CN107381834A - A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite - Google Patents
A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite Download PDFInfo
- Publication number
- CN107381834A CN107381834A CN201710735139.4A CN201710735139A CN107381834A CN 107381834 A CN107381834 A CN 107381834A CN 201710735139 A CN201710735139 A CN 201710735139A CN 107381834 A CN107381834 A CN 107381834A
- Authority
- CN
- China
- Prior art keywords
- nitrite
- immobilised enzymes
- filter plate
- water body
- reductase
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 71
- 102000004190 Enzymes Human genes 0.000 title claims abstract description 71
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 title claims abstract description 39
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 39
- 238000009395 breeding Methods 0.000 title claims abstract description 24
- 230000001488 breeding effect Effects 0.000 title claims abstract description 24
- 238000006243 chemical reaction Methods 0.000 title claims abstract description 17
- 238000012545 processing Methods 0.000 title claims abstract description 10
- 108010025915 Nitrite Reductases Proteins 0.000 claims abstract description 33
- 229920005830 Polyurethane Foam Polymers 0.000 claims abstract description 30
- 239000011496 polyurethane foam Substances 0.000 claims abstract description 30
- 239000000126 substance Substances 0.000 claims abstract description 27
- 230000004888 barrier function Effects 0.000 claims abstract description 5
- 229940088598 enzyme Drugs 0.000 claims description 66
- 239000007788 liquid Substances 0.000 claims description 19
- 239000000243 solution Substances 0.000 claims description 18
- 230000001939 inductive effect Effects 0.000 claims description 14
- 108090000854 Oxidoreductases Proteins 0.000 claims description 12
- 230000003068 static effect Effects 0.000 claims description 12
- 230000006698 induction Effects 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 8
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 8
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 8
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 8
- 102000016943 Muramidase Human genes 0.000 claims description 7
- 108010014251 Muramidase Proteins 0.000 claims description 7
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 claims description 7
- 239000004372 Polyvinyl alcohol Substances 0.000 claims description 7
- 230000004069 differentiation Effects 0.000 claims description 7
- 229960000274 lysozyme Drugs 0.000 claims description 7
- 239000004325 lysozyme Substances 0.000 claims description 7
- 235000010335 lysozyme Nutrition 0.000 claims description 7
- 229920002451 polyvinyl alcohol Polymers 0.000 claims description 7
- 241000894006 Bacteria Species 0.000 claims description 6
- IOVCWXUNBOPUCH-UHFFFAOYSA-N Nitrous acid Chemical compound ON=O IOVCWXUNBOPUCH-UHFFFAOYSA-N 0.000 claims description 6
- 239000000835 fiber Substances 0.000 claims description 6
- 239000000843 powder Substances 0.000 claims description 6
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical group CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 5
- 239000011521 glass Substances 0.000 claims description 5
- 238000002360 preparation method Methods 0.000 claims description 5
- 235000010413 sodium alginate Nutrition 0.000 claims description 5
- 239000000661 sodium alginate Substances 0.000 claims description 5
- 229940005550 sodium alginate Drugs 0.000 claims description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 229940122618 Trypsin inhibitor Drugs 0.000 claims description 4
- 101710162629 Trypsin inhibitor Proteins 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 4
- 238000004132 cross linking Methods 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 4
- 239000002504 physiological saline solution Substances 0.000 claims description 4
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Substances [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 4
- 238000001179 sorption measurement Methods 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 239000002753 trypsin inhibitor Substances 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 3
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 238000003780 insertion Methods 0.000 claims description 2
- 230000037431 insertion Effects 0.000 claims description 2
- 239000007787 solid Substances 0.000 claims 1
- 239000002351 wastewater Substances 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 abstract description 2
- 238000005265 energy consumption Methods 0.000 abstract description 2
- 239000001301 oxygen Substances 0.000 abstract description 2
- 229910052760 oxygen Inorganic materials 0.000 abstract description 2
- 241000196324 Embryophyta Species 0.000 description 10
- 244000005700 microbiome Species 0.000 description 9
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- 238000009360 aquaculture Methods 0.000 description 4
- 244000144974 aquaculture Species 0.000 description 4
- 230000015556 catabolic process Effects 0.000 description 4
- 238000006731 degradation reaction Methods 0.000 description 4
- 239000006260 foam Substances 0.000 description 4
- GQPLMRYTRLFLPF-UHFFFAOYSA-N nitrous oxide Inorganic materials [O-][N+]#N GQPLMRYTRLFLPF-UHFFFAOYSA-N 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- 241000251468 Actinopterygii Species 0.000 description 3
- 239000011324 bead Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 2
- 108090000913 Nitrate Reductases Proteins 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003344 environmental pollutant Substances 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 235000015110 jellies Nutrition 0.000 description 2
- 239000008274 jelly Substances 0.000 description 2
- 231100000719 pollutant Toxicity 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012549 training Methods 0.000 description 2
