DE10218342A1 - Novel Burkholderia microorganism aerobically degrades oils, fats and other organics over a wide temperature range and is used in waste water disposal - Google Patents

Abstract

A microorganism from the bacterial strain Burkholderia cepacia AIK, is new. An Independent claim is also included for disposal of waste water containing oil and fat by addition of a microorganism of the Burkholderia genus.

Description

This invention relates to a new microorganism and a sewage process, and its purpose is to create a new one Microorganism and a sewage process using of the new microorganism, which effective liquid and solid oils and fats and others decompose organic substances in a wide temperature range can.

Animal and vegetable oils and fats are often in large amounts in sewage systems from food factories, Restaurants and apartments available. In the usual case oils and fats contained in waste water through a flat surface Flotation device and a pressure flotation device away. In the existing physical and chemical Separation and removal processes are used in oils and fats not completely removed from the waste water. The sewage, which contains oil and fat is in a degradation container provided, and the oil and fat is through biological Disposal processes such as the plankton process, which may contain an activated sludge system, and a Biofilm processes such as that Rotary organism contact method, separated and removed.

As a biological waste disposal process, for example, which contains oil and fat is in that Official Bulletin Tokukaihei 8-197986 a proceeding in which the disposal process for the degradation of animal and vegetable oil and Fat, using a special strain of bacteria takes place, which belongs to the Pseudomonas.

Furthermore, in the official newsletter Tokukaihei 9- 85283 a disposal process for the degradation of waste water under Use of bacteria revealed which leads to the strain Burkholderia belong to the animal contained in the sewage and can break down vegetable oil and fat.

Furthermore, in the official newsletter Tokukaihei 11- 47798 a disposal process for the degradation of waste water animal and vegetable oil and fat containing, especially using a bacterial strain which belongs to the Pseudomonas strain.

However, in the aforementioned conventional ones Waste water disposal procedures some problems, such as shown below.

Oil and fat degrading bacteria found in the conventional Waste water processes can use oil and fat in reduce certain temperature ranges. So for Example the bacteria that appear in the official newsletter Tokukaihei 9-85283 are disclosed, oil and fat in one Reduce temperature range above 40 ° C. However, it is known that the temperature inside the breakdown tank, which for the biological disposal is used a strong Temperature change depending on weather conditions, seasonal changes or location changes. in the The result is shown by conventional oil and fat reducing agents Bacteria do not have a high degradability in a degradation container which is subject to a sharp change in temperature. in the The result is the water quality of the treated water, which is in the degradation container and left out is not always firm; sometimes that too treated water omitted without complete degradation.

In addition, the oil and fat-degrading bacteria, which in the aforementioned disposal processes of Waste water is used, liquid oil and fat are broken down, but they have greater difficulty in dismantling solid Oil and fat. In cases where a large amount of solid Oil and fat contained in the wastewater sometimes occur Anaerobic and bad smell on what the Water quality of the treated water is deteriorated.

With regard to the previously mentioned subject, it was called Result of an investigation, one on Burkholderia cepacia belonging bacterial strain due to its high Degradability to break down liquid and solid oil and fat found in a wide temperature range.

Accordingly, it is a general object of the present Invention to the disadvantages of the prior art overcome.

That is, the present invention relates to the Bacterial strain Burkholderia cepacia AIK (FERM-BP-7308).

The present invention relates at least to one preferred waste water disposal method, in which the Wastewater and burkholderia come into contact in such a way that the microorganism can break down oil and fat.

The present invention relates to a Waste water disposal process in which a Microorganism can break down activated sludge.

The present invention relates to a Waste water disposal process in which a Microorganism is used, which is generally for Burkholderia strain belongs, and more precisely the bacterial strain Burkholderia cepacia AIK (FERM-BP-7308) is.

Fig. 1 is a graph showing the result of Example 3.

FIG. 2 is a graph showing the result of Example 7.

Figure 3 is a graph showing the average temperatures of each month in a digester.

Fig. 4 is a graph showing the mean values of each month of an n-hexane extract in wastewater in a digester.

The microorganism to which the the present invention relates.

The microorganism to which the present invention relates is a bacterium, which by sampling and Pre-selection of land from Ishigaki Island in the prefecture Okinawa was obtained and it was observed that he was not just excellent oil mining capabilities and has fat in a wide temperature range, but also degradability in relation to organic substances such as fatty acids, proteins, hydrocarbons and Has saccharides.

At least one preferred isolation method for the new one Microorganism to which the present invention relates is explained below.

