KR20010097621A - Bacterium removing chemical softener and chemical dextrin, and manufacture method of its immobilized cells - Google Patents

Abstract

The present invention isolates 33 kinds of bacteria decomposing organopolysiloxane (OPS), which is a chemical softener, from plant wastewater producing chemical softeners and chemical glues, and neighboring soils. Nebacterium pseudodiphthericum W3712 was identified and named.

The bacteria best removed the chemical softener at 37 ° C and the removal efficiency was 66.0. In particular, the bacteria were able to remove OPS even in anaerobic conditions without modifying the wastewater storage tanks used by SMEs.

In order to compensate for the loss of biological activity when the cells of Corynebacterium pseudoditericum W3712 were directly used in the wastewater containing the chemical softener, the removal efficiency of the chemical softener was improved by using the microbial cell immobilization method.

The immobilized carrier used in the present invention used bentonite, a natural polymer having low toxicity and low cost to microorganisms, and a method for preparing immobilized cells is as follows.

At room temperature, 16 ml of this culture medium is added to 20 g of bentonite, and the cells are adsorbed and fixed into the porous bentonite to prepare immobilized cells. 10.0 g of the bacteria immobilized on bentonite were removed at least 95.0 because 100 ml of 12.000 μg / L of chemical softener (OPS) wastewater was incubated at room temperature for 10 days.

The immobilized cells that immobilized the Corynebacterium pseudoditericum W3712 bacteria in bentonite as described above have the ability to purify plant wastewater containing high concentration of chemical softener with high efficiency without modifying factory wastewater tank of SMEs. It can be purified.

Description

Bacterium removing chemical softener and chemical dextrin, and manufacture method of its immobilized cells

The present invention relates to a bacterium capable of removing chemical softeners with high efficiency and a method for producing immobilized cells thereof. Specifically, the present invention provides a low-cost, low-cost microbial culture of Corynebacterium pseudodiphthericum W3712, which the inventors have identified and isolated from plant wastewater and chemicals that produce chemical softeners and chemical inhibitors. The present invention relates to a method for purifying wastewater by immobilization to bentonite, a nontoxic natural polymer.

The chemical structural formula of organopolysiloxane (OPS), a chemical softener,

For example, Al (R) and aldehyde (R ') are alkyl groups, and any of the alkyl groups is a secret of each manufacturing plant and is not disclosed. However, no matter which group the eggs and aldehydes are made of, there is no effect on the removal efficiency of the organopolysiloxane, which is a chemical softener by microorganisms.

This product is a slightly yellow transparent liquid, weak ionic / nonionic, easily dispersed in water, used for all chemical and natural fibers, and gives a soft touch to the fiber. The product is dispersed in water in the form of a microemusion, which not only softens the treated fiber products but also gives them elasticity.

Effluents from chemical softeners and chemicals producing plants have very high chemical oxygen demand (COD). That is, to produce one item in a chemical reactor and to produce another item of product, the reactor is washed to produce another product. At this time, the reactor should be washed with as little water as possible to reduce the amount of factory wastewater.

Since these producers do not treat industrial wastewater directly, they are consigned to a wastewater treatment company, so the treatment cost is calculated according to the COD load of the wastewater and the amount of wastewater. Therefore, the amount of wastewater should be minimized and the COD load of the wastewater should be minimized. . The CODMn load of industrial wastewater from chemical softener production plants was 4,000 to 15,000 μg / l, which was high concentration industrial wastewater.

Therefore, wastewater discharged from textile dyeing factories, pretreatment preparation factories, such as hoisting and chemical softeners, are struggling to treat industrial wastewater due to very high BOD and COD loads.

In particular, the wastewater discharged from the textile dyeing plant is a problem in the chemical treatment and biological treatment because the water quality of the wastewater is greatly varied due to the variety of pollution sources by the inflow of wastewater, such as chemical softeners and chemical inhibitors as well as dyes.

For this reason, anaerobic treatment is performed as a pretreatment process, but the treatment efficiency is low due to the short-term treatment due to the quantitative problem of influent wastewater. In addition, in the case of various small-scale manufacturing plants, various wastewaters are stored and requested for disposal, including chemical softeners and chemical decontamination wastes, but the treatment cost according to the load of waste liquids becomes a problem. Therefore, the development of various microbial preparations capable of aerobic, anaerobic and anaerobic anaerobic treatment is urgently required for application in small reservoirs.

