CN110106162A - A kind of living microorganism immobilized microspheres and its preparation method and application - Google Patents
A kind of living microorganism immobilized microspheres and its preparation method and application Download PDFInfo
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- CN110106162A CN110106162A CN201910464257.5A CN201910464257A CN110106162A CN 110106162 A CN110106162 A CN 110106162A CN 201910464257 A CN201910464257 A CN 201910464257A CN 110106162 A CN110106162 A CN 110106162A
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
Abstract
The present invention discloses a kind of living microorganism immobilized microspheres and its preparation method and application.This method includes that microbial fixed carrier, bacterium solution suspension colonize, microbial activity particle and proliferation are further cultured for.For microballoon of the invention in spherical, there are hole, 3.0~4.0mm of diameter, 1.10~1.20g/cm of specific gravity in inside3, alkalescent has good settling property and mechanical strength.The microballoon can not only play carrier to the chemisorption of cadmium, moreover it is possible to improve the growth and breeding environment of active microorganism, act on to improve activated bacterial the extracellular absorption of cadmium and intracellular accumulation.In initial Cd2+In the following range of concentration 250mg/L, maximum adsorption-accumulation is up to 158.77mg/g.The present invention has that raw material are cheap and easy to get, process operability is strong, microspheres product is environmentally friendly and easy recycling, the advantages that regulation effect is significant, has broad application prospects during the biological treating of heavy metal-polluted water.
Description
Technical field
The invention belongs to environmental pollution microbial treatment technical field, be related to a kind of living microorganism immobilized microspheres and its
A kind of preparation method and application, and in particular to activated bacterial process for fixation and immobilized bacterium with heavy metal cadmium resistance
Application of the ball in biological wastewater treatment.
Background technique
Heavy metal cadmium has very strong bio-toxicity and is not easy to be biodegradable, and can be entered in the environment by food chain
Human organ, seriously threatens human health at the effects of leading to teratogenesis, carcinogenic, mutagenesis.It is main for heavy metal in water cadmium pollution
There are physical-chemical process and bioanalysis, including chemical precipitation, ion exchange, UF membrane, adsorbing separation, plant absorption and microorganism
Absorption accumulation etc..Currently, microorganism immobilization method, as one of the new technology that heavy metal pollution of water body is administered, main feature
It is that free functional microorganism is fixed on carrier using chemically or physically method, passes through absorption-accumulation of active microorganism
Effect is to improve treatment effect.Wherein, the activity for keeping microorganism is the key that the technology in waste water treatment application.
Charcoal is that biomass (such as straw and animal wastes) is pyrolyzed generation under complete or partial anoxia condition
The highly stable carbonaceous material of one kind.It can provide the place inhabited with breeding for microorganism, it is also possible to provide nutrients
Matter promotes the growth and breeding of microorganism.In China, rice straw raw material sources are abundant, and every about 300,000,000 tons of annual generation, and
And rice straw charcoal is considered as a kind of efficient heavy metal absorbent.Therefore, rice straw charcoal can be used as a kind of ideal micro-
Biological fixing carrier, it not only can reduce Heavy Metals in Waters concentration by itself suction-operated to protect microorganism, and
Good habitat can be provided to promote microbial growth, so that microorganism reduces a huge sum of money by absorption-summation
Belong to pollution level.Charcoal immobilized microorganism technique reported at present is in terms of heavy metal pollution of water body improvement, processing effect
Fruit is bad, is primarily due in wastewater treatment process most of microorganism and loses activity, it is difficult to play absorption-accumulation work
With.
Common process for fixation has absorption method, investment, cross-linking method and covalent coupling method, and wherein absorption method has operation
Simply, the advantages that at low cost, small on cell activity influence, but the disadvantage is that carrier and microbes resultant force are lower, effect is unstable.
Therefore, a kind of living microorganism immobilization product preparing simple, environmental-friendly type, working well is researched and developed, in a water body huge sum of money
It is with a wide range of applications in terms of belonging to pollution control.
Summary of the invention
The shortcomings that in order to overcome living microorganism immobilization technology in the prior art and deficiency, primary and foremost purpose of the invention exist
In providing a kind of preparation method of living microorganism immobilized microspheres.It can maintain microorganism that there is greater activity by this method.
