CN102586160A

CN102586160A - Stenotrophomonas maltophilia DS4

Info

Publication number: CN102586160A
Application number: CN2012100666333A
Authority: CN
Inventors: 郑丹; 鲍建国
Original assignee: China University of Geosciences
Current assignee: China University of Geosciences
Priority date: 2012-03-14
Filing date: 2012-03-14
Publication date: 2012-07-18
Anticipated expiration: 2032-03-14
Also published as: CN102586160B

Abstract

The invention discloses Stenotrophomonas maltophilia DS4. The Stenotrophomonas maltophilia DS4 is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO:M2010147. The Stenotrophomonas maltophilia DS4 has a strong growing ability and a good ability of removing organic pollutants from saponin wastewater in high-concentration sulfate ion environments; and the strain can remove chemical oxygen demand (COD) at the concentration of 2,930mg/L from the saponin wastewater within 72 hours, and the removal efficiency reaches 62.34 percent. The invention provides a useful bacterial source for degrading high-concentration organic pollutants in the saponin wastewater, broadens the application of functions of Stenotrophomonas maltophilia, and has relatively high application value.

Description

Germ oligotrophy unit cell DS4

Technical field

The invention belongs to biological technical field, relate to a kind of germ oligotrophy unit cell (Stenotrophomonas maltophilia) DS4.

Background technology

The chroma in waste water that produces in saponin (like the diosgenin) production process is big, acidity big, the Organic pollutants substrate concentration is high, chemical oxygen demand (COD) is high, biodegradability is poor, is a kind of very unmanageable waste water.Regulation should be lower than 300mg/L from COD quantity discharged in 2009, still in " saponin industrial water pollution thing emission standard " (GB 20425-2006); Saponin hydrolysising original liquid COD concentration reaches 20000-40000mg/L at present, and comprehensive wastewater COD concentration is 4000-12000mg/L, if directly enter water body; Can cause the organic contamination substrate concentration of water body to increase considerably; Thereby cause the rivers and lakes severe contamination, not only directly influenced people's living environment, also caused the massive losses of national economy.

Contain a large amount of organic pollutants in the saponin waste water, wherein part toxicity aldehyde, ketone, aldehydes matter are inhibited to the mikrobe in the mud in the Sewage treatment systems, and general mikrobe can not normal growth.Simultaneously, sulfate concentration is high in the saponin waste water, is about about 8000mg/L, and in the biological treatment system of high salinity waste water, microbial cell film and endobacillary enzyme are destroyed, and causes sludge activity to descend.In addition; Mikrobe has certain adaptive faculty to environment; Mikrobe is through the osmotic pressure in the osmoregulation mechanism statocyte of self or protect intracellular protoplasma in certain salinity range; Regulate self metabolism to be suitable for the variation of salinity, active sludge after taming after a while, will have certain salt tolerance in saline environment.

At present the processing of saponin waste water is adopted the method for physics or chemistry more, also do not had microbial degradation method in the prior art and the report of the bacterial strain of the saponin waste water organic pollutant that is used to degrade.

Summary of the invention

The purpose of this invention is to provide a kind of germ oligotrophy unit cell DS4, it can effectively be removed and contain high concentration sulphate ionic saponin waste water organic pollutant.

Above-mentioned purpose of the present invention realizes through following technical scheme:

A kind of germ oligotrophy unit cell DS4 is preserved in Chinese typical culture collection center, and preserving number is CCTCC NO:M2010147.

The colonial morphology of described germ oligotrophy unit cell DS4 is faint yellow, opaque, circular, smooth surface, center protrusion, neat in edge; Strict aerobic, Gram-negative straight-bar bacterium extremely gives birth to many flagellums, oxidase negative, DNA enzyme, proteolytic enzyme, urase, the lysine decarboxylase positive, MR, VP feminine gender, H ₂S is negative; The righttest growth pH value is 6.5～7.5, and optimum growth temperature is 25-35 ℃.

The invention has the beneficial effects as follows: germ oligotrophy unit cell DS4 has the ability of organic pollutant in stronger energy for growth and the good removal saponin waste water in the high concentration sulphate ionic environment; This bacterial strain can be removed the COD 2930mg/L in the saponin waste water within 72 hours, remove efficient and reach 62.34%.The present invention has widened the application to the germ oligotrophy unit cell function aspects for degraded saponin waste water high concentration organic contaminant provides useful bacterium source, has stronger using value.

Embodiment

Below in conjunction with specific embodiment the present invention is further specified.

The screening method of embodiment 1 germ oligotrophy unit cell DS4

(1) material is prepared:

1, experiment waste water and pool bottom sludge are taken from the biochemical treatment system that Shiyan City, Hubei Province saponin factory produces waste discharge.

2, substratum:

Isolation medium: K ₂HPO ₄0.5g/L, MgSO ₄7H ₂O 0.5g/L, NH ₄C1 1.0g/L, CaCl ₂0.01g/L, KH ₂PO ₄0.5g/L, glucose 0.5g/L, pH value 7.0.

Screening culture medium: K ₂HPO ₄0.5g/L, MgSO ₄7H ₂O 0.5g/L, NH ₄C1 1.0g/L, CaCl ₂0.01g/L, KH ₂PO ₄0.5g/L, dissolve in the 1L saponin waste water, pH value 7.0.

LB liquid nutrient medium: peptone 10.0g/L, yeast extract 5.0g/L, sodium-chlor 10.0g/L, zero(ppm) water 1000mL, pH7.0.

The LB solid medium: peptone 10.0g/L, yeast extract 5.0g/L, sodium-chlor 10.0g/L, zero(ppm) water 1000mL, agar 15g/L (Gu), pH7.0.

