CN105273040A

CN105273040A - Method for extracting 0092 type filamentous bacterium high-expansion sludge close integration type extracellular polymeric substances

Info

Publication number: CN105273040A
Application number: CN201510824005.0A
Authority: CN
Inventors: 高春娣; 樊士信; 焦二龙; 田烨; 李任飞; 彭永臻
Original assignee: Beijing University of Technology
Current assignee: Beijing University of Technology
Priority date: 2015-11-24
Filing date: 2015-11-24
Publication date: 2016-01-27

Abstract

The invention discloses a method for extracting a 0092 type filamentous bacterium high-expansion sludge close integration type extracellular polymeric substances, and relates to the field of biological treatment on sewage. In a 0092 type filamentous bacterium high-expansion sludge system, TB layer protein and polysaccharide in 0092 type filamentous bacterium high-expansion sludge EPS are extracted by using a common ultrasonic method or a formaldehyde and ultrasonic method, the ultrasonic intensity is 96-288W, and the time is 10 minutes. Filamentous bacterium high-expansion sludge TB-EPS that a 0092 type filamentous bacterium is superior can be rapidly and efficiently extracted, and a stable and efficient extraction method for later accurate quantification of protein, polysaccharide and DNA is provided.

Description

A kind of 0092 type thread fungus height bulking sludge is combined closely type extracellular polymeric (TB-EPS) extracting method

Technical field

The present invention relates to field of biological sewage treatment, there is provided a kind of to combine closely the optimization method that type extracellular polymeric (TB-EPS) extracts for 0092 type thread fungus height bulking sludge, can extract quickly and efficiently with 0092 type thread fungus for advantage thread fungus height bulking sludge TB-EPS, the accurate quantitative analysis for subsequent protein, polysaccharide and DNA provides the extracting method of stability and high efficiency.

Background technology

Extracellular polymeric (extracellularpolymericsubstances, EPS) be microorganism under certain environmental conditions, the polymer be enclosed in outside cell walls of metabolic process secretion, EPS mainly comprises the polymkeric substance such as protein, polysaccharide and nucleic acid, it is generally acknowledged that EPS accounts for 50% ~ 90% of active sludge total organic matter.The bundle water capacity of EPS to mud flco has great effect, and in mud, most water is combined in EPS, and therefore EPS is considered to the main factor affecting dewatering performance of sludge.Meanwhile, there are some researches show that the content of the main component protein, carbohydrate etc. in EPS all becomes positive correlation with sludge volume index (SVI), the existence of EPS changes sludge surface charge, thus directly affects the settleability of mud.Therefore, precise and high efficiency extracts and quantitatively EPS is of great significance research sludge settling property and sludge bulking situation tool.

The method extracted EPS in mud at present mainly contains heating extractions, centrifugal extraction, alkalescence extraction, extraction using alcohol, Zeo-karb extraction, sonication exchange with CER the extracting method combined.The key element that EPS extracts is that it not only can separate from cell surface by selected method exactly, and it is enough gentle, avoid the cracking and the dissolving that cause cell, thus cause the interior polymers of cell to leak into outside born of the same parents, cause the error of EPS quantity and composition analysis, generally take the relative content of DNA in EPS as the standard of inspection cell relative breakdown degree.

But, for the mud of different qualities, the layering of fixing mud extracellular polymeric and extracting method are to the high difference of its extraction efficiency, current shortage develops different extracting method for different qualities mud, especially the mud for there is filamentary discharges is lacked, its sludge characteristics is completely different from the characteristic of common sludge, more needs efficient stable to the extraction of protein and polysaccharide in its EPS, thus analyzes its impact on bulking sludge settling character.

Summary of the invention

For the weak point of above-mentioned research, the invention provides and to combine closely the optimization method that type extracellular polymeric (TB-EPS) extracts for 0092 type thread fungus height bulking sludge, the material such as protein and polysaccharide in 0092 thread microbial high bulking sludge TB-EPS can be extracted quickly and efficiently, but not broken born of the same parents, accurate quantitative analysis for subsequent protein, polysaccharide and DNA provides the extracting method of stability and high efficiency, provides basis for analyzing it to sludge settling property.

To combine closely for 0092 type thread fungus height bulking sludge the optimization method of protein and Polyose extraction in type extracellular polymeric (TB-EPS), it is characterized in that:

There is 0092 type thread fungus height bulking sludge system, ordinary ultrasonic method is adopted to extract for TB layer protein in this 0092 type thread fungus height bulking sludge EPS and polysaccharide, the 1 times of volume PBS solution of the mud substrate after loose type extracellular polymeric (LB-EPS) will be extracted and recovered original volume, then ultrasonic, ultrasound intensity is 96-288W (preferred 96W), and the time is 10min.