Another object of the present invention is to provide the living microorganism immobilization being prepared by above-mentioned preparation method is micro-
Ball.
A further object of the present invention is to provide the applications of above-mentioned living microorganism immobilized microspheres.
The purpose of the invention is achieved by the following technical solution:
A kind of preparation method of living microorganism immobilized microspheres, includes the following steps:
(1) microbial fixed carrier: rice straw after drying, crushing and sieving, simultaneously grind by the pyrolysis of limit oxygen, cooling
Sieving, obtains microbial fixed carrier；
(2) bacterium solution suspension colonizes: bacillus cereus (Bacillus cereus) RC-1 being activated, cell suspension is obtained；
5~20mL bacterial suspension and 0.2~1.5g microbial fixed carrier is taken to mix, mixed serum A is made in 1~2h of shaken cultivation；
(3) sodium alginate soln of the mass fraction 2%~6% of sterilizing microbial activity particle: is added to mixing slurry
In liquid A, mixed serum B is obtained；Then mixed serum B is added dropwise to the CaCl of sterilizing2In solution, solidify 2~10h, is formed micro-
It is biological active granulated；Wherein, dosage of the microbial fixed carrier in mixed serum B is 0.2~1.5g/100mL；
(4) proliferation is further cultured for: the microbial activity particle that step (3) is obtained is in fresh beef cream peptone Liquid Culture
Base proliferation is further cultured for 1~4h, behind sterile water immersion treatment surface, obtains living microorganism immobilized microspheres.
Preferably, the preparation method of microbial fixed carrier described in step (1), includes the following steps: rice straw
Stalk is placed in 300~700 DEG C of lower limit oxygen in pyrolysis oven and is pyrolyzed 2~4h, cooling is simultaneously ground by natural air drying and after shredding grinding
Be sieved 60~100 mesh, obtains microbial fixed carrier；
It is furthermore preferred that the preparation method of microbial fixed carrier described described in step (1), including walk as follows
Rapid: rice straw is placed in 700 DEG C of lower limits in the pyrolysis oven for being first passed through 20min nitrogen by natural air drying and after shredding grinding
Oxygen is pyrolyzed 2h, and cooling is simultaneously ground up, sieved 100 mesh, obtains microbial fixed carrier；
Described bacillus cereus (Bacillus cereus) RC-1 is by from cadmium pollution soil (1.4Cdmg/
Kg separation screening obtains in), morphological feature are as follows: it is rod-shaped, in pairs or catenation, it is raw in gemma or end is raw partially, ellipse or
Stake shape, without parasporal crystal, bacterium colony circle milky, rough surface belong to Gram-positive.The bacterial strain RC-1 can be applied to
During heavy metal in water cadmium pollution is administered.
The preservation information of described bacillus cereus (Bacillus cereus) RC-1 is as follows: depositary institution: China is micro-
Biological inoculum preservation administration committee common micro-organisms center (CGMCC), preservation date: on 03 27th, 2017, preservation address:
Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, deposit number: CGMCC NO:13930.
Preferably, the OD of cell suspension described in step (2)600=1.2~1.5, i.e. increased logarithmic phase cell suspension；
Preferably, the dosage of bacterial suspension described in step (2) is 5mL；
Preferably, the time of shaken cultivation described in step (2) is 2h；
Preferably, the mass fraction of sodium alginate soln described in step (3) is 4%；
Preferably, the speed of instillation described in step (3) is 5~10mL/min；More preferably 7mL/min；
Preferably, CaCl described in step (3)2Solution is the CaCl that mass fraction is 2~10%2Solution；More preferably
The CaCl that mass fraction is 5%2Solution.
Preferably, dosage of the microbial fixed carrier in mixed serum B described in step (3) is 1.5g/
100mL；
Preferably, the cured time described in step (3) is 2~5h；More preferably 2h；
Preferably, the formula of beef extract-peptone fluid nutrient medium described in step (4) are as follows: 10g/L peptone, 3g/L
Beef extract, 5g/L NaCl2, pH value is 7.0 ± 0.2；
Preferably, it is 4h that the time being further cultured for is proliferated described in step (4)；
A kind of living microorganism immobilized microspheres, are prepared by above-mentioned preparation method.
