JPS5929235B2 - How to decompose surfactants - Google Patents

Description

Detailed Description of the Invention The present invention uses sulfate-based anionic surfactants and polyoxyethylene higher alcohol ether-based nonionic surfactants as Pseudomonas LF 2102nov, sp.
- (Microorganism Accession No. 3795).

More specifically, the microorganisms are cultured in a medium, and the grown microorganisms are decomposed using bacterial cells, crushed bacterial cells treated by a conventional method, or a structure in which the bacterial cells or crushed bacterial cells are immobilized by a conventional method. Regarding how to.

Conventionally, most household and industrial surfactants have been treated by the decomposition ability of microorganisms in nature, and some of them have been biologically treated by equipment at sewage treatment plants and the like.

However, problems with the biological treatment method include bulking phenomenon, complicated operation management, narrow treatment condition range, generation of sludge, and foaming phenomenon.

In order to eliminate or improve these various problems, equipment such as pre-treatment processes, regulating ponds, and sludge fire tanks are required, and the reality is that the number of treatment processes and equipment scale have to be increased. .

This is a common problem in commonly known mixed microorganism systems such as activated sludge, and is also considered to indicate the limits of improvement in known methods of this type.

Therefore, the present inventors have conducted extensive research into methods that can efficiently (in a short time) decompose various surfactants in wastewater even at high concentrations, and that can also simplify and downsize treatment equipment.

The results showed that microorganisms belonging to a new species of Pseudomonas have a remarkable ability to decompose sulfate-based anionic surfactants and polyoxyethylene higher alcohol ether-based nonionic surfactants, and that the intracellular products of this microorganism have remarkable They discovered that they have similar resolution, and completed the present invention.

The present inventors first isolated a large number of bacteria from sewage water, repeated plate separation three times to obtain a single colony, and then used a liquid medium containing 1100 pp of surfactant as a carbon source.
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A loopful of these isolated bacteria was inoculated into 300 m7 of SO4.7H200,025%, and cultured by shaking at 25°C.

However, the surfactants include (1) sulfate-based anionic surfactants such as dodecyl alcohol sulfate and dodecyl alcohol ethoxysulfate (
(2) Polyethylene glycol decyl ether (EO2P=7) was used as a polyoxyethylene higher alcohol ether nonionic surfactant.

The decomposed dust was measured by the methylene blue method (JISK3363) for sulfate-based products, and the cobalt thiocyanate method (JISK3364) for polyoxyethylene higher alcohol ether-based products.

In order to isolate microorganisms that exhibit a wide range of degrading abilities for the anionic and nonionic surfactants mentioned above, the above-mentioned isolated bacteria were investigated, and it was found that a bacterium named LF2102 had a particularly excellent degrading range. Served.

When the mycological properties of LF2102 were examined, it was unexpectedly confirmed that it belonged to a new bacterial species.

Therefore, I decided to use this LF2102 as a base/manual/
Its morphological and physiological properties were examined with reference to the method of Determinative Bacteriology, 8th Edition and other literature.

The results are as follows.

Morphological properties Form Bacillus flagella Single hair Motile Cloth Durham staining Negative bacterial body axis Straight Line Solid medium surface colony (gravy agar medium) Round, central convex stripe, surface smooth, viscous (7 days culture), edges Smooth edges, opaque slanted medium (gravy agar medium) Growing, colorless, uniform shape, smooth edges, odorless surface, puncture culture growth level Cloth, surface growth cloth, growth status linear, Liquid medium (gravy agar medium) Medium) Moderate growth, no precipitation, heat of gas generation, odorless, heat of color, uniform turbidity, heat of surface growth Oxygen-requiring aerobic growth temperature Maximum 37-38°C Optimal 37°C growth pH 5
, 0 to 9.0 Optimal 7.0 Pigment production No gelatin No liquefaction, physiological properties Nitrate reduction + (N2 generation) Nitrate respiration + (N2 generation) Oxidative breakdown of glucose + (weak) No fermentation, oxidase + Catalase + Indole production Hydrogen monosulfide generation - Urea breakdown - Carotenoid production - Starch hydrolysis - Sugar breakdown (acid production) Glucose + Xylose - Lactose - Sucrose + Starch - Glycerin - Availability of glucose as sole carbon source + Starch -xylose
- Glycerin - Lactose - Trehalose - Sucrose + May4nocyto-/L/ -β-alanine + DL-
Arginine + L-valine + As described above LF 2102 bacteria are Gram-negative bacilli, move with extremely single hairs, and decompose glucose to produce acid, although weakly aerobically.

