JPS59213493A - Treatment of water - Google Patents

Abstract

PURPOSE:To remove efficiently colloidal materials of protein, org. materials, etc. by adding a flocculating agent contg. the flocculation active material obtd. by culturing the flocculation active material generating bacteria belonging to the dematiacea of the black bacterium family as an effective component to source water. CONSTITUTION:A medium contg. the flocculation active material generating bacterium belonging to the dematiacea of the black bacterium family or the medium obtd. by removing physically and chemically the bacteria by sterilizing by heating the medium is used as a flocculating agent contg. the flocculation active material or a material contg. the flocculation active material as an essential component or said agent is prepd. by refining such medium with an org. solvent. Such flocculating agent is added to the source water to flocculate and settle protein, org. materials, living bacteria, etc. Said agent is added to the source water in the form of liquid or solid at about several ppm - several tens ppm. The flocculation is accelerated if the flocculating agent is added to the source water while stirring at 60rpm/min and if aluminum sulfate is added thereto several minutes after, according to need.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to the use of flocculants by microorganisms, more specifically, to the use of flocculants produced by culturing all the flocculant-active substance-producing bacteria belonging to the genus Dematium of the family Dernatiaceae. This invention relates to a method of coagulating and sedimenting not only proteins but also organic substances, inorganic substances, mineral substances, biological bacteria, etc. using

Conventionally, methods for producing protein aggregation active substances using microorganisms have been proposed, for example, in JP-A-51-861.89 and JP-A-51-115993. A wide range of colloidal substances in which organic substances, inorganic substances, mineral substances, biological cells, etc. are suspended, dispersed, or suspended in liquids, as well as proteins, by making full use of the flocculating active substances obtained using microorganisms. The present invention provides a complete method for achieving excellent flocculation and sedimentation.

The microorganism used in the method of the present invention has the number NFL4.
Black fungi (L) ematiacea, including all bacteria No. 257
e) is a flocculating active substance producing bacterium belonging to the genus Dematium (J) emaHum).

Hereinafter, the present invention will be referred to as
1 (hereinafter referred to as t-acting sterilization) sound is used. Implemented bacteriostatic total carbon (glucose 5u)
When cultured in liquid or solid state on microbial cells, a large amount of the flocculating active substance is produced outside the bacterial cells.
1 inorganic and organic insoluble suspended substances in hot water and sewage water;
The present invention provides a method for aggregating colloidal substances, insoluble and soluble proteins, etc., which is used as a flocculant for coagulation and removal.

The agglutinating active substance or the aggregating active substance-containing substance used in the present invention is a culture solution containing bacterial cells, a culture solution obtained by heating and sterilizing this culture solution to physically or chemically remove the bacterial cells, or a culture solution containing this culture solution in an organic solvent. means the solid substance obtained in Purification 1~. Further, in carrying out the present invention, if the hot water is alkaline, or after making the hot water completely alkaline, calcium ions and the flocculating active substance or the flocculating active substance-containing substance can be added. This flocculating substance is added to hot water in liquid or solid form, and the amount added varies depending on the quality of the wastewater, but is several pp.
m-1&+pprr. The addition method is hot water F60
Stir at r p m/WrR, add this flocculating substance to the solution, and if necessary, add aluminum sulfate after a few minutes and stir. Coagulation will occur in a few minutes (1 to 2 minutes) and the formation of flocs. It is fast and the settling property is very good. The number of cloudy fish in the supernatant liquid is 72.
T% was measured at 0 nμ and compared against a total distilled water control.

The color value was measured at 420 nμ, and the pH of the treatment solution was varied depending on the quality of the treated water, and the amount of aluminum sulfate added was also varied depending on the quality of the treated water.

Specific examples will be described below, and the mycological characteristics of Example α will be described below for reference.

Mycological characteristics of the A0 isolate The colony initially grew like yeast with a smooth gray-white, slimy, and glossy oil droplet shape (fat-like), and filamentous fungal bodies grew radially from the periphery and became curly. It is filamentous and develops in a dendritic manner. These filamentous fungi grow well not only on the surface of the medium but also inside the medium.

After a while, light brown spots appear on the surface of the colony, which gradually turn into black spots until the entire surface becomes dark black. This filamentous fungus harbors numerous pale brown oval or oval conidia. These conidia easily break apart. On the other hand, conidia are also attached to the surface of the oil droplet-shaped colony.

The culture solution containing all the sugars becomes very viscous and produces thick colonies of leathery black moss on the surface of the solution. The optimal growth temperature is 2
At 0 to 25°C, all organic acids are produced from sugars such as glucose and sucrose before alcohol.1-1 It also has a unique aroma.