The living microorganism immobilized microspheres are in spherical, and there is a hole in inside, 3.0~4.0mm of diameter, and specific gravity 1.10~
1.20g/cm3, alkalescent has good settling property and mechanical strength.
The weakly alkaline pH range is 7.66~8.76.
Application of the living microorganism immobilized microspheres in the improvement for heavy metal pollution；
Further, application of the living microorganism immobilized microspheres in heavy metal pollution of water body improvement.
Preferably, the heavy metal is cadmium (Cd).
Specifically comprise the following steps:
Containing initial concentration 250mg/L or less Cd2+Beef extract-peptone fluid nutrient medium (pH be 7.0 ± 0.2) in,
The above-mentioned living microorganism immobilized microspheres that solid-to-liquid ratio is 0.2g/L are added, shaken cultivation is realized to Cd2+Absorption.
Preferably, the Cd2+Initial concentration be 5~250mg/L；Further preferably 100~220mg/L；Again into
One step is preferably 110~200mg/L；More preferably 150~180mg/L；Most preferably 180mg/L.
The condition of the shaken cultivation is 28 ± 2 DEG C, 90~150r/min shaken cultivation, 5~30h；Preferably 28 ± 2
DEG C, 5~30h of 150r/min shaken cultivation；Further, incubation time is 16~30h；Further, incubation time 19
~26h；It is furthermore preferred that incubation time is 19~26h；Most preferably, incubation time be 22~for 24 hours.
The present invention has the following advantages and effects with respect to the prior art:
(1) living microorganism immobilized microspheres produced by the present invention have both passive adsorption and the micro- life of activity of biological high-area carbon
Active absorption-summation of object, obvious processing effect.Immobilized microspheres are 7.0 ± 0.2,28 ± 2 DEG C of temperature, 150r/ in pH
Min shaken cultivation for 24 hours, to initial Cd2+Concentration is that the adsorbance of 180mg/L is up to 158.77mg/g.
(2) living microorganism immobilized microspheres produced by the present invention have useless while maintaining compared with high absorption capacity
Waste resource Environmental Safety, is easily recycled, the advantages that microbial activity is high, can be widely applied to heavy metal-polluted water and
The biological treating of soil.
(3) living microorganism immobilized microspheres produced by the present invention can not only play change of the microbial fixed carrier to cadmium
Learn suction-operated, moreover it is possible to improve the growth and breeding environment of active microorganism, thus improve activated bacterial to the extracellular absorption of cadmium and
Intracellular accumulation effect.In initial Cd2+In the following range of concentration 250mg/L, maximum adsorption-accumulation is up to 158.77mg/g.This
It is excellent that invention has that raw material are cheap and easy to get, process operability is strong, microspheres product is environmentally friendly and easy recycling, regulation effect are significant etc.
Point has broad application prospects during the biological treating of heavy metal-polluted water.
Detailed description of the invention
Fig. 1 is the preparation flow material object schematic diagram of living microorganism immobilized microspheres in embodiment of the present invention method 1.
Fig. 2 is the scanning electron microscope (SEM) photograph of the immobilized microspheres product of embodiment 1.
Fig. 3 be in embodiment 2 different initial pH to the influence of immobilized microspheres absorption-accumulation；Wherein, rice straw microballoon is
Refer to living microorganism immobilized microspheres made from embodiment 1；Chicken manure microballoon refers to that using chicken manure be carrier material referring to embodiment 1
The immobilized microspheres that preparation method is prepared；Sludge microballoon refers to the preparation side using sludge as carrier material referring to embodiment 1
The immobilized microspheres that method is prepared.
Fig. 4 is different initial Cd in embodiment 32+Concentration to immobilized microspheres absorption-accumulation (a), bacterial number (b) and
The influence of ATP enzyme (c) living.
Fig. 5 is different initial Cd under cell metabolic inhibitor DCC in embodiment 42+Concentration is to immobilized microspheres absorption-accumulation
The influence of amount.
