparation of pRS313-TRP-TEF1-SynCaMAA45-CYC1t recombinant vector

[0060] Use SexA1 and Asc1 to perform double enzymes on p-SynCaMAA45 (synthesized by the company, as shown in the 431-1879 position of sequence 2 in the sequence listing) and plasmid pRS313-TRP-TEF1-MAA45-CYC1t (recorded in Chinese patent 201611037629.9) respectively Cutting; Purify the target gene SynCaMAA45 and the large vector backbone fragment pRS313-TRP-TEF1- / -CYC1t ...

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Abstract

The invention discloses a protein from centella asiatica and an application of protein in the preparation of maslinic acid and corosolic acid. The protein provided by the present invention is any oneof the following: A1) a protein having an amino acid sequence as sequence 1; A2) a protein obtained by substituting and / or deleting and / or adding one or more amino acid residues to the amino acid sequence represented by the sequence 1 and having same functions; A3) a protein having 99%, 95%, 90%, 85% or more than 80% homology with the amino acid sequence defined above and having the same function;and A4) a fusion protein obtained by connecting a tag at a N-terminus and / or C-terminus of any protein as defined above. Experiments prove that CaMAA45 can prepare maslinic acid and / or corosolic acidby taking oleanolic acid and / or ursolic acid as substrates, can prepare maslinic acid and corosolic acid by using CaMAA45 and its coding gene, and also catalyze the triterpene 2 position alpha-hydroxylization by CaMAA45.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a protein derived from centella asiatica and its application in preparing maslinic acid and corosolic acid. Background technique [0002] 2-position α-hydroxylated medicinal triterpenes represented by Maslinic acid (Formula 1), Corosolic acid (Formula 2) and Asiatic acid (Formula 3) The active ingredients are all synthesized by medicinal plants such as hawthorn (Crataegus pinnatifida Bunge), loquat (Eriobotrya japonica), crape myrtle (Lagerstroemia speciosa (L.) Pers.) and Centella asiatica (L.) Urb.) Trace secondary metabolites, both maslinic acid and corosolic acid are pentacyclic triterpenic acids, which belong to β-balsamic resin type or α-balsamic resin type pentacyclic triterpenoids, maslinic acid and corosolic acid are the same Isomers, in plants, the two often co-exist. Maslinic acid, corosolic acid and Asiatic acid are widely used in the fields of anti-cancer, anti-HIV, beau...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N1/19C12P33/06C12R1/865
CPCC12N9/0071C12P33/06
Inventor 张学礼戴住波马晓琳王冬
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI