CN106587378A - Magnetic composite paint flocculant based on chitosan - Google Patents

Abstract

The invention discloses a magnetic composite paint flocculant based on chitosan. The magnetic composite paint flocculant is prepared through the following steps of 1 strain separating and purifying, 2 strain screening, 3 strain fermenting culturing and 4 composite magnetic microsphere preparing. The magnetic composite paint flocculant has a good deslagging effect on paint wastewater, is high in flocculation efficiency, can achieve water purification and sterilization, does not corrode water circulating equipment and is wide in raw material source, low in cost, capable of easily achieving large-scale application and popularization and worthy of popularization.

Description

A kind of magnetic coupling Paint flocculant based on shitosan

Technical field

The present invention relates to Paint flocculant technical field, more particularly to a kind of magnetic coupling paint flocculation based on shitosan Agent.

Background technology

With the development of the industries such as automobile industry and furniture manufacturing industry, the consumption of spray paint is more and more, therefore has The coating cloud that much can not be coated on workpiece just becomes " cross and spray " coating, crosses discharge rate and reaches 25-60%.Containing big in these coating Amount volatile organic solvent, endanger the health of the mankind, but also the environment and air around severe contamination.Spraying is to apply Paint is uniformly dispersed into tiny granule, most by spray gun or dish-style nebulizer under pressure or centrifugal action by material Coating cloud granule is just applied to the process of body surface afterwards.The coating cloud for crossing spray normally enters recirculated water, by the tank of indoor and outdoor Take recirculated water out of outdoor, thus staff's body harm can be reduced, and water can be recycled." cross spray " coating by Just very big in the viscosity of itself, therefore will stick on the wall of tank, water pipe and water pump, Jing accumulates over a long period, will be formed compared with Big paint block causes line clogging, thus the work efficiency of blood circulation will be reduced.Mechanical curettage method, chemical agent absorption are solidifying Poly- partition method and biology say that solution etc. is the most frequently used several method of spray painting room processing cycle water, wherein being being widely used Learn coacervation.Due to the long-term addition of chemical type flocculating agent, in causing recirculated water, content of organics is progressively accumulated, and COD progressively rises Height, water quality are deteriorated therewith, cause the recirculated water replacement cycle to shorten, and the sediment in waste water resource, and circulation is too many, Solid waste amount is big, and processes now solid waste mainly by the method burned and fill, therefore increased the pollution to environment, non- But the pollution problem of paint slag is fundamentally solved, the burden of environment is also add, therefore is researched and developed green non-poisonous dirt The little flocculant of dye, not only can be greatly lowered solid waste amount, and can be with reduces cost.

Microbial flocculant is biodegradable by the safety non-toxic of output in various microorganism growth process Natural product, it can make flocculation, the precipitations such as somatic cells in water, particle and colloidal particle, with good ring The border compatibility.Yang Shuanshuan exists《Flocculation of the microbial flocculant to paint waste water is studied》Pass through a series of bolter in one text Choosing, obtains one plant of preferably flocculation bacterial strain FL-2, by carrying out single factor experiment to bacterial strain FL-2, it is determined that optimal condition of culture 30 DEG C, pH is 9, shaking speed is 160r/min, in the composition of culture medium charcoal source be glucose, mixed nitrogen be sodium nitrate, Ammonium chloride and carbamide, inorganic salt are magnesium sulfate, and good flocculating effect is reached when compounding with bacillusmusilaginosiengineering, but Effect when two kinds of microbial bacterias are compounded with inorganic flocculating agent is bad.A series of bacterium for above being obtained by separation screening steps Strain FL -2 have good flocculating effect, but from from Large-scale culture conditions, relatively costly, needs find and can substitute Charcoal source and the culture medium of nitrogen source culture, reduces cost, are advantageously implemented economic benefit, and control are needed when paint waste water is processed Amount and the addition sequence of compounding strain that system is added, cause complex steps, improve cost, can by bio-immobilized technology by Strain is fixed on carrier, is realized continuous operation, is reduced the purpose of production cost, reaches economic benefit and environmental benefit doulbe-sides' victory Effect.

The content of the invention

The object of the invention is exactly for the defect for making up prior art, there is provided a kind of magnetic coupling based on shitosan is painted Flocculant.

