JP3525165B2 - Novel bacterial strain Y-104 - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel Y which has the ability to rapidly absorb saccharides under aerobic conditions and to accumulate polysaccharide at a high concentration in cells. -104 strains. 2. Description of the Related Art Conventionally, in the activated sludge method in water treatment, the sugars in the wastewater are absorbed by the sludge and accumulated in the sludge relatively early after the wastewater is added to the sludge. It has been known. This is thought to be due to the presence of microorganisms having the property of specifically absorbing and accumulating sugars in sludge. Therefore, by separating and culturing such microorganisms, saccharides are accumulated in the microorganisms, and by separating the accumulated saccharides from the microorganisms, various saccharides can be recovered, or such saccharides can be recovered. The accumulated microorganisms can be used for feeds and the like, but such microorganisms have not yet been isolated from sludge. [0003] Therefore, the present inventors have
As a result of screening various types of sludge to isolate microorganisms with excellent saccharide accumulation ability from the sludge for the purpose of separating and recovering saccharides in solution in wastewater treatment, etc. This was obtained by isolating a novel bacterium strain Y-104 from sludge. [0004] That is, the present invention has the ability to absorb saccharides under aerobic conditions, accumulate the absorbed saccharides at high concentrations in the cells, and have the ability to form spores. A novel bacterium Y-104, which is a gram-positive coccus which is oxygen demanding and has catalase activity, and which does not belong to the genera Micrococcus, Planococcus, Dinococcus, Staphylococcus, and Stematococcus. Strain (Deposit No. FERM
P-14709). The novel Y-104 strain of the present invention will be described below in more detail. The novel strain Y-104 of the present invention is obtained by collecting sludge from an activated sludge tank (Tsukuba City, Ibaraki Prefecture, A sewage treatment plant) that performs sewage treatment operation by an anaerobic / aerobic method, and colonies are simply isolated by a dilution plate method. It was separated by a separating method. Also,
At this time, the culture medium shown in Table 1 was used as the plate medium for separation, which was solidified with 1.5% agar and used. [Table 1] [0007] In the colony isolation method by the dilution plate method, it is considered that a microorganism having a high colony forming ability which appeared early inhibits the colony formation of a microorganism having a slow colony formation. Culture was carried out while removing the agar, and colonies that appeared after 10 days of culture were isolated. As a result, a novel bacterium having the ability to absorb various carbohydrates in excess of the cell weight and to accumulate as polysaccharide in the cell was successfully isolated. The bacteriological properties of this strain are as follows. [Table 2] [Table 3] [Table 4] As described above, based on the mycological properties shown in Tables 2 to 4, Bergey's Manual of Systematic Bacteriology, Volume 2, 1986 (Bergey's Manual
ofSystematic Bacteriology, Volume 2, 1986). As a result, Gram-positive cocci having no spore-forming ability were classified into 15 genera, and 10 genera having oxygen demand for endogenous growth of the 15 genera. Furthermore, those having catalase activity are five genera of the genus Micrococcus, Planococcus, Dinococcus, Staphylococcus, and Stematococcus, and Dinococcus is a special fungus having radiation resistance. Because of this, Planococcus sp. Has motility and Smatococcus sp. Has a capsular membrane. Therefore, this strain is considered to belong to the genus Micrococcus or Staphylococcus. However, in the cell wall diamino acid type and the quinone type, this strain was different from these two genera. In addition, recently as a new bacterial classification index,
The base sequence of 16s-ribosomal RNA has attracted attention, but the base sequence database created by the European Institute for Molecular Biology
In comparison with (DNASIS EMBL), it was clear that this strain had low homology to existing bacteria. Therefore, the present strain is a novel strain in which no genus is classified according to the conventional bacterial classification index. [0012] The novel strain of the present invention is characterized by
No. 04 strain, and deposited with the Institute of Biotechnology, Institute of Biotechnology, National Institute of Bioscience and Biotechnology, No. 2120 (FERM P-147).