• 1. Nährstoffkulturmedium mit 5% Olivenöl in der Zusammensetzung; ein Kulturmedium, dass entsprechend einer Zusammensetzung in der später beschriebenen Tabelle 1 eingestellt war, wurde in Reinstwasser gelöst und ergab 1000,01 ml, es wurden 1,0 g/L Hefeextrakt bzw. Casaminosäure hinzugegeben.
• 2. Kulturnährmedium mit 10% Olivenöl.
• 3. Kulturnährmedium mit 10% Olivenöl. Ferner erfolgte dreimal eine Subkultivierung.
• 4. Tatsächliches Abwasser.
• 5. Kulturnährmedium mit 2% Schweinefett.
• 6. Kulturnährmedium mit 2% Salatöl.
• 7. Kulturnährmedium mit 10% Schweinefett.
• 8. Kulturnährmedium mit 1% Salatöl. Ferner erfolgte dreimal eine Subkultivierung.
• 9. Kulturnährmedium mit 2% Talg.

First, 126 soil samples collected on Ishigaki Island in Okinawa Prefecture were cultured in a culture medium (1). Only samples that grew well in the culture medium (1) were then cultivated in the culture medium (2). In the same way, only by culturing well-growing samples in the following culture media, finally six (6) samples were obtained.

• 1. Nutrient culture medium with 5% olive oil in the composition; a culture medium, which was set according to a composition in Table 1 described later, was dissolved in ultrapure water and gave 1000.01 ml, 1.0 g / L yeast extract or casamino acid were added.
• 2. Culture medium with 10% olive oil.
• 3. Culture medium with 10% olive oil. Subcultivation was also carried out three times.
• 4. Actual wastewater.
• 5. Culture medium with 2% pork fat.
• 6. Culture medium with 2% salad oil.
• 7. Culture medium with 10% lard.
• 8. Culture medium with 1% salad oil. Subcultivation was also carried out three times.
• 9. Culture medium with 2% sebum.

Regarding the six (6) through the previous procedure isolated samples, the amount of degradation and lipase activity measured in wastewater containing oil and fat, and a sample was isolated which had the best results.

In this way, the bacterial strain Burkholderia cepacia AIK from Ishigaki Island in Okinawa Prefecture preserved soil isolated.

• 1. Morphologie
  • a) Form und Größe der Zelle; Bacillus mit einer Länge von 1,0~1,5 µm, Breite 0,8 µm
  • b) Gramfärbung; -
  • c) Zystoide Sporen; -
  • d) Beweglichkeit; +
  • e) Erscheinungsbild der Kolonie; kugelförmig, gewellter Rand, leicht konvex, gestreifte Oberflächenstruktur und gelb (wasserlösliches Pigment)
  • f) Epiphytische Flagellen; polare Flagellation
• 2. Erscheinungsbild in der Kultur
  • a) Flüssigbouillonkultur; Bildung einer Oberflächenhaut
  • b) Lackmusmilch; unverändert
• 3. Physiologische Eigenschaften
  • a) Katalase; +
  • b) Oxidase; +
  • c) O/F-Untersuchung; -
  • d) Nitratreduktion; +
  • e) Indolproduktion; -
  • f) Glucoseansäuerung; -
  • g) Arginindihydrolase; +
  • h) Urease; -
  • i) Äskulinhydrolyse; +
  • j) Gelatinhydrolyse; -
  • k) β-Galactosidase; -
  • l) Denitrifizierungsreaktion; -
  • m) MR-Untersuchung; +
  • n) VP-Untersuchung; -
  • o) H₂S-Produktion; -
  • p) Stärkehydrolyse; -
  • q) Verwertung von Zitronensäure; Koser/+, Christensen/+
  • r) Verwertung von anorganischen Stickstoffquellen; NaNO₃/+, (NH₄)₂/SO₄/+
  • s) Pigmentproduktion; wasserlösliches gelbes Pigment
  • t) Anaerobes Wachstum; -
  • u) Abbau von Tween 80; +
  • v) Abbau von Poly-β-Hydroxybuttersäure; -
  • w) Substratassimilationsfähigkeit; Glucose, L- Arabinose, D-Mannose, D-Mannitol, N-Acetyl-D-glucosamin, Kaliumgluconat, N-Caprinsäure, Adipinsäure, DL- Äpfelsäure, Natriumcitrat und Phenylacetat werden assimiliert. Maltose wird nicht assimiliert.
  • x) Bildung von Säure aus Sacchariden; Glucose, Xylose, Mannitol, Lactose und Malzzucker bilden Säure.
  • y) Bildung von Säure und Gas im OF-Kulturmedium; (Säure/Gas); L-Arabinose (-/-), D-Xylose (+/-), D- Glucose (+/-), D-Mannose (-/-), D-Fructose (-/-), D- Galactose (+/-), Maltose (+/-), Saccharose (+/-), Lactose (+/-), Trehalose (+/-), D-Sorbitol (-/-), D- Mannitol (+/-), Inositol (-/-), Glycerin (-/-), Stärke (-/-).