The present invention is achieved by the following technical solutions：

A kind of magnetic coupling Paint flocculant based on shitosan, comprises the following steps：

（1）Strain separating purification

The paint waste water for processing later stage is loaded in aseptic triangular flask, 4 DEG C of preservations are placed in, seed culture is based on into 121 DEG C Lower sterilizing, takes 100 μ L waste water and is inoculated in the seed culture medium of 100ml after cooling, in 37 DEG C of constant temperature amplification culture 24h, then Culture fluid is diluted to 10-1,10-2,10-3,10-4,10-5,10-6 times, then the bacteria suspension coating by each after diluting respectively On isolation medium, 37 DEG C of constant temperature culture 24h, good bacterium colony of choosing form complete initial gross separation, then will be singled out each Bacterium is inoculated in seed culture medium, 37 DEG C of constant temperature culture 24h, line is diluted and carried out to the culture fluid of each bacterium and is isolated and purified, The purification single bacterium colony for finally obtaining is stored on isolation medium；

（2）Bacteria selection

By step（1）The bacterium separated is inoculated in seed culture medium with inoculating loop respectively, at 30 DEG C, the condition of 160r/min Lower culture 72h, tests the flocculating rate of strain with the aqueous suspension ofkaolin of 4g/L, and strain of the flocculating rate more than 50% is retained, and Repeatedly passed on, by the culture presevation for filtering out；

（3）The fermentation culture of strain

A, beer waste water and sugaring fermentation waste water are centrifuged into 20min respectively under 5000rpm, remove suspended impurity, wherein refining sugar Fermentation waste water total sugar content is 19.85g/L, and total nitrogen content is 3263.3mg/L, and beer waste water total sugar content is 196.1g/L, always Nitrogen content is 1796.8mg/L, then according to total sugar concentration 7g/L, charcoal source and nitrogen source compare 50:1 is diluted, and adds phosphoric acid Sodium dihydrogen and disodium hydrogen phosphate, wherein phosphate dihydrogen sodium concentration are 2g/L, and disodium hydrogen phosphate concentration is 5g/L, adds magnesium sulfate The pH for adjusting fermentation medium is 9, and autoclaving is standby；

B, by step（2）In be deposited in the producing strains strain on inclined-plane and be inoculated in fermentation medium, in 30 DEG C, 150r/min constant temperature 3h is cultivated on agitator, is made each bacterial strain be in exponential phase, is made strain suspension, be inoculated with according to the inoculum concentration of 5-10% In the fermentation medium prepared to 100ml steps a, under the conditions of being 160r/min in 30 DEG C, rotating speed, 72h is cultivated；

C, fermentation liquid obtained in step b is placed on 4 DEG C of High speed refrigerated centrifuges in 8000r/min be centrifuged 30-40min, will To supernatant in add 2 times amount dehydrated alcohol stand 24h at 4 DEG C, then proceed under the conditions of 4 DEG C in 8000r/min Centrifugation 30-40min, by the precipitation for obtaining dehydrated alcohol cyclic washing, is placed in bag filter the 48h that dialyses, after lyophilization Microbial flocculant；

(4) preparation of composite magnetic microsphere

D, carrageenan is added in deionized water the solution for making that concentration is 5g/L, heated and stirred is cooled to until be completely dissolved The ferric chloride solution and ferrous sulfate of 0.2mol/L is added when 40 DEG C and 15-25min is stirred, appropriate 1mol/L hydrogen is subsequently adding Sodium hydroxide solution, regulation solution ph are 10.5-11.5, are filtered, and precipitation is standby in neutral post-drying with distilled water wash；

E, shitosan is added in the acetic acid solution that volume fraction is 1%, stirring at normal temperature is until be completely dissolved into volume fraction 0.1% Chitosan solution, by tourmaline powder be diluted with distilled water into mass concentration be 2-2.5% solution, be added to above-mentioned shitosan molten In liquid, magnetic agitation 0.4-0.6h then proceedes to 25% glutaraldehyde solution is slowly added under stirring, after room temperature ageing 24h Microbial flocculant prepared by step c is added to continue magnetic agitation 1-1.5h, the pH value for adjusting mixture is 4.8-5.8, then plus Enter the product of step d preparation in 40 DEG C of insulation reaction 2-3h, reaction terminates rear Magneto separate, product dehydrated alcohol and distillation washing Wash several times, be obtained after being dried.

The formulation ratio of described isolation medium is：Carnis Bovis seu Bubali cream 3-4g/L, peptone 10-12g/L, Sodium Chloride 5-6g/ L, agar 15-20g/L, plus distilled water to 1000ml；PH value is adjusted to 7.0-7.2, sterilize 20min in 121 DEG C；Seed culture medium Recipe ratio is：Carnis Bovis seu Bubali cream 3-4g/L, peptone 10-13g/L, Sodium Chloride 5-7g/L, plus distilled water to 1000ml, adjust pH value to 7-7.2, sterilize 20min in 121 DEG C.

