CN103436518B - The preparation method of a kind of immobilization algal toxin degradation bacterium and application thereof - Google Patents
The preparation method of a kind of immobilization algal toxin degradation bacterium and application thereof Download PDFInfo
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- CN103436518B CN103436518B CN201310299001.6A CN201310299001A CN103436518B CN 103436518 B CN103436518 B CN 103436518B CN 201310299001 A CN201310299001 A CN 201310299001A CN 103436518 B CN103436518 B CN 103436518B
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Abstract
The present invention discloses preparation method and the application thereof of a kind of immobilization algal toxin degradation bacterium, immobilization algal toxin degradation bacterium selects activated carbon fiber as immobilization material, described immobilization material is through washing, boil, after the pre-treatment step such as salt acid soak and oven dry, join in LB substratum by certain ratio, and then inoculate degrading microcystic toxins bacterial strain, being fixed cultivation being fixed algal toxin degradation bacterium, immobilization algal toxin degradation bacterium of the present invention is in the application, while carbon fiber adsorption and catalytic combustion Algae toxins, the algal toxin degradation bacterium being fixed in activated carbon fiber can be degraded Algae toxins, can significantly shorten the time of degraded MCs, activated carbon fiber can be carried out bio-regeneration simultaneously. the bacterial strain of degrading microcystic toxins of the present invention, can reach more than 91% to the degradation rate of MC-LR. it is particularly useful for blue-green alga bloom extracts in phycobiliprotein process the Microcystin degraded produced.<!--1-->
Description
Technical field
The present invention relates to water treatment, field of environment pollution control, particularly a kind of for removing, by biochemical method, immobilization algal toxin degradation bacterium and its preparation method that blue-green alga bloom extracts in phycobiliprotein process the Microcystin produced.
Background technology
In recent years, " wawter bloom " based on microcystic aeruginosa is all broken out in Chaohu every year, sometimes several centimeters are reached thick, after corrupt decomposition, send stench, havoc water body and surrounding environment, pollute for eliminating Chaohu blue-green algae, a large amount of blue-green algaes is salvaged to come up, and the recycling of the blue-green algae salvaged has become emphasis research topic. Containing abundant Phycocyanins, C-in bloom blue algae, Phycocyanins, C-is the natural pigment of a kind of sapphire, it is widely used in food, beverage, medicine, cosmetic industry, but mainly from the spirulina of freshwater aquiculture, extract Phycocyanins, C-both at home and abroad at present, its cost is expensive, limits the application development of Phycocyanins, C-. Simultaneously, owing to the bloom blue algae of Chaohu outburst is based on microcystic aeruginosa, containing Microcystin in microcystic aeruginosa, although Microcystin is present in cell, but extracting the necessary broken wall of Phycocyanins, C-from blue-green algae, once break, Microcystin is just discharged in water body, Microcystin stable in properties, it is that one causes liver cancer toxin, human body is had great harm. Microcystin must be removed by the suitability for industrialized production therefore utilizing blue-green algae to extract Phycocyanins, C-.
The minimizing technology of current Microcystin mainly contains physical method, chemical process etc., and MCs (Microcystin) is ring-type seven peptide compounds, has interval double bond, Stability Analysis of Structures, and traditional water technology is difficult to remove it from water. The method eliminating Microcystin in water body at present mainly contains the technology such as charcoal absorption, photodegradation and photochemical catalytic oxidation, chemical oxidation, membrane filtration, biological degradation. But physics method and chemical method also exist certain limitation in actual applications separately, as chemical oxidization method can produce secondary pollution, the cost of membrane filter method is more high.
Summary of the invention
The object of the present invention mainly is to provide preparation method and the application thereof of a kind of immobilization algal toxin degradation bacterium, not only cost is low but also to Microcystin degraded thoroughly, the Microcystin degradation effect that blue-green alga bloom particularly extracts generation in phycobiliprotein process is particularly evident for this immobilization algal toxin degradation bacterium.
For achieving the above object, the present invention by the following technical solutions:
A preparation method for immobilization algal toxin degradation bacterium, its feature is to comprise the steps:
1) fixation support preparation: using activated carbon fiber as fixation support, described activated carbon fiber pre-treatment step is: activated carbon fiber material ultrapure water is swung eccysis decon gently, put into beaker, add ultrapure water and boil 1.0h, then the salt acid soak 24h of functional quality concentration 10%, take out, baking oven is put into after being washed till neutrality with ultrapure water, dry to constant weight at 140 DEG C of temperature, take out, be placed in dry encloses container stand-by;
2) prepared by immobilization algal toxin degradation bacterium: select LB substratum to be the nutritional medium of degrading microcystic toxins bacterium; Add through step 2 to LB substratum in 2.0-10g/L ratio) activated carbon fiber material that processes, sterilizing 20min under 121 DEG C of high temperature, treating that temperature is down to room temperature, the bacterial strain of inoculation degradable Microcystin, cultivates 3d in shaking table, shaking speed 150r/min, temperature is 30 DEG C, after bacterial classification is fully fixed on activated carbon fiber material, takes out, with PBS washing, obtain immobilization algal toxin degradation bacterium.