- 238000009777 vacuum freeze-drying Methods 0.000 description 2
- PAWQVTBBRAZDMG-UHFFFAOYSA-N 2-(3-bromo-2-fluorophenyl)acetic acid Chemical compound OC(=O)CC1=CC=CC(Br)=C1F PAWQVTBBRAZDMG-UHFFFAOYSA-N 0.000 description 1
- 241000272525 Anas platyrhynchos Species 0.000 description 1
- 241000193755 Bacillus cereus Species 0.000 description 1
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 1
- 235000017491 Bambusa tulda Nutrition 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 101000841301 Homo sapiens Utrophin Proteins 0.000 description 1
- 208000005016 Intestinal Neoplasms Diseases 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 206010068052 Mosaicism Diseases 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 241001597008 Nomeidae Species 0.000 description 1
- 244000082204 Phyllostachys viridis Species 0.000 description 1
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 1
- 208000005718 Stomach Neoplasms Diseases 0.000 description 1
- 102100029092 Utrophin Human genes 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229960000892 attapulgite Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 239000011425 bamboo Substances 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000011449 brick Substances 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000021121 fermented vegetables Nutrition 0.000 description 1
- 238000007667 floating Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 206010017758 gastric cancer Diseases 0.000 description 1
- 201000002313 intestinal cancer Diseases 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- XKLJHFLUAHKGGU-UHFFFAOYSA-N nitrous amide Chemical compound ON=N XKLJHFLUAHKGGU-UHFFFAOYSA-N 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229910052625 palygorskite Inorganic materials 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 210000003765 sex chromosome Anatomy 0.000 description 1
- 201000011549 stomach cancer Diseases 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
-
- 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
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2203/00—Apparatus and plants for the biological treatment of water, waste water or sewage
- C02F2203/006—Apparatus and plants for the biological treatment of water, waste water or sewage details of construction, e.g. specially adapted seals, modules, connections
Abstract
A kind of application the invention discloses immobilised enzymes biological reaction apparatus and its in processing breeding water body nitrite, the device is rectangular box, including left and right ends face and front and rear four tank walls up and down, both ends of the surface set water inlet and delivery port respectively, some immobilised enzymes filter plates are vertically arranged in device interior edge water (flow) direction, the immobilised enzymes filter plate is by netting the fixed filter plate formed by polyurethane foam suspended substance；Barrier layer is provided between the immobilised enzymes filter plate.Nitrite reductase is fixed on polyurethane foam suspended substance by invention, directly can efficiently handle nitrite breeding water body, can prevent nitrite reductase from being broken up by waste water to take away；Reduce the content of oxygen in breeding water body using the physics water seal effect of nitrite breeding water body current simultaneously, so as to substantially increase the reaction efficiency of nitrite reductase.The plant investment is few, and floor space is small, simple to operate, has the advantages of low-energy-consumption high-efficiency, is adapted to industrialized production.
Description
Technical field
The invention belongs to aquaculture and field of environment protection, and in particular to a kind of immobilised enzymes biological reaction apparatus and its
In the application of processing breeding water body nitrite.
Background technology
The problem of water pollution is whole world extensive concern.In the prevalence of nitrous acid in the environment and food of human lives
Salt.In recent years, nitrite breeding water body has caused the attention of field of Environment Protection and global range, many studies have shown that nitrite
The precursor of inferior ammonium nitrate, nitrosamine is a kind of strong carcinogen, can induce a variety of cancerations in digestive system, as stomach cancer,
Intestinal cancer and liver cancer.In view of aquaculture system nitrite is exceeded, and normal breeding water body nitrite is usually no more than
0.1mg/L is advisable, and after nitrite accumulation is too high, will suppress the oxygen carrying capability of Fish Blood, cause fish slow poisoning,
The reduction of fish amount, gill tissue is caused lesion occur, expiratory dyspnea, disturbance are uneasy, and fulminant death then occurs when serious, seeks
Look for the effectively method of control or degrading nitrite imperative.Carry out in terms of domestic and international nitrite water body processing more
Research.Especially most of researcher obtains larger progress, nitrous in enzymatic treatment nitrite breeding water body research
Nitrite-oxidizing can be reduced to nitrogen by hydrochlorate reductase as catalyst." aquaculture system is in situ for Chinese invention patent
Purification complex microorganism film and its application method (CN201310172159.7) " discloses a kind of aquaculture system purified in situ
Complex microorganism film and its application method, its drip irrigation device are carrier selection, bacterium solution preparation, and microbial immobilized wait walks
Suddenly, complex microorganism film is arranged in breeding water body, beneficial microbe starts activation procreation.Although the immobilized microorganism film
It is simple in construction, but complex microorganism film device is highly prone to the influence of external environment, is unfavorable for mass producing.
