JPS6070088A - Production of isopropanol - Google Patents

Abstract

PURPOSE:To produce isopropanol, by culturing a microbial strain belonging to Hyphomicrobium genus in a medium containing methanol as a carbon source. CONSTITUTION:A microbial strain belonging to Hyphomicrobium genus and capable of converting methanol to isopropanol, e.g. Hyphomicrobium variable 42-3- 1 (FERM-P No.7019) is cultured aerobically in a medium containing methanol as a carbon source, and isopropanol is separated from the cultured liquid.

Description

[Detailed description of the invention] The present invention relates to a method for producing isopropanol.

As a result of various studies on the production method of isopropanol using methanol as a carbon source, the present inventors discovered that microorganisms belonging to the genus Hyphomicrobium have the ability to convert methanol into isopropanol. reached.

That is, the gist of the present invention is that a microorganism belonging to the genus Hyphomicrobium and having the ability to produce isopropanol is cultured using methanol as a carbon source, and isopropanol is obtained from the culture. There is a method for producing isopropanol.

The present invention will be explained in detail below.

Hyhomicrobium Barrierville 42-3-1 (Hv4
2-3-1 (Hvpho variable 442
-3-1) (FERP-7019).

The above-mentioned Hyphomicrobium variabil 42-3-1 was isolated from soil collected in Australia in December 1981, and its bacteriological properties are as follows.

(1) Microscopic characteristics Agar plate medium containing 1.5% methanol, culture properties at 30°C for 5 days (colony morphology) a) External shape: round mouth) Size: 1. 0~2.01lIc) Surface ridges: Convex lenticular shape 2) Surface shape: Smooth E) Gloss: None) Color tone: Creamy to pale yellow G) Transparency 2) Transparent C) Periphery; Liquid medium containing 1.5% methanol and agar plate medium,
Morphological properties at 30°C for 2 to 5 days a) Cell size: 0.5 to 1. O
× Za 1, 0 to 3.0 Il c) Polymorphism: None 2) Motility: Yes, single polar benzo e) Sporulation
: None) Durham staining: Negative G) Acid fasting: Negative H) Division mode: Proliferates by buddina. Its germination mode is reminiscent of the germination of mold spores.

The tips of germ tube-like hyphae are swollen. As a result, daughter cells are formed.

(2) Growth status in each medium a) Growth status in slant medium containing 1.5% methanol at 30°C for 5 days.

Vigorous growth, growing uniformly along the inoculation line. Colony color is cream to pale yellow. The surface is smooth. The surrounding area is completely smooth. Opacity.

Mouth) Growth condition in high-rise medium containing 1.5% methanol at 30°C for 5 days.

It grows only along the inoculation line. Growth on the surface is vigorous.

c) Liquid static culture containing 1.5% methanol, 30°C, 5
Daily growth status.

Medium growth. It becomes cloudy. No film was formed.

2) Gelatin medium containing 1.5% methanol, 30°C, 5
Daily growth status.

Medium growth. Does not liquefy gelatin.

e) Growth status on broth agar slant medium at 30°C for 5 days.

Growth conditions are similar to those on methanol-containing slants.

f) Growth condition in gravy layer medium, 30°C, 5 days.

3- Same as growth on methanol-containing strata medium.

g) Meat juice liquid static culture, growth condition at 30°C for 5 days.

Same as growth in methanol-containing liquid culture.

h) Growth condition in meat juice/gelatin medium at 30°C for 5 days.

Same as growth in gelatin medium containing methanol. Does not liquefy gelatin.

4- (3゛) Physiological properties (λ4'J Presence or absence of acid and gas generation from weighing sugars (5)
Assimilation of sugars 7- (7) Chemical composition analysis of [)terminative Bacteri
ology 8th ed.

<1974)) Hyphomicrobium (
It was found that it belonged to the genus Hyphomicrobium.

Clobium (Hyphomicrobium),
Three genera of HyphOIOnaS and pedomicrobium have been described.

Hyphontonas uses C, compounds, and Pedomtcrobiul does not use C, compounds and has a polymorphic form, so H
yphon+retail identified from the genus Icrobium.

Species-level identification The genus Hyphomicrobium includes H, Neptinium (Taka
da, 1974) are known. It is known as a resistant bacterium.

When this strain, 42-3-1, was compared with the original description of H. coagulans, it was found that the growth in peptone water was weak, and the growth in peptone water was weak.
It was distinguished from H. coagulance by not forming ll+018> and by its pattern of carbon source assimilation.

This strain, 41-3-1 and To1゜ Barrier Hill standard strain (
When we compared various microbial properties of NCIB 10517), as shown in test items (3) to (8), both strains were superior in terms of carbon source assimilation and other physiological properties. They were very similar. Furthermore, when the GC content of the DNA of both strains was measured, it was 60.2 to 60.7 mol% for H. barrier leech (NGIB 10517), and 60.2 to 60.7 mol% for this strain 42-. No. 3-1 showed a measured value of 60.2 to 60.5 mol%, and both teeth matched well in this respect as well. Therefore, this bacterium ~1. Wholesale 1 - Not limited.

1,'i 1j:j As the carbon source, in addition to methanol, various carbon salts, urea, etc. can be used.

Furthermore, various phosphates, 11-sulfur mti, etc. can be used as inorganic substances, and various organic nutrients can also be added as necessary.

Cultivation is usually carried out under aerobic conditions for about 12 hours to 10 days, but part of the culturing (the latter stage) can also be carried out under anaerobic conditions.

The pH of the medium is selected from 4-101, and the temperature is selected from about 20-40°C.

In the production of isopropanol, both proliferating bacterial cells and dormant bacterial cells can be used.

When collecting and purifying isopropanol from culture,
Methods generally used for collecting and purifying organic compounds can be employed.

The present invention will be further explained below with reference to Examples.

In addition, the identification of substances in the examples is Gaschroma 12-thiamine (1 mg), riboflavin (1 mo), sodium D-pantothenate (11110), folic acid (0.01
n+o), Fe So, 71-1. O(1n+g
), Zn So, -7H20Nn+o), Cu So
,・5H,O(0,1111(+),Mn C12-4
Dissolve H,O(0,04n+a), 1) Dispense 50ml of medium (A > 50ml) adjusted to H7,0 into a 5001 capacity kolben and sterilize it at 120°C for 10 minutes.Agar 20ml was added to this medium A. Hyphomicrobium barrierville 42-3-1 bacteria was cultured on a methanol-containing agar slant medium supplemented with !+1.e at 30°C for 4 days, a loopful of the culture was inoculated into the above-mentioned Kolben, and the culture was incubated in a reciprocating shaker at 30°C. Culture was carried out for 6 days by applying a rotation speed of 112 times/min.
By statically culturing at 0°C for 4 days, 1 g of isopropanol 4II was obtained (at the same time, ethanol, ace1i = bacteria, and further suspended in 10 ml of the same medium A, applicant: Agency of Industrial Science and Technology Nagagawa 1) Yube 15 -697-

Claims (1)

[Claims] 1. A method for producing isopropatool, which comprises culturing the isopropatool and obtaining isopropatool from the culture.