Or the 0092 type thread fungus height bulking sludge system of generation, formaldehyde is adopted to add ultrasonic extracting method for TB layer protein in this 0092 type thread fungus height bulking sludge EPS and polysaccharide, for: the 1 times of volume PBS solution of the mud substrate after loose type extracellular polymeric (LB-EPS) will be extracted and recovered original volume, then formaldehyde is added, and in 4 DEG C of condition stored refrigerated 1h, then ultrasonic, ultrasound intensity is 96-288W (preferred 288W), and the time is 10min.

Above-mentioned high bulking sludge general DSVI value is 1800-2000mL/g (preferred 2000mL/g), average MLSS is 1500-3500mg/L (preferred 1500mg/L),

Compared with prior art, the present invention has the following advantages:

(1) combine closely optimization method that type extracellular polymeric (TB-EPS) extracts of 0092 type thread fungus height bulking sludge provided by the invention is specially for extracting the method occurred with type extracellular polymeric of combining closely in the 0092 type thread fungus high bulking sludge that is dominant bacteria, extracts have more specific aim to TB-EPS in this type of bulking sludge.

(2) be 2000mL/g for average DSVI value, average MLSS is 1500mg/L, in 0092 type thread fungus height bulking sludge EPS of class, TB layer protein and Polyose extraction ordinary ultrasonic best practice are: ultrasound intensity is 96W, time is 10min, extract protein, polysaccharide and DNA in type extracellular polymeric (TB-EPS) of combining closely and be respectively 56.14,6.74 and 3.8mg/L, DNA percentage composition is 5.70%, does not occur seriously breaking born of the same parents.

(3) the present invention each provides ordinary ultrasonic method and formaldehyde and adds the optimization method that ultrasonic method extracts type extracellular polymeric of combining closely with 0092 type thread fungus height bulking sludge, makes to extract have selectivity and specific aim for the type extracellular polymeric of combining closely of mud in this type of mud.Be 2000mL/g for average DSVI value, average MLSS is that in the 0092 type thread fungus height bulking sludge EPS of 1500mg/L, TB layer protein and Polyose extraction formaldehyde add ultrasonic best practice and is: ultrasound intensity is 288W, time is 10min, extract protein, polysaccharide and DNA in type extracellular polymeric (TB-EPS) of combining closely and be respectively 66.23,14.68 and 3.5mg/L, DNA percentage composition is 4.15%, does not occur seriously breaking born of the same parents.

Accompanying drawing explanation

Fig. 1 DSVI value is the 0092 type thread fungus FISH picture (1000 times) of 2000mL/g

The mud TB-EPS that the 0092 type thread fungus height of Fig. 2 is DSVI value of the present invention to be 2000mL/g, MLSS be 1500mg/L expands extracts result figure.

Embodiment

Below in conjunction with accompanying drawing and specific implementation method, the present invention is described in further detail, but the present invention is not limited to following examples.

Get test mud sample and remove the materials such as coarse sand by the mesh screen of 1.2mm, the former mud sieved precipitates 1.5 hours at 4 DEG C, with siphon pipe, upper solution is collected as supernatant layer EPS; By under throw out whizzer 2000g condition centrifugal 15 minutes, upper liquid is collected as slime layer EPS, substrate 1*PBS solution recovers original volume, suspension under 5000g condition centrifugal 15 minutes, upper liquid is collected as LB-EPS, and substrate mud is the mud substrate after having extracted loose type extracellular polymeric (LB-EPS).

Embodiment

1, be advantage filamentary discharges with 0092 type thread fungus (Fig. 1) mud carries out the former step extraction process of EPS to DSVI value to be 2000mL/g, MLSS be 2000mg/L;

2, the mud substrate 1*PBS solution after loose type extracellular polymeric (LB-EPS) will have been extracted and recovered original volume;

3, above-mentioned sample is divided into 9 parts, every part of 10ml;

4, the 1st increment product are set to control group (as a comparison case), and collecting upper liquid after adopting the centrifugal 20min of 20000g rotating speed is that control group extracts TB-EPS;

5, the 2nd increment product are set to heating group (as a comparison case), and adopting the centrifugal 20min of 20000g rotating speed to collect upper liquid after adopting constant temperature 70 DEG C heating 30min is that heating group extracts TB-EPS;

