JPS59143598A - Production of polysaccharide - Google Patents

Abstract

PURPOSE:To obtain an antiviral polysaccharide suitable for treating viral hepatitis, etc. at a low cost, by fermenting gramineous vegetalbe fibers in an aqueous solution containing ammonia nitrogen, nitrate nitrogen, soluble phosphoric acid and soluble potassium. CONSTITUTION:Gramineous vegetable fibers, e.g. rice straw, are finely cut to chips of about 5-30mm. size, and mixed with a basic culture fluid consisting of water, ammonia nitrogen, nitrate nitrogen, soluble phosphoric acid and soluble potassium, put into a cultivation fermenter, closed and cultivated for about 120-240 days. Previously prepared protozoans, e.g. of the family Monadidae, may be added to the basic cultivating fermentation solution to shorten the cultivation period. After completing the cultivating fermentation, fibers are pressed and filtered to remove precipitates from the resultant filtrate by centrifugation. The filtrate is then ultrafiltered to remove metallic ions as impurities, and the resultant purified substance is sterilized to afford the aimed low-molecular weight polysaccharide.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing a polysaccharide, and more particularly to a method for producing an antiviral polysaccharide obtained by fermenting grass fiber.

Conventionally, most of the polysaccharides used as pharmaceuticals are low molecular weight polysaccharides, and one known method for obtaining them is an enzymatic method in which cellulose is degraded by cellulase.

However, when treating vegetable fibers such as wood chips, wheat straw, rice straw, etc., there is a drawback that decomposition cannot be promoted unless the fibers are mechanically, chemically or biologically destroyed in advance.

The purpose of this invention is to directly decompose natural cellulose, which is a grass fiber, and preferably to directly decompose rice straw, which is least utilized, to easily and inexpensively produce low-molecular polysaccharides and their derivatives. It's in the manufacturing method.

That is, in the method for producing the polysaccharide according to the present invention, grass fiber is made into chips of 5 to 30 m in length, the ratio of grass fiber chips to water is 1:15 to 1:60, and ammonia nitrogen is added. 1.0%, nitrate nitrogen 5.5%, soluble phosphoric acid 6.0%, soluble processed 19.0%, the concentration of the basic culture solution is 0.01% to 0.5% and mixed in a culture fermentation tank. and keep it in a sealed state. However, the material may be any material as long as it satisfies the conditions for sunlight to enter the fermentation tank. The culture fermentation period is 120 to 240 days, and the pH is 7.
The end point is 0 to 9.0. In order to shorten the culture fermentation time, by adding a solution containing protozoa, preferably Monas, prepared in advance into the basic culture fermentation solution and maintaining the temperature of the culture fermentation solution in the range of 10 ° C to 30 ° C. Proliferation of Monas species is promoted. After completion of culture fermentation, the fibers are squeezed and filtered, and the filtrate is centrifuged to remove precipitates and ultrafiltrated to remove metal ions as impurities.

The purified product is sterilized at high temperature and high pressure, and then the polysaccharide is confirmed by high-speed chromatography and stored.

EXAMPLES The present invention will be explained in more detail with reference to Examples below.

Example I Put 1,000 g of a 0.1x concentration solution of the basic culture solution described above into a fermentation tank, add 4 g of Monas liquid, and mix uniformly. Next, 40 g of dried reed chips cut to a length of about 15 mm are added and the chips are thoroughly immersed in the culture solution. Then cover with a transparent plastic sheet to seal it. This culture fermentation tank was placed outdoors, exposed to sunlight, and heated to 10°C.
Natural culture fermentation at ~30°C. During this time Pl+
Observe the state of change and check the fermentation status. The Pl+ measurement results are shown in FIG.

After culturing, the fibers are squeezed and filtered, and the filtrate is centrifuged to remove precipitates and ultrafiltrated to remove impurities such as ions.

Pressure 1.2 kg/crA, high temperature high pressure sterilization at 121°C for 60 minutes, high performance liquid chromatography Hitachi 635
In the mold, [;olum 5hodex Ionpak s'
800p+s'804+s'801Sample 1
0μβ Detector: R14x io” RLUFS
'Pressure: 25 kg/cntEl
uent: R20 Flow Rate: 1. OJ 7m1nCha
rt 5peed 5 m/minColumn t
emp: Analyzed under 60'CRT conditions to confirm polysaccharides. The results of polysaccharide confirmation are shown in FIG. 2 by gel chromatography.

Example (2) 1,000 g of a 0.1x concentration solution of the basic culture solution described above is placed in a fermentation tank, and 4 g of Monas liquid is added and mixed uniformly. Next, add 40 g of dried wheat straw chips cut to about 15 mm in length and thoroughly immerse the chips in the culture solution.

After that, put the transparent vinyl lid on to seal it. Place this culture fermenter outdoors and expose it to sunlight.
Natural fermentation is carried out in the range of °C to 30 °C. During this time PH
Observe the state of change and check the fermentation status. l] The I+ measurement results are shown in FIG.

After completion of the culture, the fibers are squeezed and filtered, and the filtrate is centrifuged to remove precipitates and ultrafiltrated to remove impurities such as ions.

Pressure 1.2 kg/cni'/! 60 at A degree 121°C
Sterilized at 21°C, high performance liquid chromatography Hitachi 6
35 type, Column 5hodex 1onpak s'8
00+s'804+s'801Sample: IQ
μR Detector: R14X10 RLUFS'Pr
Essure: 25kg/cJEluent
: 1120 FlowRate: 1.0? lJ2/m1nChart
5peed: 5 in/minColumn t
emp: Analyzed under 60° CRT conditions to confirm polysaccharides. The results of polysaccharide confirmation are shown in FIG.