Fig. 6 be in embodiment 5 reaction time to the influence of immobilized microspheres absorption-accumulation；Wherein, a:20mg/L Cd2 +, b:50mg/L Cd2+, c:100mg/L Cd2+。
Fig. 7 be in embodiment 6 different mechanisms to the contribution of living microorganism immobilized microspheres absorption-accumulation.
Specific embodiment
Present invention will now be described in further detail with reference to the embodiments and the accompanying drawings, but embodiments of the present invention are unlimited
In this.For not specifically specified technological parameter, routine techniques progress can refer to.
1 immobilized microspheres preparation flow of embodiment and material morphology and property
Referring to Fig. 1, microspheres product preparation step is as follows:
The first step, rice straw are placed in by natural air drying and after shredding grinding and are passed through 20 minutes heat of nitrogen in advance
It solves in furnace, is pyrolyzed 2h in 700 DEG C of lower limit oxygen of temperature, 100 mesh are ground up, sieved after being cooled to room temperature, obtain microbial immobilized load
Body.
Bacillus cereus RC-1 is chosen 1~2 ring of bacterium and is inoculated in beef extract-peptone body fluid nutrient medium by second step,
At 30 DEG C, (OD after 8~10h of culture activation under 150r/min600=1.2~1.5) cell suspension, is obtained；Take 5mL cell suspension
It is mixed with 1.5g microbial fixed carrier, mixed serum A is made in shaken cultivation 2h；By the seaweed of the mass fraction 4% of sterilizing
Acid sodium solution is added in mixed serum A to 100mL, obtains mixed serum B.
Third step, by the resulting mixed serum B of second step by peristaltic pump with 7mL/min speed instillation be containing mass fraction
5% CaCl2In solution, microbial activity particle is formed after solidifying 2h.
Microbial activity particle access fresh beef cream peptone fluid nutrient medium proliferation is further cultured for 4h, used by the 4th step
Behind sterile water immersion treatment surface, living microorganism immobilized microspheres are obtained.
The present embodiment prepares the preparation flow material object schematic diagram of living microorganism immobilized microspheres as shown in Figure 1, microballoon
For grain in spherical, there are hole, 3.0~4.0mm of diameter, 1.10~1.20g/cm of specific gravity in inside3, alkalescent has good sedimentation
Performance and mechanical strength (99% ± 1%).
Wherein, mechanical strength measuring method: 100, immobilized bacterium ball and 50mL deionized water are put into 100mL conical flask, with
300r/min vibrates, and the complete number of bacterium ball is observed after 48h, and calculate the ratio that intact bacterium ball number accounts for opportunistic pathogen ball sum,
Indicate the strength factor of immobilized microspheres.
The scanning electron microscope (SEM) photograph of living microorganism immobilized microspheres made from the present embodiment is as shown in Fig. 2, microbial immobilized
The a large amount of microorganisms of pore interior and area load of carrier.
2 pH of embodiment is to the influence of immobilized microspheres absorption-accumulation
As initial Cd2+Under the conditions of concentration is 100mg/L, for the microbial fixed carrier of three kinds of different raw material preparations
(rice straw, chicken manure and sludge), take solid-to-liquid ratio be 0.2g/L immobilized microspheres (2,3,4,5,6 and 7) different pH beef extract
In peptone fluid nutrient medium (pH is 7.0 ± 0.2), after 150r/min shakes fast shaken cultivation for 24 hours under room temperature, centrifuging and taking supernatant
(10000rpm, 15min) measures Cd2+Concentration.
From figure 3, it can be seen that as initial pH is in 2~5 variation range, absorption-accumulation of three kinds of immobilized microspheres
It quicklys increase, then gradually slows down, be finally reached balance.When initial pH is 5~7 range, absorption-product of immobilized microspheres
Tired amount range is 91.84~124.05mg/g, and especially in pH=7, it is solid that maximum adsorption-accumulation is respectively as follows: rice straw charcoal
Surely change microballoon (124.05mg/g) > chicken manure charcoal immobilized microspheres (107.67mg/g) > sludge organism charcoal immobilized microspheres
(95.37mg/g).In addition, the pH is also very favorable for the growth and breeding of viable bacteria.