1.Culture 11￥￥撼（c） ■Solid medium: Potato, glucose agar medium-”2.
At first, the colony has a smooth, transparent, glossy oil drop-like, viscous, grayish-white yeast-like appearance, and from the periphery of the colony radially curled, filament-like, exactly dendritic 1T solids grow, and this filament-like fungal body grows. grows well not only on the surface of the medium but also in the medium. Eventually, some parts of the tree-like branches turn blackish brown. After 3 to 4 days of culturing, pale dark brown spots appear on the surface of the colony, but after that, they gradually become pale dark black and spread over the entire surface, and finally turn black as a whole (after 7 days of culturing).

Although growth is slow on Zapek agar medium, the culture characteristics are as described above. It takes about 3 weeks for the surface of the colony to become entirely black.

@Liquid culture: Potato and glucose medium, with bacterial cells appearing in a floating state from the Yth day onwards (3rd day of culture), the number of colonies gradually increases, and eventually (7 days of culture) the liquid is filled with viscous colonies. Then, dark black fungal moss appeared on the tube wall and gradually formed on the liquid surface (15 days of culture). This closure is thick with a gelatinous consistency.

In addition, V cells grow in the same way in Zuapek's medium, but their bodies are small and a considerable black fungus moss is formed on the liquid surface after about 3 weeks of culture.3, 2. Morphological characteristics: Young cells, 11 'Transparent filamentous, scattered dendritic,
The fungal bodies (filamentous) are black, oval, and spore-like in some places. In addition, the oil drop-shaped colonies have black spore-like substances attached to them. This will fall apart when subjected to full impact.

3. Physiological characteristics: The optimal growth temperature is 20-25°C, which produces viscous substances such as glucose and sucrose, and has a unique aroma that converts sugars such as glucose to alcohols and organic acids.

Written by George SMITH, Applied Dutch Studies Guidelines (4n)
1ntroduction to industry
Based on ia1mycology81)68-97) Applied Mechanical Biology (p83-87).

B. Isolation of bacteria and detection of flocculation. Prepare a 5% solution of raw sugar as a bacterial isolation medium, sterilize it using a conventional method, and use 100m7! Dispense 20ml into an Erlenmeyer flask and adjust the pH.
After adjusting it to be slightly acidic, sterilize it again and add 1ml of the stock solution described below to this liquid medium. The cells were cultured by a static method (room temperature 25 to 30° C.), and samples were sampled every - day to examine aggregation. That is, a 1% bath solution of kaolin (special grade reagent manufactured by Kurade Pharmaceutical Co., Ltd.), which is normally used in aggregation tests, was prepared and the pH was made slightly acidic to prepare a test solution.

The above stock solution is granulated sugar, the separation of high molecular weight polysaccharides contained in raw sugar, and the viscosity of a dilute solution of raw sugar (a solution dialyzed with a 24A0 (oral) cellulose membrane) that was left in a beaker for a long time during analysis. After noticing that the blood pressure had increased, I filtered it completely in order to undergo dialysis again. In its operation, when a small amount of diatomaceous earth or activated carbon is added, this lip fluid has a very remarkable flocculating property, i.e., diatomaceous earth or activated carbon instantly solidifies at the bottom of the beaker, which is not seen with other commercial flocculants. I discovered that there is a cohesive force. Furthermore, various substances such as kaolin, bentonite, etc. whose main component is aluminum silicate, or inorganic substances such as calcium carbonate, barium sulfate,
AJT'c was qualitatively confirmed that significant agglomeration occurred when neutral salts such as silver chloride and organic substances were added [7]. Is this sample solution suitable for this agglutination test? :Take 50 test tubes,
The culture solution was added with 1 me of 7JIIL and stirred 10 times, then allowed to rest for 3 minutes, and the total concentration of the supernatant was measured. In addition, as a result of measuring the kaolinka soil remaining in the supernatant liquid using the total weight method and investigating the cohesive sound, it was found that the initial J? 3. The agglomeration property of the nutrient solution is remarkable. This small amount of this flocculating active substance metabolized in the medium has a flocculating effect in minute amounts (44) ``C Suggestion'' 7. As the culture progresses (96 hours), acetone and butanol become stronger. Therefore, the total culture time was set to 72 hours.
This culture '(rlO) is repeated several times to produce a strong cohesive culture solution with no acetone or buknol odor, and a unique aroma (
Honkan's pure separation was carried out from the culture solution k iAbiko having a rose flower aroma.

