JPS633598B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing an oxygen-containing compound from a cellulose-containing material, and more particularly to a method for producing an oxygen-containing compound containing butanol as a main component from a cellulose-containing material in a one-step process by a fermentation method. . In the present invention, a cellulose-containing substance refers to cellulose and a substance containing cellulose as a main component. Attempts to produce useful substances using cellulose-containing substances as fermentation raw materials have been made for a long time, and methods for producing alcohols and the like are known. However, the number of microorganisms that can assimilate cellulose-containing substances is relatively limited, so when using cellulose-containing substances as fermentation raw materials, the cellulose-containing substances are first decomposed by cellulose-degrading bacteria or hydrolyzed, and the resulting A two-step process was common in which the decomposition products were used to produce desired useful substances. However, since the cellulose-containing substance was not efficiently decomposed in the first stage, the target substance could not be produced in good yield in the second stage of fermentation. Furthermore, such a two-step process requires a large amount of equipment and is therefore impractical. In order to overcome the drawbacks of such a two-step process method, a method has been proposed in which cellulase and alcohol-producing bacteria are allowed to act simultaneously on a cellulose-containing material. Although this method appears to be a one-step process at first glance, it requires a step of preparing cellulase as a pre-step, and is essentially a two-step process. Furthermore, another method has been proposed in which organic fuel is produced by culturing at least two types of Clostridium microorganisms in combination in a medium using a cellulose-containing substance as a carbon source. However, the product obtained by this method has a low butanol content and is not suitable as a material for blending into automobile gasoline, which has recently been in increasing demand. The object of the present invention is to provide a process for producing a product with a high butanol content from cellulose-containing materials in a one-step process by fermentation. The present invention uses anaerobic cellulose-degrading bacteria selected from the group consisting of Clostridium cellobiparum, Clostridium thermocellum, Ruminococcus albus, and Cellulomonas flavisiena and Clostridium satucaloperbutylacetate in a medium containing a cellulose-containing substance as a main carbon source. This is a method for producing an oxygen-containing compound from a cellulose-containing material, which is characterized by producing an oxygen-containing compound containing butanol as a main component in a one-step process by culturing Nicum. Any known anaerobic cellulolytic bacteria can be used; for example, Clostridium cellulobiparum ( Clostridium
cellobioparum) ATCC 15832, Clostridium thermocellum ( C. thermocellum ) ATCC
27405, Clostridium dissolvens ( C.
Bacteria belonging to the genus Clostridium such as C. dissolvens ), C. cellulosolvens , bacteria belonging to the genus Ruminococcus such as Ruminococcus albus ATCC 27211, and the facultative anaerobe Cellulomonas flavigena ) ATCC 482; 484;
There are bacteria belonging to the genus Cellulomonas such as 486; 488; 491. These act on cellulose-containing substances to assimilate cellulose and produce decomposition products such as glucose. Next, Clostridium satucaloperbutylacetonicum ( C.
saccharoperbutylacetonicum) as ATCC
27021; 27022, etc., and this bacterium uses the above-mentioned cellulose decomposition products to produce useful oxygen-containing compounds in high yield, and is particularly characterized by the high content of butanol in the fermentation products. . In the present invention, the microorganisms described above are cultured in a medium containing a cellulose-containing substance as the main carbon source. Here, the cellulose-containing substances include pine, cedar, beech,
Wood such as poplar; stems, leaves, and bark such as hemp, honeysuckle, rice straw, bagasse, and rice hulls; seed wool such as cotton; waste paper such as newspapers, magazines, and waste cardboard; other fibrous waste; pulp, Examples include cellulose powder, and it is desirable to use these as a carbon source after subjecting them to pulverization or other pretreatment as necessary. These cellulose-containing substances are used so as to be contained as cellulose in the medium at a rate of about 0.5 to 10%. The types and amounts of other components of the medium, such as nitrogen sources, inorganic salts, and other substances necessary for fermentation, may be appropriately determined by conventional methods. Furthermore, the culture medium is sterilized according to a conventional method before use. Cultivation is performed under anaerobic conditions, and conditions such as temperature and pH should be set within a range that allows the microorganisms used to grow well and the desired product to accumulate sufficiently. ℃ temperature, PH range of 4-9. The culture period is set until the desired oxygen-containing compound is sufficiently produced and accumulated, and is generally 2 to 20 minutes.
