JP3404499B2 - Novel bacterial strain Y-73 - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for rapidly absorbing saccharides under aerobic conditions, having the ability to accumulate polysaccharide at a high concentration in cells, and having a self-sustaining ability. The present invention relates to a novel agglutinating Y-73 strain. 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, carbohydrates are accumulated in the microorganisms, and various kinds of sugars can be recovered by separating the accumulated carbohydrates from the microorganisms. 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 kinds of sludge to isolate microorganisms having excellent saccharide-accumulating ability as described above from sludge for the purpose of separating and recovering saccharides in solution in wastewater treatment and the like, This was obtained by isolating a novel bacterium strain Y-73 from sludge. [0004] Means for Solving the Problems That is, the present invention absorbs sugars under aerobic conditions, have a and self-aggregation properties have the ability to accumulate polysaccharide at a high concentration in the cells, Has sporulation ability
Gram-positive cocci that have an oxygen demand and
No enzyme activity, Streptococcus sp., Leuconos
Toccus, Peococcus, Aerococcus, Jae
A new bacterium strain Y-73 which does not belong to the genus Mera (accession number FER)
M P-14449). The novel Y-73 strain of the present invention will be described below in more detail. The novel strain Y-73 of the present invention collects 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. In addition, the plate medium for separation at this time uses the medium solution shown in Table 1,
This was solidified with 1.5% agar and used. [0006] In the method of isolating colonies by the dilution plate method, it is considered that microorganisms having an early colony forming ability and inhibiting colony formation of microorganisms having a slow colony formation may be considered. Culture was performed 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 cells was successfully isolated. In addition, this strain, which is a new bacterium, can not only accumulate various carbohydrates as polysaccharides in the cells at a high concentration, but also has strong self-aggregating properties, which makes it extremely easy to recover the cells from the culture solution. is there. The bacteriological properties of this strain are as follows. [0008] [0009] [0010] 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 sporulation ability were classified into 15 genera, and 10 genera having oxygen demand for ingrowth of the 15 genera were included. Further, those having no catalase activity are the five genera of Streptococcus, Leuconostoc, Pediococcus, Aerococcus, and Gemera, but recently, GC of DNA has become an important indicator in the classification of bacteria.
This strain was completely different from these five genera in molar content, cell wall diamino acid type and quinone type. Recently, the base sequence of 16s-ribosomal RNA has attracted attention as a new classification index for fungi, but the base sequence database (DNASIS EM) created by the European Institute for Molecular Biology has been attracting attention.
BL), it was clear that this strain had low homology with other 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
Named 3 strains and joined the Institute of Biotechnology and Industrial Technology,
Deposited under the microorganism deposit number of Life Science Bacterial Deposit No. 1342 (FERM P-14449). Strain of the present invention, the sugar under aerobic conditions
Absorbs kind, and have the ability to accumulate polysaccharide at a high concentration in the cells have a self-cohesive, no spore-forming ability grams
Positive cocci with oxygen demand and catalase activity
Non-sensitive, Streptococcus, Leuconostoc
Genus, pecoccus, aerococcus, gemera
It is a novel strain that does not belong to the group , and this strain makes it possible to separate and recover saccharides in a solution in wastewater treatment or the like. 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-73 of the present invention and cultured with shaking for 3 days. After the culture, the cells were collected by centrifugation, and the cells were washed with distilled water. 0.1 g of the washed cells was placed in a 100 ml graduated cylinder, and distilled water was added thereto to adjust the cell concentration to 1.0 g / L. 0.1 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 concentration of glucose in the culture solution were measured at predetermined time 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, measure the absorbance at an absorption wavelength of 490 nm using an absorptiometer (U-3210, manufactured by Hitachi, Ltd.). Alternatively, the glucose concentration in the test cells was determined. Example 2 Using the washed cells of the bacterium Y-73 of the present invention used in Example 1, sorbitol 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 sorbitol (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 sorbitol, the amount of sorbitol taken up into the cells and the sorbitol concentration in the culture solution were measured at predetermined time intervals, and the ability of sorbitol to accumulate in the cells was evaluated. First, 1 ml of the culture suspension was taken, and the bacterial cells and the supernatant were separated by filtration. The cells were then resuspended in 1 ml of distilled water. The total organic carbon concentration of each liquid was measured using a total organic carbon meter (TOC500, manufactured by Shimadzu Corporation), and the amount of sorbitol was determined from the measurement results. The amount of sorbitol in the cells was measured in advance by determining the total amount of organic carbon of the cells in a state where sorbitol was not accumulated, and using the difference in the amount of total organic carbon after accumulation. FIG. 2 shows the concentration of sorbitol accumulated in the cells and the concentration of sorbitol in the culture solution. EFFECT OF THE INVENTION By utilizing the strain of the present invention, not only can these be rapidly and at high concentration be separated from a solution containing various sugars and sugar alcohols, but also cells containing polysaccharide can be recovered. This can be reused for feed and the like.
In addition, since the present strain has self-aggregating properties, it is advantageous because the cells can be easily separated from the culture solution, and the activated sludge treatment by water treatment is easy.

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-73 of the present invention. FIG. 2 is a graph showing the results of accumulation of sorbitol into cells by the novel bacterial strain Y-73 of the present invention.

Continuing on the front page (72) Inventor Mamoru Kawaharazaki 1-3-1, Higashi, Tsukuba-shi, Ibaraki Pref. Institute of Biotechnology and Industrial Technology (72) Inventor Yukihiko Yoshimi 2nd, Midoricho, Beppu-cho, Kakogawa-shi, Hyogo Taki Chemical Co., Ltd. In-house (56) References JP-A-3-56102 (JP, A) JP-A-2-4802 (JP, A) JP-A-61-173796 (JP, A) (58) Fields investigated (Int. . ^7, DB name) C12N 1/00 MEDLINE (STN)

Claims (1)

(57) [Claims] [Claim 1] It has the ability to absorb saccharides under aerobic conditions, accumulate polysaccharide at a high concentration in cells, and have self-aggregating properties.
Gram-positive cocci that do not have the ability to form spores.
With no elemental requirement and no catalase activity
Genus Coccus, Leuconostoc, Pseudococcus,
A novel bacterium strain Y-73 which does not belong to the genus Aerococcus or Gemera (Accession No. FERM P-14449).