CN106893702A - Low-temperature plasma treatment method for improving activity of beta-mannase - Google Patents
Low-temperature plasma treatment method for improving activity of beta-mannase Download PDFInfo
- Publication number
- CN106893702A CN106893702A CN201611219440.1A CN201611219440A CN106893702A CN 106893702 A CN106893702 A CN 106893702A CN 201611219440 A CN201611219440 A CN 201611219440A CN 106893702 A CN106893702 A CN 106893702A
- Authority
- CN
- China
- Prior art keywords
- beta
- mannase
- temperature plasma
- enzymatic activity
- low temperature
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 67
- 238000009832 plasma treatment Methods 0.000 title claims abstract description 15
- 230000000694 effects Effects 0.000 title abstract description 16
- 230000008569 process Effects 0.000 claims abstract description 34
- 239000011148 porous material Substances 0.000 claims abstract description 11
- 238000012545 processing Methods 0.000 claims abstract description 9
- 239000007789 gas Substances 0.000 claims description 80
- 230000002255 enzymatic effect Effects 0.000 claims description 66
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 20
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 20
- 229910052786 argon Inorganic materials 0.000 claims description 10
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 10
- 229910052757 nitrogen Inorganic materials 0.000 claims description 10
- 239000001301 oxygen Substances 0.000 claims description 10
- 229910052760 oxygen Inorganic materials 0.000 claims description 10
- 239000000835 fiber Substances 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 5
- 229920000914 Metallic fiber Polymers 0.000 claims description 2
- 239000012528 membrane Substances 0.000 claims description 2
- 239000000126 substance Substances 0.000 abstract description 7
- 230000014759 maintenance of location Effects 0.000 description 23
- 108090000790 Enzymes Proteins 0.000 description 17
- 102000004190 Enzymes Human genes 0.000 description 17
- 239000000243 solution Substances 0.000 description 13
- 238000006243 chemical reaction Methods 0.000 description 12
- 238000012360 testing method Methods 0.000 description 11
- 238000005406 washing Methods 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 230000000149 penetrating effect Effects 0.000 description 10
- 238000007789 sealing Methods 0.000 description 10
- 238000006073 displacement reaction Methods 0.000 description 9
- 230000004044 response Effects 0.000 description 9
- 238000005201 scrubbing Methods 0.000 description 9
- 108010001682 Dextranase Proteins 0.000 description 6
- 239000007853 buffer solution Substances 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- 238000006555 catalytic reaction Methods 0.000 description 5
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 229920002752 Konjac Polymers 0.000 description 4
- GZCGUPFRVQAUEE-KVTDHHQDSA-N aldehydo-D-mannose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O GZCGUPFRVQAUEE-KVTDHHQDSA-N 0.000 description 4
- 239000012496 blank sample Substances 0.000 description 4
- 238000009835 boiling Methods 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 150000002500 ions Chemical class 0.000 description 4
- LUEWUZLMQUOBSB-FSKGGBMCSA-N (2s,3s,4s,5s,6r)-2-[(2r,3s,4r,5r,6s)-6-[(2r,3s,4r,5s,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5s,6r)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-4,5-dihydroxy-2-(hydroxymethyl)oxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@@H](O[C@@H]2[C@H](O[C@@H](OC3[C@H](O[C@@H](O)[C@@H](O)[C@H]3O)CO)[C@@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O LUEWUZLMQUOBSB-FSKGGBMCSA-N 0.000 description 3
- LWFUFLREGJMOIZ-UHFFFAOYSA-N 3,5-dinitrosalicylic acid Chemical compound OC(=O)C1=CC([N+]([O-])=O)=CC([N+]([O-])=O)=C1O LWFUFLREGJMOIZ-UHFFFAOYSA-N 0.000 description 3
- 241000233866 Fungi Species 0.000 description 3
- 229920002581 Glucomannan Polymers 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 229940046240 glucomannan Drugs 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 description 3
- 241000186046 Actinomyces Species 0.000 description 2
- 244000247812 Amorphophallus rivieri Species 0.000 description 2
- 235000001206 Amorphophallus rivieri Nutrition 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000000252 konjac Substances 0.000 description 2