6,3-5 increment product are set to ordinary ultrasonic group, total power is adopted to be 20% of 480w respectively, the ultrasound intensity of 40%, 60% carries out 10min supersound process, and adopting the centrifugal 20min of 20000g rotating speed to collect upper liquid is extract TB-EPS under the different ultrasound intensity of ordinary ultrasonic group;

7,6-8 increment product are set to ultrasonic group of formaldehyde, the amount adopting mass percentage 36.5% formaldehyde to recover the mud substrate of original volume with 60ul/10ml is added in sample, and with 4 DEG C of condition stored refrigerated 1h, adopt 20% respectively afterwards, 40%, the ultrasound intensity of 60% carries out 10min supersound process, and collecting upper liquid after adopting the centrifugal 20min of 20000g rotating speed is extract TB-EPS under the different ultrasound intensity of ultrasonic group of formaldehyde;

8, the 9th increment product are set to formaldehyde+NaOH group (as a comparison case), the amount first adopting 36.5% formaldehyde to recover the mud substrate of original volume with 60ul/10ml is added in sample, and in 4 DEG C of condition stored refrigerated 1h, to add in sample and 4 DEG C of condition stored refrigerated 3h with the NaOH of 1mol/L afterwards, collecting upper liquid after adopting the centrifugal 20min of 20000g rotating speed is that formaldehyde+NaOH organizes and extracts TB-EPS;

9, by the membrane filtration of each group of TB-EPS extracted with 0.45 μm;

10, folin-phenol method protein graticule is set up, and protein content in working sample;

11, pentanoic method DNA graticule is set up, and DNA content in working sample;

12, anthrone method polysaccharide graticule is set up, and polysaccharide content in working sample;

13, the data results obtained is carried out com-parison and analysis (Fig. 2), obtain DSVI value for 2000mL/g, MLSS be 1500mg/L with 0092 type thread fungus for the optimum extracting method of advantage filamentary discharges mud TB-EPS.

Ordinary ultrasonic best practice is: ultrasound intensity is 96W, time is 10min, extract combine closely protein, polysaccharide and DNA in type extracellular polymeric (TB-EPS) be respectively 56.14,6.74 and 3.8mg/L, DNA percentage composition be 5.70%, do not occur seriously breaking born of the same parents; Formaldehyde adds ultrasonic best practice: ultrasound intensity is 288W, time is 10min, extract combine closely protein, polysaccharide and DNA in type extracellular polymeric (TB-EPS) be respectively 66.23,14.68 and 3.5mg/L, DNA percentage composition be 4.15%, do not occur seriously breaking born of the same parents.

The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.

Claims

1. 0092 type thread fungus height bulking sludge is combined closely the optimization method that type extracellular polymeric (TB-EPS) extracts, and it is characterized in that, comprises the following steps:

There is 0092 type thread fungus height bulking sludge system, ordinary ultrasonic method is adopted to extract for TB layer protein in this 0092 type thread fungus height bulking sludge EPS and polysaccharide, the 1 times of volume PBS solution of the mud substrate after loose type extracellular polymeric (LB-EPS) will be extracted and recovered original volume, then ultrasonic, ultrasound intensity is 96-288W, and the time is 10min;

Or the 0092 type thread fungus height bulking sludge system of generation, formaldehyde is adopted to add ultrasonic extracting method for TB layer protein in this 0092 type thread fungus height bulking sludge EPS and polysaccharide, for: the 1 times of volume PBS solution of the mud substrate after loose type extracellular polymeric (LB-EPS) will be extracted and recovered original volume, then formaldehyde is added, and in 4 DEG C of condition stored refrigerated 1h, then ultrasonic, ultrasound intensity is 96-288W, and the time is 10min.

2. according to the method for claim 1, it is characterized in that, the DSVI value of high bulking sludge is 1800-2000mL/g, and average MLSS is 1500-3500mg/L.

3. according to the method for claim 1, it is characterized in that, is 2000mL/g for DSVI value, and average MLSS is the 0092 type filamentary discharges mud of 1500mg/L, and when adopting ordinary ultrasonic method, ultrasound intensity is 96W; When adopting formaldehyde to add ultrasonic extracting method, ultrasound intensity is 288W.

4. according to the method for Claims 2 or 3, it is characterized in that, the amount adopting mass percentage 36.5% formaldehyde to recover the mud substrate of original volume with 60ul/10ml is added.