It is days. There are no particular restrictions on the inoculum amount or ratio of cellulose-degrading bacteria and oxygen-containing compound-producing bacteria during culture, but usually the former: the latter = 1 to 9:
Inoculate at a ratio of 9 to 1, preferably 3 to 9:7 to 1. As the oxygen-containing compound obtained by the present invention, n
-Butanol is the main one, but other substances include ethanol, acetone, acetic acid, and butyric acid. Known means may be applied to collect these compounds from the culture. According to the present invention, oxygen-containing compounds can be obtained from cellulose in a one-step process, and the yield is large, and the content of n-butanol in the product is high. Therefore, it has excellent compatibility as an automobile blended gasoline, and is expected to be used as a fuel.
In addition, the oxygen-containing compounds obtained by the present invention are useful as solvents, chemical raw materials, and the like. In addition, the method of the present invention has features such as not requiring a pre-process for preparing cellulase, allowing stable culture, and allowing the culture solution to be used as it is as a starter for the next fermentation. . Next, the present invention will be explained by examples. Example 1 Cellulose 1%, powdered yeast extract 0.2%, polypeptone 0.5%, ammonium sulfate 0.13%, monopotassium phosphate 0.15%, dipotassium phosphate 0.3%,
Magnesium chloride (hexahydrate) 0.1%, calcium chloride 0.015%, ferrous sulfate (heptahydrate) 1.25mg/,
Prepare a medium (PH7.8) containing 0.05% cysteine hydrochloride and 2.0 mg/resazurin sodium salt,
Sterilization treatment was carried out using conventional methods. On the other hand, as a seed culture solution, add cellulose-degrading bacterium Clostridium cellobioparum to 100 ml of the above medium.
ATCC 15832 was inoculated and cultured anaerobically at 37°C for 5 days (seed culture solution A), and 20 ml of the above medium with glucose added instead of cellulose was incubated with the oxygenated compound producing bacterium Clostridium saccharoperbutylacetonicum ATCC. 27022 was inoculated and cultured anaerobically at 37°C for 3 days (seed culture solution B). Add 20ml of the above medium to 20ml of seed culture solution B and seed culture solution.
After centrifuging 100 ml of A and removing the supernatant, the cells were inoculated and statically cultured at 37°C under anaerobic conditions for 13 days. The production amount and product composition of oxygenated compounds in the culture were determined by gas chromatography (carrier chromosolve).
101, 2m glass column, 190°C). The results are shown in Table 1. Example 2 200 ml of the culture solution from Example 1 was used as a seed culture to inoculate 200 ml of the medium shown in Example 1, and incubated at 37°C for 14 hours.
Anaerobic culture was performed for 1 day. Table 1 shows the results of analysis conducted in the same manner as in Example 1. Example 3 The same procedure as in Example 1 was carried out except that 20 ml of the seed culture solution A was used without concentration treatment and the culture was carried out for 11 days. The results are shown in Table 1. Comparative Example The same procedure as in Example 3 was carried out except that Clostridium acetobutylicum IFO 13948 was used instead of Clostridium acetobutylicum ATCC 27022. The results are shown in Table 1. 【table】

Claims (1)

[Claims] 1. Anaerobic cellulose-degrading bacteria selected from the group consisting of Clostridium cellobiparum, Clostridium thermocellum, Ruminococcus albus and Cellulomonas flavisiena and Clostridium satucaloperbutylacetonicum are added to a medium containing a cellulose-containing substance as the main carbon source. A method for producing an oxygen-containing compound from a cellulose-containing material, which comprises producing an oxygen-containing compound containing butanol as a main component in a one-step process by culturing.