Yeast extract (manufactured by Kurade Pharmaceutical Co., Ltd.) K 0.2% was added, agar (0.16%) was added, and the mixture was heat sterilized by a conventional method and dispensed into petri dishes to prepare a separation medium. Add culture solution to this with cocoon killing water.
00 times, 200 times, 500 times K f@95<L 1-dispense into a teschial dish L Cultured at 30°C. As a result, all three types of colonies were detected, and by repeating slant culture and liquid culture, three types of dental sounds were isolated. J @ Colonies in the early stage of cultivation (culture time around 48 hours) are all cream colored, but as time passes, the center of the surface of the colony becomes black and as time passes, black hyphae appear on the back of the colony at the center of the colony. In (-), it is a tooth-like isolate I that grows like a seed root in the medium.The cream-colored colony does not change color with the passage of time, but the number of tooth-like isolates rises to 9. Let's call it Isolated Bacteria 1■.

The three types of isolated bacteria were statically cultured in a culture solution with the composition shown in Figure 2, and the cohesive force of the culture solution was examined using kaolin 1φf:'# solution. The results showed that -9-aCt aggregation was observed, with the colony surface turning black over time, and black hyphae extending in a seed-like manner on the L~ surface. The apparent viscosity of the liquid increased with the incubation time, and furthermore, a butyric acid odor was detected in the case of isolate ①, which is characteristic of this bacterium.
1) ematiaceae ) (/J dematium (1
) ematium) belongs to the genus Aggregation Active Substance.

Culture using C8 isolate I and raw Mal of aggregate ψ↓
Purely isolated bacteria 1 (1) ematiaceae l
) e-matium spp.) production of agglomerate storage using all
The results of the moxibustion culture are as follows. Using a carbon source (~), hexoses such as glucose, fructose, galactose, disaccharides such as sucrose, polysaccharides such as starch, and adding 0.2% of yeast extract i; One week later, the flocculation of the culture solution was
However, in both cases, culture fluids with flocculating properties were obtained.

Also, carbon? 14F, aggregates are produced in the entire culture solution by simply adding glucose, etc. As mentioned above, it was found that flocculent substances were produced in the culture solution with a simple medium composition consisting of all carbohydrates. For example, if raw sugar is used as a total carbon source, it is not necessary to add all other nutrients (N bed temperature minerals, etc.). As a result of culturing at a carbon source medium concentration of 5 to 20%, it was found that as the concentration increased, the amount of aggregated substances produced relative to the substrate decreased, and that around 5% was preferable. That is, since the viscosity of the wood aggregate Jtff is extremely high, it was determined that the growth of the fungus would be physically inhibited when this substance reached a certain concentration.

P I-1 does not require particularly strict adjustment if u and j are adjusted to be slightly acidic during the w period. The acidity decreases to some extent during the course of culture. Although both static and shaking culture produce aggregates Tt, it has been found that shaking culture produces faster aggregates. The yield of this flocculating substance based on the substrate (carbon source) is 10% or more and is inversely proportional to the substrate concentration.

Culture conditions, medium composition Carbon Garlic Glucose Concentration 5% Fructose J% Granulated sugar 5% EC sugar 5% N bed temperature Powdered yeast as raw material; 0.2% added to J9 soil Pl-i 5.0 with Hct Adjustment temperature 28~
30°C Inoculation 1 ¥1 is a culture solution f obtained by shaking culture of the main Bacterium I for 7 days
f: l ml was added.

D1 Distribution of the flocculated substance produced by this isolated bacterium I 111 Production method 1 Add the molecule jlf1s'i + (
Completely heat the culture medium inoculated and cultured with Black Dentistry Dema tium (100°C for 15 minutes), and centrifuge treatment 3.
,000 r pIn/n = 1° to remove or filter and separate the bacterial cells. ] Ethanol and culture? A thin film forms on the liquid surface of 1 yen. When stirred, a fibrous or cotton-like substance 1↓ is instantly produced in one place. The aggregated substance is centrifuged or adhered to a stirring rod. The aggregated substance is obtained by drying it in water, adding ethanol to re-agglomerate it, and drying it under reduced pressure after separation. It is inferred that this substance is a homogeneous high-molecular-weight substance because it coagulates instantly with low concentration of ethyl alcohol.

