Method for producing oleanolic acid by white birch suspension culture

Abstract

A process for using white birch suspending culture for producing oleanolic acid. The method includes: taking the dormant bud, leaf and seed of one-year-old branches of white birch as materials, inoculating in 1mg / L of WPM+6-BA medium to obtain white birch aseptic seedling, and then taking the stem and leaf of the aseptic seedling as explants, inducing on the medium IS to get stable and rapid growing callus, and finally culturing the suspension cell lines in a B5 medium in which the hormones are 0.1 - 0.4mg / L 6-BA and 0.01 - 0.1mg / L TDZ, the sucrose is 20g / L, the pH value is 6.0 - 6.5, the fresh weight of inoculating amount is 30 - 50g / L. Stable white birch suspending culture cell lines are obtained by multi-times of successive transfer culture at 23 - 25 DEG C, 110 - 130 turn / min, lighting 8 - 10h / d and light intensity 1400 - 1700Lx, 15 days / successive transfer. Tested by a high performance liquid chromatography, the oleanolic acid content in the obtained white birch suspending culture cells can attain 0.217 - 0.714mg / g (dry weight). The present invention produces the oleanolic acid in the cell engineering method form white birch plants in the first time, which has short period, low cost, largely saved manpower and material and raised labor productivity, to provide a new way to solve the the shortage of natural plant drugs sources and industrial mass production of oleanolic acid drugs.

Description

technical field [0001] The invention belongs to the field of bioengineering, and in particular relates to a method for producing oleanolic acid by cultivating white birch by a suspension method. Background technique [0002] It is recorded in "Compendium of Materia Medica" that birch bark can be used for the treatment of jaundice, breast pain, scabies and other diseases. In the folk, birch bark is also used to treat chronic dysentery, clear heat and detoxify, relieve cough, and relieve asthma. According to research, the main medicinal components of birch bark are secondary metabolite triterpenoids, and betulin is the most abundant, accounting for about 15% to 35% of the bark dry weight. Many studies have confirmed that birch triterpenes can inhibit the growth of tumor cells such as rectal cancer, lung cancer, leukemia, lymphoma, prostate cancer, and ovarian cancer (Zhukova, 2005). Oleanolic acid is a component of birch triterpenes, which can reduce alanine aminotransferase...

AI Technical Summary

Problems solved by technology

Although there is a report (2003) containing dammarane in birch leaves and suspension culture cells, there is no report on the ability to synthesize oleanolic acid using calli induced by aseptic seedlings from birch and suspension culture cells

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