Enzymatic treatment nitrite is also paid attention at present, such as a kind of Chinese invention patent " Lactobacillus casei rhamnose
The preparation method (CN201410437877.7) of the nitrite reductase of subspecies " and " a kind of bacillus cereus and its making
Application (CN201410390980.0) in standby nitrite reductase " discloses a kind of side for preparing nitrite reductase
Method, obtained NiR purity is high, and NiR is to solve in 3 fields such as fermented vegetable, meat products and breeding water body containing nitrite
Problem provides a kind of new method.Easily operated and the advantages that can be recycled although NiR preparation conditions are simple, enzyme is consolidated
Fixedization and enzyme adaptation sex chromosome mosaicism are so as to causing to be unable to industrial treatment nitrite breeding water body.And in the form of immobilised enzymes
Grain bead accounts for the overwhelming majority, but bead bag bacterium amount obtained by immobilized spherule method is low, small volume, is easily washed etc. and can not meet Asia
The actual treatment situation of nitrate breeding water body.
The content of the invention
To solve the shortcomings that prior art and weak point, primary and foremost purpose of the invention is to provide a kind of immobilised enzymes life
Thing reaction unit and its application in processing breeding water body nitrite.Using polyurethane foam suspended substance as nitrite
The fixation support of reductase, it is further fixed with fishing net, multilayer class fishing net formula immobilised enzymes polyurethane foam suspended substance
Ordered arrangement forms a set of immobilised enzymes biological reaction apparatus, using polyurethane foam suspended substance fix nitrite reductase into
Ball is so as to helping to handle the nitrite in breeding water body.
The purpose of the present invention is achieved through the following technical solutions：
A kind of immobilised enzymes biological reaction apparatus, the device are rectangular box, including left and right ends face and front and rear up and down four
Individual tank wall, both ends of the surface set water inlet and delivery port respectively, and some immobilised enzymes filter plates are vertically arranged in device interior edge water (flow) direction,
The immobilised enzymes filter plate is by netting the fixed filter plate formed by polyurethane foam suspended substance；Between the immobilised enzymes filter plate
Provided with barrier layer.
Preferably, the upper tank wall of described device is dismountable transparency glass plate.
Preferably, the barrier layer is cobblestone and plant leaf (such as bamboo branch leaf, weeds leaf).
Preferably, the front and rear tank wall is provided with concave neck, the immobilised enzymes filter plate detachable insertion card
In groove.
Preferably, described device is handling the application of breeding water body nitrite, specifically by embedding medium and nitrous acid
Salt reductase crude enzyme liquid fully mixes, and is drawn into polyurethane foam suspended substance, after polyurethane foam suspended substance adsorption saturation,
Insert saturation CaCl21-5min is rotated in solution, in 0~4 DEG C of 12~48h of crosslinking curing, is produced with normal saline flushing twice
Load the polyurethane foam suspended substance of enzyme；Nitrite breeding water body will be contained again and be passed through said apparatus.
Preferably, described embedding medium is sodium alginate (SA) or polyvinyl alcohol (PVA).
