CN103205412B - The method of immobilized microorganism preparation and employing said preparation process meat product processing waste water - Google Patents
The method of immobilized microorganism preparation and employing said preparation process meat product processing waste water Download PDFInfo
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- CN103205412B CN103205412B CN201310144749.9A CN201310144749A CN103205412B CN 103205412 B CN103205412 B CN 103205412B CN 201310144749 A CN201310144749 A CN 201310144749A CN 103205412 B CN103205412 B CN 103205412B
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Abstract
The invention discloses immobilized microorganism preparation, it adopts following steps preparation: the fermentation culture of S1, bacterial classification, by subtilis, bacillus megaterium, bacterium nitrobacter and Nitrosomonas, is inoculated in enrichment medium respectively and cultivates; The mixing of S2, thalline, obtains thalline mixed solution; The immobilization of S3, PVA-sodium alginate, after PVA gel and thalline mixed solution are mixed, instillation low speed and continuously stirring containing CaCl
2boric acid solution in, take out after immobilization is crosslinked, with deionized water wash, namely lyophilize obtains immobilized microorganism preparation; Also disclose the method adopting immobilization biological preparation process meat product processing waste water.The invention has the beneficial effects as follows: microbe species is many, can multiple pollutant in degrading waste water, and degradation speed is very fast, ability is strong; Have and adapt to water quality and pH changing capability; Sludge yield is less, and product separation is easy, good stability, and technological operation is simple, and production cost is lower.
Description
Technical field
The present invention relates to a kind of method of meat product processing waste water treatment process technical field, particularly immobilized microorganism preparation and employing said preparation process meat product processing waste water.
Background technology
In recent years, along with improving constantly of Chinese society expanding economy and living standards of the people, the consumption of people to meat product also increases day by day, and meanwhile, meat product processing industry have also been obtained very fast development.But while meat product processing industry develops rapidly, the pollution problem of the sector particularly water pollution problems more and more causes the attention of people.Compare with sanitary wastewater with commonly industrial wastewater, meat product processing industry factory effluent has the advantages that Pollutant levels are high and unit product wastewater discharge is large, adopts common waste water treatment process to be usually difficult to obtain desirable treatment effect.Just can cause serious environmental pollution if processed not in time, this is not only harmful to the health of people and animals, also can cause secondary pollution in meat product production process simultaneously, affect the quality of product.
Summary of the invention
The object of the invention is to the shortcoming overcoming prior art, a kind of microbe species is more, degradation speed is fast, processing efficiency is high immobilized microorganism preparation is provided and adopts the method for said preparation process meat product processing waste water.
Object of the present invention is achieved through the following technical solutions: immobilized microorganism preparation, and it adopts following steps preparation:
The fermentation culture of S1, bacterial classification, by subtilis, bacillus megaterium, bacterium nitrobacter and Nitrosomonas, be inoculated into respectively in enrichment medium, at 28 ~ 32 DEG C, 22 ~ 26h is cultivated, to obtain the logarithmic phase cell of each thalline after the centrifugation 10min of 5000rpm under 150 ~ 170rpm condition;
The mixing of S2, thalline, by the cell in step S1 with after phosphate buffered saline buffer washing 2 ~ 3 times, by volume percentages, get 40 ~ 60% subtilises respectively, 20 ~ 30% bacillus megateriums, 10 ~ 15% bacterium nitrobacters, 5 ~ 15% nitrosification bacillus mixing, be suspended in physiological saline, obtain thalline mixed solution;
The immobilization of S3, PVA-sodium alginate, it comprises following sub-step:
The preparation of S31, PVA gel, get PVA to be added to the water, dissolve completely under 80 DEG C of conditions, the weight/volume of PVA and water is 1:9 ~ 11, add sodium alginate, silicon-dioxide and calcium carbonate after cooling and mix, PVA: sodium alginate: silicon-dioxide: the weight ratio of calcium carbonate is: 10:2 ~ 4:4 ~ 6:0.3 ~ 0.5, obtains PVA gel;
S32, thalline adsorb: add in the thalline mixed solution obtained in step s 2 with mixeding liquid volume than the gac being 5%, mixing and absorption;
The preparation of S33, immobilization composite bacteria PVA-sodium alginate particle: after above-mentioned obtained PVA gel and the thalline mixed solution adding gac are mixed, obtain thalline-PVA-sodium alginate suspension, by thalline-PVA-sodium alginate suspension instillation low speed, continuously stirring contains in the boric acid solution of 1.5 ~ 2.5%CaCl2, described boric acid solution pH is in neutral, take out after the crosslinked 22 ~ 26h of 4 ~ 6 DEG C of immobilizations, with deionized water wash 2 ~ 3 times, namely lyophilize obtains immobilized microorganism preparation.
