ES2221786B1 - PRODUCTION OF GIBERELINAS, GA1 AND GA3 THROUGH FERMENTATIONS WITH WILD STRAPS OF HONGO GIBBERELLA FUJIKUROI. - Google Patents

Abstract

Production of gibberellins, GA1 and GA3 by fermentation with wild strains of the Gibberella Fujikuroi fungus. The object of this invention is a method of producing gibberellins (especially GA1 and GA3) by wild strains of Gibberella fujikuroi, without the need for prior genetic manipulation of the fungus. The method is independent of the wild strain of the Gibberella fujikuroi fungus that is used. In a first step, the lines are inoculated in a complex medium and the fermentation is carried out with orbital agitation at a temperature ranging from 22 to 32 ° C and in a period of time ranging from 5 to 15 days. Once the fermentation is finished, the culture broth is filtered, acidified and extracted with organic solvents. After evaporation of the solvent there is a residue containing a mixture of gibberellins in an amount around 2 g per liter of medium, with a richness in GA1 + GA3 greater than 70% and in an approximately equivalent proportion.

Description

Method of production of gibberellins, GA1 and GA3 by fermentation with wild strains of the Gibberella Fujikuroi fungus.

Object

The object of this invention is a method of producing gibberellins GA 1 and GA 3, using wild Gibberella Fujikuroi lines, without the need for prior genetic manipulation of the fungus.

Gibberellins (GAs) are natural phytohormones that regulate the growth and flowering of plants among other physiological activities. These phytohormones are at the level of traces in plants but their exogenous application produces spectacular results so they have a great interest in intensive agriculture and the beer industry. More than 100 gibberellins have been described, among which GA3 ( 1 ), also known as gibberellic acid, has traditionally been considered the most active gibberellin in plants (MacMillan, J, Natural Product Report , 1997 , 221). However, recent studies have shown that GA 1 ( 2 ) has an important biological activity, even higher than GA 3 in plants such as rice (Kobayashi M, MacMillan J, Phinney B, Gaskin P, Spray CR, Hedden P , Phytochemistry , 2000 , 55 , 317), tomato (Grünzwig JM, Rabinowitch HD, Katan J, Wodner M, Ben-Tal Y, Phytochemistry 1997 , 46 , 811), cherry tree (Huanpu M, Blake PS, Browning G; Taylor JM, Phytochemistry , 2001 , 56 , 67) or the pea (Yaxley JR, Ross JJ, Sherriff LJ, Reid JB, Plant Physiol , 2001 , 125 , 627).

one

Background of the invention

The filamentous fungus Gibberella Fujikuroi is an organism capable of producing gibberellins in sufficient quantities for its industrial exploitation.

The production of gibberellins by different wild or mutant strains of Gibberella Fujikuroi in various culture media has been described several times. Specifically, the production of GA_ {3} ( 1 ) has been covered under different patents (GB844341, GB850018, GB886551, US2918413) as well as the production of the active mixture of gibberellins GA_ {4} + GA_ {(US6287800, CN1222575 ). However, the production of mixtures of GA1 + GA3 at levels that may be useful in the agrochemical industry has not been previously described, and precisely the invention presented in this patent application consists of a method of gibberellin production that results in a mixture of GA_ {1} + GA_ {3} in a wealth of 70%.

Description of the invention

This patent application describes a method for the production of gibberellins (especially GA1 and GA3) by wild strains of Gibberella Fujikuroi , without the need for prior genetic manipulation of the fungus.

The method is independent of the wild strain of the Gibberella Fujikuroi fungus that is used. In a first step, the lines are inoculated in a complex medium and the fermentation is carried out with orbital agitation at a temperature ranging from 22 to 32 ° C and in a period of time ranging from 5 to 15 days. After fermentation, the culture broth is filtered, acidified and extracted with organic solvents. After evaporation of the solvent, there is a residue containing a mixture of gibberellins in an amount taken at 2 g per liter of medium, with a richness in GA1 {GA3} greater than 70% and in a proportion approximately equivalent.

Description of a preferred embodiment

To illustrate the embodiment of the invention, the line FKMC1995 of the Gibberella Fujikuroi fungus belonging to the University of Kansas Collection has been taken as a starting point.

To carry out the fermentation, the complex culture medium was distributed in Erlenmeyer flasks, which were inoculated with a spore suspension of the wild strain FKMC1995 from Gibberella Fujikuroi . The flasks, covered with fatty cotton, were incubated at a temperature of 28-30 ° C with an orbital shaking of approximately 200 rpm. After 8 days the fermentation was stopped and the culture broth was filtered, acidified to pH 1.5-3 with HCl, and extracted with ethyl acetate. Evaporation of the solvent under vacuum and subsequent degreasing of the extract generated a residue (2.3 g per liter of culture medium) consisting mostly of gibberellins. The majority gibberellins of this mixture were GA1 and GA3 (in approximately equivalent proportions) among which they added about 1.5 g per liter of culture medium.

Claims (2)

1. Method of production of gibberellins GA 1 and GA 3 by fermentation with wild strains of the Gibberella Fujikuroi fungus characterized in that it comprises the following stages:

to)

Inoculation by suspension in a half complex

b)

Incubation and fermentation of the culture with orbital agitation at a temperature between 22ºC-32ºC and for a period of time between 5-15 days

C)

Filtration, acidification and extraction with organic solvents from the culture broth

d)

Evaporation of the broth solvent culture.

2. Residue obtained from the application of the method according to claim 1, characterized in that it contains a mixture of gibberellins in an amount around 2 g / l of cultivated medium and with a richness GA 1 + GA 3 greater than 70% and in an approximately equivalent proportion.