This example demonstrates that rice straw charcoal immobilized microspheres are to water body Cd2+Absorption-accumulation ability is best, is it not
Guarantee is provided with the application under pH environmental condition.
The initial Cd of embodiment 32+The concentration influence living to immobilized microspheres absorption-accumulation, micro organism quantity and ATP enzyme.
In different initial Cd2+Concentration (5,10,20,50,80,100,110,120,150,180,200,210,220,
In beef extract-peptone fluid nutrient medium (pH is 7.0 ± 0.2) 250mg/L), the embodiment 1 that solid-to-liquid ratio is 0.2g/L is added
The living microorganism immobilized microspheres of preparation, under room temperature at the uniform velocity shaken cultivation for 24 hours after (150r/min), centrifuging and taking supernatant
(10000rpm, 15min) measures Cd2+Concentration.In addition, the immobilized microspheres separated are placed in 20mL buffer solution
(0.05mol/L Na2CO3With 0.02mol/L citric acid), shaken cultivation 1h carries out microorganism plate after microballoon is completely dissolved
It counts and ATP enzyme is lived and measured.
It can be seen that from Fig. 4 a with initial Cd2+Absorption-accumulation of the increase of concentration, immobilized microspheres quicklys increase；
Work as Cd2+When concentration is greater than 110mg/L, absorption-accumulation gradually slows down and tends to balance, especially Cd2+Concentration is 180mg/L
When, maximum value 158.77mg/g；As initial Cd2+When concentration is greater than 200mg/L, absorption-accumulation is gradually reduced, even if initially
Cd2+When rising to 250mg/L, absorption accumulation also maintains high value 123.62mg/g.At the same time, Fig. 4 b shows initial Cd2+
Influence of the concentration to bacterial number, with Cd2+The increase (10~250mg/L) of concentration, micro organism quantity is from 2.5 × 1012CFU/
ML drops to 2.8 × 107CFU/mL.Similarly, Fig. 4 c shows initial Cd2+Influence of the concentration to bacterial activity, with Cd2+Concentration
Increase (0~250mg/L), bacterial ATP enzymatic activity drops to 1.35U/mgprot from 11.53U/mgprot.These show with
Initial Cd2+The increase of concentration, the number and activity of bacterium are significantly affected, even if initial Cd2+When concentration maximum
(250mg/L), microorganism still keeps a degree of activity, to improve treatment effect.
This example demonstrates that in different initial Cd2+Under concentration conditions, absorption-accumulation of immobilized microspheres is higher, is
It provides good basis and guarantee in practical applications.
Different initial Cd under 4 cell ATP enzyme inhibitor DCC (dicyclohexylcarbodiimide) of embodiment2+Concentration is to immobilization
The influence of microballoon absorption-accumulation.
Under the conditions of inhibitor DCC (1mmol/L), different initial Cd2+Concentration (5,10,20,50,80,100,110,120,
150,180,200,210,220,250mg/L) beef extract-peptone fluid nutrient medium (pH be 7.0 ± 0.2) in, solid-liquid is added
Than the living microorganism immobilized microspheres of the preparation of embodiment 1 for 0.2g/L, under room temperature at the uniform velocity shaken cultivation for 24 hours after, centrifuging and taking
Supernatant measures Cd2+Concentration.
From fig. 5, it can be seen that even if under the conditions of existing for the cell metabolic inhibitor DCC, with Cd2+The increase of concentration is inhaled
Attached-accumulation is still continuously increased, and then progressivelyes reach balance, as initial Cd2+When concentration is greater than 180mg/L, absorption-accumulation
It reduces therewith.In initial Cd2+When concentration is 150mg/L, maximum adsorption-accumulation is 131.43mg/g.This shows that charcoal carries
Physical efficiency is effectively protected the activity of bacterium, and in initial Cd2+Concentration is within the scope of 100~180mg/L, the suction of immobilized microspheres
Attached-accumulation is still higher, maintains 113.02~131.43mg/g range.
This example demonstrates that even if immobilized microspheres are to Cd under cell metabolic inhibitor2+Absorption-still with higher
The application that accumulation is it in adverse environment condition provides guarantee.