The present inventors separated the bacteria decomposing OPS, a chemical softener having a chemical structure as shown in Table 1, from the wastewater and the adjacent soil of the wastewater storage tank of a factory producing a chemical softener, a chemical inhibitor, and the like. The bacterium W3712 was selected.

Table 1

Chemical Structure of Organopolysiloxane (OPS), a Chemical Softener

R and R 'represent an alkyl group.

The taxonomy of selected strains of W3712 was identified and identified as Corynebacterium pseudodiphthericum W3712.

Wastewater containing 2,500 μg / L of chemical softener was treated for 10 days at 37 ° C. so that the chemical softener could be clarified with removal efficiency of 65.2-67.9.

There are a number of problems with using these bacteria directly in plant wastewater. In other words, in order to solve problems such as high COD load of industrial wastewater and long-term use of bacteria, the biological activity of bacteria is lost, and the storage capacity of bacterial cells is low. I wanted to increase.

The present inventors immobilized the bacterium by the immobilization method to maintain the biological activity for a long time and increase the shelf life of the bacterium. In other words, the immobilization method for removing the chemical softener from the industrial wastewater of the chemical softener with high efficiency was examined and the efficiency of the industrial wastewater was removed.

Bentonite (about 200 mesh) was used as the immobilization carrier of the bacteria.

10.0 g of the immobilized cell immobilized with 16 ml of this culture medium in 20 g of bentonite was used to incubate 100 ml of 12.000 µg / L OPS wastewater at room temperature for 10 days to remove OPS with high efficiency of 95.0 or more.

As a result, the immobilized cells could be purified without modification of the storage tank of the plant wastewater.

As a result, the main object of the present invention is a wastewater storage tank of a small and medium-sized company that produces chemical softeners and chemical inhibitors using immobilized cells in which the inventors identified and identified Corynebacterium pseudodifericum W3712 bacteria in bentonite. It is to provide a way to purify waste water over 95.0 without remodeling.

In order to achieve the above object, the present inventors identified the bacteria that purify the OPS, which is a chemical softener, well from the wastewater of a factory producing chemical softeners and chemical inhibitors in Korea, and identified it as Corynebacterium pseudodifericum W3712.

The characteristics of this bacterium are grown at 37 ° C. as Gram-positive, spore-free, non-motor bacilli, as shown in Table 2 below and FIG. 1, and positive for catalase and oxidase test. And negative in MR, VP, OF, indol test.

Table 2

Physiological and Morphological Properties of Corynebacteriumpseudiphthericum W3712 Isolated by Inventors

Furthermore, it was a bacterium that did not produce gas and acid from glucose but reduced nitrate.

The strain hydrolyzes gelatin, but does not break Tween 80 and casein, and uses citric acid as a carbon source.

This bacterium has an optimal growth pH at pH 7.0-7.5 and is best grown at 37 ° C., as well as aerobic anaerobic bacteria.

In particular, the bacterium was cultured at 37 ° C. for 10 days, so that 1,200 ㎍ / ℓ of chemical softener was removed by more than 66.0, and the removal efficiency of chemical softener was also very high.

As a result, Corynebacterium pseudoditericum W3712 bacteria can be purified without agitation of waste water or air supply when purifying chemical softeners, because air supply is not essential for bacterial growth and purification of chemical softeners. Therefore, not only is it economical for the purification of wastewater, but it is also a simple bacterium to operate the purification of wastewater.

Cells were immobilized by immobilization in order to increase the shelf life and increase the biological activity of Corynebacterium pseudoditericum W3712, an organopolyoxysiloxane (OPS) degradation bacterium.

The immobilization carrier of this bacterium is a non-toxic and inexpensive natural polymer material. The carrier is made of bentonite (about 200 mesh), powdered zeolite (about 80-100 mesh) and zeolite. Zeosil (about 360 mesh), which is a synthetic zeolite, was used.

The physical adsorption efficiency of OPS by immobilized carrier () was treated with 10 g of immobilized carrier for 10 days at 37 ° C by adding 1 g of immobilized carrier to 100 ml of standard wastewater of 1,200 ㎍ / ℓ CODMn. () Is shown. Bentonite was also excellent in the immobilization ability of the cells as shown in Table 3 below.