- 235000010485 konjac Nutrition 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- TWNIBLMWSKIRAT-RWOPYEJCSA-N (1r,2s,3s,4s,5r)-6,8-dioxabicyclo[3.2.1]octane-2,3,4-triol Chemical compound O1[C@@]2([H])OC[C@]1([H])[C@@H](O)[C@H](O)[C@@H]2O TWNIBLMWSKIRAT-RWOPYEJCSA-N 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 102100032487 Beta-mannosidase Human genes 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 244000182067 Fraxinus ornus Species 0.000 description 1
- 235000002917 Fraxinus ornus Nutrition 0.000 description 1
- 229920000926 Galactomannan Polymers 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000237852 Mollusca Species 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241001655322 Streptomycetales Species 0.000 description 1
- 241000223259 Trichoderma Species 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 206010047400 Vibrio infections Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229940101006 anhydrous sodium sulfite Drugs 0.000 description 1
- 108010055059 beta-Mannosidase Proteins 0.000 description 1
- 239000011942 biocatalyst Substances 0.000 description 1
- 108010089934 carbohydrase Proteins 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000003851 corona treatment Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 239000008236 heating water Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229920000592 inorganic polymer Polymers 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229920000620 organic polymer Polymers 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- LJCNRYVRMXRIQR-OLXYHTOASA-L potassium sodium L-tartrate Chemical compound [Na+].[K+].[O-]C(=O)[C@H](O)[C@@H](O)C([O-])=O LJCNRYVRMXRIQR-OLXYHTOASA-L 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 235000011083 sodium citrates Nutrition 0.000 description 1
- 235000011006 sodium potassium tartrate Nutrition 0.000 description 1
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical compound [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000004753 textile Substances 0.000 description 1
- 230000005619 thermoelectricity Effects 0.000 description 1
- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical class [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2477—Hemicellulases not provided in a preceding group
- C12N9/2488—Mannanases
- C12N9/2491—Beta-mannosidase (3.2.1.25), i.e. mannanase
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01025—Beta-mannosidase (3.2.1.25), i.e. mannanase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Enzymes And Modification Thereof (AREA)
- Chemical Or Physical Treatment Of Fibers (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to a low-temperature plasma treatment method for improving activity of beta-mannase, which comprises the following steps: arranging a support table in a treatment chamber of the low-temperature plasma treatment device, arranging a porous material on the support table, and placing beta-mannase on the porous material; and (3) evacuating the processing chamber, introducing working gas, and performing discharge processing for 2-12min, wherein the gas pressure is 20-95Pa, and the discharge power is 50-300W. The method provided by the invention can effectively improve the activity of the beta-mannase and improve the treatment effect, has high treatment efficiency, belongs to a dry state treatment method in a low-temperature plasma treatment process, does not use chemicals, is green and environment-friendly, saves energy and reduces emission, and has very wide market prospect.
Description
Technical field
The present invention relates to enzyme modified working process technical field, more particularly to a kind of raising beta-mannase enzymatic activity
Low Temperature Plasma Treating method.
Background technology
'beta '-mannase is manna oligosacchride, mannocarolose (sweet dew of the hydrolysis with β -1,4-D- mannopyranoses as main chain
Glycan, glucomannan, galactomannans) endo hydrolysis enzyme.The source of 'beta '-mannase is wide, including thin
Bacterium, fungi, actinomyces, plant and mollusk etc..Wherein, microorganism is the main source for producing 'beta '-mannase.Bacterium
In bacillus, pseudomonad, vibrios, in the aspergillus of fungi, trichoderma, yeast, mould, bracket fungus, sclerotinite and actinomyces
Streptomycete all be produce 'beta '-mannase common monoid.