5 reaction time of embodiment is to the influence of immobilized microspheres absorption-accumulation.
Containing 20,50 and 100mg/L Cd2+Beef extract-peptone fluid nutrient medium (pH be 7.0 ± 0.2) in, be added
Living microorganism immobilized microspheres prepared by the embodiment 1 that solid-to-liquid ratio is 0.2g/L, in different time (0,1,2,3,5,7,10,
13,16,19,22,26,30h) centrifuging and taking supernatant is carried out, and measure wherein Cd2+Concentration.
Fig. 6 a can be seen that as initial Cd2+When concentration is 20mg/L, with the increase in reaction time, the suction of immobilized microspheres
Attached-accumulation is continuously increased, and is tended to balance in 22h, and maximum adsorption-accumulation is 30.51mg/g.Fig. 6 b shows when initial
Cd2+Under the conditions of concentration is 50mg/L, as in reaction time 16h, absorption-accumulation of immobilized microspheres is gradually increased, when anti-
Between seasonable within the scope of 16~22h, absorption-accumulation is quicklyd increase, when reacted between reach balance, maximum adsorption-product after 22h
Tired amount is 81.66mg/g.Fig. 6 c is shown, in initial Cd2+When concentration is 100mg/L, the increase at any time of absorption-accumulation and it is fast
Speed increases, and balance is reached in reaction time 22h, and maximum adsorption-accumulation is 141.37mg/g.
This example demonstrates that immobilized microspheres are to Cd under the conditions of various concentration2+Absorption-accumulation equilibration time it is shorter,
Absorption-accumulation is also higher, and heavy metal pollution of water body is quickly administered for it and provides technology guarantee.
Embodiment 6
In initial Cd2+The beef extract-peptone fluid nutrient medium that concentration is 20,50 and 100mg/L (pH is 7.0 ± 0.2)
In, be added solid-to-liquid ratio be 0.2g/L embodiment 1 prepare living microorganism immobilized microspheres, at the uniform velocity shaken cultivation for 24 hours after, from
The heart takes supernatant to measure Cd2+Concentration calculates total absorption-accumulation Q.Liquid is discarded supernatant, the ddH of equivalent is added2O, oscillation training
2h is supported, is centrifuged and measures Cd in supernatant2+Concentration, gained Q1As electrostatic attraction acts on contributed absorption-accumulation, accordingly
Contribution proportion be Q1/Q.Liquid is discarded supernatant, the 1.0mol/L NH of equivalent is added4NO3Solution, shaken cultivation 2h measure supernatant
Cd in liquid2+Concentration obtains Q2, as ion-exchange mechanism contribution absorption-accumulation, contribution proportion Q2/Q.Then it abandons again
Supernatant is removed, the 0.1mol/L EDTA-Na of equivalent is added2Solution measures Cd in supernatant after shaken cultivation 2h2+Concentration Q3, i.e.,
Absorption-the accumulation contributed by functional group's chelation, contribution proportion Q3/Q.At the same time, to sediment in centrifuge tube
Carry out resolution measurement Cd2+Concentration Q4, Q4As absorption-accumulation of cell interior accumulation contribution, contribution proportion Q4/Q.Finally,
Total absorption-accumulation Q obtains Q after subtracting absorption-accumulation of above four kinds of mechanism5, as precipitation mechanism contribution absorption-product
Tired amount, contribution proportion Q5/Q。
Fig. 7 can be seen that three kinds of initial Cd of difference2+Under concentration conditions, ion exchange is immobilized microspheres absorption-product
Tired main mechanism, chelation mechanism take second place.Especially it is noted that active somatic cell inside Cd accumulation (Q4) initial at three kinds
It is respectively 1.2,6.7 and 6.3mg/g under concentration, accounts for microballoon always absorption-accumulation 3.7%, 8.2% and 5.3%, this table respectively
Bright immobilization technology can effectively keep the activity of functional microorganism, and the active absorption and inner accumulation for playing outside are made
With.
This example demonstrates that living microorganism immobilized microspheres can play microbe carrier chemisorption and
Absorption-summation of living microorganism can be improved, it is common to provide technical guarantee effectively to administer heavy metal pollution of water body.