The step（4）In each raw material according to weight be：Iron chloride 8.1-8.6, ferrous sulfate 0.4-0.6, card Draw glue 0.6-0.8, shitosan 5-5.6, tourmaline powder 0.4-0.6,25% glutaraldehyde 0.2-0.3, microbial flocculant 1.8-2.2, 1mol/L Sodium hydroxide q. s, appropriate dehydrated alcohol, appropriate acetic acid, distillation appropriate amount of water.

It is an advantage of the invention that：The present invention from paint waste water screening, extracting and developing, be purified into microbial flocculant produce Raw bacterium, optimization culture improve its flocculation activity, and the production of microbial flocculant is reduced using cheap substrate fermentation culture Cost so as to can large-scale production culture, and the resource to discarding reuses, with good economic benefit and Social benefit, environmental protection is that all many conditions when improving the single use of microbial flocculant or compounding use are limited, the present invention Which is made to possess good absorption and magnetic behavior using the magnetic microsphere loading microorganisms flocculant for preparing, and it is easily separated, can Repetitive cycling is utilized, and carrageenan is a kind of widely distributed linear polysaccharide with sulfate group in nature, due to special Chemical constitution, strong charging property is widely used in gellant, thickening agent and emulsifying agent, shitosan be it is a kind of extensively point The Natural polycations linear polysaccharide of cloth, due to amino, hydroxyl and acetyl group, more as albumen, dyestuff and metal from The adsorbent of son, the present invention can pass through molecular entanglement, ionic bond, hydrogen bond using the contrary electric charge of carrageenan and shitosan band Or hydrophobic forces form complex, shitosan forms hydrogel cladding microbial flocculant in the presence of cross-linking agent, then It is incorporated on carrageenan magnetic nano-particle by Electrostatic Absorption, Stability Analysis of Structures and the flocculation of microorganism can be effectively improved Rate, and also the effect that can be purified water and sterilize, are also capable of achieving sharp separation and recovery, flocculant pair prepared by the present invention Paint waste water slagging-off effect is good, and flocculation efficiency is fast, purifies water and sterilizes, corrosion-free to water circulating apparatus, and raw material sources are wide, into This is low, easily realizes large-scale promotion application, is worthy to be popularized.

Specific embodiment

A kind of magnetic coupling Paint flocculant based on shitosan, comprises the following steps：

（1）Strain separating purification

The paint waste water for processing later stage is loaded in aseptic triangular flask, 4 DEG C of preservations are placed in, seed culture is based on into 121 DEG C Lower sterilizing, takes 100 μ L waste water and is inoculated in the seed culture medium of 100ml after cooling, in 37 DEG C of constant temperature amplification culture 24h, then Culture fluid is diluted to 10,10,10,10,10,10 times respectively, then each bacteria suspension after diluting is coated into isolation medium On, 37 DEG C of constant temperature culture 24h, good bacterium colony completes initial gross separation to choose form, then each bacterium that will be singled out is inoculated in seed In culture medium, 37 DEG C of constant temperature culture 24h dilute and carry out line and isolate and purify, by what is finally obtained to the culture fluid of each bacterium Purification single bacterium colony is stored on isolation medium；

（2）Bacteria selection

By step（1）The bacterium separated is inoculated in seed culture medium with inoculating loop respectively, at 30 DEG C, the condition of 160r/min Lower culture 72h, tests the flocculating rate of strain with the aqueous suspension ofkaolin of 4g/L, and strain of the flocculating rate more than 50% is retained, and Repeatedly passed on, by the culture presevation for filtering out；

（3）The fermentation culture of strain

A, beer waste water and sugaring fermentation waste water are centrifuged into 20min respectively under 5000rpm, remove suspended impurity, wherein refining sugar Fermentation waste water total sugar content is 19.85g/L, and total nitrogen content is 3263.3mg/L, and beer waste water total sugar content is 196.1g/L, always Nitrogen content is 1796.8mg/L, then according to total sugar concentration 7g/L, charcoal source and nitrogen source compare 50:1 is diluted, and adds phosphoric acid Sodium dihydrogen and disodium hydrogen phosphate, wherein phosphate dihydrogen sodium concentration are 2g/L, and disodium hydrogen phosphate concentration is 5g/L, adds magnesium sulfate The pH for adjusting fermentation medium is 9, and autoclaving is standby；

B, by step（2）In be deposited in the producing strains strain on inclined-plane and be inoculated in fermentation medium, in 30 DEG C, 150r/min constant temperature 3h is cultivated on agitator, is made each bacterial strain be in exponential phase, is made strain suspension, be inoculated into according to 5% inoculum concentration In fermentation medium prepared by 100ml steps a, under the conditions of being 160r/min in 30 DEG C, rotating speed, 72h is cultivated；