The morphology of the isolated bacterial strain Burkholderia cepacia AIK is explained below. Furthermore, an investigation of the morphology and based on the description by E. Yabuuchi et al., Microbiology and Immunology, 36, 1251-1275, 1992 was carried out.

• 1. Morphology
  • a) shape and size of the cell; Bacillus with a length of 1.0 ~ 1.5 µm, width 0.8 µm
  • b) Gram staining; -
  • c) cystoid spores; -
  • d) mobility; +
  • e) appearance of the colony; spherical, wavy edge, slightly convex, striped surface structure and yellow (water-soluble pigment)
  • f) epiphytic flagella; polar flagellation
• 2. Appearance in culture
  • a) liquid broth culture; Formation of a surface skin
  • b) litmus milk; unchanged
• 3. Physiological properties
  • a) catalase; +
  • b) oxidase; +
  • c) O / F examination; -
  • d) nitrate reduction; +
  • e) indole production; -
  • f) acidification of glucose; -
  • g) arginine dihydrolase; +
  • h) urease; -
  • i) Aesculin hydrolysis; +
  • j) gelatin hydrolysis; -
  • k) β-galactosidase; -
  • l) denitrification reaction; -
  • m) MR examination; +
  • n) VP examination; -
  • o) H ₂ S production; -
  • p) starch hydrolysis; -
  • q) utilization of citric acid; Koser / +, Christensen / +
  • r) utilization of inorganic nitrogen sources; NaNO ₃ / +, (NH ₄ ) ₂ / SO ₄ / +
  • s) pigment production; water soluble yellow pigment
  • t) anaerobic growth; -
  • u) degradation of Tween 80; +
  • v) degradation of poly-β-hydroxybutyric acid; -
  • w) substrate assimilation ability; Glucose, L-arabinose, D-mannose, D-mannitol, N-acetyl-D-glucosamine, potassium gluconate, N-capric acid, adipic acid, DL-malic acid, sodium citrate and phenylacetate are assimilated. Maltose is not assimilated.
  • x) formation of acid from saccharides; Glucose, xylose, mannitol, lactose and malt sugar form acid.
  • y) formation of acid and gas in the OF culture medium; (Acid / gas); L-arabinose (- / -), D-xylose (+/-), D-glucose (+/-), D-mannose (- / -), D-fructose (- / -), D-galactose (+ / -), maltose (+/-), sucrose (+/-), lactose (+/-), trehalose (+/-), D-sorbitol (- / -), D-mannitol (+/-), Inositol (- / -), glycerin (- / -), starch (- / -).

From the morphological properties and from the previous ones the above-mentioned documents both catalase and oxidase, a mobile, gram-negative bacillus, and acid production from glucose judging that the new microorganism of present invention as a microorganism identify who belongs to Burkholderia.

Furthermore, the new microorganism on which the The present invention relates to acid from glucose, xylose, Mannitol, lactose and maltose and shows arginine dihydrolase Activity, and it is concluded that he is a Is microorganism belonging to Burkholderia cepacia, and it is called the bacterial strain Burkholderia cepacia AIK designated.

The bacterial strain Burkholderia cepacia AIK was developed under the Filing number for microorganisms FERM-BP-7308 at National Institute of Bioscience and Human-technology of Industrial Science and Technology Ministry of International Trade and Industry deposited on September 25, 2000.

For the cultivation of the Burkholderia cepacia bacterial strain culture medium used is not particularly limited, and ordinary liquid and solid media are used. As nutrients are nutrients like a carbon source and an inorganic salt for the growth of the bacteria necessary. Glucose, cane sugar, Honey and starch can be used. As a nitrogen source can be both organic and inorganic Nitrogen sources are used. For the purpose of Illustration, but not limitation, can be used as organic nitrogen sources, peptone, yeast extract, Malt infusion, meat extract, soybean breakdown product and Urea can be used and as an inorganic Nitrogen sources can be ammonium nitrate and sodium nitrate be used. As an inorganic salt can, but are not limited to phosphate, magnesium sulfate, potassium, Sodium, calcium, copper, manganese, zinc, calcium chloride and iron (II) sulfuric acid can be used.