The LB slant medium: peptone 10.0g/L, yeast extract 5.0g/L, sodium-chlor 10.0g/L, zero(ppm) water 1000mL, agar 20g/L (Gu), pH7.0.

3, laboratory apparatus and equipment:

State China SHA-B constant temperature oscillator, the biochemical incubator of eastern device SPX, vertical pressure steam sterilizer, opticmicroscope (Olympus ltd), pH meter (sarporiusPD-10), Bechtop, PTC200 type PCR appearance, electrophoresis apparatus and electrophoresis chamber, UVP gel ultraviolet visualizer.

(2) screening and separating of bacterial strain and purifying:

1, screening and separating:

(1) take by weighing the pool bottom sludge sample of acidulated pool tail region in the 2 gram saponin factory Waste Water Treatments, add in the triangular flask of the 250mL that the 100mL sterilized water is housed that sterilization is good in advance, 30 ℃ of constant-temperature shaking 200rpm smash zoogloea, mud mixture, subsequent use;

(2) mud mixture is changed in the 500mL Erlenmeyer flask that the 100mL isolation medium is housed, 30 ℃ of constant temperature leave standstill and are cultured to the substratum muddiness, obtain bacterial suspension A;

(3) get the 5mL bacterial suspension A and in the 250mL Erlenmeyer flask of the fresh screening culture medium of dress 100mL, cultivate a week, get bacteria suspension 5mL repetitive operation 5-6 time, the bacteria suspension B that obtains;

2, purifying: with aseptic pipette, extract 0.5mL bacteria suspension B, dilution is coated on the LB solid medium.37 ℃ of constant temperature culture 3 days; Picking has single bacterium of notable difference on colony characteristics, on the LB solid medium streak culture repeatedly three times until obtaining single bacterial strain, and on the LB slant medium, preserve; Obtain a strain bacterial strain, i.e. germ oligotrophy unit cell DS4 bacterial strain.

(3) colony morphology characteristic and physio-biochemical characteristics

The colonial morphology of bacterial strain DS4 is: bacterium colony is faint yellow, opaque, circular, smooth surface, center protrusion, neat in edge; Strict aerobic, Gram-negative straight-bar bacterium extremely gives birth to many flagellums, oxidase negative, DNA enzyme, proteolytic enzyme, urase, the lysine decarboxylase positive, MR, VP feminine gender, H ₂S is negative.The righttest growth pH value: 6.5～7.5, optimum growth temperature: 25-35 ℃.

Adopt the BLAST analytical method that 16S rRNA gene order and the comparative analysis of GenBank DB of bacterial strain DS4 are found that the homology of bacterial strain DS4 and germ oligotrophy unit cell is up to 99.0%.

The Physiology and biochemistry and the Molecular Identification result of comprehensive bacterial strain, bacterial strain DS4 called after germ oligotrophy unit cell DS4.Consult pertinent data, still do not have the report that relevant germ oligotrophy unit cell belongs to has degraded saponin waste water organic pollutant ability.Germ oligotrophy unit cell DS4 is new bacterial strain, delivers Chinese typical culture collection center (the being called for short CCTCC) preservation that is positioned at Wuhan City, Hubei Province Wuhan University on June 13rd, 2010, and preserving number is CCTCC NO:M2010147.

This bacterial strain that screening and separating of the present invention goes out has the function of the saponin waste water organic pollutant of degrading preferably; This has widened the applied research thinking of people to the germ oligotrophy unit cell function aspects; And useful bacterium source and technology is provided for degrading high concentration organic pollutant saponin waste water, have stronger actual application value.

The application of embodiment 2 germ oligotrophy unit cell DS4

The single bacterium colony of picking germ oligotrophy unit cell DS4; Overnight cultures in the LB solid medium; Volume ratio by germ oligotrophy unit cell DS4 after cultivating and saponin waste water (promptly test waste water, take from the biochemical treatment system of Shiyan City, Hubei Province saponin factory waste discharge) is the ratio inoculation of 0.5-2:100,30-35 ℃ of constant temperature culture; The pH value is 6-8, reacts 48-72 hour.

Described LB solid medium is: peptone 10.0g, and yeast extract 5.0g, NaCl10.0g, zero(ppm) water 1000mL, agar 15g (Gu), pH7.0.

Measure the mikrobe that filters out degradation efficiency to organic pollutant in the saponin waste water.

Picking list bacterium colony respectively, in isolation medium before overnight cultures, get 1mL bacterium liquid and be inoculated in 9mL saponin waste water (about the initial COD 4700mg/L of depickling water, pH1.28) and add respectively in the test tube, stay the blank contrast of a flag.Regulate water sample pH value, placed 37 ℃, 150rpm shaking culture 3 days, calculate the COD removal situation of bacterial strain waste water.This bacterial strain can be removed the COD 2930mg/L in the saponin waste water within 72 hours, remove efficient and reach 62.34%.

Claims

1. a germ oligotrophy unit cell (Stenotrophomonas maltophilia) DS4 is preserved in Chinese typical culture collection center, and preserving number is CCTCC NO:M2010147.

2. germ oligotrophy unit cell DS4 as claimed in claim 1 is characterized in that colonial morphology is faint yellow, opaque, circular, smooth surface, center protrusion, neat in edge; Strict aerobic, Gram-negative straight-bar bacterium extremely gives birth to many flagellums, oxidase negative, DNA enzyme, proteolytic enzyme, urase, the lysine decarboxylase positive, MR, VP feminine gender, H ₂S is negative; The righttest growth pH value is 6.5～7.5, and optimum growth temperature is 25-35 ℃.