CN101475931B - Preparation of embedding immobilized effective microorganism gel bead - Google Patents

Preparation of embedding immobilized effective microorganism gel bead Download PDF

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CN101475931B
CN101475931B CN2009101109528A CN200910110952A CN101475931B CN 101475931 B CN101475931 B CN 101475931B CN 2009101109528 A CN2009101109528 A CN 2009101109528A CN 200910110952 A CN200910110952 A CN 200910110952A CN 101475931 B CN101475931 B CN 101475931B
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embedding
effective microorganism
bead
gel bead
gel
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CN101475931A (en
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熊小京
李博
郑天凌
赵志新
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厦门大学
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Abstract

A preparation method of embedding immobilized effective microorganism gel pellets relates to a microbial cell immobilization method for saprobe denitrification, specifically to a method taking polyvinyl alcohol and sodium alginate as the embedding agent, calcium chloride and boric acid as a cross linker and shell powder as additives to embed the effective microorganism. The invention provides a preparation method of embedding immobilized effective microorganism gel pellets which can not only effectively resolve the defects in the preparation and use process of the gel pellet, but also can effectively improve the biological denitrification effect by taking the effective microorganism as embedding thalli. The method includes: mixing uniformly the embedded agents and additives, dissolving in water, completely dissolving and mixing uniformly and then cooling to below 40 DEG C, mixing with the centrifuged effective microorganism concentrated bacterium liquor, stirring, and then adding into the cross-linker to form the gel pellets.

Description

A kind of preparation method of embedding immobilized effective microorganism gel bead
Technical field
The present invention relates to a kind of microbial cell immobilization method that is used for bio-denitrifying sewage, especially relating to a kind of employing polyvinyl alcohol and sodium alginate is embedding medium, with calcium chloride and boric acid is linking agent, with shell powder as additive, the method of embedding effective microbe (Effective Microorganisms is called for short EM).
Background technology
Microorganism embedded immobilization technology is newer technology in the biological wastewater treatment process field, and it is that microorganism is enclosed in natural macromolecule amylose class or the synthetic macromolecule gel, thereby makes microbial immobilized.Compare common activated sludge process, the embedded immobilization technology has the following advantages: prevent that microorganism is run off, can reach higher microorganism concn, antitoxin thing and impact load in the reactor, settling property is good, help solid-liquid separation etc.
The key problem in technology of entrapping method is to select suitable embedding medium, mainly contains agar, polyacrylamide (ACAM), gelatin, sodium alginate (CA) and chitin and polyvinyl alcohol (PVA) etc.There are some researches show, agar strength is relatively poor, polyacrylamide gel is toxic to biology, though the gelatin internal structure is closely knit, mass-transfer performance poor ([1] Duan Mingfeng, Wu Weixia, Xiao Junxia, Li Lianghong. utilize immobilized cell technology to handle waste water progress [J]. oil-gas field environment protection, 2004,14 (3): 17-20; [2] CarberyJB.Model of algalbacterial clay wastewater treatment system[J] .Water Science and Technology, 1992,26 (7-8)); By contrast, alginate calcium (CA) and polyvinyl alcohol (PVA) gel physical strength and mass-transfer performance are all better, to biological nontoxic, and anti-Biodegradable is good, be comparatively suitable fixed cell carrier ([3] Jiang Yuhong, Huang Xia, Yu Yuxin. the comparison of several fixed cell carriers [J]. environmental science, 2001,14 (2)).
Alginates is the intracellular product of brown algae, and it is made up of two kinds of dissimilar monose, promptly 1, and 4-beta-D-mannuronic acid (M) and 1,4-α-L-guluronic acid (G).Alginates can not form the thermal reversibility gel, normally adds divalent ion in its sodium salt solution, as Ca 2+, pass through Ca 2+Replace Na +Form gel network.The character of gel is relevant with the type (mainly being the proportion of composing of two kinds of monose) of the divalent cation of introducing and sodium alginate.Divalent cation has bigger influence to the physical strength of gel, and the second main group metal ion of the periodic table of elements is in the gel of sodium alginate salt formation, and its intensity order is as follows: Ba 2+>Sr 2+>Ca 2+>>Mg 2+Utilize sodium alginate gel immobilized microorganism method not only safety, fast, preparation is simple, reaction conditions is gentle, with low cost, and be applicable to immobilization ([4] Men Xuehu of most of microbe, Li Yanfeng, Zhou Lincheng. the preparation of polyvinyl alcohol carrier and applied research progress [J]. Gansu science journal, 2004,16 (3): 30-35).