Preferably, the preparation method of the nitrite reductase crude enzyme liquid, comprises the following steps：
(1) freeze-dried powder of Lactobacillus plantarum (Lactobacillus plantarum) is placed in MRS culture mediums, in 30
DEG C~40 DEG C of static gas wave refrigerator 18h~36h after, seed liquor is made；
(2) with 1%~10% percent by volume, above-mentioned seed liquor is seeded in inducing culture A, it is static in 40 DEG C
Fiber differentiation 18h~24h, obtain induction working stock culture；The inducing culture A formulas are NaNO2The μ g/ml of 5 μ g/ml~20
MRS culture mediums；
(3) with 5%~15% percent by volume, induction working stock culture is seeded in inducing culture B, in 30 DEG C~
40 DEG C of static Fiber differentiation 24h~36h, obtain the nutrient solution containing nitrite reductase；The inducing culture B is formulated：Contain
Derivant NaNO2The μ g/ml of 10 μ g/ml~50 MRS culture mediums；
(4) nutrient solution of the reductase containing nitrite is centrifuged, centrifuged after being cleaned with physiological saline, obtained containing Asia
The thalline of nitrate reductase；
(5) add and put forward enzyme buffer liquid, it is 150mg/ml~400mg/ml to make the cell concentration containing nitrite reductase, is added
Enter lysozyme and (be purchased from Sigma-Aldrich, Shanghai, enzyme activity：70000U/mg) 10mg/ml, 1h~5h after uniform broken wall,
Supernatant of the broken wall solution after centrifugation is nitrite reductase crude enzyme liquid.
Preferably, the concentration of described lysozyme is 10mg/ml~50mg/ml.
Preferably, the described enzyme buffer liquid that carries contains trypsin inhibitor and dithiothreitol (DTT), and its concentration is respectively 2 μ
The μ g/ml and 2.0 μm of ol/ml~20.0 μm ol/ml of g/ml~10.
Described polyurethane foam suspended substance refers to the urethane foam microorganisms fixation support that can be suspended in liquid
(noun can be found in the nano-attapulgite clay compounded hydrophilic urethane foam microorganisms immobilizations of CN 200910117393 load
Body), but it is not limited to spherula, brick body, the irregular body prepared by polyurethane foam for main material.
The principle of present apparatus processing nitrite is as follows：Nitrite breeding water body is by the primary settling of early stage and pre-
Processing procedure flows into class fishing net formula immobilised enzymes biological reaction apparatus, with the nitrous acid in the polyurethane foam suspended substance in device
Salt reductase and the class fishing net abundant haptoreaction of formula immobilised enzymes filter plate of certain fixation is played to reach degraded nitrous
The effect of hydrochlorate.Its other cobble is that using it there is certain weight can prevent that plant leaf is washed away, plant
Branches and leaves can obstruct the bulky grain discarded object in waste water, while can slow down water velocity so as to which assistant degradation nitrite cultivates
Other pollutants in water body.The glass plate of top can observe the disposition of water, while convenient extraction glass plate, and it is fixed to realize
Phase clears up, and changes the effect of plant leaf.
The invention has the advantages that and beneficial effect：
(1) the class fishing net formula immobilised enzymes biological reaction apparatus used in the present invention is created using breeding water body itself water seal
The almost environment of anaerobic, make nitrite reductase service efficiency higher.
(2) nitrite reductase is fixed on polyurethane foam suspended substance by the present invention, directly can efficiently be handled
Nitrite breeding water body, it can prevent nitrite reductase from being broken up by waste water to take away；Nitrite reductase is embedded in outstanding
Nitrite reductase mixing can be made abundant in floating body.
(3) make the features such as the excellent physical and mechanical properties of poly- ammonia foam suspension body, heat-insulating property, ageing-resistant performance obtained
Immobilised enzymes bead has the characteristics that mechanical strength is good, stability is strong, body surface area is big, bag enzyme amount is more, nontoxic.
(4) present invention uses original cobblestone and plant leaf in nature, and plant leaf can be obstructed in waste water
Bulky grain discarded object, while water velocity can be slowed down so as to other pollutants in assistant degradation nitrite breeding water body.
Compared with large purification equipment, a large amount of expenses are on the one hand saved, on the other hand with the characteristics of simple to operate, energy consumption is low.
Lactobacillus plantarum of the present invention is the (Lactobacillus of Lactobacillus plantarum DMDL 9010
PlantarumDMDL9010), in August in 2011 to be preserved in China Committee for Culture Collection of Microorganisms within 19th commonly micro-
Bio-Centers, abbreviation CGMCC, deposit number are CGMCC NO.5172, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3,
Institute of Microorganism, Academia Sinica.The bacterial strain disclosed in Chinese patent CN102978134A, belongs to prior art.
Brief description of the drawings
Fig. 1 is poly- ammonia foam suspension body schematic diagram.
Fig. 2 is class fishing net formula immobilised enzymes filter plate schematic diagram.