The above embodiment is a preferred embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present invention,
It should be equivalent substitute mode, be included within the scope of the present invention.
Claims (10)
1. a kind of preparation method of living microorganism immobilized microspheres, characterized by the following steps:
(1) microbial fixed carrier: after drying, crushing and sieving, limit oxygen is pyrolyzed, is cooling and ground rice straw
Sieve, obtains microbial fixed carrier；
(2) bacterium solution suspension colonizes: bacillus cereus (Bacillus cereus) RC-1 being activated, cell suspension is obtained；Take 5
Mixed serum A is made in~20mL bacterial suspension and the mixing of 0.2~1.5g microbial fixed carrier, 1~2h of shaken cultivation；
(3) sodium alginate soln of the mass fraction 2%~6% of sterilizing microbial activity particle: is added to mixed serum A
In, obtain mixed serum B；Then mixed serum B is added dropwise to the CaCl of sterilizing2In solution, solidify 2~10h, forms microorganism
Active particle；Wherein, dosage of the microbial fixed carrier in mixed serum B is 0.2~1.5g/100mL；
(4) proliferation is further cultured for: the microbial activity particle that step (3) is obtained increases in fresh beef cream peptone fluid nutrient medium
It grows and is further cultured for 1~4h, behind sterile water immersion treatment surface, obtain living microorganism immobilized microspheres；
Described bacillus cereus (Bacillus cereus) RC-1, was preserved in China Microbiological on 03 27th, 2017
Culture presevation administration committee common micro-organisms center, preservation address: section, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China
Institute of microbiology, institute, deposit number: CGMCC NO:13930.
2. the preparation method of living microorganism immobilized microspheres according to claim 1, it is characterised in that:
The preparation method of microbial fixed carrier described in step (1) includes the following steps: rice straw by natural wind
After doing and shredding grinding, it is placed in 300~700 DEG C of lower limit oxygen in pyrolysis oven and is pyrolyzed 2~4h, it is cooling to be simultaneously ground up, sieved 60~100
Mesh obtains microbial fixed carrier.
3. the preparation method of living microorganism immobilized microspheres according to claim 1, it is characterised in that:
The OD of cell suspension described in step (2)600=1.2~1.5；
The speed of instillation described in step (3) is 5~10mL/min；
CaCl described in step (3)2Solution is the CaCl that mass fraction is 2~10%2Solution；
The cured time described in step (3) is 2~5h.
4. the preparation method of living microorganism immobilized microspheres according to claim 3, it is characterised in that:
The dosage of bacterial suspension described in step (2) is 5mL；
The time of shaken cultivation described in step (2) is 2h；
The mass fraction of sodium alginate soln described in step (3) is 4%；
The speed of instillation described in step (3) is 7mL/min；
CaCl described in step (3)2Solution is the CaCl that mass fraction is 5%2Solution；
Dosage of the microbial fixed carrier in mixed serum B described in step (3) is 1.5g/100mL；
The cured time described in step (3) is 2h；
It is 4h that the time being further cultured for is proliferated described in step (4).
5. the preparation method of living microorganism immobilized microspheres according to claim 1,2,3 or 4, it is characterised in that:
The formula of beef extract-peptone fluid nutrient medium described in step (4) are as follows: 10g/L peptone, 3g/L beef extract, 5g/L
NaCl2, pH value is 7.0 ± 0.2.
6. a kind of living microorganism immobilized microspheres, it is characterised in that: pass through the described in any item preparation sides of Claims 1 to 5
Method is prepared.
7. living microorganism immobilized microspheres according to claim 6, it is characterised in that:
The living microorganism immobilized microspheres are in spherical, and there is a hole in inside, 3.0~4.0mm of diameter, and specific gravity 1.10~
1.20g/cm3, alkalescent；
The weakly alkaline pH range is 7.66~8.76.
8. application of the living microorganism immobilized microspheres in the improvement for heavy metal pollution described in claim 6 or 7.
9. application according to claim 8, it is characterised in that:
Application of the living microorganism immobilized microspheres in heavy metal pollution of water body improvement.
10. application according to claim 8 or claim 9, it is characterised in that:
The heavy metal is cadmium.
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