JPH0362392B2 - - Google Patents

Description

[Detailed description of the invention]

[Industrial Application Field] The present invention relates to a method for producing sorbic acid, and more particularly to a method for producing sorbic acid by allowing a specific microorganism to act on 2,4-hexadienal. [Problems to be solved by the conventional technology and the invention] In producing sorbic acid, two to four raw materials are
- When hexadienal is oxidized by organic chemical methods, the formation of by-products is unavoidable. Therefore, in order to obtain highly pure sorbic acid, purification operations are necessary, but these operations are complicated;
It was difficult to obtain sorbic acid of desired purity. Therefore, the present inventors used microorganisms to
- When we investigated to develop a method for producing sorbic acid from hexadienal, we discovered that by selecting and using specific microorganisms, it was possible to produce highly pure sorbic acid from 2,4-hexadienal. The present invention was completed based on this knowledge. [Means for Solving the Problems] That is, the present invention is directed to the use of Arthrobacter genus, Bacillus genus,
Bacteridium, Brevibacterium, Citrobacter, Corynebacterium, Enterobacter, Escherichia, Flavobacterium , Klebsiella spp., Micrococcus spp., Microbacterium spp., Nocardia spp., Paracoccus spp.
Genus, Proteus, Pseudomonas, Serratia,
One or more microorganisms belonging to the genus Rhodotorula and the genus Saccharomycopsis that have the ability to oxidize 2,4-hexadienal were added to 2,4-hexadienal. The present invention relates to a method for producing sorbic acid, which comprises bringing it into contact with dienal. The microorganisms that can be used in the present invention belong to the various genera mentioned above and have the ability to oxidize 2,4-hexadienal. Specifically, Arthrobactor oxydans IFO 12138, Arthrobactor aurescens
aurescens) IAM 12340, Bacillus cereus (Bacillus cereus) IFO 3131, Bacteridium sp. R 341 (Bacteridium sp. R 341) CBS
496.74, Brevibacterium Iactofermentum ATCC
21420, Brevibacterium flavum ATCC 13826, Brevibacterium roseum
roseum) ATCC 13825, Brevibacterium
Ammoniagenes (Brevibacterium)
ammoiagenes) ATCC 13746, Corynebacterium glutamicum (Corynebacterium
glutamicum) ATCC 13032, Corynebacterium herculis
ATCC 13868, Corynebacerium sp. ATCC21341, Enterobacter cloacae
IFO3320, Escherichia coli
coli) ATCC 11303, ATCC 9723E, ATCC
ATCC9723D, Flavobacterium suaveolens IFO 3752,
Klebsiella pneumonia
pneumoniae) IFO 3318, Micrococcus sp. A111 (Micrococcus sp. A111) CBS497.74,
Microbacterium ammoniaphilum ATCC
15354, Nocardia erythropolis IFO 12321, Nocardia sp. ATCC 21145,
Paracoccus denitrificans IFO 13301, Proteus mirabilis IFO
3849, Pseudomonas pseudoalcaligenes
ATCC12815, Pseudomonas aeruginosa ATCC 15524, Pseudomonas aeruginosa
acidovorans) ATCC 15668, Serratia marcescens (Serratia marcescens) IAM 1205, Rhodotorula minuta (Rhodotorulaminuta) IFO
387, Saccharomycopsis lipolytica IFO 746, etc., and these can be used alone or in combination of two or more. Microorganisms can be used in various forms, such as cells in the growth phase, cells in the resting phase, immobilized cells, etc., and even extracts from the cells. It may be. Here, the immobilization of microbial cells is carried out by carrier binding method, crosslinking method,
This can be done by applying conventional immobilization techniques such as the blanket method. Further, as an extraction method, a method can be employed in which a suspension of microbial cells is crushed using ultrasound, a French press, a high-pressure homogenizer, etc., and then a soluble extract is obtained by centrifugation or the like. On the other hand, 2,4-hexadienal is also called sorbic aldehyde, and is converted into sorbic acid by an oxidation reaction by the above-mentioned microorganisms. As a medium for culturing the above-mentioned microorganisms, one containing a carbon source, a nitrogen source, etc., and in which these microorganisms can grow is used. As a carbon source, any compound can be used as long as it does not inhibit the sorbic acid production activity of microorganisms, such as glucose, sucrose, ethanol, ethylene glycol, propylene glycol, 1,4-butanediol, glycerin, acetaldehyde, acetic acid, Examples include propionic acid. Further, as the nitrogen source, meat extract, peptone, cornstarch liquor, urea, ammonium sulfate, ammonium chloride, sodium nitrate, etc. can be used.