The alginates method is compared with polyvinyl alcohol-borate method, though physical strength is better, mass-transfer performance is excellent, but absorption property is not as good as the polyvinyl alcohol gel ball, and polyvinyl alcohol-borate method list easily becomes flock with polyvinyl alcohol and the crosslinked balling-up difficulty of boric acid, and after the balling-up, the spheroid physical strength is relatively poor.Though the polyvinyl alcohol gel survival time is longer than alginates entrapping method, but polyvinyl alcohol gel is not sometimes because crosslinked thorough, a small amount of organic carbon composition stripping, and polyvinyl alcohol gel sclerosis required time is longer, the condition that immobilization requires comparatively complicated ([5] Liu Lei. the embedded material in the bio-immobilized technology [J]. water purification technology, 2005,24 (1)).
Polyvinyl alcohol-boric acid embedded immobilization method compares that other embedded immobilization method has physical strength height, the height of anti-the aeration intensity, anti-Biodegradable is good, cost is low and advantage such as long service life, but some problems also occurred in it is used.Polyvinyl alcohol is a kind of high viscosity substance, and should be anti-slower with boric acid, when colliding, drop can bond together, and progressively fused agglomerating, promptly polyethenol pellets has very strong agglomeration tendency, makes polyvinyl alcohol gel balling-up difficulty.When polyvinyl alcohol and boric acid react fixedly, 3 hydroxyls on the boric acid only partly react, also residual on the superpolymer gel that reaction back forms have hydrophilic-OH, make immobilized spherule in application, have very big water-swellable, growth along with duration of service, intensity weakens greatly, and is totally unfavorable in actual applications.Also there are some problems in the alginates embedding process, as the K that has high density +, Mg 2+, when phosphoric acid salt and other valent metal ions, the structure of calcium alginate gel can be damaged.In addition, because the pore dimension of calcium alginate gel network is too big, enzyme may leak from network, therefore the immobilization of uncomfortable synthase.
In addition, in embedding medium, add an amount of gac, CaCO 3Powder, Ca (OH) 2Powder, iron powder, SiO 2Powder can improve the character of gelled pill, the performances such as intensity of raising immobilized spherule as additive.
The biochemical process denitrogenation comprises 3 reaction process: aminating reaction, nitration reaction and anti-nitration reaction.In the municipal effluent secondary treatment yielding water, nitrogen mainly exists with the form of ammonia-state nitrogen, and then requiring to comprise nitrification in 3 grades of processing is main denitrification process.In addition, owing to all contain the ammonia nitrogen of higher concentration in the waste water of all conglomeraties such as iron and steel, oil refining, chemical fertilizer, inorganic chemical industry, iron alloy, glass manufacturing, meat processing and fodder production.Therefore nitration reaction is as the important step in the biochemical denitrification process, for the ammonia nitrogen of removing effectively in the waste water, subdues the ammonia nitrogen discharging pollution of water body is had great importance.
Nitration reaction divides two processes to finish under the effect of two kinds of autotrophic bacterias.At first by Nitrosomas ammonia nitrogen is converted into nitrite, and then by the nitrification bacterium nitrite-oxidizing is become nitrate, its reaction equation is as follows:
By above-mentioned reactive chemistry metering-type as can be known, under the competent condition of oxygen, the NH of every removal 1mol 4 +-N produces the H of 2mol +, the NH of promptly every removal 1g 4 +-N need consume the basicity of 7.14g (with CaCO 3Meter).