Table 3

Removal efficiency of OPS by immobilized cells in which bacteria cells used in the invention were immobilized on various kinds of immobilized carriers

As a result of measuring the removal efficiency of OPS of the standard wastewater by adsorbing and immobilizing the bacteria cells on the immobilized carrier, the standard wastewater of 1,200 ㎍ / ℓ CODMn was treated for 10 days as shown in Table 3. Bentonite was the best.

The removal efficiency of OPS by the immobilized cells in which the culture medium of bacteria used in the present invention was fixed to bentonite differently, that is, the cell weights were fixed differently was measured.

As shown in Table 4 below, the treatment was performed for 10 days with immobilized cells immobilized with 4 ml of bacterial culture solution in 5 g of bentonite in a 12,000 μg / L CODMn standard wastewater. It was best to immobilize the amount (ml) of bacterial culture.

Table 4

This is a chart measuring the removal efficiency of OPS by varying the amount of cells to be immobilized when immobilized on bentonite.

And the removal efficiency of OPS by immobilized cells had a synergistic effect of about 40 removal efficiency than treatment with immobilized carrier only. The immobilized cells were prototypes of immobilized cells that were stable at room temperature for 60 days.

Furthermore, as a result of measuring the removal efficiency of OPS from the standard wastewater of 12.000 μg / L CODMn by immobilized cell mass, as shown in Table 5, the removal efficiency of OPS increased with increasing carrier amount.

Table 5

Removal efficiency of OPS according to the amount of immobilized cells immobilized with Corynebacterium pseudodifericum W3712 on bentonite ()

10 g of immobilized cells immobilized on bentonite removed 95.0 or more of OPS from 100 ml of high concentration wastewater (12.000 µg / L CODMn) containing OPS.

The immobilized cells immobilized on bentonite retained biological activity at least about 87.5 when stored at room temperature for 60 days.

As a result, the immobilized cell that immobilized Corynebacterium pseudoditericum W3712 on bentonite was found to be a good way to remove the chemical softener organopolyoxysiloxane (OPS) with high efficiency. It is a very useful method for wastewater treatment because it can be used without modification.

1: Electron micrograph of the Corynebacterium pseudodifericum W3712 identified and separated by the present inventors

Hereinafter, an embodiment of the present invention will be described in detail.

These examples are only for illustrating the present invention, it will be appreciated by those skilled in the art that the scope of the present invention is not limited by these examples.

Example 1

[Isolation and Characterization of Chemical Softener, Organophospholipid Degrading Bacteria]

The strains used in the present invention were collected from the wastewater storage tanks and the adjacent soils of the chemical softeners and chemical inhibitors located in Korea, and bacteria were isolated and selected. The collected soil sample and waste water were suspended in a test tube containing 5 ml of autoclaved sterilized water and 1 g of soil sample and 1 ml of wastewater, respectively. After the suspension was allowed to stand for about 3 minutes, the supernatant was taken up with 100 ml of LB agar medium [1 peptone, 0.5 salt (NaC) l, 0.5 yeast extract containing a chemical softener (4,000 ㎍ / ℓ CODMn). ), 1.5 agar (agar), pH 7.0] was isolated from the bacterial colonies produced by incubation at 37 ℃ was separated from the organophosphoric acid (OPS) decomposition bacteria, a chemical softener as shown in Table 1 below.

Table 1

Chemical Structure of Organopolysiloxane (OPS), a Chemical Softener

R and R 'represent an alkyl group.

Bacteria degrading OPS from 25 strains and 8 strains from the plant wastewater were isolated. The culture characteristics of five strains with excellent OPS-degrading activity were examined.

In order to examine the effect of initial pH on the proliferation of selected 5 strains, inoculated so that the culture solution of the preculture strain was inoculated to 1 standard wastewater with different pH, and shaken at 30 ° C for 5 days to measure the proliferation of the bacteria. Strains, S3721 strains, W2823 strains and W2811 strains were well grown at 37 ℃ and W3712 strains.

Morphological and physiological properties were investigated to determine the taxonomic characteristics of the strain W3712, which had the best ability to remove OPS. The morphology of this strain was observed by light microscopy and electron microscopy.

As shown in Table 2 and FIG. 1, the W3712 strain having excellent chemical softening ability, grows well at 37 ° C. as Gram-positive, spore-free, non-motor bacilli, and catalase and oxidase tests ( oxidase test) and negative in MR, VP, OF, and indole test. Moreover, it was a bacterium that did not produce gas and acid from glucose but reduced nitrate. The strain hydrolyzes gelatin, but does not break Tween 80 and casein, and uses citric acid as a carbon source.