The bacterial strain of degradable Microcystin of the present invention, is selected from the bacterial strain that preserving number is CCTCCM2013265.
Another object of the present invention is the application protecting the immobilization algal toxin degradation bacterium prepared as stated above to remove Microcystin in blue-green alga bloom phycobiliprotein extraction purification process.
Compared with the prior art, the useful effect of the present invention is embodied in:
1, immobilization algal toxin degradation bacterium of the present invention selects activated carbon fiber as immobilization material, this being fixed of bacterial strain is cultivated, while carbon fiber adsorption and catalytic combustion Algae toxins, the algal toxin degradation bacterium being fixed in activated carbon fiber can be degraded Algae toxins, the degradation rate of Microcystin can be increased considerably, improve algal toxin degradation bacterium to the tolerance of unfavorable factor, activated carbon fiber is carried out bio-regeneration simultaneously.
2, the present invention adopts activated carbon fiber as the carrier of degradation bacteria, owing to activated carbon fiber has flourishing microvoid structure, excellent absorption property, higher physical strength and good biocompatibility, the immobilization algal toxin degradation bacterium therefore prepared by this carrier has the advantages such as microbe density height, reaction is rapid, microorganism loss is few.
3, the present invention from the settling of Chaohu, filter out a strain can the bacterial strain of degrading microcystic toxins, through qualification belong to wax genus bacillus, the immobilization algal toxin degradation bacterium prepared with this bacterial strain is to MC-LR(microcapsule algae toxin) degradation rate reach more than 90%.
The bacterial strain of the degradable Microcystin that the present invention uses screens to obtain from the settling of Chaohu, and its name is called wax genus bacillus M9; Preserving number: CCTCCNO:M2013265; Depositary institution: China typical culture collection center; Preservation date: on June 16th, 2013; Preservation address: Wuhan University of China
Embodiment
Below by way of specific embodiment, technical solution of the present invention is further explained explanation.
Activated carbon fiber specific surface area in following examples is 1600-1800m2/ g; Degradation bacteria is preserving number is the bacterial strain of CCTCCM2013265.
Embodiment 1: the preparation of immobilization algal toxin degradation bacterium
The first step: fixation support activated carbon fiber pre-treatment: activated carbon fiber material ultrapure water swings gently washes some times with except attachment removal impurity, put into beaker, add ultrapure water and boil 1.0h, then functional quality concentration 10% hydrochloric acid soaked overnight, take out, put into baking oven after being washed till neutrality with ultrapure water, dry to constant weight at 140 DEG C of temperature, take out, it is placed in dry encloses container stand-by.
2nd step: immobilization algal toxin degradation bacterium: configuration nutritional medium, nutritional medium selects LB substratum, and it consists of Tryptones 10.0g, yeast extract 5.0g, sodium-chlor 10.0g, distilled water 1000mL, regulates pH to be 7.0; The activated carbon fiber material processed through the first step taking certain mass (in the ratio of 10g/L) adds in this nutritional medium, sterilizing 20min under 121 DEG C of high temperature, treats that temperature is down to room temperature, inoculation algal toxin degradation bacterium, inoculum size is 10%, cultivating 3d in shaking table, shaking speed 150r/min, temperature is 30 DEG C, after bacterial classification is fully fixed on activated carbon fiber material, take out, with PBS washing, obtain immobilization algal toxin degradation bacterium.
3rd step: immobilization algal toxin degradation bacterium is added in the water sample containing Algae toxins, timing sampling measurement processing effect.
Embodiment 2 immobilization algal toxin degradation bacterium removes the application of Microcystin in blue-green alga bloom phycobiliprotein extraction purification process.
1) from eutrophication water, salvaging blue-green algae is gathered, through freeze thawing, saltout, aqueous two-phase extraction, ultrafiltration and concentration purifying obtains Phycocyanins, C-, adopts SPE HPLC to measure the waste liquid in Phycocyanins, C-extraction purification process, the content of contained Microcystin in waste residue, detected result display Microcystin is mainly present in the filtrate of ultrafiltration, and dense, and wherein MC-LR reaches 11.2mg/L.
2) the immobilization algal toxin degradation bacterium of embodiment 1 gained is added in step 1) containing, in the ultrafiltrated of microcapsule algae toxin, after 3d, sampling and measuring processes effect. Result is as follows:
1, the immobilization algal toxin degradation bacterium that prepared by different activities Carbon fibe consumption is to the removal effect of microcapsule algae toxin
Test water sample: microcapsule algae toxin starting point concentration is 6.8mg/L, and pH value is 8, temperature 30 DEG C.
When activated carbon fiber consumption consumption is 2.0g/L-10.0g/L, microcapsule algae toxin clearance along with the increase of activated carbon fiber consumption and rise (77.3%-92.9%); After consumption is more than 10.0g/L, microcapsule algae toxin clearance starts on a declining curve.
