Triterpene-2-alpha-hydroxylase MAA45, related biological materials thereof and application of triterpene two-bit alpha-hydroxylase MAA45 and related biological materials in preparing maslinic acid and corosolic acid

A technology of α-hydroxylase, biomaterials, applied in the field of biology

Active Publication Date: 2016-07-27
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there have been relevant advances in the research on the biosynthetic pathway of triterpenoids, including cyclases forming the basic skeleton of each triterpene and performing 6-, 11-, 16-, 22-, 23-, 24- , 28-oxidized P450 enzyme, but the enzyme that undergoes α-hydroxylation at the 2 position (a P450 enzyme) has not been excavated

Method used

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  • Triterpene-2-alpha-hydroxylase MAA45, related biological materials thereof and application of triterpene two-bit alpha-hydroxylase MAA45 and related biological materials in preparing maslinic acid and corosolic acid
  • Triterpene-2-alpha-hydroxylase MAA45, related biological materials thereof and application of triterpene two-bit alpha-hydroxylase MAA45 and related biological materials in preparing maslinic acid and corosolic acid
  • Triterpene-2-alpha-hydroxylase MAA45, related biological materials thereof and application of triterpene two-bit alpha-hydroxylase MAA45 and related biological materials in preparing maslinic acid and corosolic acid

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Embodiment 1

[0082] Example 1, MAA45 can be used to prepare maslinic acid and corosolic acid

[0083] The present invention provides a protein from hawthorn (Crataegus pinnatifida Bunge), whose name is maslinic acid / corosolic acid-related protein (MAA45). MAA45 can catalyze the α-hydroxylation of the 2-position of triterpene, and belongs to the 2-position α-hydroxylation of triterpene. The enzyme, the amino acid sequence of MAA45 is shown in sequence 1 in the sequence listing, which can be encoded by the MAA45 gene shown in the 795-2225th position of sequence 2.

[0084] 1. Preparation of recombinant vector

[0085] Using the restriction endonuclease SacII, insert the DNA fragment shown in the 7-2540th position of sequence 2 between the SacII recognition sequence of the vector pRS313, keep the other sequences of the vector pRS313 unchanged, obtain the recombinant vector, and name the recombinant vector is pRS313-HIS-PGK1-MAA45-CYC1t.

[0086]Among them, the 1-6th and 2541-2546th position...

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Abstract

The invention discloses triterpene-2-alpha-hydroxylase MAA45, related biological materials thereof and application of the triterpene two-bit alpha-hydroxylase MAA45 and the related biological materials in preparing maslinic acid and corosolic acid. The triterpene two-bit alpha-hydroxylase MAA45 is the following protein shown as A1), or A2) or A3), wherein A1) represents protein with an amino acid sequence being the sequence 1 in a sequence table, A2) represents protein which has the same function and is obtained by substituting or missing or adding one or more amino acid residues to the amino acid sequence shown in the sequence 1, and A3) represents fused protein obtained by connecting labels to the N terminal or / and C terminal of A1) or A2). It is proved through experiments that maslinic acid can be prepared from MAA45 with oleanolic acid as a substrate, corosolic acid can be prepared from MAA45 with ursonic acid as a substrate, MAA45 and coding genes thereof can be used for preparing maslinic acid and corosolic acid, and MAA45 can be used for catalyzing triterpene two-bit alpha-hydroxylation.

Description

technical field [0001] The invention relates to triterpene 2-position α-hydroxylase MAA45 and related biological materials in the field of biotechnology and their application in preparing maslinic acid and corosolic acid. Background technique [0002] The 2-position α-hydroxylated medicinal triterpenes represented by Maslinicacid (Formula 1), Corosolic acid (Formula 2) and Asiatic acid (Formula 3) are all It is a trace secondary metabolite synthesized by medicinal plants such as hawthorn (Crataegus pinnatifida Bunge), loquat (Eriobotrya japonica), Lagerstroemia speciosa (L.) Pers.) and Centella asiatica (L.) Urb. Both maslinic acid and corosolic acid are pentacyclic triterpenic acids, which belong to β-balm type or α-balm type pentacyclic triterpenoids. Maslinic acid and corosolic acid are isomers. In vivo, the two often co-exist. Maslinic acid, corosolic acid and Asiatic acid are widely used in the fields of anti-cancer, anti-HIV, beauty and liver protection, and have bee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C07K19/00C12N15/53C12N15/80C12N1/19C12P33/06C12R1/865
CPCC07K2319/20C07K2319/21C12N9/0004C12P33/06
Inventor 张学礼戴住波刘芸王冬张丽丽李守连
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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