From the above taxonomic characteristics, the strain W3712 was identified and identified as Corynebacterium as Gram-positive spore bacillus, which is non-motile, catalase-positive and does not produce indole. The genus Corynebacterium does not produce acid from glucose, fructose, maltose, trehalose, etc., does not break down esculin, reduces nitrate, and is positive for urea reaction. Corynebacterium pseudodiphthericum W3712 was named red because it was negative in red and had an optimal growth temperature of 37 ° C.

Example 2

Removal Efficiency of OPS in Wastewater by Separating Bacteria

OPS removal efficiency () by five strains (W2811 strain, W2823 strain, W3712 strain, S3721 strain, and S3723 strain) excellent in OPS removal efficiency () used in the present invention was measured.

The standard wastewater used in the present invention was diluted with deionized distilled water by Silisoft DH-ES (35OPS), a product of a chemical softener, so that the CODMn values were 1,200 µg / l and 12,000 µg / l. Prepared. Dilution of Sirisoft DH-S products degrades slowly, so standard wastewater was prepared before the experiment.

COD was measured according to the standard method, and the commonly used potassium permanganate (KMnO4) method was used. This experiment measured the chemical oxygen demand by potassium permanganate at acidic 100 ℃. In other words, take a suitable amount of the sample in a 300 ml round bottom flask, add water to make 100 ml, add 10 ml of 32.3 sulfuric acid (H2SO4) and shake about 1 g of silver sulfate (Ag2SO4) powder. After leaving for several minutes, 10 ml of 0.025 N potassium permanganate solution is correctly added, a cooling tube is attached to the round bottom flask, and heated for 30 minutes in a water bath where the water surface of the water bath is higher than the water surface of the sample. Rinse the cooling tube with a small amount of water through the end of the cooling tube, remove the cooling tube, add 10 ml of 0.025 N sodium oxalate (Na2C2O4, sodium oxalate) correctly, and hold 0.025 N while maintaining 60-80 ° C. The CODMn value was determined by titrating with potassium permanganate solution until the color of the reaction solution became pale red. The control test was carried out under the same conditions using 100 ml of water.

The reduction of the chemical softener by decomposition or biological adsorption of the chemical softener wastewater by the separation strain was expressed as wastewater removal efficiency ().

The removal efficiency of the wastewater was calculated by comparing the CODMn values before and after the treatment by inoculating a constant amount of separated strains into the standard wastewater.

Removal efficiency () = A-B × 100 A

A: COD value before treatment of standard wastewater with separate strain

B: COD value after treatment of standard wastewater with separate strain

All experiments were carried out through three or more repeated experiments.

The pH of the effluents of chemical softeners and chemical hopping plants is 4.85 and 6.64, indicating acidic or slightly acidic pH depending on the production plant. In order to examine the effect of pH on the removal of chemical softeners according to the wastewater characteristics by isolated strains, the pH of standard wastewater containing 2,500 ㎍ / ℓ chemical softener was adjusted to 5.0, 7.0 and 8.0, respectively. Was inoculated to 1.

The culture temperature of each strain was W2811, S3721, S3723 and W2823 strains were shaken at 30 ℃, W3712 strains were cultured at 37 ℃ for 5 days to determine the removal efficiency of CODMn was calculated.

The control group without the isolated strain showed a natural removal efficiency of 4.2 as 5 days of culture at pH 7.0. However, the experimental group treated with the isolate showed the best removal efficiency at pH 7.0 and the removal efficiency at 62.0 to 65.6. In particular, W3712 strain, which had the best growth of bacteria at 37 ℃, removed 65.6 chemical softeners at 5 days of culture at pH 7.0 and showed removal efficiency of 64.0 and 65.6 at acidic pH 5.0 and weak alkaline pH 8.0, respectively.

This result shows that the isolate is excellent in degradability even under acidic and weak alkaline conditions, and is suitable for the decomposition of chemical softeners and wastewater with pH in various ranges.

To examine the effect of temperature on the purification of chemical softeners by this strain, inoculate the pre-culture solution of each strain with 1,500 μg / L of chemical softener (pH 7.0) at 25 ℃, 30 ℃ Shake culture was carried out for 5 days at 37 ℃.