'beta '-mannase as a kind of biocatalyst, with efficient selectivity, reaction condition it is gentle, to environment without dirt
The remarkable advantages such as dye, are employed for multiple fields.Such as in feed industry, 'beta '-mannase is added as a kind of green feed
Agent, digestion and absorption for improving feed, reduces cost promotes growth of animal etc..Locate before conduct one kind in textile industry
Reason biology enzyme, is refined to bafta, and degumming process etc. is carried out to bast fiber fabrics, efficiently avoid traditional chemical degumming institute
The chemicals consumption that brings is big, and energy consumption water consume is high, the problems such as environmental pollution is serious.However, as biological enzyme formulation, β-sweet dew
Dextranase there is also catalysis activity and receives the process conditions such as extraneous pH value, temperature greatly in use, and Activity and stabill is inadequate
Height, to the low problem of degradation of substrates speed.Thus, how further to improve, improve the catalysis activity of 'beta '-mannase and steady
It is qualitative, to strengthening its catalytic efficiency, expand its range of application and use condition, it is significant.
Low temperature plasma is the state of material the 4th after solid-state, liquid, gaseous state, when applied voltage reach gas
When thermoelectricity is pressed, gas molecule is breakdown, produces including electronics, ion, atom and mixture etc. free radical.Low temperature plasma belongs to
In the upper state for exciting, ionizing, the negative electrical charge of its electronics and the positive charge sum of ion are equal, do not show electricity externally macroscopically
Property, in neutrality.Although discharge of plasma in low temperature process electron temperature is very high, heavy particle temperature is very low, and whole system is presented
Low-temperature condition, so referred to as low temperature plasma, is also nonequilibrium condition plasma.
The content of the invention
In order to solve the above technical problems, it is an object of the invention to provide a kind of low temperature for improving beta-mannase enzymatic activity etc.
Gas ions processing method, the method that the present invention is provided can effectively improve the activity of 'beta '-mannase, improve treatment effect,
And treatment effeciency is high, and Low Temperature Plasma Treating process belongs to dry state facture, does not use chemicals, environmental protection, section
Energy emission reduction, with boundless market prospects.
A kind of Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity of the invention, comprises the following steps：
(1) about one penetrating horizontal stand platform, support are set in the processing chamber housing of apparatus for low-temperature plasma treatment
Platform is provided with the porous material in thin, planar shape, and 'beta '-mannase is put on the porous material；
(2) processing chamber housing is emptied, is passed through working gas, and discharge process 2-12min, wherein gas pressure intensity are
20-95Pa, discharge power is 50-300W.
Further, in step (1), porous material is perforated web or porous membrane.
Further, in step (1), the material of porous material is macromolecular fibre or metallic fiber.Macromolecular fibre
It is organic polymer fiber or inorganic polymer fiber.
Further, in step (1), 'beta '-mannase is solid-state 'beta '-mannase.
Further, in step (1), mannosan thinly, is equably laid on the porous material.
Further, in step (1), in step (2), discharge type is glow discharge.
Further, in step (2), working gas elder generation gas washing 2-3 times is used.After gas washing terminates, intake valve is opened, treated
After steady air current, electric discharge device is opened.
Further, in step (2), working gas is one or more in oxygen, nitrogen and argon gas.
Further, in step (2), when working gas is oxygen, gas pressure intensity is 65-95Pa, and discharge power is 50-
100W, the discharge process time is 2-10min.
Further, in step (2), when working gas is nitrogen, gas pressure intensity is 65-80Pa, and discharge power is 50-
250W, the discharge process time is 8-10min.
Further, in step (2), when working gas is argon gas, gas pressure intensity is 20-65Pa, and discharge power is 50-
300W, the discharge process time is 8-12min.
Further, it is further comprising the steps of after step (2)：
Collection step (2) treatment after 'beta '-mannase, by it at 2-8 DEG C sealing preserve.