Example (2) 1,000 g of a 0.1x concentration solution of the basic culture solution described above is placed in a fermentation tank, and 4 g of Monas liquid is added and mixed uniformly. Next, add 40 g of dried rice straw chips cut to about 15 mm in length and thoroughly immerse the chips in the culture solution.

Then cover with a transparent vinyl sheet to seal it. This culture fermentation tank was placed outdoors and exposed to sunlight.10
Natural fermentation is carried out in the range of °C to 30 °C. During this time P
Observe the state of H change and check the fermentation status. The PH measurement results are shown in FIG.

After the culture is completed, the fibers are squeezed and filtered, and the filtrate is centrifuged to remove precipitates and ultrafiltrated to remove impurities such as ions.

Pressure 1.2 kg/cffl Temperature 60 at 121°C
High-temperature autoclave sterilization for minutes, high-performance liquid chromatography Hitachi 6
35 type, Column 5hoclex 1onpak s'
800+s'804+s'801Sample: 1
0μ71! Detector: R14X lo RLUFS
'Pressure: 25 kg/cfflI
Miuent: HzO Flow Rate: 1.047m1nCba
rt 5peed: 5 III/minC
Olum temp: Analyze at 60°C RT to confirm polysaccharides and use clinically. The results of polysaccharide confirmation and clinical results are shown in FIGS. 6 to 10.

For clinical administration, 100 μl of the polysaccharide was extracted with hot water and divided into approximately 11 portions, divided into three doses per day and taken daily. The clinical subject is a 50-year-old man with HB (hepatitis B).

Moreover, FIG. 11 shows an infrared absorption spectrum, which shows that it is a polysaccharide.

Among the grass fibers, rice straw is the best, and the yield of polysaccharides is significantly improved.

When a polysaccharide is used clinically using rice straw, a polysaccharide that has an extremely excellent pharmacological effect against type 13 hepatitis virus has been obtained. In particular, Figures 8 to 11 show that the drugs were administered to patients with hepatitis B, and showed very remarkable effects, with no side effects.They were discharged from the hospital in 50 days, whereas they usually require hospitalization for more than 4 months. As a result, we were able to obtain an antiviral polysaccharide that is thought to have an extremely excellent immune antibody production effect by using Ifuwara, which is said to have the least useful value.

In this way, the resultant product produced by the manufacturing method according to the present invention is
Since it is a polysaccharide containing amino sugars, it has the effect of producing immune antibodies, and may be used as a medicine for viral diseases such as viral hepatitis, herpes, and viral leukemia.

It can also be used in health foods as a food additive for miso, bread, sweets, etc.

Furthermore, the remaining rice straw has been decomposed into lignin and can be used as feed and fertilizer.

Although the present invention has been variously explained above using preferred embodiments, the present invention is not limited to these embodiments (
Of course, many modifications can be made without departing from the spirit of the invention.

[Brief explanation of the drawing]

Figure 1 shows the fermentation status of Example I; pH over time.
Figure 2 is a gel chromatography chart showing the presence of polysaccharides, Figure 3 is a graph of pH changes over time showing the culture conditions of Example H, and Figure 4 is a gel chromatography chart showing the presence of polysaccharides. Chromatography chart, FIG. 5 is a gel chromatography chart showing the culture conditions of Example ①, FIG. 7 is a graph showing changes in GPT and GOT, and FIG. 8 is a graph showing changes in LAP and r-GTP. , Figure 9 is a graph showing changes in yellowness index and platelet count, Figure 10 is a graph showing hemadcrit value, hemoglobin level, red blood cell count, and white blood cell count, and Figure 11 is an infrared graph showing that it is a polysaccharide. It is a chart of an absorption spectrum. Patent applicant Kokichi Hanaoka Drawing 1 Figure 214 J 20' 30
'1, -11 side Figure 3 0 30 system) ri 0 1
2Q 150 1 light 1 210 Number of units 10'20'30' Figure
Figure 5 Change in Pl+ over time (1: Hl fifl ReQ
12f'1 150 18F+ 2
1 (11) Number 61 Peel 1 Kelk's Madography Drawing Plane No. 7I71 0 1020 30 4F1 50 (io
7f + Number of days Figure 8 fHH1 00F □ □ 60i+ 1- o 10 20 3OAo 5060 70 Number of days 91 Number of days Figure 10 0 10 20 30 /lO50fi (1 Days procedure amendment 11 Town 58 June 29, 2007 Patent Office Director Kazuo Wakasugi 1, Indication of the case 11 total, 1958 Patent Application No. 17866 2, Name of the invention Process for producing polysaccharide 3.1iIi Correct person Relationship with the case Patent applicant 4, A2 Wang Rijin, May 31, 1980 6, Number of inventions increased by amendment 7, correct target 8, Contents of amendment ■) Page 9, lines 6 to 7 of the specification: `` Figure 5 is a gel chromatography chart showing the culture conditions of Example ①.'' has been corrected as follows. [Figure 5 is a graph of PH change over time showing the culture status of Example ①, Figure 6 is a chart of gel chromatography showing the presence of polysaccharide, j 2) Figures 1 to 6 are attached. Correct as follows. lA detective] 1st interval □ Knee 1 'l □ 217th □ □ □ □ 1st U Lshi〈1 □ Yl th = lin □ \ □ (

Claims (1)

[Claims] 1. A method for producing a polysaccharide obtained by fermenting grass fiber in an aqueous solution containing ammonia nitrogen, nitrate nitrogen, soluble phosphoric acid, and soluble processing. 2. The method for producing a polysaccharide according to claim 1, wherein the grass fiber is rice straw.