The isolated strain W3712 showed the highest removal efficiency at the optimum growth temperature of 37 ℃ and the removal efficiency of 66.0. The strains W2811, W2823, S3721 and S3723 also showed the highest removal efficiency at the optimum growth temperature of 30 ℃.

This result was consistent with the physiological characteristics of selected strains by the ability of OPS, which is a chemical softener, to be consistent with the optimal temperature of physiological growth. In particular, five strains of the selected strains used in the present invention exhibited an optimum removal efficiency even at a relatively low temperature of 25 ℃, and a relatively high removal efficiency even at 37 ℃.

The isolated strain was considered to be a purified strain having a very high practical ability to remove OPS because of its ability to remove OPS over a relatively wide temperature range.

In order to examine the effect of shaking on the purification of chemical softeners by this strain, W3712 strains were inoculated in a standard wastewater containing chemical softeners (2,500 ㎍ / ℓ CODMn, pH 7.0) so that the total culture solution of each strain was 1 At 4 ° C, the remaining 4 strains were cultured and shaken at 30 ° C for 5 days to compare and examine the decrease in COD value.

Removal efficiency of the chemical softener by the strain 5 used in the present invention did not show a significant effect by shaking culture. That is, the strain W3712 removed 65.8 of chemical softeners because of stationary culture for 5 days, but showed a slightly high removal efficiency of 66.3 because of shaking culture.

This result suggests that the characteristics of the strains used in the present invention are close to breathable anaerobic, and can be easily purified without agitation or aeration in the decomposition of chemical softeners and chemical waste liquids. Suitable for direct use on.

Example 3

[Selection of Immobilized Carriers and Immobilization of Cells for Immobilization of Isolated Bacteria]

The experimental strain was immobilized on the immobilized carrier by physical adsorption method. A constant amount of the culture medium was added to the immobilized carrier and left at 30 ° C. for 30 minutes to allow the cells to be sufficiently adsorbed.

In order to immobilize the Corynebacterium pseudodiphthericum W3712 strain to select an excellent immobilization carrier from the removal efficiency of OPS, which is a chemical softener by immobilized cells, 1 ml of the culture strain of the experimental strain was used as a 1 g immobilization carrier. Immobilized in zeolite, bentonite, powdered zeolite and small particle zeolite, respectively.

The immobilized cells were added to 100 ml of the standard wastewater of 1,200 ㎍ / ℓ CODMn containing OPS, and the samples were taken after 30 minutes, and the CODMn value was measured. For 10 days at room temperature (25-30 ° C), The decrease in the CODMn value was measured.

The removal efficiency () of OPS was expressed as a percentage () of the decrease in CODMn values before and after treatment by treating the standard wastewater with immobilized cells for 10 days. As shown in Table 3, the immobilized cells immobilized in zeolite, small particle zeolite and powdered zeolite showed 81.4, 87.0 and 92.0 removal efficiencies, respectively, and the immobilized cells immobilized in bentonite were 1,200 ㎍ / ℓ chemical softener. Bentonite was the best as an immobilization carrier by removing 94.1 standard wastewater.

The immobilized carrier used in the present invention was good in the order of bentonite, zeolite in powder form, zeolite in small particle state and zeolite.

Example 4

Removal Efficiency of OPS in Wastewater by Immobilized Cells Immobilized on Bentonite of Corynebacterium Pseudoditericum W3712

In order to purify the high concentration (12,000 ㎍ / ℓ CODMn) wastewater containing OPS, the strain of the present invention was immobilized to bentonite, an immobilized carrier, by varying the cell weight. That is, 1, 2, 3, and 4 ml of the culture solution of the strain were immobilized to 5 g of the carrier bentonite, respectively, and added to 100 ml of standard wastewater (12,000 µg / L CODMn) and removed at room temperature. Immediately after the immobilized cells were treated in the standard wastewater (30 minutes), the CODMn value was measured and the CODMn value was measured over time from day 1 to 10, and the residual CODMn value was calculated as the removal efficiency of the initial CODMn value. .

When immobilized on the carrier, 3 ml of water was added when 1 ml of the culture medium of this strain was used to fix the cells to 4 ml of the bacterial culture and the amount of water, and 4 ml of water was absorbed into the carrier without using the culture medium. As a control.

As a result of measuring the removal efficiency of OPS by keeping the fixed immobilized carrier volume constant and changing the immobilized microbial mass, the immobilized cells were treated in standard wastewater (12,000 ㎍ / ℓ CODMn) as shown in the following table. Regardless, all experiments showed removal efficiencies of over 50.

