JPS63115804A - Production of plant growth promoter - Google Patents

Abstract

PURPOSE:To produce a plant growth promoter extremely useful in terms of industry, by extracting brassinolide from a deodorized distillate of edible vegetable oils as an industrial waste, separating and purifying. CONSTITUTION:A deodorized distillate, a bad-smell component removed in processes of squeezing oil from soybean oil, colza corn, etc., and purifying the oil, is saponified with an alkali and then extracted with an ether to give an unsaponified soybean oil. Brassinolide is separated from the unsaponified soybean oil by solvent fractionation through partition operation between solvents and column chromatography of silica gel and is used as a plant growth promoter.

Description

[Detailed Description of the Invention] (Industrial Application Field) This invention relates to a method for producing a plant growth promoting substance,
More specifically, it relates to a manufacturing method for extracting brassinolide from the deodorized distillate produced during oil extraction of soybeans.

(Prior Art) Auxins, gibberellins, cytokinins, apsisic acid, ethylene, and brassinolides have been isolated as growth-promoting substances present in plants. Plascinoids are plant growth-promoting substances discovered in rapeseed pollen extracts, but are also known to exist in other higher plants.

(The Mano point that the invention seeks to solve) The inventor of the present invention discovered that the content of 1-length promoting substances is generally high in immature seeds and pollen, and that the extraction of rapeseed oil from canola led to the discovery of brassinolide. They focused on soybean oil among a variety of products and discovered that brassinolide was contained in the deodorized distillate from the process of refining extracted crude oil into edible oil.

The feature of this invention is that brassinolide is extracted from the deodorized distillate of edible vegetable oils such as soybean oil, or in other words, brassinolide is extracted from the components that are discarded as malodorous components of edible vegetable oils such as soybean oil. It can be said that it has extremely great industrial significance in that it extracts plant growth promoting substances from the desired components as raw materials.

(Means for Solving the Problems) The method for producing a plant growth promoting substance according to the present invention involves extracting oil from soybeans, rapeseed, corn, etc., and extracting brassicas from deodorized distillate, which is a malodorous component removed in the refining process. This is to separate and purify the ride.

Crude oil extracted as soybean oil or the like is refined through the processes of degumming, deacidification, decolorization, and deodorization to make it into edible oil. The deodorizing distillate produced in the deodorizing process is removed from the edible oil as a malodorous component.

When a deodorized distillate such as soybean oil is saponified with an alkali, most of the lipids turn into soap, but there are substances that do not form soap, and these are called unsaponifiable substances.

The present invention involves saponifying a deodorized distillate such as soybean oil and then extracting brassinolide from the unsaponifiable matter of soybean oil extracted with ether using solution fractionation through a partition operation between solvents and silica gel column chromatography. Separation vI is produced.

The inventor has already clarified that the composition of unsaponifiable soybeans consists of hydrocarbons, vitamin E (tocopherol), (terpene) alcohol, sterols, and pigments, but the present invention has further revealed that it contains prasinoid. revealed.

(Example) The present invention will be explained in more detail below with reference to Examples.

First, the method for producing brassinolide from soybean oil deodorized distillate is as follows.

Figure 1 shows the manufacturing method.

About 1 kg of soybean oil deodorized distillate is saponified with IN methanol potash and then extracted with ether. 450 g of the ether extract was obtained as a soybean oil unsaponifiable product.

The unsaponifiables are carbon tetrachloride: methanol; water (136
: 68 : 9 V/V) solution fractionation using a mixed solvent system was carried out five times, and the methanol layer was collected and dried to obtain about 18 g of a brown rubbery substance.

Then silica gel column chromatography (solvent benzene:methanol:acetic acid 90:16:8V/
The eluate from the column was fractionated into 10ml portions and the transmittance at 400 nm was measured using a spectrophotometric needle (UV). After a minimum was observed, the two fractions that showed 80% or more were combined again. After drying, approximately 1.5 g of substance was obtained.Furthermore, silica gel column chromatography was performed once again in the same manner to increase the precision, and the active substance brassinolide was 0.
．． 3g was obtained.

The active substance was then identified by instrumental analysis.

The results showed that when the absorbance of this active substance was measured in the ultraviolet region using a +11 spectrophotometer, a maximum was observed at 290 nm.
Those with aldehyde or ketone groups.

(2) When the melting point was measured using a field-of-field micro melting point measuring device, it was found that some crystals melt at 110-130°C and others melt at 280-290°C.
Crystals were observed to melt.

(3) When looking at IR absorption using an infrared spectrophotometer, it was found to be a compound with hydroxyl groups, ketone groups, etc., as well as ester bonds and glycosidic bonds.

(4) Based on the results of (3), we attempted to compare it with m-classes such as rhamnose and xylose by paper chromatography, but the Rf values did not match at all, so it was not a saccharide.

(5) Four peaks were observed in gas chromatography GLC (adsorbent silicon ov-i).The relative retention times are similar to those of β-tocopherol and Todoria conchun.Figure 2 shows the results of gas chromatography GLC. However, the existence of congeners is expected.

From these facts, the molecular weight of this active substance is approximately 400.
It was found that the number of carbon atoms was around 30.

Next, the physiological effects of this substance were investigated by bioassay.

Tables 1 and 2 show the elongation test for common beans.

Table 1 Table 2 The results are as follows: +11 When a very small amount of the active substance obtained by the manufacturing method shown in Figure 1 was applied to the second side of a green bean, which was 5 degrees wide, and lanolin was used as a solvent, Approximately 2.5 times the elongation was observed compared to the control group. In addition, some individuals were observed to enlarge as well as elongate.

(2) When leaf blades of wheat (Norin No. 69) grown for 6 days in the dark were cut and soaked in a methanol solution containing a small amount of active substance, leaf blades expanded by 48% in 24 hours and 66% in 48 hours. (10% and 40% in the control group, respectively).

As a result of (1) and (2), it was found that this active substance was succumbing to auxin, gibberellin, or brassinolide, or was a similar substance, based on these physiological activities.

Identification using paper chromatography and gas chromatography revealed that it did not match auxin or gibberellin.

Based on the results of the above-mentioned instrumental analysis and bioassay, it is predicted that the active substance separated and purified from the deodorized distillate of soybean oil is brassinolide, which is a plant growth promoting substance.

(Effects of the Invention) The present invention has extremely great industrial significance in that a plant growth promoting substance has been separated and purified using the deodorized distillate of soybean oil, which should be treated as industrial waste as a malodorous component of soybeans, as a raw material.

Although the present invention has been variously explained above with reference to preferred embodiments, the present invention is not limited to these embodiments, and it goes without saying that many modifications can be made without departing from the spirit of the invention. That's true.

[Brief explanation of the drawing]

Figure 1 shows a method for producing prasinoid from a deodorized distillate of soybean oil. Figure 2 shows identification by gas chromatography GLC.

Claims (1)

[Claims] 1. A method for producing a plant growth promoting substance, which involves extracting brassinolide from the deodorized distillate produced when extracting edible vegetable oil from soybeans.