The pH of the culture medium is 2.0 ~ 12.0 and a pH of 5.0 ~ 8.0 is preferred, 6.0 ~ 8.0 is more preferred, and 7.0 ~ 8.0 is most preferred.

The temperature of the culture medium can also fluctuate, as long as the temperature is kept in a range that the bacterial strain Burkholderia cepacia AIK grow, in particular the temperature is preferably between 15 ~ 45 ° C, and a temperature between 20 ~ 35 ° C is more prefers.

The bacterial strain Burkholderia cepacia AIK can contain oil and fat break down effectively. The temperature at which the Bacterial strain Burkholderia cepacia AIK breaks down oil and fat, is not particularly limited, but a range between 15- ~ 45 ° C is preferred and between 20 ~ 35 ° C is more prefers. Usually the temperature fluctuates Mining container in the range of 20 ~ 35 ° C, a range in which the bacterial strain Burkholderia cepacia AIK outstanding Has mining capabilities in a mining container.

The bacterial strain Burkholderia cepacia AIK can also be found both animal and vegetable oil and fat effectively break down, furthermore the bacterial strain Burkholderia cepacia AIK not just salad oil, olive oil and Break down sesame oil that is liquid at normal temperatures, but can also use pork fat, tallow and coconut oil dismantle that are solid at normal temperature. Furthermore, as described below, the Burkholderia bacterial strain cepacia AIK the waste water down to an extremely low amount dismantle in a short time.

The bacterial strain Burkholderia cepacia AIK can contain oil and fat degrade in the presence of detergents. It means that the bacterial strain Burkholderia cepacia AIK oil and fat can degrade even if the oil and fat in one actual sewage system is what a mixture including a detergent.

In addition, the bacterial strain Burkholderia cepacia AIK suitable, in addition to oil and fat, fatty acids such as Lauric acid, myristic acid and oleic acid in addition to organic substances such as proteins, Mine hydrocarbons and saccharides. The temperature is not special for the degradation of these organic substances limited, and is sufficiently similar to that of oil and fat, and the temperature range is between 15 ~ 45 ° C and one The range of 15 ~ 40 ° C is preferred and is of 20 ~ 35 ° C more preferred.

In the following, the wastewater process is explained the present invention relates. The Sewage process to which the present invention relates brings back bacteria belonging to Burkholderia Wastewater in contact.

The selection of bacteria is not limited to the Burkholderia genus, and any bacterium can be used as long as it can break down oil and fat, but bacteria that can break down oil and fat over a wide temperature range are preferred. The reason for this is that the temperature in a mining container is not constant and it fluctuates due to many conditions such as weather, seasonal fluctuations or the location of the container. With reference to FIG. 3, a graphic representation is provided which shows a change in the average temperature in the degradation container. In order to measure the effects of the temperature changes in the degradation container in relation to the degradability of the microorganisms, the water quality of the treated water in the degradation container was monitored. The degradability of the microorganism was observed to decrease over time, and sometimes treated water containing incompletely degraded oil and fat was discharged.

Referring to Fig. 4, a graphical representation shows the amount of n-hexane extract from the digester, i.e. a measurement of the amount of oil and fat measured in the treated water drained from the digester. In other words, using bacteria that have high degradability in a wide temperature range to which the degradation container is exposed, particularly between 15 ~ 40 ° C, 20 ~ 35 ° C is more preferable even if a temperature change occurs in the degradation container degradability does not decrease, and stable and effective disposal can be achieved.

Furthermore, using microorganisms which are used for Genus Burkholderia belong to, not just the breakdown of liquid Oil and fat, but also solid oil and fat in normal Temperatures reached. The reason for this is that with normal Temperatures of both liquid and solid oil and fat in the wastewater flowing in a degradation container is contained. Bacteria belonging to the genus Burkholderia can in addition to oil and fat, organic substances, such as Fatty acids, proteins, hydrocarbons and saccharides dismantle. The reason for this is that by breaking down organic substances the biological oxygen demand (BOD) and the chemical oxygen demand (COD) in waste water is reduced can be.

Bacteria that meet the aforementioned conditions can belong to the species Burkholderia cepacia, and a specific example of the bacterium is the bacterial strain Burkholderia cepacia AIK (FERM-BP-7308).

