JPH0866179A - New bacillus sp. bacillus carbophilus - Google Patents

Abstract

PURPOSE: To provide Bacillus carbophilus capable of being used as a sensor detecting a sound wave relating to cell growth of microorganisms, plants and animals. CONSTITUTION: This new Bacillus sp. Bacillus carbophilus has the following mycological characteristics; the Bacillus sp. is aerobic and gram-positive rod, forms ellipsoidal spores and sporangia without swelling, has 38.0+0.2mol% of (G+C) content and is not DNA-homologous with Bacillus megaterium, Bacillus flexus, Bacillus simplex and Bacillus lentus and highly requiring π electrons.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a novel microorganism Bacillus carbophyllus.

[0002]

2. Description of the Related Art Carbon graphite (graphite) and activated carbon have a large amount of active π electrons because of their highly conjugated double bond structure. They tend to absorb visible light and are therefore black and warm in nature. These carbon substances have been used successfully for culturing plants and bacteria and have been successful, but it has been thought that the effect is that harmful substances are removed by the adsorption action of them. However, in fact, it was found that this was due to two direct effects on cell proliferation by the carbon substance and the acoustic signal emitted from the flora. The present inventors require a carbon substance rich in π electrons for growth under lethal conditions such as high salt concentration, and efficiently detect a certain acoustic wave signal emitted by the same or different species of bacteria. I found a fungus.

[0003]

The present invention is a novel strain of Bacillus carbofilus (Bacillus carbo).
philus) and has the following mycological characteristics. That is, the bacterium is an aerobic, Gram-positive bacillus, forms an elliptic spore, does not swell the sporangium, has a G + C content of 38.0 ± 0.2 mol%, and has Bacillus megaterium (Bacillus megaterium).
um), Bacillus flexus, (Bacillus
flexus), Bacillus simplex (Baci)
It has no DNA homology with illsimplex and Bacillus lentus, and has a π-electron-demanding mycological feature.

The present inventors have newly isolated a strain from the air of Numazu City, Shizuoka Prefecture [Kasumi 6,
1, 2, 3, 4 and 7 strains] have the following mycological properties.

(A) Morphological properties

(B) Physiological properties

(C) Physiological properties

(D) G + C content and DNA homology

From the above results, 6 strains are considered to be the same species. These are aerobic (non-anaerobic) Gram-positive bacilli that form elliptical spores and do not swell the sporangium. With such properties, the G + C content is 38.0 ±
0.2 mol% of Bacillus bacteria is B. meg
aterium, B.A. flexus, B.A. simple
x, B. There is lentus. Therefore, the value of DNA homology with these reference strains was determined.

[0010] From the above results, these strains are known to be Bacillus.
It was considered to be a new species different from the s genus bacterium.

Therefore, this new species was named Bacillus carbophilus. Bacillus, a strain belonging to this species
carbophilus Kasumi 6 has been deposited as FERMP-14455 at the Institute of Biotechnology, Institute of Industrial Science and Technology. [Consignment date (August 1, 1994)] For culturing strains of Bacillus carbophyllus, a general method for culturing microorganisms is applied as it is. That is, it is produced by growing it in a nutrient medium containing an assimilable carbon source and a nitrogen source under aerobic (eg, shaking culture, submerged culture) conditions. The medium may be synthetic, semi-synthetic or natural.

[0012] Preferred carbon sources are glucose, mannose, glycerin, molasses, starch, starch hydrolysis and the like, and preferred nitrogen sources are meat extract, casein hydrolyzate, peptone, gluten powder, cornmeal and cottonseed flour. , Soybean powder, corn steep liquor, dry yeast, phosphoric acid, ammonium, ammonium sulfate, urea and the like. Inorganic salts such as phosphates, chlorides and other metal salts such as disodium hydrogen phosphate, potassium dihydrogen phosphate, calcium carbonate, ferrous sulfate, magnesium sulfate, copper sulfate, zinc sulfate, manganese chloride, chloride Magnesium or the like may be added. When foaming is remarkable during fermentation, an appropriate amount of a defoaming agent such as vegetable oil such as soybean oil and linseed oil, higher alcohol such as octadecanol may be added. Culturing is preferably performed at about 30 ° C. for 30 to 100 hours. The above conditions will select the optimal combination of conditions according to the characteristics of the microorganism used.