It was identified as belonging to a species of the genus Pseudomonas based on its oxidase and calatase positivity.

Further, a species search was conducted, and eight bacterial species belonging to Pseudomonas, which are considered to be related to LF2102, were selected and various physiological properties were compared and examined.

The results of this comparison are shown below.

+: 90 or more positive -: 90ql or more negative d
:Positive among 1O-90 staff *Microorganism Deposit Number 3299, Institute of Microbial Technology, Agency of Industrial Science and Technology Among the mycological properties of LF 2102 mentioned above, it is particularly important to have nitrate respiration ability, exhibit denitrification effect, and growth Maximum temperature is 38-3
It is characterized by a temperature of 9°C, slow oxidative decomposition of glucose in the OF test, and weak production of melanin pigment.

Also, the oxidative breakdown of glucose is slow in the OF test.

Ps, malt, and ophilia are DNase negative and do not liquefy gelatin.
effaciens do not liquefy gelatin, and DNase
The difference was that the results were negative.

In the above points, the characteristics of LF2102 do not match the previously described bacterial species, so LF2102 was identified as a new bacterial species, and Pseudomonas LF2102nov, sp.
- (Tentative name: FEIKEN Entrustment No. 3795).

In carrying out the present invention, (1) LF acclimated in a liquid medium to which one or more of the various surfactants described above are added;
2102, or LF 21 grown in broth medium.
02 cells, immobilized cells, culture solution, etc., are brought into contact with a solution containing various surfactants and reacted, or (2)
The bacterial cells are treated by conventional methods, such as washing, ultrasonic treatment, freeze-thaw treatment, homogenizer treatment, and other crushing extraction treatments, and then separated or not separated, and then subjected to immobilization treatment, such as agar. It may be fixed with a polyacrylamide gel or the like, and brought into contact with various surfactant-containing solutions.

The decomposition dust of the surfactant was measured by the methylene blue method, the cobalt thiocyanate method, and the gas chromad analysis, and was expressed as the residual rate of the surfactant in the solution.

The above-mentioned treated product was purified by conventional methods to obtain active substances with increased activity, and various enzymatic properties were also investigated.

Of course, these active substances can also be used in the decomposition treatment method of the present invention.

Thus, the various surfactants mentioned above could be efficiently decomposed by the method of the present invention.

Examples are shown below.

Example I Surfactant decomposition region LF of strain LF2102
Strain 2102 was transferred from the slant medium to a 10 mJ broth liquid medium and precultured at 30°C for 18 hours.

Using this as a seed culture solution and using various surfactants as a carbon source,
It was added to 300 g of a liquid medium containing 100 pp, and the presence or absence of a decomposition reaction was examined.

The results are as follows.

*The remaining amount in the liquid medium after 3 days of contact is shown as a percentage.

Example 2 Surfactant concentration loading limit of LF2102 strain The surfactant degradable range of LF2102 strain was investigated from the viewpoint of concentration.

First, the surfactant loading concentration was started from 1100 pp,
For those samples for which completion of decomposition was confirmed at that concentration, the loading amount was sequentially increased by loop ppm and acclimatization was performed.

The results are as follows.

*Load limit is indicated as the highest concentration that can reduce the residual rate of surfactant to below factor 3 within 3 days when it comes into contact with bacteria.

Example 3 Decomposition reaction LF21 in surfactant mixed system
In order to find out the decomposition ability of strain 02 in a surfactant mixed system, a study was conducted using a liquid medium containing 100 ppm of various surfactants, and the results are as follows.

*Measurement results after 3 days of reaction As shown in the table above, it was found that the decomposition ability was hardly inhibited even in the surfactant mixed system.

Claims (1)

[Claims] 1 Sulfate-based anionic surfactant and polyoxyethylene higher alcohol ether-based nonionic surfactant characterized by using Pseudomonas -LF2102nov, sp- (Microorganism Accession No. 3795) or a processed product of its bacterial cells How to disassemble.