The degradation process for waste water according to the invention can from Plankton processes, such as the activated sludge process, the Rotational organism contact method, the Trickle filter process, the catalytic oxidation process, the biofilm process, such as the aerobic filter process, or the method with immobilized microorganisms to be selected. This means that the bacterium that is responsible for Genus Burkholderia belongs as a preferred Microorganism comprising an activated sludge is used can be, and also as a group of Microorganisms can be used, the biological Mucus can break down on the surface of a disc adhere, furthermore they can be a group of microorganisms can be used, which can be obtained in a vector.

The sanitation process of the present invention can break down oil and fat without being affected by environmental conditions such as about season, weather condition and location of the Mining container to be affected as that Wastewater disposal process sufficient liquid and solid Oil and fat can degrade without changing temperature to be influenced in the degradation container. Furthermore, that Waste water treatment process the occurrence of bad Odors by suppressing the multiplication of Anaerobes are diminished and there can be one at any time have high degradability, consequently the Water quality of the treated water that comes from the Removal container is disposed of and left out, almost constant being held.

Based on the following examples, the present one Invention explained in detail, but it is to be understood that the detailed explanations for the purpose of Are illustrative and not limiting.

Preparation of the sample and the comparison sample

The bacterial strain Burkholderia cepacia AIK was developed in one cultivated as described in Example 1 below. The bacterium Super H (brand name) was also produced by KONDO FRP) as an approach to the comparative sample in Example 1 cultivated, activated sludge (cultivated by the National Institute of Bioscience and humantechnology of Industrial Science and Technology Ministry of International Trade an Industry) was developed as a preparation of the Comparative Example 2 cultivated.

example 1 Degradation of lard

The composition shown in Table 1 was in 1000.0 ml Pure water dissolved, then they were removed through the culture medium set to the 500 ppm pork fat and 10 ppm detergent was added in a sterile environment. The Ultrapure water mixture was added by shaking at 16 hours manufactured at a speed of 150 rpm simulate artificial sewage. It has been confirmed that the artificial wastewater similar properties as a has actual sewage.

Next, 20 ml batches (including cells with a dry weight of 0.06 g), which in an LB Culture medium were pre-cultivated with the set artificial sewage washed, to 200 ml of artificial Given wastewater and with shaking at 30 ° C and 150 rpm cultured.

Eventually, the existing in the artificial wastewater Pig fat concentration at fixed intervals measured. Concentration was used as a type of measurement an n-hexane extract of the artificial wastewater with a Extraction efficiency of 98.6% measured (there were three Controls and these were measured). Three samples were made per Measurement taken and their mean value determined. Further was the same experiment using approaches of Comparative Example 1 performed.

The result is shown in Table 2. Table 1 Potassium hydrogen phosphate 0.2 g dipotassium 1.6 g ammonium sulfate 1.0 g Ammonium sulfate 7-hydrate 0.2 g Iron sulfate 7-hydrate 0.01 g Calcium chloride-2-hydrate 0.02 g Sodium manganate (IV) 0.5 mg manganese sulfate 0.5 mg sodium chloride 0.1 g Table 2

As shown in Table 2, the invention shows Bacterial strain Burkholderia cepacia AIK compared with the Comparative Example 1 has superior degradability. The The bacterial strain of the present invention builds solid oil and Fat to about a quarter of its initial concentration in 4 Hours and about a tenth of the initial concentration of solid oil and fat in 9 hours. He also builds in 24 Hours concentrating on an almost nonexistent one Level down.

Example 2 Degradation of salad oil

500 ppm were added to the culture medium used as in Example 1 Salad oil and 10 ppm detergent added and sterilized. The ultrapure water mixture was made by shaking for 16 hours at a speed of 150 rpm made to simulate artificial sewage. It it was confirmed that the artificial sewage was similar Properties like actual sewage.

Next, 20 ml batches (including cells with a dry weight of 0.06 g) in LB culture medium were pre-cultivated with the set artificial Wastewater washed, to 200 ml of artificial wastewater given, and with shaking at 30 ° C and 150 rpm cultured.

Finally, the concentration of the in the artificial Waste water from existing salad oil at fixed intervals measured. As a type of measurement, the concentration of an n-hexane extract of the artificial waste water with one Extraction efficiency of 97.5% measured (there were three Controls and these were measured). There were three samples taken per measurement, and their average is determined. Further was the same experiment using approaches of Comparative Example 1 and Comparative Example 2 carried out.

The result is shown in Table 3. Table 3

As shown in Table 3, the bacterial strain shows Burkholderia cepacia AIK compared with the Comparative Examples 1 and 2 a remarkably superior one Degradability. The bacterial strain according to the invention builds solid oil and fat to about a seventh of his Initial concentration in 4 hours, and about a tenth the initial concentration of solid oil and fat in 9 hours from. It also builds up the concentration of oil after 24 hours and fat to an almost nonexistent level.