2, under condition of different pH immobilization algal toxin degradation bacterium to the removal effect of microcapsule algae toxin
Immobilization algal toxin degradation bacterium: activated carbon fiber consumption 10g/L
Test water sample: microcapsule algae toxin starting point concentration is 6.8mg/L, temperature 30 DEG C.
When pH is 5-9, the Algae toxins clearance of immobilization algal toxin degradation bacterium is 87.2%-91.1%; In acid condition, the algal toxin degradation activity of immobilized bacterium is still higher, and microcapsule algae toxin clearance still remains on higher level, and when pH is 8, the microcapsule algae toxin clearance of immobilization Algae toxins bacterium reaches maximum 91.1%.
3, under condition of different temperatures immobilization algal toxin degradation bacterium to the removal effect of microcapsule algae toxin
Immobilization algal toxin degradation bacterium: activated carbon fiber consumption 10g/L
Test water sample: microcapsule algae toxin starting point concentration is 6.8mg/L, and pH value is 8.
When temperature is at 10-35 DEG C, immobilization algal toxin degradation bacterium is less by the impact of temperature, it is then 79.1-92.7% to the clearance of microcapsule algae toxin, under cryogenic, the microcapsule algae toxin degrading activity of immobilized bacterium group is still higher, and microcapsule algae toxin clearance still remains on higher level.
4, under different Algae toxins starting point concentration immobilization algal toxin degradation bacterium to the removal effect of Algae toxins
Immobilization algal toxin degradation bacterium: activated carbon fiber consumption 10g/L
Test water sample: temperature 35 DEG C, pH value is 8.
When Algae toxins starting point concentration is 1.7-13.6g/L, the clearance of immobilization algal toxin degradation bacterium is 78.8%-93.6%, and Algae toxins clearance is overall upward trend along with the rising of Algae toxins concentration.
Claims (2)
1. the preparation method of an immobilization algal toxin degradation bacterium, it is characterised in that comprise the steps:
1) fixation support preparation: using activated carbon fiber as fixation support, described activated carbon fiber pre-treatment step is: activated carbon fiber material ultrapure water is swung eccysis decon gently, put into beaker, add ultrapure water and boil 1.0h, then the salt acid soak 24h of functional quality concentration 10%, take out, baking oven is put into after being washed till neutrality with ultrapure water, dry to constant weight at 140 DEG C of temperature, take out, be placed in dry encloses container stand-by;
2) prepared by immobilization algal toxin degradation bacterium: select LB substratum to be the nutritional medium of degrading microcystic toxins bacterium; Add through step 1 to LB substratum in 2.0-10g/L ratio) pretreated fixation support activated carbon fiber, sterilizing 20min under 121 DEG C of high temperature, treating that temperature is down to room temperature, the bacterial strain of inoculation degradable Microcystin, cultivates 3d in shaking table, shaking speed 150r/min, temperature is 30 DEG C, after bacterial classification is fully fixed on activated carbon fiber material, takes out, with PBS washing, obtain immobilization algal toxin degradation bacterium;
The bacterial strain of described degrading microcystic toxins, belong to wax genus bacillus, called after wax genus bacillus (Bacilluscereus) M9, described wax genus bacillus (Bacilluscereus) M9 is preserved in China typical culture collection center, and preserving number is: CCTCCM2013265.
2. the immobilization algal toxin degradation bacterium prepared by claim 1 method removes the application of Microcystin in blue-green alga bloom phycobiliprotein extraction purification process.
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| CN103834621A (en) * | 2013-12-30 | 2014-06-04 | 贵州大学 | Production method for laccase |
| CN104404025A (en) * | 2014-11-14 | 2015-03-11 | 河南工业大学 | Immobilized cell and application thereof |
| CN107602680A (en) * | 2016-07-11 | 2018-01-19 | 河南工业大学 | A kind of technique for extracting high-purity wheat embryo globulin |
| CN107739722A (en) * | 2017-09-15 | 2018-02-27 | 常州大学 | A kind of method that algal toxin degradation bacterium is screened in the turbulent waves fish guts from Taihu Lake |
| CN109231488B (en) * | 2018-10-19 | 2021-06-18 | 浙江海洋大学 | Method for synchronously treating nitrogen and phosphorus inorganic nutrients and organic pollutants in water body |
| CN110172456B (en) * | 2019-05-28 | 2023-02-28 | 中南大学 | Biological material with functions of dissolving algae, degrading algal toxins and removing nitrogen and phosphorus, and preparation method and application thereof |
| CN112080488B (en) * | 2020-09-24 | 2022-04-05 | 安徽大学 | Immobilization method of wild thatch mushroom mycelium and application of wild thatch mushroom mycelium in dye decoloration |
| CN114196664B (en) * | 2021-12-14 | 2023-12-01 | 重庆大学 | Methane-oxidizing bacteria-porous adsorbent compound and application thereof |
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