Separation and purification method■ It was found that the flocculating substances in the culture solution were significantly flocculated when aluminum ions were added in acidic conditions, and were flocculated by calcium ions in alkaline conditions. The aluminum ions to be added are aluminum sulfate or its polymer, and the calcium ions are chlorides, lime, etc. Based on this property, we established a method to completely separate aggregated substances in the culture solution. That is, after heat-treating the culture solution (100°C for 75 minutes), bacterial cells are removed by filtration or centrifugation, and 0.05 to 0.10% of inorganic ions, i.e., liquid 'ff:
When the mixture is acidified, an aluminum compound is added, and when the mixture is alkaline, a calcium compound is added and stirred, and the agglomerated substance is completely agglomerated. This can be separated by filtration or centrifugation, and dried to obtain an aggregated substance in the form of a solid powder.

Separation and purification method ■ After complete heat treatment of the culture solution (100℃ 15 minutes), remove the bacterial cells,
Solution 'ff' that does not contain any bacterial cells: Concentrate (approximately 10%) and use as a flocculant. When used industrially as a coagulant, it is very rational to handle it as a liquid for both use and production, as this substance is a very stable substance and there is no need for common understanding when using it. The table below shows all separation and purification methods for this substance (agglomerated substance).

Separation of this substance by adding inorganic salts, separation method nr
In order to react with rr, this substance and 15 salts; -1, the concentration of this substance in the culture solution is determined and the calculated amount of inorganic salt is added. Since the residual liquid from separation method (2) contains some residual carbon source, it is adjusted to a pH suitable for culture and used repeatedly. The added inorganic ions such as aluminum do not act as a culture inhibitor for this bacteria.

E. Use or utilization as a flocculant Isolated Bacteria 1, namely of the family Black Mycoceae]) ema ti ur
It was obtained by culturing N. genus and using the above separation method. Liquids, solids, and solid aggregates containing inorganic ions are present in very small amounts, i.e., 0.000% relative to the amount of liquid. It was discovered that by adding lppm-3'pPm, Denne has the property of completely coagulating and precipitating organic and inorganic substances or biological bacteria that are dispersed, suspended, colloidal, or floating in water or a liquid containing water. The flocculating effect of this substance is significantly stronger than that of commercially available flocculants (inorganic and organic).Also, since it is a metabolite of microorganisms, it has the advantage that there is no problem of secondary contamination caused by flocculants. ing.

Conditions for aggregation are: (1) The optimum pH range is from 1q- to 100% acid; alkaline conditions do not provide sufficient cohesive force. ■The reaction temperature ranges from room temperature to high temperature, and is not related to cohesive force.

It is advantageous to carry out gentle agitation after addition of the agglomerated material. ■The amount used as a flocculant is 0. ], ppm~3, O
It may be expressed in ppm, and the amount used is not related to substances to be aggregated except for special substances. On the acidic side, for aqueous solutions of substances that do not agglomerate with this agglomerating active substance, such as cellulose powder and starch particles, add this aggregating active substance and stir, then add aluminum ions of 1/30 to i/4offi of the added aggregating active substance. It was discovered that when added and stirred, cellulose powder, starch particles, etc. instantly coagulated and precipitated. As a result, all organic and inorganic substances that are suspended, dispersed, floating, or colloidal in water on the acidic side were able to coagulate and precipitate.

Although the cohesive force of this agglomerating active substance in Alka 1J1111 is very weak, it has been discovered that by adding calcium ions, the cohesive force becomes similar to the cohesive force of this substance on the acidic side. The amount of calcium ions added is greater than the amount of aluminum ions added on the acidic side, and the amount added is 20 to 30 times the amount of aggregated substances, that is, 40 to 80.
Requires all ppm.

From the results of this experiment, it was found that in all PL regions, this flocculating active substance coagulates and precipitates the organic and inorganic substances suspended, dispersed, floating, and colloidally present in water. The flocculating effect of Genuine T↓ is that it has an extremely uniform high molecular weight and has an affinity for water.
Because the force is so high (this inference can be made from the fact that alcohol coagulates instantaneously with low concentrations of alcohol), it is as if fine-grained silk was packed in water.
1. However, there are also charged fine particles 1:i;Q
When K l substances enter, the dispersed net becomes 'r! There are times when you lose your balance and become agglomerated. - Reasoning that you can catch matter like you catch fish with a net. This (7-)-li becomes clear when a small amount of aluminum ion is added to the water bath solution of the flocculating active substance [7], and the entire solidification is observed by a rabbit.

Furthermore, when a solution of this agglomerating active substance is exposed to ethanol at a total saturation of 1, a thin film is formed on the interface, similar to the formation of colotion, and it can be observed that many such boats are shaped into cars. In addition, this flocculating substance is used as a water bath liquid at the time of use. Further, the aluminum-containing flocculating substance and calcium-containing flocculating active substance obtained by purification method H are used as an alkaline or acidic aqueous solution.