The present invention uses the conditions such as different experiments atmosphere, gas pressure intensity, process time and discharge power, to beta-mannase
Enzyme carries out low temperature plasma dry state treatment, using serial energetic ion, electronics isoreactivity material and β in low temperature plasma-
Mannosan enzyme effect, is capable of achieving to modify the macromolecular of 'beta '-mannase and its Lian Shang functional groups, or even can play sky
Between conformation etc. change, from its activity and stability can be caused to be obviously improved, be conducive to further developing β-sweet dew poly-
Application potential of the carbohydrase in fields such as weavings.
By such scheme, the present invention at least has advantages below：
1st, the invention provides a kind of Low Temperature Plasma Treating method for improving 'beta '-mannase Activity and stabill,
By 'beta '-mannase enzymatic activity after the before processing for contrasting low temperature plasma, β-sweet dew after Low Temperature Plasma Treating is found
Enzyme and stability are significantly improved；
2nd, the inventive method is the modified working process to standard biologic enzyme, corona treatment cycle is short, it is simple to operate,
Convenient and swift, processing cost is low；
3rd, the inventive method belongs to dry state treatment, and use chemicals useless is environmentally safe, efficiently avoid biography
The chemicals consumption that is brought of system chemical Degumming is big, and energy consumption water consume is high, the problems such as environmental pollution is serious.
Described above is only the general introduction of technical solution of the present invention, in order to better understand technological means of the invention,
And can be practiced according to the content of specification, below with presently preferred embodiments of the present invention and coordinate accompanying drawing describe in detail as after.
Brief description of the drawings
Fig. 1 is the standard curve of 'beta '-mannase active testing of the present invention；
Fig. 2 is 'beta '-mannase result figure in the embodiment of the present invention 1,2,3；
Fig. 3 is 'beta '-mannase result figure in the embodiment of the present invention 4,5,6；
Fig. 4 is 'beta '-mannase result figure in the embodiment of the present invention 7,8,9.
Specific embodiment
With reference to the accompanying drawings and examples, specific embodiment of the invention is described in further detail.Hereinafter implement
Example is not limited to the scope of the present invention for illustrating the present invention.
The reagent and its compound method that the present invention is used are as follows：
DNS reagents：Weigh 3,5- dinitrosalicylic acids 3.15g and be dissolved in 500mL water, 45 DEG C of water-bath is placed in after stirring 5s
In.The sodium hydroxide solution of 100mL 0.2g/mL is then gradually adding, while being stirred continuously, it is ensured that temperature is not higher than 48 DEG C, directly
It is as clear as crystal to solution.Rochelle salt 91.0g, phenol 2.50g and anhydrous sodium sulfite 2.50g are gradually added again.After
Continue 45 DEG C of heating water baths, while adding water 300mL, be stirred continuously, until the material for adding is completely dissolved.Stop heating, cooling
To room temperature, add water constant volume 1000mL.Filtered with fritted glass filter.Filtrate is taken, is stored in brown bottle, kept in dark place.
Can be used after depositing 7 days at room temperature, the term of validity is 6 months.
Buffer solution (pH=4.8,0.1mol/L)：Take 0.1mol/L citric acid solutions 460mL and 0.1mol/L sodium citrates
Solution 540mL is well mixed, and adjusts pH value to 4.8 standby.
D-MANNOSE solution (5mg/mL)：Weigh and the mannose 0.25000g until constant weight is dried at 105 DEG C, be accurate to
0.0001g, with the citric acid-trisodium citrate buffer solution of pH=4.8, the constant volume in 50mL volumetric flasks.
Konjaku sol solution (2mg/mL)：Weigh konjac glucomannan 0.2000g, be accurate to 0.0001g, with the citric acid of pH=4.8-
Trisodium citrate buffer solution, is settled to 100mL.