Fig. 3 is immobilised enzymes biological reaction apparatus structural representation.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Embodiment 1
As shown in Figure 1, 2, 3, immobilised enzymes biological reaction apparatus of the present invention is rectangular box, including left and right ends
Face and front and rear four tank walls, both ends of the surface up and down set water inlet 1 and delivery port 2, are vertically arranged in device interior edge water (flow) direction respectively
Some immobilised enzymes filter plates 5, cobblestone and plant leaf are provided between the immobilised enzymes filter plate 5.The upper tank wall of described device
For dismountable transparency glass plate.The front and rear tank wall is provided with concave neck, and the immobilised enzymes filter plate 5 is detachable embedding
Enter in the neck.
Described immobilised enzymes filter plate 5 is by 20~60 polyurethane foam suspended substances 3 and long 1m~5m, wide 10cm~20cm
Rectangle fishing net forms；Its annexation is that the 3 crisscross fixation of polyurethane foam suspended substance is formed class fishing by two layers of rectangle fishing net
Net formula immobilised enzymes filter plate 5.
Embodiment 2
The fish pond waste water nitrite in certain city is 0.25mg/L, is flowed into by primary settling and after being pre-processed
Class fishing net formula immobilised enzymes biological reaction apparatus, the reaction unit processing pond are made up of 100 layers of class fishing net formula immobilised enzymes filter plate,
The long 6.00m of single class fishing net formula immobilised enzymes filter plate, wide 0.08m, high 1.80m, effective active area 10.80m2, whole place
It is 10800m to manage the total effective active areas of Chi2.Contain cobblestone, weeds between each two class fishing net formula immobilised enzymes filter plate
Leaf layer, it can integrally play a part of assistant degradation nitrite.It is fixed with polyurethane foam suspended substance from resistance to nitrous acid
The nitrite reductase extracted in bacterium.
Nitrite reductase described in this example is made by following steps：(1) by Lactobacillus plantarum DMDL9010 jelly
Dry powder is placed in MRS culture mediums, and after 30 DEG C~40 DEG C static gas wave refrigerator 24h, seed liquor is made.(2) with 6% volume basis
Than above-mentioned seed liquor is seeded in inducing culture A, in 40 DEG C of static Fiber differentiation 24h, obtains induction working stock culture；Institute
It is NaNO to state inducing culture A formulas210 μ g/ml MRS culture mediums；(3) with 10% percent by volume, by induction work fermentation
Agent is seeded in inducing culture B, in 37 DEG C of static Fiber differentiation 24h, obtains the nutrient solution containing nitrite reductase；It is described to lure
Leading culture medium B formulas is：NaNO containing derivant230 μ g/ml MRS culture mediums；(4) by the training of the reductase containing nitrite
Nutrient solution is centrifuged, and is centrifuged after being cleaned with physiological saline, obtains the thalline containing nitrite reductase；(5) add and carry enzyme buffer
Liquid, it is 300mg/ml to make the cell concentration containing nitrite reductase, addition lysozyme (Sigma-Aldrich is purchased from,
Shanghai, enzyme activity：70000U/mg) 10mg/ml, 4h after uniform broken wall, supernatant of the broken wall solution after centrifugation is nitrous acid
Salt reductase crude enzyme liquid, the thick enzyme powder of nitrite reductase is obtained through vacuum freeze drying.The concentration of described lysozyme is 40mg/
ml.The described enzyme buffer liquid that carries contains trypsin inhibitor and dithiothreitol (DTT), and its concentration is respectively 8 μ g/ml and 10.0 μ
mol/ml。
Immobilised enzymes polyurethane foam suspension production procedure described in this example is as follows：By a certain amount of embedding medium and nitrous
Hydrochlorate reductase crude enzyme liquid fully mixes, and is drawn into polyurethane foam suspended substance, treats polyurethane foam suspended substance adsorption saturation
Afterwards, saturation CaCl is inserted21min is rotated in solution, in 4 DEG C of crosslinking curing 24h in refrigerator, with normal saline flushing twice,
Polyurethane foam suspended substance immobilization compound bacteria (as shown in Figure 1) can be obtained, it is standby.Described embedding medium includes but is not limited to sea
Mosanom (SA), polyvinyl alcohol (PVA) etc..After treatment, content of nitrite is down to 0.07mg/L.
Embodiment 3
Certain duck culturing pond waste water nitrite is 0.20mg/L, is flowed into by primary settling and after being pre-processed
Class fishing net formula immobilised enzymes biological reaction apparatus, the reaction unit processing pond are made up of 60 layers of class fishing net formula immobilised enzymes filter plate,
The long 5.00m of single class fishing net formula immobilised enzymes filter plate, wide 0.08m, high 2.00m, effective active area 10.00m2, whole place
It is 10000m to manage the total effective active areas of Chi2.Contain polyurethane foam between each two class fishing net formula immobilised enzymes filter plate
Suspended substance, cobblestone, weeds leaf layer, can integrally play a part of assistant degradation nitrite.Polyurethane foam suspended substance
On be fixed with the nitrite reductase extracted from resistance to Nitrosomas.