Because nitration reaction need consume a large amount of basicity, basicity deficiency in sewage, and ammonia nitrogen loading can make the pH of mixed value in the treatment unit reduce to below 6.5 when higher, nitration reaction speed is reduced and nitrifier is suppressed, so need carry out the basicity accounting during engineering design.At present, the mode of additional basicity mainly is to add NaHCO 3Or NaOH, this can increase the difficulty of running cost and management undoubtedly.
Shell is as a kind of marine products rubbish, and the generation in China's Coastal Areas every year is huge.For the treatment process of shell, generally take the form of air storage, the coastland environment has been caused immense pressure.Though shell is of a great variety, come in every shape, the color difference, chemical constitution is similar, mainly contains the lime carbonate and a spot of conchiolin that account for full shell 95%.It is reported, behind the product mussel shell dried and crushed powdered of Yantai, Shandong, survey its elemental composition with atomic absorption spectrophotometer, wherein major element K, Na, Ca, Mg massfraction are respectively: 0.01%, 0.35%, 15.1% and 0.17%, and micro-massfraction is respectively (mg/kg): Fe 206.0, Zn 453.3, Se 0.85, I 2.3, Cu 10.7.Other shell is because of the difference of originating, each massfraction slightly difference ([6] Li Jinzhi. Comprehensive Utilization of Shells [J]. Huaihai Institute of Technology journal .2001,10 (monographs): 22-23).Shell powder can dissolve gradually with the reduction of pH in waste water, for biological denitrification process provides basicity, can effectively improve the effect of Microbial denitrogenation.
The biological denitrificaion of waste water is in the actually operating operational process, to having relatively high expectations of processing condition, but many sewage works can not satisfy the requirement of bio-denitrifying sewage well to the setting of processing condition, some back yard industry sewage works particularly, because its water quality characteristics and operational condition are nonconforming, degraded to pollutent is difficult to reach emission request, trace it to its cause and mainly contain, anaerobism and aerobic section are provided with unreasonable, sludge concentration is low excessively in the Waste Water Treatment, and the too short and pH value of sludge age does not meet the denitrogenation requirement.
Effective microbe is than good a kind of multifunctional biological agent according to professor's husband development by Japanese Ryukyu university.It has comprised that mainly 10 of 5 big classes (the thread flora of photosynthetic bacteria group, lactobacillus, yeast flora, Gram positive actinomycetes group and fermentation system) belong to more than 80 kind of microorganism, make their symbiotic co-existences, form a strong functions colony, improve the physiological activity sexual function of object.There are some researches show that EM can effectively remove pollution substances such as COD in the waste water, nitrogen, phosphorus, and more traditional activated sludge process is more efficient.But because EM bacterium liquid is liquid, directly add when disposing of sewage, the EM flora lacks carrier, runs off with water outlet easily, makes it not high to the pollution substance clearance.The phase in epoch of nitrobacteria is long, and the enrichment and growth that this more is unfavorable for denitrifier among the EM is unfavorable for the removal of nitrogen.
Summary of the invention
The objective of the invention is in order to overcome in the embedding process, gelled pill difficult forming and sclerosis are slower, and gelled pill intensity after the embedding, work-ing life, the easy defective of water absorption and swelling, and the relatively poor difficult problem of wastewater biological denitrificaion effect, provide a kind of and can not only effectively solve gelled pill preparation and the shortcoming when using, and with effective microbe (EM) as the embedding thalline, can effectively improve the preparation method of the embedding immobilized effective microorganism gel bead of Nitrogen removal effect.
The technical solution adopted for the present invention to solve the technical problems is:
Embedding medium and additive are mixed, soluble in water, dissolve fully and mix after be cooled to below 40 ℃, concentrate bacterium liquid and mix with effective microbe (EM) after centrifugal, stir, add in the linking agent formation gelled pill then.
Described embedding medium is polyvinyl alcohol and sodium alginate, and additive is a shell powder, and linking agent is the saturated boric acid solution that contains calcium chloride.