The collocation method of described enrichment medium is: in 1L water, add following each composition and stirring and evenly mixing, and the weight ratio that each composition accounts for water is: peptone 1%, yeast powder 0.5%, NaCl0.5%, KH
2pO
40.06%, MgSO
4h
2o0.03%, pH7.0-7.2, be then 0.15Mpa at pressure, temperature is sterilizing 20min under the condition of 121 DEG C, and it is stand-by that sterilizing wild Oryza species puts into sterilisable chamber cooling.
Adopt the method for immobilized microorganism preparation process meat product processing waste water, it comprises following operation: by after immobilized microorganism preparation in 3 ~ 5%(W/V) ratio join in meat product processing waste water, at 28 ~ 32 DEG C, under the condition of 160rpm, process 24h.
Before adding immobilized microorganism preparation, also comprise the step pH value of meat product processing waste water being adjusted to 6.8 ~ 7.2.
The present invention has the following advantages:
The method that the present invention prepares immobilized microorganism preparation is easy, and by cell coacervation together with polymkeric substance, cell is embedded among the polymkeric substance of formation, and cell is easy to leak not, good stability; The immobilized microorganism preparation of preparation has higher cell-volume, and in preparation, cell content can reach 50% ~ 70%
Utilize treatment based on immobilized microorganisms meat product processing waste water compared to the method for physics and chemistry process meat product processing waste water, there is the advantages such as processing efficiency is high, stability strong, reaction is easy to control, bacterial classification is high-purity efficiently, sludge quantity is few, good effect of separating solid from liquid, loss of activity can be recovered, economy is better.
In the immobilized microorganism utilizing the present invention to obtain, microbe species is more, can multiple pollutant in degrading waste water, and degradation speed is very fast, and ability is strong, and processing efficiency is high.
It is high that the immobilized microorganism utilizing the present invention to obtain has bacterial classification concentration, has and adapt to water quality and the strong advantage of pH changing capability.
The sludge yield that the immobilized microorganism utilizing the present invention to obtain produces for the treatment of meat product processing waste water is less, and product separation is easy, good stability, and technological operation is simple, and production cost is lower.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, and protection scope of the present invention is not limited to the following stated:
embodiment 1:
Immobilized microorganism preparation, it adopts following steps preparation:
The fermentation culture of S1, bacterial classification, by subtilis (BacillussubtilisATCC14579), bacillus megaterium (BacillusmegateriumATCC13578), bacterium nitrobacter (NitrobacterATCC14123) and Nitrosomonas (NitrosomonaseuropaeaATCC25978), be inoculated into respectively in enrichment medium, 30 DEG C, 24h is cultivated, to obtain the logarithmic phase cell of each thalline after the centrifugation 10min of 5000rpm under 160rpm condition;
The mixing of S2, thalline, after the cell phosphate buffered saline buffer in step S1 is washed 2 times, by volume percentages, get 40% subtilis respectively, 30% bacillus megaterium, 15% bacterium nitrobacter, 15% nitrosification bacillus mixing, is suspended in physiological saline, obtains thalline mixed solution;
The immobilization of S3, PVA-sodium alginate, it comprises following sub-step:
The preparation of S31, PVA gel, get PVA to be added to the water, dissolve completely under 80 DEG C of conditions, the weight/volume of PVA and water is 1:10, add sodium alginate, silicon-dioxide and calcium carbonate after cooling and mix, PVA: sodium alginate: silicon-dioxide: the weight ratio of calcium carbonate is: 10:3:5:0.4, obtains PVA gel;
S32, thalline adsorb: add in the thalline mixed solution obtained in step s 2 with mixeding liquid volume than the gac being 5%, mixing and absorption;