Nitrite reductase described in this example is made by following steps：(1) by Lactobacillus plantarum DMDL9010 jelly
Dry powder is placed in MRS culture mediums, and after 37 DEG C of static gas wave refrigerator 24h, seed liquor is made.(2), will be upper with 8% percent by volume
State seed liquor to be seeded in inducing culture A, in 37 DEG C of static Fiber differentiation 24h, obtain induction working stock culture；The induction training
It is NaNO to support base A formulas28 μ g/ml MRS culture mediums；(3) with 6% percent by volume, induction working stock culture is seeded to
In inducing culture B, in 37 DEG C of static Fiber differentiation 24h, the nutrient solution containing nitrite reductase is obtained；The inducing culture
B is formulated：NaNO containing derivant234 μ g/ml MRS culture mediums；(4) nutrient solution of the reductase containing nitrite is carried out
Centrifugation, centrifuges after being cleaned with physiological saline, obtains the thalline containing nitrite reductase；(5) add and put forward enzyme buffer liquid, make containing Asia
The cell concentration of nitrate reductase is 300mg/ml, adds lysozyme 10mg/ml, 3h after uniform broken wall, broken wall solution pass through from
Supernatant after the heart is nitrite reductase crude enzyme liquid, and the thick enzyme powder of nitrite reductase is obtained through vacuum freeze drying.It is described
The concentration of lysozyme be 40mg/ml.The described enzyme buffer liquid that carries contains trypsin inhibitor and dithiothreitol (DTT), its concentration
Respectively 9 μ g/ml and 12.0 μm of ol/ml.
Immobilised enzymes polyurethane foam suspension production procedure described in this example is as follows：By a certain amount of embedding medium and nitrous
Hydrochlorate reductase crude enzyme liquid fully mixes, and is drawn into polyurethane foam suspended substance, treats polyurethane foam suspended substance adsorption saturation
Afterwards, saturation CaCl is inserted21min is rotated in solution, in 4 DEG C of crosslinking curing 24h in refrigerator, with normal saline flushing twice,
Polyurethane foam suspended substance immobilization compound bacteria (as shown in Figure 1) can be obtained, it is standby.Described embedding medium includes but is not limited to sea
Mosanom (SA), polyvinyl alcohol (PVA) etc..After treatment, content of nitrite is down to 0.08mg/L.
Claims (10)
1. a kind of immobilised enzymes biological reaction apparatus, it is characterised in that the device is rectangular box, including left and right ends face is with before
Four tank walls, both ends of the surface set water inlet and delivery port, some fixations are vertically arranged in device interior edge water (flow) direction respectively up and down afterwards
Change enzyme filter plate, the immobilised enzymes filter plate is by netting the fixed filter plate formed by polyurethane foam suspended substance；The immobilization
Barrier layer is provided between enzyme filter plate.
2. device according to claim 1, it is characterised in that the upper tank wall of described device is dismountable clear glass
Plate.
3. device according to claim 1 or 2, it is characterised in that the barrier layer is cobblestone and plant leaf.
4. device according to claim 3, it is characterised in that the front and rear tank wall is provided with concave neck, described solid
Surely in the change enzyme filter plate detachable insertion neck.
5. Claims 1 to 4 any one described device is in the application of processing breeding water body nitrite.
6. application according to claim 5, it is characterised in that embedding medium and nitrite reductase crude enzyme liquid is fully mixed
It is even, it is drawn into polyurethane foam suspended substance, after polyurethane foam suspended substance adsorption saturation, inserts saturation CaCl2In solution
1-5min is rotated, in 0~4 DEG C of 12~48h of crosslinking curing, produces the polyurethane foam of load enzyme with normal saline flushing twice
Suspended substance；Nitrite breeding water body will be contained again and be passed through said apparatus.
7. application according to claim 6, it is characterised in that described embedding medium is sodium alginate or polyvinyl alcohol.