The massfraction of described polyvinyl alcohol is preferably 10% (W/V Embedding medium), the massfraction of sodium alginate is preferably 1% (W/V Bag Bury agent), the massfraction of shell powder is preferably 0.2% (W/V Embedding medium), the massfraction that contains the calcium chloride in the saturated boric acid solution of calcium chloride is preferably 3% (W/V Linking agent).
Water temperature soluble in water is preferably 80~90 ℃.
Cross-linking agent solution is cooled to 4 ℃ earlier before using, and when making bead, uses magnetic stirrer.With the polyvinyl alcohol of this massfraction and sodium alginate as the saturated boric acid solution of embedding medium, this massfraction calcium chloride as linking agent, can solve the agglomeration problem of gelled pill preferably, strengthen the balling-up ability of gel, and keep excellent mass transferring performance.In line with the principle of the treatment of wastes with processes of wastes against one another, shell powder is joined in the embedding medium as additive, can improve every performance such as intensity, swelling property, work-ing life of gelled pill.This gelled pill also can constantly provide basicity owing to the dissolving of shell powder in the process of wastewater biological denitrificaion, the space that the dissolving back produces has also been created favourable condition for bead to the absorption of pollution substance in the waste water; While also provides an outlet for processing and the disposal of seashell.For solving the relatively poor difficult problem of Nitrogen removal effect, as the embedding bacterium, microorganism cells is fixed on gelled pill inside with EM, prevent that it runs off, and for denitrifying bacteria wherein provides a metastable anaerobic zone, the optimal efficacy of performance EM.
The invention has the beneficial effects as follows, strengthened the balling-up ability when gelled pill prepares, improved the intensity and the work-ing life of bead, and the Nitrogen removal effect of waste water is significantly improved, the while also provides a new way for the processing and the disposal of shell.
Description of drawings
The PVA bead denitrification effect of Fig. 1 embedding EM.In Fig. 1, X-coordinate be the cycle of operation/time, ordinate zou is NH 4 +-N clearance/%.
Fig. 2 water outlet NH 3-N concentration and turbidity be curve over time.In Fig. 2, X-coordinate is time/h, and left ordinate zou is water outlet NH 3-N concentration/mgL -1, right ordinate zou is delivery turbidity/NTU; ▲ be concentration curve, ■ is the turbidity curve.
The contrast of Fig. 3 different ingredients PVA bead denitrification effect.In Fig. 3, X-coordinate be the cycle of operation/time, ordinate zou is NH 3-N clearance/%; ▲ for shell, EM centrifugate are arranged; △ is no shell, EM centrifugate; is no shell, EM rejuvenation liquid.
The contrast of the different EM bacterium of Fig. 4 liquid embedding amount PVA bead denitrification effect.In Fig. 4, X-coordinate be the cycle of operation/time, ordinate zou is NH 3-N clearance/%; ▲ 10, ◆ 30, ■ 50, ● 80.
Embodiment
1.EM the preparation of bacterium liquid
The preparation of EM rejuvenation liquid: by volume, EM-1: molasses: the ratio preparation EM of water=3: 3: 94 enlarges liquid.The room temperature lower seal stores, and treats that the pH value drops to below 3.5 and after sour-sweet smell is arranged, promptly obtains EM rejuvenation liquid.
The preparation of EM centrifugate: with the centrifugal 20min of the speed of 5000r/min, abandoning supernatant is collected the concentrated thalline of EM of centrifuge tube bottom enrichment with the rapid EM rejuvenation liquid for preparing of previous step.The EM that every 10mL EM rejuvenation liquid obtains after centrifugal concentrates thalline and contains the bacterium amount that is equivalent to 10mL EM rejuvenation liquid.
2. the preparation of shell powder
With medicinal herb grinder oyster shells is pulverized, crossed 80 mesh sieves, promptly obtain shell powder.