Fig. 1 is the standard curve of 'beta '-mannase active testing of the present invention, and its preparation method is as follows：
5mg/mlD- mannose solution 0ml, 1.0ml, 1.5ml, 2.0ml, 2.5ml, 3.0ml and 3.5ml are drawn respectively, point
Be not settled to 50ml with the citric acid-trisodium citrate cushioning liquid of pH=4.8, make concentration for 0,0.10mg/ml,
0.15mg/ml, 0.20mg/ml, 0.25mg/ml, 0.30mg/ml and 0.35mg/mlD- mannose standard liquid.
7 25ml color-comparison tubes numberings 1~7 are taken, the D-MANNOSE standard liquid for drawing above-mentioned concentration series is each
2.0ml, is sequentially added into colorimetric cylinder, then is separately added into 3.0mlDNS reagents, shakes up, and 5min is heated in boiling water bath.Then
Take out colorimetric cylinder and be cooled to room temperature, add water and be settled to 25ml, stood after shaking up.It is blank sample with No. 1 pipe solution,
On TU-1810 type spectrophotometry instrument in the range of 400~700nm sequentially determining and record each sample absorbance
Value.Then with D-MANNOSE concentration C as ordinate, the absorbance A of its correspondence maximum characteristic wave strong point (λ max/485nm) is
Abscissa, sets up standard working curve.
Embodiment 1
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using argon gas reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From argon gas as working gas, condition of work is for this implementation：Gas pressure intensity 20Pa, discharge power
100W, process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, unlatching is put
Electric switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum are closed
Valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, opens cavity door, and collect the β-sweet after treatment
Dew dextranase, the sealing preserve at 2-8 DEG C.
Enzymatic activity is carried out using following methods and enzymatic activity retention rate is tested：
Beta-mannase enzyme solutions (1.0mg/mL)：The 'beta '-mannase 0.05000g after above method treatment is weighed,
0.0001g is accurate to, with the citric acid-trisodium citrate buffer solution of pH=4.8,50mL is settled to.
2mg/mL konjaku sol solution 1mL are added in color-comparison tube, are put into 45 DEG C of thermostat water baths and are balanced 10min,
Same equilibrated 1.0mg/mL beta-mannase enzyme solutions 1mL in 45 DEG C of water-baths are added, is shaken up, while stopwatch is at once
Start timing.Colorimetric cylinder is taken out after accurate reaction 10min, 3mLDNS reagents are rapidly joined, boiling in boiling water is put into after shaking up at once
Boil 5min.Cooled down with cold water after taking-up, plus deionized water constant volume 25mL, shake up.'beta '-mannase in said process is replaced
On behalf of buffer solution, blank sample is prepared using same method.After 2mL buffer solutions and 3mL DNS boilings are boiled into 5min constant volumes 25mL
Solution is baseline, determines the absorbance that sample and blank sample are tested at 485nm.
Enzymatic activity is calculated as follows：
Enzymatic activity is defined：Under the conditions of 45 DEG C, pH are for 4.8,1g 'beta '-mannases 1min hydrolysis konjac glucomannan generations 1ug
D-MANNOSE is 1 catalysis activity unit, is represented with U/g.Simultaneously according to the fitting side of above-mentioned experiment gained standard working curve
Journey, the computing formula of its catalysis activity is as follows：
In formula, X represents the 'beta '-mannase catalysis activity in sample, and unit is U/g；AEIt is test specimen through β-sweet dew
The absorbance of glycan enzyme catalyzed hydrolysis liquid；ABIt is the absorbance of correspondence blank sample；K and b are respectively the slopes of standard curve
And intercept；V is the volume of enzyme digestion reaction liquid, units/ml；T is the beta-mannase enzyme catalyzed hydrolysis time, and unit is min；m
It is the 'beta '-mannase quality added in reaction, unit g.
After Low Temperature Plasma Treating, beta-mannase enzymatic activity retention rate R can be calculated according to equation below
Arrive：
In formula, R is enzymatic activity retention rate, X0And X1Refer to the work of 'beta '-mannase before and after Low Temperature Plasma Treating respectively
Property, unit U/g.