Table 4

This is a chart measuring the removal efficiency of OPS by varying the amount of cells to be immobilized when immobilized on bentonite.

The results showed that the chemical softener in the standard wastewater rapidly adsorbed to the immobilized carrier. The longer the treatment time, the higher the removal efficiency was in proportion to the cell weight.

In the control group that did not fix the cells, the chemical softener was removed from the standard wastewater by physical adsorption to the bentonite, which is an immobilization carrier, but the treatment group treated with immobilized cells fixed with 4 ml of culture medium for 10 days showed 94.9 removal efficiency. There was a synergistic effect of removal efficiency of about 39.7.

Example 5

[Efficient Removal of OPS from Wastewater by the Amount of Prototype with Corynebacterium Pseudoditericum W3712 Immobilized on Bentonite and Storage of Prototype]

Prototypes in which 16 ml of the culture strain of the experimental strain was fixed to 20 g of the bentonite carrier were inoculated into 100 ml of standard wastewater (12,000 µg / L CODMn) from 0.1 g to 10.0 g, respectively. The treatment time was measured immediately after the immobilized cells (30 minutes), the CODMn value was measured and the CODMn value was measured over time for 10 days, and the reduction rate of the remaining CODMn value to the initial CODMn value was calculated as removal efficiency.

As shown in Table 5 below, the removal efficiency was increased by increasing the amount of the immobilized carrier for 30 minutes by treating the immobilized cells. When 0.1 g of the immobilized cells were treated in 100 ml of standard wastewater, the removal efficiency was hardly removed. As it increased, the removal efficiency increased. In particular, 0.1 g of the immobilized cells was treated for 10 days, resulting in an increase in the removal efficiency of 1.0 rather than the natural removal efficiency of 3.2. However, treatment with 2.0 g of immobilized cells resulted in a removal efficiency of 57.6. When treated with 5.0 g of immobilized cells, 74.2 and 10 g of immobilized cells were treated with 100 ml of a high concentration of standard wastewater containing 12,000 ㎍ / ℓ CODMn containing OPS. Adsorption is performed, and after 9 days of incubation at room temperature, the removal of 95.0 is considered to be useful for high concentration wastewater purification.

Table 5

Removal efficiency of OPS according to the amount of immobilized cells immobilized with Corynebacterium pseudodifericum W3712 on bentonite ()

The immobilized cells were used as a prototype to be useful for the purification of OPS, a chemical softener.

In order to use commercially available prototypes of the immobilized cells prepared by the present inventors, storage capacity, that is, biological elimination capacity should be maintained at room temperature for a long time, so that the prepared immobilized cells were left at room temperature to measure the maintenance of the purification ability.

The prepared immobilized cells maintained the removal ability of the immobilized cells when stored for 30 days or more, and maintained about 87.5 when stored for 60 days.

As a result, the chemical softener removal ability of the prepared immobilized cells was maintained to be high and commercially available.

Claims (4)

Bacteria that remove chemical softeners and chemical inhibitors from the plant wastewater, characterized by allowing the cells to be adsorbed on the carrier by adding an appropriate amount (g) of immobilized carrier to the culture solution (mL) of OPS (organopolysiloxane) decomposed bacteria. sieve. The mixing ratio of the culture medium (ml) and the immobilized carrier (g) is 1: 1, wherein the culture medium is Corynebacterium pseudodiphthericum W3712, and the immobilized carrier is bentonite. ), And the leaving condition is a bacterium that removes chemical softeners and chemical inhibitors from the factory wastewater, which is produced by leaving it for 30 minutes at 30 ° C. Immobilization to remove chemical softeners and chemical inhibitors from the plant wastewater characterized by allowing the cells to be adsorbed on the carrier by adding an appropriate amount (g) of immobilized carrier to the culture medium (mL) of OPS (organopolysiloxane) decomposed bacteria. Production method of the cells. The mixing ratio of the culture medium (ml) and the immobilized carrier (g) is 1: 1, wherein the culture medium is Corynebacterium pseudodiphthericum W3712, and the immobilized carrier is bentonite. Method of manufacturing immobilized cells to remove chemical softeners and chemical inhibitors from the factory wastewater, characterized in that left to stand for 30 minutes at 30 ℃.