Example 3 Temperature dependence of oil and oil degradation fat

500 ppm were added to the culture medium used as in Example 1 Salad oil and 10 ppm detergent added and sterilized. The ultrapure water mixture was shaken for 16 hours manufactured at a speed of 150 rpm simulate artificial sewage. It has been confirmed that the artificial wastewater similar properties as a has actual sewage.

Next, 20 ml batches (including cells with a dry weight of 0.06 g), which in an LB Culture medium were pre-cultivated with the set artificial sewage washed, to 200 ml of artificial Given waste water and with shaking at a fixed Temperature and cultured at 150 rpm for 4 hours.

Eventually the rate of degradation of lettuce oil in the artificial sewage measured over a period of time. As Art the concentration of an n-hexane extract in the artificial wastewater with an extraction efficiency of 97.5% measured (there were three controls and these were measured).

Three samples were taken per measurement and their mean determined.

Furthermore, the rate of degradation of the salad oil using the Approaches of comparative example 1 measured.

The results are in Table 4 and Figure. 1 shown. In Fig. 1, the solid line shows the current example in the graphical representation, and the broken line shows the results of Comparative Example 1. Table 4

As shown in Table 4 and Fig. 1, the bacterial strain Burkholderia cepacia AIK according to the invention shows degradability at low temperatures of less than 15 ° C and at high temperatures above 40 ° C. Furthermore, the present strain shows effects in a temperature range of about 20-35 ° C, and has an excellent degradability, in which more than 80% degradation can always be demonstrated.

Example 4 Influence of the amount of bacteria in the breakdown of salad oil

500 ppm were added to the culture medium used as in Example 1 Salad oil and 10 ppm detergent in a sterile environment given. The ultrapure water mixture was shaken for 16 hours at a speed of 150 rpm made to simulate artificial sewage. It it was confirmed that the artificial sewage was similar Properties like actual sewage.

Next, 20 ml batches (including cells with a dry weight of 3.0 g in 1 liter of Culture medium), which is pre-cultivated in LB culture medium with the set artificial sewage washed to 200 ml of the given artificial wastewater and with shaking at 30 ° C and 150 rpm for 4 hours.

Finally, the concentration of the salad oil was in measured artificial sewage. As a kind of measurement the concentration of an n-hexane extract in the artificial Wastewater measured with an extraction efficiency of 97.5%, (There were three controls and these were measured). Three samples were taken per measurement and their mean determined.

The results are shown in Table 5. Table 5

As shown in Table 5, the invention Bacterial strain Burkholderia cepacia AIK Oil and fat in one dismantle for a short time, even if it is in very small quantities is added.

Example 5 Breakdown of oil and fat in one actual sewage

After actual sewage was collected, it became sterilized and filtered. The amount of an n-hexane extract in this wastewater was 15 ppm.

Next, salad oil was added, making the amount of the n-hexane extract was 500 ppm, and below for 16 hours Shake cultivated.

After 20 ml batches (including cells with a Dry weight of 0.06 g) in an LB culture medium were pre-cultivated with the set artificial Waste water was washed, they became 200 ml of the added artificial sewage and with shaking Cultivated at 30 ° C and 150 rpm for 4 hours.

Finally the concentration of the n-hexane extract in the wastewater measured at fixed intervals.

Three samples were taken per measurement and their mean value was determined. The result is shown in Table 6. Table 6

As shown in Table 6, the inventive one Bacterial strain Burkholderia cepacia AIK almost the same Ability on how with regard to artificial sewage in Table 3, with the associated explanations in Shown individually.

Example 6 Influence of peptone in the degradation of salad oil

To the culture medium used as in Experiment 1 were 500 ppm salad oil, 10 ppm detergent and 0.1% peptone added and sterilized. The pure water mixture was by shaking for 16 hours at a rate of 150 rpm produced to produce an artificial sewage simulate.

Next, the concentration of the n-hexane extract in the artificial wastewater was measured at the specified intervals by the same method as previously described with respect to Experiment 2. The result is shown in Table 7. Table 7

As shown in Table 7, the inventive one Bacterial strain Burkholderia cepacia AIK almost the same Ability to break down oil and fat in the presence of Peptone as without peptone (based on the results of the previous experiment shown in Table 3 in the Absence of peptone).