Beta-mannase enzymatic activity is 33451.52U/g before using the above method to measure Low Temperature Plasma Treating, and this
The treated beta-mannase enzymatic activity of embodiment is 35224.25U/g, and enzymatic activity retention rate is 105.30%.Result such as Fig. 2
Shown in middle a.It can be seen that enzymatic activity retention rate is more than 100% under the treatment conditions of embodiment 1, the enzyme of 'beta '-mannase is illustrated
Activity is better than before processing enzymatic activity, and enzymatic activity is improved.
Embodiment 2
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using nitrogen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From nitrogen as working gas, condition of work is for this implementation：Gas pressure intensity 65Pa, discharge power
100W, process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, unlatching is put
Electric switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum are closed
Valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, opens cavity door, and collect the β-sweet after treatment
Dew dextranase, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 2
Shown in b.Result shows that the present embodiment beta-mannase enzymatic activity is 34530.57U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 103.23%.As can be seen that under the treatment conditions of the present embodiment 'beta '-mannase enzymatic activity and enzymatic activity
Retention rate is obtained for and significantly improves.
Embodiment 3
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using oxygen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From oxygen as working gas, condition of work is for this implementation：Gas pressure intensity 95Pa, discharge power
100W, process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, unlatching is put
Electric switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum are closed
Valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, opens cavity door, and collect the β-sweet after treatment
Dew dextranase, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 2
Shown in c.Result shows that the present embodiment beta-mannase enzymatic activity is 34453.50U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 103.00%.It can be seen that the enzymatic activity of 'beta '-mannase and enzymatic activity are protected under the treatment conditions of the present embodiment
Rate is stayed to be obtained for raising.
Embodiment 4
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using oxygen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From oxygen as working gas, condition of work is for this implementation：Gas pressure intensity 65Pa, discharge power 50W,
Process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, open electric discharge and open
Put row glow discharge into, countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum valve are closed, so
After close intake valve, after question response cavity air pressure returns to atmospheric pressure, open cavity door, and collect treatment after beta-mannase
Enzyme, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 3
Shown in a.Result shows that the present embodiment beta-mannase enzymatic activity is 38384.34U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 114.75%.As can be seen that under the treatment conditions of the present embodiment 'beta '-mannase enzymatic activity and enzymatic activity
Retention rate is all improved.
Embodiment 5
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using nitrogen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From nitrogen as working gas, condition of work is for this implementation：Gas pressure intensity 65Pa, discharge power
250W, process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, unlatching is put
Electric switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum are closed
Valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, opens cavity door, and collect the β-sweet after treatment
Dew dextranase, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 3
Shown in b.Result shows that the present embodiment beta-mannase enzymatic activity is 35147.18U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 105.07%.
Embodiment 6
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using argon gas reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From argon gas as working gas, condition of work is for this implementation：Gas pressure intensity 35Pa, discharge power
300W, process time 4min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, unlatching is put
Electric switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum are closed
Valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, opens cavity door, and collect the β-sweet after treatment
Dew dextranase, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 3
Shown in c.Result shows that the present embodiment beta-mannase enzymatic activity is 36226.23U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 108.29%.
Embodiment 7
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using oxygen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From oxygen as working gas, condition of work is for this implementation：Gas pressure intensity 65Pa, discharge power 50W,
Process time 2min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, open electric discharge and open
Put row glow discharge into, countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum valve are closed, so
After close intake valve, after question response cavity air pressure returns to atmospheric pressure, open cavity door, and collect treatment after beta-mannase
Enzyme, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 4
Shown in a.Result shows that the present embodiment beta-mannase enzymatic activity is 36149.16U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 108.06%.