3. the preparation of immobilized spherule
The preparation of embedding medium: be dissolved in after polyvinyl alcohol, sodium alginate and shell powder mixed in 80~90 ℃ the water.In the dissolution process, the vessel surface that holds is added a cover, and stirs frequently.After treating that it dissolves fully, be cooled to below 40 ℃, with 30% (V The corresponding EM rejuvenation liquid of bacterium amount/ V Embedding medium) EM concentrate thalline and mix, make the ultimate density of PVA, sodium alginate and shell powder be respectively 10% (W/V Embedding medium), 1% (W/V Embedding medium) and 0.2% (W/V Embedding medium).
The preparation of linking agent: with concentration is 3% (W/V Linking agent) CaCl 2Be dissolved in the saturated boric acid solution, be cooled to about 4 ℃.
Immobilization: the eminence of embedding medium from about 10cm that mixes splashed into the linking agent linking agent magnetic stirrer with syringe.It is bead about 3mm that embedding medium and linking agent are cross-linked to form diameter.Immobilized spherule is immersed in continues in the refrigerator that is stored in 4 ℃ in the linking agent about sclerosis 24h.Clean 3 times with physiological saline before using.
Fig. 1 provides the clearance of the SBR treatment system ammonia nitrogen water outlet that moves continuously through 8 cycles.
After the gelled pill (hereinafter to be referred as the A bead) of embedding EM is handled, ammonia nitrogen removal frank is totally in rising trend, move 8 all after date ammonia-N removal rate and rise to 86.3% by initial 10.5%, the microorganism of embedding is through after a while recovery with adapt to the degradation capability that shows gradually ammonia nitrogen in the bead.
The turbidity that the dissolving of gelled pill produces can exert an influence to the removal effect of ammonia nitrogen.The gelled pill of any microorganism of embedding does not concern over time the removal effect of ammonia nitrogen and delivery turbidity and sees Fig. 2.
After the absorption of 0.5h, ammonia nitrogen concentration drops to 38.4mg/L by 61.7mg/L to gelled pill in the water sample in the simulation ammonia nitrogen waste water, and turbidity rises to 43.47NTU by 1.43NTU.After 14h in, ammonia nitrogen concentration fluctuates in the 40-50mg/L scope all the time in the water sample; Turbidity is totally in rising trend, but lift velocity is slower, and the concentration of ammonia nitrogen also maintains in the more stable scope.Before turbidity influenced the water outlet ammonia nitrogen concentration, gelled pill was about 1-2mg (NH to the adsorptive power of ammonia nitrogen in the waste water 3-N)/g (gelled pill).Delivery turbidity begins obvious increase after vibration 14h, rises to 128 to the 24h turbidity, and ammonia nitrogen concentration drops to 2.77mg/L.This shows that gelled pill is dissolving to some extent along with the prolongation of duration of oscillation, the turbidity of water sample increases gradually, and ammonia nitrogen concentration then begins obvious reduction, and the concentration of ammonia nitrogen and turbidity have confidential relation in the water outlet.In sum, gelled pill has certain adsorption effect to the ammonia nitrogen in the waste water, and colloid that its dissolving produces also has adsorption to ammonia nitrogen, makes the gelled pill of embedding EM not to the removal of ammonia nitrogen unusual effect be arranged also.
In embedding medium, add a certain amount of shell powder, have or not help with intensity and the raising denitrification effect that increases gelled pill; Use EM centrifugate and rejuvenation liquid (consumption all is equivalent to 10mL EM rejuvenation liquid) to compare, with the contrast molasses to the proterties of gelled pill and embedding after the active issuable influence of EM.Prepared 4 kinds of different beads thus, the prescription of different gelled pills sees Table 1.
Table 1
In the preparation process of bead, A, C, D bead be globulate well, B bead shape irregular.Behind the sclerosis 24h, the hardness of A bead is significantly better than C, D bead, shows that shell powder in the preparation helps to improve the hardness of gelled pill; A, C bead outward appearance are pure white, and B, D bead outward appearance are light brown, show in the bead of embedding EM rejuvenation liquid to contain molasses.All break after the operation of B bead through 3 cycles, run off with water outlet.Through the continuous operation in 6 cycles, A, C, 3 kinds of different beads of D are seen Fig. 3 to the removal effect of ammonia nitrogen.