[0013]

EFFECT OF THE INVENTION Bacillus carbophil
Since us has a high requirement of π-electrons and can efficiently detect a sound wave signal emitted by a cell, it can be used as a sensor for detecting a sound wave signal involved in cell growth of bacteria, plants and animals. For example, it is said that it is difficult to grow an animal cultured cell or a plant protoplast from a single cell, but it is thought that a certain kind of sound wave trigger facilitates this. And as a means for detecting sound waves in the wavelength range required for it, a Bacillus car
It is conceivable to utilize boophilus to define its sonic range and to create a generator that emits it to grow cells that are difficult to culture.

Further, from a more practical point of view, it has been reported that good results can be obtained by listening to music for making sake or cultivating plants. For example, it can be applied to a wide range of industries and medical fields related to the growth control of animal cells such as cancer cells, by using it as a means for detecting a sound wave range that acts on microbial cells or cells of cultivated plants.

Example 1 Bacillus carbophilus Kasu
From slant of mi 6, Trypto-soya-B
100 ml of roth medium (Nissui Pharmaceutical Co., Ltd.) (500 ml Erlenmeyer flask) was inoculated with one platinum loop, and then at 30 ° C. for 48 hours.
When cultured on a rotary shaker (250 rpm), it grows to an absorbance of OD 600 nm up to 3.0,
By centrifugation at 000 rpm for 10 minutes, the bacterial cells of the bacterium could be obtained.

Example 2 Bacto Antibiotic Medium 3
(Trade name) (Difco; abbreviated as BAM3) or a medium (double abbreviated as BAM3 / 2) obtained by doubling it with water to give agar (Wako Pure Chemical Industries) at 1.5% to obtain a medium use. In addition, in the case of BAM3 / 2, KCl was used by adding it to 1% (abbreviated as BAM3 / 2 + KCl). Agar was added to BAM3 or BAM3 / 2 + KCl medium and sterilized at 120 ° C. for 20 minutes, and then 15 to 20 cc was dispensed into a plastic petri dish having a diameter of 9 cm to solidify.
Bacillus carbophilus Kasu
Although it was not possible to grow by smearing mi 6 on such a medium and culturing at 44 ° C, growth was observed by sprinkling an appropriate amount of carbon graphite (Toyo Carbon) or activated carbon on the culture. Was given. This effect was not observed even when alumina powder and glass powder were used in the same manner. The strain is BAM3 or BAM3 / 2 +
It is a strain that has been isolated by sprinkling an appropriate amount of carbon graphite powder on an agar Petri culture medium of KCl and culturing.

Example 3 BAM3 or BAM3 / 2 agar medium was used for the signal-producing bacterium side, and BAM3 / 2 + KCl for the signal-receiving bacterium side.
Using agar medium, put two plastic petri dishes with a diameter of 9 cm back to back (with the lids on top of each other)
Overlap. Bacillus subtilis Marb was placed on the agar medium of the petri dish on the lower side (signal generating bacterium side).
urg 188 (signal-producing bacterium) is smeared, and Baci is placed on the upper side (signal-receiving bacterium side) of the petri dish agar medium.
Smear the illuscarbophilus Kasumi 6 As a control, one without smearing bacteria was set on the lower side (signal-producing bacteria side).

As a result, Bacillus carbo was added to the upper plate only when the bacteria were well grown on the lower plate.
Growth of philus Kasumi 6 was observed. On the other hand, as a control, in the case where the lower plate was not smeared with the bacteria, the growth of the bacteria was not observed in the upper plate. These are due to the signal emitted from the strain that has become able to grow even under lethal conditions due to the action of carbon graphite, and the signal emitted from the strain that is actively growing under normal conditions.
It also means that a strain in the vicinity, which was also unable to grow under lethal conditions, was changed to viable. This signal can pass through air plastics, plus glass, thickness 0.5m
Since it can pass through an iron plate of m, it is a sound wave rather than a heat transfer or electromagnetic wave.

Example 4 As a signal-producing bacterium, Bacillus subtil
The same test as in Example 3 was performed using is ATCC 6633, and the result similar to that in Example 3 was obtained.

Claims (1)

[Claims] 1. An aerobic, Gram-positive bacillus, which forms an oval spore, does not swell the sporangium, and has a G + C content of 38.0 ±.
At 0.2 mol%, Bacillus megaterium, Bacillus
A new strain of Bacillus carbophyllus, which has no DNA homology with Flexus, Bacillus simplex, and Bacillus lentus, and has mycological characteristics of highly demanding π electrons.