C, fermentation liquid obtained in step b is placed on 4 DEG C of High speed refrigerated centrifuges in 8000r/min be centrifuged 30min, by what is obtained Add the dehydrated alcohol of 2 times of amounts 24h to be stood at 4 DEG C in supernatant, then proceed to be centrifuged in 8000r/min under the conditions of 4 DEG C 30min, by the precipitation for obtaining dehydrated alcohol cyclic washing, is placed in bag filter the 48h that dialyses, and microorganism wadding is obtained after lyophilization Solidifying agent；

(4) preparation of composite magnetic microsphere

D, carrageenan is added in deionized water the solution for making that concentration is 5g/L, heated and stirred is cooled to until be completely dissolved The ferric chloride solution and ferrous sulfate of 0.2mol/L is added when 40 DEG C and 15min is stirred, appropriate 1mol/L hydrogen-oxygens are subsequently adding Change sodium solution, it is 10.5 to adjust solution ph, is filtered, and precipitation is standby in neutral post-drying with distilled water wash；

E, shitosan is added in the acetic acid solution that volume fraction is 1%, stirring at normal temperature is until be completely dissolved into volume fraction 0.1% Chitosan solution, tourmaline powder is diluted with distilled water into into the solution that mass concentration is 2%, is added in above-mentioned chitosan solution, Magnetic agitation 0.4h, then proceedes to 25% glutaraldehyde solution is slowly added under stirring, adds step c after room temperature ageing 24h The microbial flocculant of preparation continues magnetic agitation 1h, and the pH value for adjusting mixture is 4.8, adds the product of step d preparation In 40 DEG C of insulation reaction 2h, reaction terminates rear Magneto separate, and product dehydrated alcohol and distilled water wash several times, can be obtained after being dried Arrive.

The formulation ratio of described isolation medium is：Carnis Bovis seu Bubali cream 3g/L, peptone 10g/L, Sodium Chloride 5g/L, agar 15g/L, plus distilled water to 1000ml；PH value is adjusted to 7.0, sterilize 20min in 121 DEG C；Seed culture medium recipe ratio is：Beef Cream 3g/L, peptone 10g/L, Sodium Chloride 5g/L, plus distilled water to 1000ml, adjust pH value to 7, sterilize 20min in 121 DEG C.

The step（4）In each raw material according to weight portion（Kilogram）Proportioning is：Iron chloride 8.1, ferrous sulfate 0.4, OK a karaoke club Glue 0.6, shitosan 5, tourmaline powder 0.4,25% glutaraldehyde 0.2, microbial flocculant 1.8,1mol/L Sodium hydroxide q. s, nothing Appropriate water-ethanol, appropriate acetic acid, distillation appropriate amount of water.

In order to embody the effectiveness of the performance of the Compound paint flocculant prepared in embodiment, configuration paint waste water enters Row performance verification, process are as follows：

100ml cold running waters are added in the open bottles equipped with magnetic stirring bar, and it is 12 pH to be adjusted with the sodium hydroxide of 1mol/L, Turn on agitator adds the commercially available automotive paints of 6 drops, stands and obtain paint waste water after stirring, and concentration is about 30mg/L, to The Compound paint flocculant of waste water total amount mass fraction 0.3%, high-speed stirred 1min, observation knot after standing are added in paint waste water Really.It has been observed that：Bottle wall does not have obvious bur, and water quality is limpid, and the floccule of formation is compact in cellular and to bubble through the water column, Flocculating rate is 92.8%.

Claims (3)

1. a kind of magnetic coupling Paint flocculant based on shitosan, it is characterised in that comprise the following steps：

（1）Strain separating purification

The paint waste water for processing later stage is loaded in aseptic triangular flask, 4 DEG C of preservations are placed in, seed culture is based on into 121 DEG C Lower sterilizing, takes 100 μ L waste water and is inoculated in the seed culture medium of 100ml after cooling, in 37 DEG C of constant temperature amplification culture 24h, then Culture fluid is diluted to 10-1,10-2,10-3,10-4,10-5,10-6 times, then the bacteria suspension coating by each after diluting respectively On isolation medium, 37 DEG C of constant temperature culture 24h, good bacterium colony of choosing form complete initial gross separation, then will be singled out each Bacterium is inoculated in seed culture medium, 37 DEG C of constant temperature culture 24h, line is diluted and carried out to the culture fluid of each bacterium and is isolated and purified, The purification single bacterium colony for finally obtaining is stored on isolation medium；