The preparation of S33, immobilization composite bacteria PVA-sodium alginate particle: after above-mentioned obtained PVA gel and the thalline mixed solution adding gac are mixed, obtain thalline-PVA-sodium alginate suspension, peristaltic pump and thalline-PVA-sodium alginate suspension is connected with thin plastic pipe, under the conveying of peristaltic pump, thalline-PVA-sodium alginate suspension through diameter be the thin plastic pipe instillation low speed of 1.5mm and continuously stirring containing 2%CaCl
2boric acid solution in, described boric acid solution pH, in neutral, takes out after 5 DEG C of immobilizations are cross-linked 24h, and with deionized water wash 2 times, namely lyophilize obtains immobilized microorganism preparation.
The collocation method of described enrichment medium is, adds following each composition and stirring and evenly mixing in 1L water, and the weight ratio that each composition accounts for water is: peptone 1%, yeast powder 0.5%, NaCl0.5%, KH
2pO
40.06%, MgSO
4h
2o0.03%, pH7.0-7.2, be then 0.15Mpa at pressure, temperature is sterilizing 20min under the condition of 121 DEG C, and it is stand-by that sterilizing wild Oryza species puts into sterilisable chamber cooling.
Adopt the method for immobilization biological preparation process meat product processing waste water, it comprises following operation: measure 200mL wastewater sample in the triangular pyramidal bottle of 500mL, waste water ph is regulated to be 7.0, by above-mentioned obtained immobilized microorganism preparation in 3%(W/V) ratio join in Erlenmeyer flask, put into shaking table at 30 DEG C, under the condition of 160r/min, cultivate 24h.
The handled former water-quality COD of meat product processing waste water is 2200mg/L, and after process, the COD of water quality is 80mg/L.
embodiment 2:
Immobilized microorganism preparation, immobilized microorganism preparation, it adopts following steps preparation:
The fermentation culture of S1, bacterial classification, by subtilis (BacillussubtilisATCC14579), bacillus megaterium (BacillusmegateriumATCC13578), bacterium nitrobacter (NitrobacterATCC14123) and Nitrosomonas (NitrosomonaseuropaeaATCC25978), be inoculated into respectively in enrichment medium, 28 DEG C, 26h is cultivated, to obtain the logarithmic phase cell of each thalline after the centrifugation 10min of 5000rpm under 170rpm condition;
The mixing of S2, thalline, after the cell phosphate buffered saline buffer in step S1 is washed 3 times, by volume percentages, get 60% subtilis respectively, 25% bacillus megaterium, 10% bacterium nitrobacter, 5% nitrosification bacillus mixing, is suspended in physiological saline, obtains thalline mixed solution;
The immobilization of S3, PVA-sodium alginate, it comprises following sub-step:
The preparation of S31, PVA gel, get PVA to be added to the water, dissolve completely under 80 DEG C of conditions, the weight/volume of PVA and water is 1:9, add sodium alginate, silicon-dioxide and calcium carbonate after cooling and mix, PVA: sodium alginate: silicon-dioxide: the weight ratio of calcium carbonate is: 10:2:4:0.3, obtains PVA gel;
S32, thalline adsorb: add in the thalline mixed solution obtained in step s 2 with mixeding liquid volume than the gac being 5%, mixing and absorption;
The preparation of S33, immobilization composite bacteria PVA-sodium alginate particle: after above-mentioned obtained PVA gel and the thalline mixed solution adding gac are mixed, obtain thalline-PVA-sodium alginate suspension, peristaltic pump and thalline-PVA-sodium alginate suspension is connected with thin plastic pipe, under the conveying of peristaltic pump, thalline-PVA-sodium alginate suspension through diameter be the thin plastic pipe instillation low speed of 1.5mm and continuously stirring containing 2.5%CaCl
2boric acid solution in, described boric acid solution pH, in neutral, takes out after 4 DEG C of immobilizations are cross-linked 22h, and with deionized water wash 3 times, namely lyophilize obtains immobilized microorganism preparation.