Furthermore, inorganic salts such as phosphates, magnesium salts, calcium salts, iron salts, copper salts, zinc salts, and other nutritional substances necessary for the growth of microorganisms can be added to the medium as necessary. The contact between 2,4-hexadienal and microorganisms can be carried out in various ways, and it may be added all at once to the area where the microorganisms are present, or may be added in small portions in several portions. Further, the timing of addition is also arbitrary, and it may be added before inoculating the culture medium with the microorganisms, taking into account the microorganisms to be used, or may be added during culturing of the microorganisms or to the culture solution. Alternatively, after completion of the culture, the bacterial cells may be collected and made into a suspension, and then added. The reaction between the microorganism and 2,4-hexadienal is carried out under aerobic conditions and may be appropriately determined in consideration of the properties of the microorganism used. Furthermore, there are no particular restrictions on the reaction temperature, time, pH, etc., and they may be carried out with consideration given to producing the desired sorbic acid with high purity and efficiency. For example, the appropriate temperature is 5 to 80°C, preferably 10 to 50°C, and the pH is 3 to 10, preferably 5 to 8.
is appropriate. Furthermore, the reaction can be carried out in various ways, such as by combining a culture method using microorganisms in the growth phase with a reaction using resting microbial cells, or by performing a reaction using other immobilized microorganisms or a processed microbial cell extract alone or in combination with the above methods. is possible. After completion of the reaction, separation and purification of the target sorbic acid may be carried out by a conventional method after solid-liquid separation. [Example] Next, the present invention will be explained in detail with reference to Examples. Example 1 Nocardia erythropolis
A loopful of IFO 12320 bacterial cells was inoculated into a Sakaguchi flask containing 100 ml of a medium having the composition shown in Table 1, and cultured with shaking at 30°C for 48 hours.
After culturing, the culture solution was centrifuged at 11,000×g for 10 minutes at 5°C. The viable cells obtained were washed with 1/15M phosphate buffer (PH7), and then added to 10ml of 1/15M phosphate buffer. The bodies were suspended at an OD of 10. 2,
10mM of 4-hexadienal was added. The reaction was carried out at 30°C, and after 30 minutes, a portion of the reaction solution was taken and sterilized by centrifugation. Disinfecting solution, 6N
When the pH was adjusted to 2 with hydrochloric acid and quantitative analysis was performed by gas chromatography, the amount of sorbic acid produced was 56 mg/kg. The conditions for gas chromatography are as follows: Column packing material: Thermon
3000celite 545, column temperature: 160°C, nitrogen flow rate: 50 ml/min (at this time, the retention time of sorbic acid was 4 minutes). Table 1 Meat extract 5.0g Peptone 15.0g Ammonium chloride 5.0g KH ₂ PO ₄ 5.0g 1,4-butanediol 5.0g * Make up to 1 with distilled water. (PH7.0) Example 2 The reaction was carried out in the same manner as in Example 1, except that the microorganisms shown in Table 2 were used instead of the microorganisms in Example 1. The results are shown in Table 2.

【table】

〔Effect of the invention〕

According to the present invention, 2,4
- Sorbic acid can be efficiently produced from hexadienal. In particular, since no by-products are produced unlike in conventional organic chemical methods, highly pure sorbic acid can be obtained under mild conditions. In addition, the obtained sorbic acid is used as a preservative in the food industry, and is also useful as a raw material for synthetic resins.

Claims (1)

[Scope of Claims] 1. Genus Arthrobacter, Genus Bacillus, Genus Bacteridium, Genus Brevibacterium, Genus Citrobacter, Genus Corynebacterium. , Enterobacter, Escherichia, Flavobacterium, Klebsiella, Micrococcus, Microbacterium, Nocardia, Paracoccus ( Paracoccus)
Genus, Proteus, Pseudomonas, Serratia,
One or more microorganisms belonging to the genus Rhodotorula and the genus Saccharomycopsis that have the ability to oxidize 2,4-hexadienal are used to produce 2,4-hexadienal. A method for producing sorbic acid, which comprises bringing it into contact with naal. 2. The method according to claim 1, wherein the microorganism is any one of cells in a growth phase, cells in a resting phase, immobilized cells, and a processed cell extract.