F0 Physicochemical properties of this active substance This flocculated active substance separated and purified with ethanol is soluble in water, and its 0.1% aqueous solution has a specific viscosity of 4 to 5, which is 4 to 5 that of disaccharide.
Corresponds to the viscosity of the zero bath liquid.

The Anthrone, Molisch, and Biuret reactions of this flocculating active substance are all the same, and the qualitative reaction of 1000 H groups by the carbazole reaction is also shown in ■.The quantitative value is expressed as total galacturonic acid, and its content is 1 ( ) to 15%. In addition, when hydrolyzed with IN H, SO for 24 hours, undecomposed substances remained.1-The sugar composition of the hydrolyzed solution was detected using paper chromatography to detect sugars such as glucose, galactose, and mannose. In this infrared chromatography, absorption of -COOH was confirmed, but absorption of amide groups etc. was not clear.

This aggregating active substance is a flocculating active substance purified and separated from a high molecular weight culture solution containing an organic acid whose main components are glucose, galactose, etc., and has a viscosity of 0.01. 1,6
20 0, U 5 2,200 Acidic, alkaline 0.10 5,200 No change in viscosity Solvent i, oo.

30℃ 55℃ 0.01 1,620 1.6100.05
2,200 2.1000.10 5
,200 5.100 Elemental analysis of purified and separated aggregated active substance) (6,52% C41,04 N O,14 051,74 Ash O,56 (hygroscopic ■) Molecular weight
100 or more (estimated) Qualitative reaction Anthrone reaction Carbazole reaction ■ Biuret reaction Ninhydro-11 reaction e 8'l'f 'J'j Separation and solubility of flocculating active substances Commonly understood strength in ethyl alcohol (alcohol) Less than 40 fissures Soluble 40-45 No sound, egg white-like.

Water retention: 45 heads or more It becomes albumen-like or film-like (collodion film), but when agitated, it becomes a cotton-like aggregate.

Purified and separated flocculating active substance Odor Odorless Taste Tasteless Hygroscopicity Weak (rich temperature) Color Grayish brown fibrous substance
prn) with all additions of divalent or trivalent inorganic ions or heavy metal ions such as Ca++, At-1Mg, Zn, and Pd (
etc. - less than 1/10 of Kanomata C Maso) then the real thing! f completely aggregates and becomes fibrous. This reaction with ions is quantitative.

Example 1. Test results for hot water at Miyazaki City water treatment plant Table 1 □...~I (Source water VC flocculant (microbial production flocculant) is added completely and stirred for 5 minutes (60r prn/rr=) (,,5 The transmittance of the supernatant liquid that had been allowed to stand still for a minute was compared with that of the distilled water control [7 and wavelength 720
Table 1 shows the results measured in mμ.

As is clear from Table 1, the flocculant 'fd: several ppm,
That is, by adding 1 to 4 ppm to the source water, inorganic, organic, colloidal substances or suspended substances in hot water can be instantly turned into flocs with good sedimentation properties, and the permeability of muddy water is 999, that is, equivalent to distilled water. It was found that the transmittance was increased. This shows that the flocculant produced by the microorganisms of the present inventors can be advantageously used as a flocculant for removing colloidal substances or suspended substances in hot water such as water purification treatment. Normally, in water purification treatment (industrial water, tap water), in order to reduce the concentration to less than 2 degrees f: ippm (j
F acid aluminum (sulfuric acid band) is added in an amount of 20 to 30 ppm (or 100 pprn when the amount is large), and flocculated gold is formed in a settling tank, which is then sedimented and removed.

However, by fully using the flocculant produced by this microorganism, the amount of flocculant added may be 1 to 2 ppm, and the sedimentation of the formed flocs is very good, so it is presumed that the processing equipment etc. can be rationalized.

In addition, several pp+11 of this microbial flocculant was added to the source water of purified water, and 1 to 2 pp of Mj fi or aluminum was added to the source water.
When m slowing down to 11, the coca filtration rate of the treated oil becomes even better. In other words, its permeability is comparable to that of distilled water. The results are shown in the second layer.

Table 2

Claims (1)

[Scope of Claims] 1. A flocculant containing as an active ingredient a flocculant active substance obtained by culturing a flocculant active substance-producing bacterium belonging to the genus Pematium of the family Dematiaceae, added to whole source water. A water treatment method characterized by coagulation and sedimentation treatment. 2. The water treatment method according to claim 1, wherein a flocculant containing a flocculating active substance as an active ingredient is used in combination with an aluminum compound.