The collocation method of described enrichment medium is, adds following each composition and stirring and evenly mixing in 1L water, and the weight ratio that each composition accounts for water is: peptone 1%, yeast powder 0.5%, NaCl0.5%, KH
2pO
40.06%, MgSO
4h
2o0.03%, pH7.0-7.2, be then 0.15Mpa at pressure, temperature is sterilizing 20min under the condition of 121 DEG C, and it is stand-by that sterilizing wild Oryza species puts into sterilisable chamber cooling.
Adopt the method for immobilization biological preparation process meat product processing waste water, it comprises following operation: measure 200mL wastewater sample in the triangular pyramidal bottle of 500mL, waste water ph is regulated to be 7.2, by above-mentioned obtained immobilized microorganism preparation in 4%(W/V) ratio join in Erlenmeyer flask, put into shaking table at 32 DEG C, under the condition of 160r/min, cultivate 24h.
The handled former water-quality COD of meat product processing waste water is 2350mg/L, and after process, the COD of water quality is 85mg/L.
embodiment 3:
Immobilized microorganism preparation, immobilized microorganism preparation, it adopts following steps preparation:
The fermentation culture of S1, bacterial classification, by subtilis (BacillussubtilisATCC14579), bacillus megaterium (BacillusmegateriumATCC13578), bacterium nitrobacter (NitrobacterATCC14123) and Nitrosomonas (NitrosomonaseuropaeaATCC25978), be inoculated into respectively in enrichment medium, 32 DEG C, 22h is cultivated, to obtain the logarithmic phase cell of each thalline after the centrifugation 10min of 5000rpm under 150rpm condition;
The mixing of S2, thalline, after the cell phosphate buffered saline buffer in step S1 is washed 2 times, by volume percentages, get 55% subtilis respectively, 20% bacillus megaterium, 12% bacterium nitrobacter, 13% nitrosification bacillus mixing, is suspended in physiological saline, obtains thalline mixed solution;
The immobilization of S3, PVA-sodium alginate, it comprises following sub-step:
The preparation of S31, PVA gel, get PVA to be added to the water, dissolve completely under 80 DEG C of conditions, the weight/volume of PVA and water is 1:11, add sodium alginate, silicon-dioxide and calcium carbonate after cooling and mix, PVA: sodium alginate: silicon-dioxide: the weight ratio of calcium carbonate is: 10:4:6:0.5, obtains PVA gel;
S32, thalline adsorb: add in the thalline mixed solution obtained in step s 2 with mixeding liquid volume than the gac being 5%, mixing and absorption;
The preparation of S33, immobilization composite bacteria PVA-sodium alginate particle: after above-mentioned obtained PVA gel and the thalline mixed solution adding gac are mixed, obtain thalline-PVA-sodium alginate suspension, peristaltic pump and thalline-PVA-sodium alginate suspension is connected with thin plastic pipe, under the conveying of peristaltic pump, thalline-PVA-sodium alginate suspension through diameter be the thin plastic pipe instillation low speed of 1.5mm and continuously stirring containing 1.5%CaCl
2boric acid solution in, described boric acid solution pH, in neutral, takes out after 6 DEG C of immobilizations are cross-linked 26h, and with deionized water wash 3 times, namely lyophilize obtains immobilized microorganism preparation.