Example 7 pH influence in the degradation of salad oil

In the culture medium used as in Example 1, the pH of the culture medium adjusted as shown in Table 8, and by a pH meter using 0.5N HCl and and 0.5 N NaOH measured. 500 ppm salad oil and 10 ppm detergent were added and the medium was sterilized. The Mixture was shaken for 16 hours at a Speed of 150 rpm made to be an artificial To simulate wastewater. Next were 20 ml batches (including cells with a dry weight of 0.06 g in 1 liter of culture medium) in LB culture media were pre-cultivated with the set artificial Wastewater washed, to 200 ml of artificial wastewater given and with shaking at 30 ° C and 150 rpm for 4 Hours cultivated.

Finally, the concentration of the salad oil in the measured artificial sewage. As a kind of measurement the concentration of the n-hexane extract in the artificial Wastewater measured with an extraction efficiency of 97.5% (there were three controls and a special calculation). Three samples were taken per measurement and their mean determined.

The result is shown in Table 8 and Fig. 2. Table 8

As shown in Table 8 and Fig. 2, the bacterial strain Burkholderia cepacia AIK according to the invention can degrade oil and fat in a wide pH range. In particular, it has remarkably excellent degradability in a pH range between 7.0 ~ 8.0.

Example 8 Degradation of lettuce oil in tap water

500 ppm salad oil was added to tap water, then was it sterilized and shaken for artificial wastewater 16 hours at a speed of 150 rpm similar made. Next, 20 ml batches (including of cells with a dry weight of 0.06 g), which in LB- Culture media were precultivated with physiological Washed saline, to 200 ml of the adjusted given artificial wastewater and with shaking at 30 ° C and cultivated at 150 rpm for 24 hours.

Finally, the concentration of the salad oil in the measured artificial sewage. As a kind of measurement the concentration of the n-hexane extract in the artificial Wastewater measured with an extraction efficiency of 97.5% (There were three controls and these were measured).

The same experiment was also carried out using Approaches of Comparative Example 1.

Measurements were also made in a similar manner for each sample batch of the example, the comparative example 1 and control.

The results of each sample measured three times are shown in Table 9. Table 9

As shown in Table 9, the invention is Bacterial strain Burkholderia cepacia AIK active and builds oil and Fat even in an environment that is down to oil and fat is low in food.

Example 9 Influence of detergent in the breakdown of lard

To a culture medium having the same composition as in Example 1, 500 ppm of pork fat was added to tap water and then sterilized. The mixture was prepared by shaking at 150 rpm for 16 hours to simulate artificial sewage. Next, 20 ml batches (including cells with a dry weight of 0.06 g) of the sample, which had been precultivated in LB culture medium, were washed with physiological saline, added to 200 ml of the adjusted artificial wastewater and with shaking at 30 ° C and 150 rpm cultivated for 24 hours. Finally, the concentration of lard in the artificial waste water was measured. As the type of measurement, the concentration of the n-hexane extract of the artificial sewage was measured with an extraction efficiency of 98.6% over a fixed period of time (three controls were carried out and these were measured). Three samples were taken per measurement and their mean was determined. The result is shown in Table 10. Table 10

As shown in Table 10, the degradability of the bacterial strain of the invention Burkholderia cepacia AIK in Relative to solid oil and fat increases when using a detergent is added (with reference to the results of the in Table 2 example shown in which the same Experiment with the addition of detergent was made).

Example 10 Influence of detergents in the degradation of salad oil

500 ppm were added to the culture medium used as in Example 1 Salad oil added to tap water and sterilized. The Mixture was shaken for 16 hours at a Speed of 150 rpm made to be an artificial To simulate wastewater. Next were 20 ml batches (including cells with a dry weight of 0.06 g), which were precultivated in LB culture medium with washed physiological saline, to 200 ml of set artificial sewage, and at 30 ° C and Cultivated at 150 rpm for 24 hours. Finally, the Concentration of the salad oil in the artificial waste water measured. Concentration was used as a type of measurement of the n-hexane extract of the artificial wastewater Points in time with an extraction efficiency of 98.6% measured (there were three controls and these were measured).

Three samples were taken per measurement and their mean determined.

The result is shown in Table 11. Table 11

As shown in Table 11, the degradability of the bacterial strain Burkholderia cepacia AIK according to the invention in terms of liquid oil and fat decreased when detergent was added (with reference to the results of the table 3 shown example, in which the same experiment without Addition of detergent was undertaken).