（2）Bacteria selection

By step（1）The bacterium separated is inoculated in seed culture medium with inoculating loop respectively, at 30 DEG C, the condition of 160r/min Lower culture 72h, tests the flocculating rate of strain with the aqueous suspension ofkaolin of 4g/L, and strain of the flocculating rate more than 50% is retained, and Repeatedly passed on, by the culture presevation for filtering out；

（3）The fermentation culture of strain

A, beer waste water and sugaring fermentation waste water are centrifuged into 20min respectively under 5000rpm, remove suspended impurity, wherein refining sugar Fermentation waste water total sugar content is 19.85g/L, and total nitrogen content is 3263.3mg/L, and beer waste water total sugar content is 196.1g/L, always Nitrogen content is 1796.8mg/L, then according to total sugar concentration 7g/L, charcoal source and nitrogen source compare 50:1 is diluted, and adds phosphoric acid Sodium dihydrogen and disodium hydrogen phosphate, wherein phosphate dihydrogen sodium concentration are 2g/L, and disodium hydrogen phosphate concentration is 5g/L, adds magnesium sulfate The pH for adjusting fermentation medium is 9, and autoclaving is standby；

B, by step（2）In be deposited in the producing strains strain on inclined-plane and be inoculated in fermentation medium, in 30 DEG C, 150r/min constant temperature 3h is cultivated on agitator, is made each bacterial strain be in exponential phase, is made strain suspension, be inoculated with according to the inoculum concentration of 5-10% In the fermentation medium prepared to 100ml steps a, under the conditions of being 160r/min in 30 DEG C, rotating speed, 72h is cultivated；

C, fermentation liquid obtained in step b is placed on 4 DEG C of High speed refrigerated centrifuges in 8000r/min be centrifuged 30-40min, will To supernatant in add 2 times amount dehydrated alcohol stand 24h at 4 DEG C, then proceed under the conditions of 4 DEG C in 8000r/min Centrifugation 30-40min, by the precipitation for obtaining dehydrated alcohol cyclic washing, is placed in bag filter the 48h that dialyses, after lyophilization Microbial flocculant；

(4) preparation of composite magnetic microsphere

D, carrageenan is added in deionized water the solution for making that concentration is 5g/L, heated and stirred is cooled to until be completely dissolved The ferric chloride solution and ferrous sulfate of 0.2mol/L is added when 40 DEG C and 15-25min is stirred, appropriate 1mol/L hydrogen is subsequently adding Sodium hydroxide solution, regulation solution ph are 10.5-11.5, are filtered, and precipitation is standby in neutral post-drying with distilled water wash；

E, shitosan is added in the acetic acid solution that volume fraction is 1%, stirring at normal temperature is until be completely dissolved into volume fraction 0.1% Chitosan solution, by tourmaline powder be diluted with distilled water into mass concentration be 2-2.5% solution, be added to above-mentioned shitosan molten In liquid, magnetic agitation 0.4-0.6h then proceedes to 25% glutaraldehyde solution is slowly added under stirring, after room temperature ageing 24h Microbial flocculant prepared by step c is added to continue magnetic agitation 1-1.5h, the pH value for adjusting mixture is 4.8-5.8, then plus Enter the product of step d preparation in 40 DEG C of insulation reaction 2-3h, reaction terminates rear Magneto separate, product dehydrated alcohol and distillation washing Wash several times, be obtained after being dried.

2. a kind of magnetic coupling Paint flocculant based on shitosan according to claim 1, it is characterised in that described point Formulation ratio from culture medium is：Carnis Bovis seu Bubali cream 3-4g/L, peptone 10-12g/L, Sodium Chloride 5-6g/L, agar 15-20g/L, plus Distilled water is to 1000ml；PH value is adjusted to 7.0-7.2, sterilize 20min in 121 DEG C；Seed culture medium recipe ratio is：Carnis Bovis seu Bubali cream 3- 4g/L, peptone 10-13g/L, Sodium Chloride 5-7g/L, plus distilled water to 1000ml, adjust pH value to 7-7.2, sterilize in 121 DEG C 20min。

3. a kind of magnetic coupling Paint flocculant based on shitosan according to claim 1, it is characterised in that the step （4）In each raw material according to weight be：Iron chloride 8.1-8.6, ferrous sulfate 0.4-0.6, carrageenan 0.6-0.8, shell gather Sugared 5-5.6, tourmaline powder 0.4-0.6,25% glutaraldehyde 0.2-0.3, microbial flocculant 1.8-2.2,1mol/L sodium hydroxide are fitted Amount, appropriate dehydrated alcohol, appropriate acetic acid, distillation appropriate amount of water.