8. the application according to claim 6 or 7, it is characterised in that the preparation side of the nitrite reductase crude enzyme liquid
Method, comprise the following steps：
(1) freeze-dried powder of Lactobacillus plantarum (Lactobacillus plantarum) is placed in MRS culture mediums, in 30 DEG C~
After 40 DEG C of static gas wave refrigerator 18h~36h, seed liquor is made；
(2) with 1%~10% percent by volume, above-mentioned seed liquor is seeded in inducing culture A, in 40 DEG C of static inductions
18h~24h is cultivated, obtains induction working stock culture；The inducing culture A formulas are NaNO2The μ g/ml of 5 μ g/ml~20 MRS
Culture medium；
(3) with 5%~15% percent by volume, induction working stock culture is seeded in inducing culture B, in 30 DEG C~40 DEG C
Static Fiber differentiation 24h~36h, obtains the nutrient solution containing nitrite reductase；The inducing culture B is formulated：Containing induction
Agent NaNO2The μ g/ml of 10 μ g/ml~50 MRS culture mediums；
(4) nutrient solution of the reductase containing nitrite is centrifuged, centrifuged after being cleaned with physiological saline, nitrous acid must be contained
The thalline of salt reductase；
(5) add and put forward enzyme buffer liquid, it is 150mg/ml~400mg/ml to make the cell concentration containing nitrite reductase, is added molten
Bacterium enzyme 10mg/ml, 1h~5h after uniform broken wall, supernatant of the broken wall solution after centrifugation is nitrite reductase crude enzyme liquid.
9. application according to claim 8, it is characterised in that the concentration of described lysozyme is 10mg/ml~50mg/
ml。
10. application according to claim 9, it is characterised in that the described enzyme buffer liquid that carries contains trypsin inhibitor
And dithiothreitol (DTT), its concentration are respectively the μ g/ml and 2.0 μm of ol/ml~20.0 μm ol/ml of 2 μ g/ml~10.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710735139.4A CN107381834A (en) | 2017-08-24 | 2017-08-24 | A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710735139.4A CN107381834A (en) | 2017-08-24 | 2017-08-24 | A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107381834A true CN107381834A (en) | 2017-11-24 |
Family
ID=60345781
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710735139.4A Pending CN107381834A (en) | 2017-08-24 | 2017-08-24 | A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107381834A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108002651A (en) * | 2017-12-11 | 2018-05-08 | 华南理工大学 | A kind of pond culture sewage circulating treating system and sewage water treatment method |
| CN108865944A (en) * | 2018-07-18 | 2018-11-23 | 河南兰贵坊水族用品科技有限公司 | A kind of microorganism live bacteria agent for fish raising |
| US20210371845A1 (en) * | 2020-05-28 | 2021-12-02 | Nestor Maceda-Johnson | Synthesis Of Enzyme-Polymer Conjugates, Having Enhanced Activity & Stability |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2929872A1 (en) * | 1979-07-24 | 1981-03-12 | Joachim Prof. Dr. Klein | Hydrophilic immobilised enzyme preparations in gel or foam form - produced by polymerising poly:isocyanate in the presence of cells, cell fragments or enzyme to form hydrophilic polyurethane bio:catalyst |
| JPH1146754A (en) * | 1997-07-31 | 1999-02-23 | Toray Eng Co Ltd | Bioreactor |
| JP2004008923A (en) * | 2002-06-06 | 2004-01-15 | Taiho Ind Co Ltd | Metal supported microorganism-immobilized carrier and treatment method for nitrogen-containing wastewater |
| CN104212775A (en) * | 2014-08-29 | 2014-12-17 | 华南理工大学 | Preparation method of nitrite reductase of lactobacillus casei subsp rhamnosus |
| CN207511927U (en) * | 2017-08-24 | 2018-06-19 | 华南理工大学 | A kind of immobilised enzymes biological reaction apparatus |
-
2017
- 2017-08-24 CN CN201710735139.4A patent/CN107381834A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2929872A1 (en) * | 1979-07-24 | 1981-03-12 | Joachim Prof. Dr. Klein | Hydrophilic immobilised enzyme preparations in gel or foam form - produced by polymerising poly:isocyanate in the presence of cells, cell fragments or enzyme to form hydrophilic polyurethane bio:catalyst |
| JPH1146754A (en) * | 1997-07-31 | 1999-02-23 | Toray Eng Co Ltd | Bioreactor |
| JP2004008923A (en) * | 2002-06-06 | 2004-01-15 | Taiho Ind Co Ltd | Metal supported microorganism-immobilized carrier and treatment method for nitrogen-containing wastewater |