Example 11 Degradation of salad oil, inoculated with activated sludge

To a culture medium with a composition similar to in Example 1, 500 ppm salad oil and 10 ppm detergent were added Tap water added and sterilized. The mixture was by shaking for 16 hours at a Speed of 150 rpm made to be an artificial To simulate wastewater. It has been confirmed that artificial sewage the same properties as a has actual sewage.

Next, 20 ml batches (including cells with a dry weight of 0.06 g) in LB culture medium were pre-cultured with physiological saline washed, added to 200 ml of activated sludge and under Shake cultivated at 30 ° C at 150 rpm.

Finally, the concentration of the salad oil in the artificial waste water was measured after 9 hours. As a kind of measurement, the concentration of the n-hexane extract in the artificial waste water was measured after a specific period of time with an extraction efficiency of 97.5% (three controls were made and these were measured). Three samples were taken per measurement and their mean was determined. The similar experiment was also carried out. The result is shown in Table 12. Table 12

As shown in Table 12, the inventive one Bacterial strain Burkholderia cepacia AIK itself in the Presence of activated sludge related to a degradability Oil and fat on.

Example 12 Breakdown of fatty acid

An experiment to measure the ability of the Bacterial strain Burkholderia cepacia AIK for the breakdown of Fatty acid performed. The basis was the Fatty acid composition of lard and lettuce oil analyzed, and triolein, which is a single glyceride of Oleic acid is used with the largest amount in both.

In the experimental procedure, the first step was to Experiment 1 used culture medium 500 ppm triolein and 10 ppm Detergent added to tap water and sterilized. The Mixture was shaken for 16 hours at a Speed of 150 rpm made to be an artificial To simulate wastewater. Next were 20 ml batches (including cells with a dry weight of 0.06 g), which were precultivated in LB culture medium with washed physiological saline, to 200 ml of Activated sludge is added and at 30 ° C at 150 rpm Shake cultivated.

Finally, the total concentration of the n-hexane extract contained in the artificial sewage was methylated after one hour, two hours and four hours, and after adding heptadecanoic acid as an internal standard with an equal amount, each of its fixed portions was determined by gas chromatography analyzed. The peak area run through after one hour was set as standard; the peak area of oleic acid, corrected with the internal standard, is shown in Table 13. Table 13

As shown in Table 13, the invention Bacterial strain Burkholderia cepacia AIK fatty acids such as Oleic acid, but is not limited to, in a short Reduce time.

Effect of the invention

The bacterial strain Burkholderia cepacia AIK according to the invention has high degradability in terms of oil and fat in a wide temperature range. Especially in one area from about 20 ~ 35 ° C it always keeps its degradability upright. Related to the breakdown of oil and fat, not only on liquid oil and fat, but also on solid oil and Fat, the bacterial strain Burkholderia cepacia AIK has almost the same degradability for both material states.

The bacterial strain Burkholderia cepacia AIK in addition to oil and fat, organic substances such as Fatty acids, proteins, hydrocarbons and saccharides break down and he can reduce the biological oxygen demand (BOD) and the chemical oxygen demand (COD) in wastewater reduce.

The process according to the invention for the treatment of waste water, is a wastewater treatment process that contains bacteria, the oil and can effectively reduce fat. consequently can consistently mine and treat wastewater, without being affected by the temperature of the mining container his. As a result, the bacterial strain Burkholderia cepacia AIK the multiplication of anaerobes and the appearance of Prevent bad smells.

During the previous description many details contains, should not be considered as a limitation on the scope of the Invention are interpreted, but rather as examples of preferred embodiments thereof. Many other Modifications are possible without losing the essential spirit of this Deviate invention. Accordingly, the scope of the Invention not by the previously presented Embodiments are determined, but rather by the pending claims and their legal equivalents.

This invention relates to a waste water process in which is the bacterial strain Burkholderia cepacia AIK (FERM-BP- 7308) comes into contact with waste water, and the new one Microorganism aerobic liquid oil and fat and others decompose organic substances in a wide temperature range can, as well as a wastewater process that the new Microorganism used.

Claims (5)

1. Microorganism that leads to a bacterial strain Burkholderia cepacia AIK belongs. 2. Process for the disposal of waste water with oil and fat therein, by adding a microorganism to the waste water, who belongs to the genus Burkholderia. 3. Disposal method according to claim 2, wherein the Microorganism act in the presence of activated sludge can. 4. Disposal method according to claim 2, wherein the Microorganism belonging to the genus Burkholderia Bacterial strain Burkholderia cepacia AIK is. 5. mining method according to claim 3, wherein the Microorganism belonging to the genus Burkholderia, the Bacterial strain Burkholderia cepacia AIK (FERM-BP-7308) is.