The collocation method of described enrichment medium is, adds following each composition and stirring and evenly mixing in 1L water, and the weight ratio that each composition accounts for water is: peptone 1%, yeast powder 0.5%, NaCl0.5%, KH
2pO
40.06%, MgSO
4h
2o0.03%, pH7.0-7.2, be then 0.15Mpa at pressure, temperature is sterilizing 20min under the condition of 121 DEG C, and it is stand-by that sterilizing wild Oryza species puts into sterilisable chamber cooling.
Adopt the method for immobilization biological preparation process meat product processing waste water, it comprises following operation: measure 200mL wastewater sample in the triangular pyramidal bottle of 500mL, waste water ph is regulated to be 7.0, by above-mentioned obtained immobilized microorganism preparation in 5%(W/V) ratio join in Erlenmeyer flask, put into shaking table at 28 DEG C, under the condition of 160r/min, cultivate 24h.
The handled former water-quality COD of meat product processing waste water is 2100mg/L, and after process, the COD of water quality is 75mg/L.
Claims (1)
1. adopt the method for immobilized microorganism preparation process meat product processing waste water, it is characterized in that: it comprises following operation:
One, immobilized microorganism preparation is prepared:
The fermentation culture of S1, bacterial classification, by subtilis, bacillus megaterium, bacterium nitrobacter and Nitrosomonas, be inoculated into respectively in enrichment medium, at 28 ~ 32 DEG C, 22 ~ 26h is cultivated, to obtain the logarithmic phase cell of each thalline after the centrifugation 10min of 5000rpm under 150 ~ 170rpm condition; The collocation method of described enrichment medium is: in 1L water, add following each composition and stirring and evenly mixing, and the weight ratio that each composition accounts for water is: peptone 1%, yeast powder 0.5%, NaCl0.5%, KH
2pO
40.06%, MgSO
4h
2o0.03%, pH7.0-7.2, be then 0.15Mpa at pressure, temperature is sterilizing 20min under the condition of 121 DEG C, and it is stand-by that sterilizing wild Oryza species puts into sterilisable chamber cooling;
The mixing of S2, thalline, by the cell in step S1 with after phosphate buffered saline buffer washing 2 ~ 3 times, by volume percentages, get 40 ~ 60% subtilises respectively, 20 ~ 30% bacillus megateriums, 10 ~ 15% bacterium nitrobacters, 5 ~ 15% nitrosification bacillus mixing, be suspended in physiological saline, obtain thalline mixed solution;
The immobilization of S3, PVA-sodium alginate, it comprises following sub-step:
The preparation of S31, PVA gel, get PVA to be added to the water, dissolve completely under 80 DEG C of conditions, the weight/volume of PVA and water is 1:9 ~ 11, add sodium alginate, silicon-dioxide and calcium carbonate after cooling and mix, PVA: sodium alginate: silicon-dioxide: the weight ratio of calcium carbonate is: 10:2 ~ 4:4 ~ 6:0.3 ~ 0.5, obtains PVA gel;
S32, thalline adsorb: add the gac of 5% in the thalline mixed solution obtained in step s 2, after mixing and absorption;
The preparation of S33, immobilization composite bacteria PVA-sodium alginate particle: above-mentioned obtained PVA gel and the thalline mixed solution that adds gac in 1: 4 ratio mix after, obtain thalline-PVA-sodium alginate suspension, by thalline-PVA-sodium alginate suspension instillation low speed, continuously stirring contains 1.5 ~ 2.5%CaCl
2boric acid solution in, described boric acid solution pH, in neutral, takes out after the crosslinked 22 ~ 26h of 4 ~ 6 DEG C of immobilizations, and with deionized water wash 2 ~ 3 times, namely lyophilize obtains immobilized microorganism preparation;
Two, the pH value of meat product processing waste water is adjusted to 6.8 ~ 7.2, by immobilized microorganism preparation in 3 ~ 5%(W/V) ratio join in meat product processing waste water, at 28 ~ 32 DEG C, under the condition of 160rpm, process 24h.
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