| CN104212775A (en) * | 2014-08-29 | 2014-12-17 | 华南理工大学 | Preparation method of nitrite reductase of lactobacillus casei subsp rhamnosus |
| CN207511927U (en) * | 2017-08-24 | 2018-06-19 | 华南理工大学 | A kind of immobilised enzymes biological reaction apparatus |
Non-Patent Citations (3)
| Title |
|---|
| 徐凤彩等: "《酶工程》", vol. 1, 30 June 2001, 中国农业出版社, pages: 121 * |
| 李雅丽等: "《现代生物技术前沿进展》", vol. 1, 30 April 2013, 吉林大学出版社, pages: 270 * |
| 陈石根等: "《酶学》", vol. 1, 复旦大学出版社, pages: 255 - 256 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108002651A (en) * | 2017-12-11 | 2018-05-08 | 华南理工大学 | A kind of pond culture sewage circulating treating system and sewage water treatment method |
| CN108002651B (en) * | 2017-12-11 | 2023-09-26 | 华南理工大学 | Pond culture sewage circulation treatment system and sewage treatment method |
| CN108865944A (en) * | 2018-07-18 | 2018-11-23 | 河南兰贵坊水族用品科技有限公司 | A kind of microorganism live bacteria agent for fish raising |
| CN108865944B (en) * | 2018-07-18 | 2022-07-01 | 德州迈科生物技术有限公司 | Microbial live bacteria agent for fish breeding |
| US20210371845A1 (en) * | 2020-05-28 | 2021-12-02 | Nestor Maceda-Johnson | Synthesis Of Enzyme-Polymer Conjugates, Having Enhanced Activity & Stability |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101264985B (en) | Microorganism water purification slow release body and preparation method thereof | |
| Jagannath et al. | Comparative evaluation of bacterial cellulose (nata) as a cryoprotectant and carrier support during the freeze drying process of probiotic lactic acid bacteria | |
| CN107381834A (en) | A kind of immobilised enzymes biological reaction apparatus and its application in processing breeding water body nitrite | |
| CN101935105B (en) | Microbial purifying agent for aquaculture water and preparation method thereof | |
| CN101781627A (en) | Preparation method and application of sea bdellovibrio bacteriovorus ecological preparation | |
| CN106834269A (en) | A kind of immobilized microspheres of PAEs degradation bacterias and its preparation method and application | |
| CN104787899B (en) | A kind of marine aquaculture antibacterial type water quality cleansing agent and preparation method thereof | |
| CN103695353A (en) | Bacillus subtilis with capability of degrading ammonia nitrogen and application of bacillus subtilis | |
| CN101348306B (en) | Process for degrading cultivation waste water nitrite using immobilized nitrifying bacteria | |
| CN106754546A (en) | One plant of Halomonas bacterial strain and the immobilized microbial inoculum containing it of dissolving dinoflagellate | |
| CN111826324A (en) | Preparation method of lactobacillus rhamnosus powder | |
| CN101988053B (en) | Method for immobilizing pseudoalteromonas sp. SP48 by polyurethane foam | |
| CN207511927U (en) | A kind of immobilised enzymes biological reaction apparatus | |
| CN104789548B (en) | A kind of marine aquaculture carrier immobilized antibacterial type water quality cleansing agent and preparation method thereof | |
| CN107473404A (en) | A kind of water purification agent of self-forming bulk carbon carrier fixation of microbe and preparation method thereof | |
| CN107162218A (en) | It is a kind of for water quality cleansing agent of fishery cultivating and preparation method thereof | |
| CN110106162A (en) | A kind of living microorganism immobilized microspheres and its preparation method and application | |
| CN104388338B (en) | Compound bacterium preparation and application thereof | |
| CN103468622A (en) | Method for fixing algae inhibiting bacteria DH46 through sodium alginate-microporous starch | |
| CN103352010B (en) | Bacillus cereus strain CZBC1 capable of dissolving pond oscillatoria, and its application | |
| CN108585179A (en) | A kind of complex microorganism preparations of water pollution control and preparation method thereof | |
| CN109133361A (en) | A kind of biological slow-released ball and preparation method and application for black smelly river improvement | |
| CN103146672B (en) | Method for immobilizing effective microorganisms (EM) during solid fermentation | |
| CN106145382A (en) | A kind of preparation method of the microbial water-purifying agent for fishery cultivating water body | |
| CN110423714B (en) | Lactobacillus composite microbial agent and application thereof in resisting carp herpesvirus II |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20171124 |
|
| RJ01 | Rejection of invention patent application after publication |