No. 09). The strain of the present invention is a Gram-positive cocci that has the ability to absorb saccharides under aerobic conditions, accumulate the absorbed saccharides at high concentrations in the cells, and does not have spore-forming ability. It is a novel strain that has no activity and has catalase activity and does not belong to the genus Micrococcus, Planococcus, Dinococcus, Staphylococcus, or Stematococcus. Can be separated and recovered. Hereinafter, the present invention will be described in detail with reference to examples. In the examples, all percentages are by weight unless otherwise specified. (Example 1) Using the liquid medium shown in Table 1, 200 ml of this liquid medium was inoculated with the bacterium Y-104 of the present invention, and cultured with shaking for 5 days. After the culture, the cells were collected by centrifugation, and the cells were washed with distilled water. Ten
0.1 g of the washed cells was taken in a 0 ml graduated cylinder, and distilled water was added thereto to adjust the cell concentration to 1.0 g / L. 0.2 g of glucose (reagent, manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in 0.2 ml of water, added to a graduated cylinder, and agitated at a flow rate of about 0.5 ml / min. After the addition of glucose, the amount of glucose accumulated in the cells and the glucose concentration in the culture solution were measured at predetermined intervals, and the ability of glucose to accumulate in the cells was evaluated. The measurement method was as follows: first, take 1 ml of the culture suspension every predetermined time, and centrifuge (12,00
(0 rpm) to separate the cells from the supernatant. Next, the cells were washed with distilled water, suspended again in 1 ml of distilled water, and the amount of glucose was quantified by the following phenol / sulfuric acid method. FIG. 1 shows the concentration of glucose accumulated in the cells and the concentration of glucose in the culture solution. <Phenol / sulfuric acid method> The test solution or the test cells were mixed with 1 ml of a 5% aqueous phenol solution, and 5 ml of concentrated sulfuric acid was added at once and mixed well. After allowing this solution to stand for 20 minutes, the absorbance at an absorption wavelength of 490 nm was measured using an absorptiometer (U-3210, manufactured by Hitachi, Ltd.), and the test solution was prepared from a calibration curve prepared in advance. Alternatively, the glucose concentration in the test cells was determined. Example 2 Using the washed cells of the bacterium Y-104 strain of the present invention used in Example 1, sucrose was used as a saccharide, and the ability to accumulate sugar in the cells was similarly evaluated. . Take 0.1 g of the washed cells into a 100 ml graduated cylinder,
Distilled water was added thereto to adjust the cell concentration to 1.0 g / L. 0.1 g of sucrose (reagent, manufactured by Wako Pure Chemical Industries, Ltd.) was dissolved in 0.2 ml of water, added to a graduated cylinder, and agitated at a flow rate of about 0.5 ml / min. After the addition of sucrose, the amount of sucrose taken up into the cells and the concentration of sucrose in the culture solution were measured at predetermined time intervals to evaluate the ability of the cells to accumulate sucrose. The measuring method was the same as the measuring method of glucose in Example 1 (phenol / sulfuric acid method). FIG. 2 shows the concentration of sucrose accumulated in the cells and the concentration of sucrose in the culture solution. EFFECT OF THE INVENTION By utilizing the bacterial strain of the present invention, not only can these be rapidly and highly separated from a solution containing various monosaccharides and oligosaccharides, but also cells containing polysaccharide can be recovered. This can be reused for feeds and the like, and the strain of the present invention is industrially useful.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the results of accumulation of glucose in cells by the novel bacterial strain Y-104 of the present invention. FIG. 2 is a graph showing the results of accumulation of sucrose in cells by the novel bacterial strain Y-104 of the present invention.

Continuing on the front page (72) Inventor Mamoru Kawaharazaki 1-3-1, Higashi, Tsukuba, Ibaraki Pref., National Institute of Advanced Industrial Science and Technology (72) Inventor Yukihiko Yoshimi 2 Midoricho, Beppu-cho, Kakogawa-shi, Hyogo Taki Chemical Co., Ltd. In-company (56) References JP-A-3-56102 (JP, A) JP-A-2-4802 (JP, A) JP-A-61-173796 (JP, A) JP-A-1-137990 (JP, A) JP-A-8-66181 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) C12N 1/20 MEDLINE (STN) BIOSIS / WPI (DIALOG)

Claims (1)

(57) [Claims] [Claim 1] A gram that has the ability to absorb saccharides under aerobic conditions, accumulate the absorbed saccharides at high concentrations in cells, and has no spore-forming ability. A novel bacterium Y-10, which is a positive coccus, has oxygen demand and has catalase activity, and does not belong to the genera Micrococcus, Planococcus, Dinococcus, Staphylococcus, and Stomatococcus.
4 strains (deposit number FERM P-14709).