In the 1st cycle of operation; 3 kinds of gelled pills are not fine to the removal effect of ammonia nitrogen; adsorption based on bead; the A bead slightly is better than C, D bead; the D bead does not show better biological activity because containing molasses; illustrate that molasses did not significantly help microbic activity among the protection EM when bead prepared, also can in water, introduce other nutritive substance, and the physical indexs such as intensity of bead are caused negative influence.To the 6th cycle, the ammonia nitrogen removal frank of the A bead that effect is best has arrived 91.1%, illustrates that adding shell powder in embedding medium helps Microbial denitrogenation; The denitrification effect of C bead has also kept stability preferably, illustrates that embedding EM centrifugate is all favourable to the intensity and the nitrogen removal performance that improve gelled pill.3 kinds of beads have finally all reached more than 75% ammonia-N removal rate.
During operation was handled, all to a certain degree dissolving and the expansions of A, C, 3 kinds of gelled pills of D made the water outlet muddiness; The reactor water surface produces a large amount of foams, and the volumetric expansion of bead is original 2 times, and outward appearance is from initial white and the light brown canescence that progressively becomes.In the operation later stage, 3 kinds of beads all have a small amount of fragmentation, run off with water outlet.The toughness of A bead is better than C, D bead, and percentage of damage is lower, illustrates that shell powder can effectively improve physicalies such as the intensity of gelled pill and toughness.
One big advantage of microorganism embedded immobilization method is exactly to reach higher microorganism concn in reactor.Make the gelled pill of 4 kinds of different EM bacterium embedding amounts, add 10mL EM centrifugate in every 100mL embedding medium, the bacterium amount is equivalent to 10,30,50 respectively, 80mL EM rejuvenation liquid (following A, B, C, the D bead of abbreviating as respectively).Through the continuous operation in 9 cycles, the denitrification effect of 4 kinds of beads is seen Fig. 4.
In 3 initial cycles, the denitrification effect of D bead is best, and B, C bead are comparatively approaching, and A bead effect is relatively poor, and bigger to the denitrification effect influence in operation initial stage EM bacterium liquid embedding amount, the big more removal effect to ammonia nitrogen of bag bacterium amount is good more.Along with the increase of the cycle of operation, the denitrification effect of 4 kinds of beads moves closer to, and the denitrification effect of B bead is suitable with the D bead.To the operation later stage, the denitrification effect of D bead is variation gradually, and the effect of B bead is best, because the bag bacterium is measured big bead in use for some time because wherein microbial activities is comparatively remarkable, cause the gelled pill percentage of damage higher, microorganism concn reduces in the loss post-reactor, and clearance descends.As seen, from economy and treatment effect two aspects, the embedding amount of EM bacterium liquid is not The more the better, and (EM rejuvenation liquid bacterium amount)/100mL (embedding medium) is best proportioning to 30mL.

Claims (7)

1. the preparation method of an embedding immobilized effective microorganism gel bead, it is characterized in that embedding medium and additive are mixed, soluble in water, dissolve fully and mix after be cooled to below 40 ℃, mix with the concentrated bacterium liquid of effective microbe after centrifugal, stir, add then in the linking agent, form gelled pill;
Described embedding medium is polyvinyl alcohol and sodium alginate;
Described additive is a shell powder.
2. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 1 is characterized in that described linking agent is the saturated boric acid solution that contains calcium chloride.
3. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 1, the massfraction that it is characterized in that described polyvinyl alcohol is 10%W/V Embedding medium
4. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 1, the massfraction that it is characterized in that sodium alginate is 1%W/V Embedding medium
5. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 1, the massfraction that it is characterized in that shell powder is 0.2%W/V Embedding medium
6. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 2, the massfraction that it is characterized in that containing the calcium chloride in the saturated boric acid solution of calcium chloride is 3%W/V Linking agent
7. the preparation method of a kind of embedding immobilized effective microorganism gel bead as claimed in claim 1 is characterized in that water temperature soluble in water is 80~90 ℃.
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