Embodiment 8
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using nitrogen reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From nitrogen as working gas, condition of work is for this implementation：Gas pressure intensity 65Pa, discharge power 50W,
Process time 10min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, open electric discharge and open
Put row glow discharge into, countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, vacuum valve are closed, so
After close intake valve, after question response cavity air pressure returns to atmospheric pressure, open cavity door, and collect treatment after beta-mannase
Enzyme, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 4
Shown in a.Result shows that the present embodiment beta-mannase enzymatic activity is 34145.19U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 102.07%.It can be seen that the enzymatic activity of 'beta '-mannase and enzymatic activity retain under the present embodiment treatment conditions
Rate is obtained for raising.
Embodiment 9
'beta '-mannase 0.200g is weighed, about one penetrating horizontal stand platform is set in low-temperature plasma cavity,
The one thin, planar perforated web for launching is set on the trestle table, then 'beta '-mannase thinly, is equably carried out
Dispersed placement, is then shut off the cavity door of apparatus for low-temperature plasma treatment.
Above-mentioned cavity is emptied using the vavuum pump of system configuration, and using argon gas reaction cavity is carried out gas washing or
Gas displacement 2-3 times.From argon gas as working gas, condition of work is for this implementation：Gas pressure intensity 35Pa, discharge power
200W, process time 12min.After gas scrubbing terminates, intake valve is opened, after pending atmosphere pressure and steady air current, opened
Discharge switch carries out glow discharge, and countdown is started while opening discharge power.After the completion for the treatment of, discharge switch, true is closed
Empty valve, is then shut off intake valve, after question response cavity air pressure returns to atmospheric pressure, open cavity door, and collect treatment after β-
Mannase, the sealing preserve at 2-8 DEG C.
Tested using the enzymatic activity before and after the method test processes in embodiment 1 and enzymatic activity retention rate, as a result as in Fig. 4
Shown in a.Result shows that the present embodiment beta-mannase enzymatic activity is 34684.72U/g, enzymatic activity after Low Temperature Plasma Treating
Retention rate is 103.69%.
The above is only the preferred embodiment of the present invention, is not intended to limit the invention, it is noted that for this skill
For the those of ordinary skill in art field, on the premise of the technology of the present invention principle is not departed from, can also make it is some improvement and
Modification, these are improved and modification also should be regarded as protection scope of the present invention.
Claims (9)
1. it is a kind of improve beta-mannase enzymatic activity Low Temperature Plasma Treating method, it is characterised in that comprise the following steps：
(1) trestle table is set in the processing chamber housing of apparatus for low-temperature plasma treatment, and the trestle table is provided with porous material,
'beta '-mannase is placed on the porous material；
(2) processing chamber housing is emptied, is passed through working gas, and discharge process 2-12min, wherein gas pressure intensity are
20-95Pa, discharge power is 50-300W.
2. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 1, its feature exists
In：In step (1), the porous material is perforated web or porous membrane.
3. according to claim 1 or 3 raising beta-mannase enzymatic activity Low Temperature Plasma Treating method, its feature
It is：In step (1), the material of the porous material is macromolecular fibre or metallic fiber.
4. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 1, its feature exists
In：In step (1), the 'beta '-mannase is solid-state 'beta '-mannase.
5. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 1, its feature exists
In：In step (1), in step (2), discharge type is glow discharge.
6. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 1, its feature exists
In：In step (2), the working gas is one or more in oxygen, nitrogen and argon gas.
7. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 6, its feature exists
In：In step (2), when the working gas is oxygen, gas pressure intensity is 65-95Pa, and discharge power is 50-100W, electric discharge
Process time is 2-10min.
8. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 6, its feature exists
In：In step (2), when the working gas is nitrogen, gas pressure intensity is 65-80Pa, and discharge power is 50-250W, electric discharge
Process time is 8-10min.
9. the Low Temperature Plasma Treating method for improving beta-mannase enzymatic activity according to claim 6, its feature exists
In：In step (2), when the working gas is argon gas, gas pressure intensity is 20-65Pa, and discharge power is 50-300W, electric discharge
Process time is 8-12min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611219440.1A CN106893702B (en) | 2016-12-26 | 2016-12-26 | Low-temperature plasma treatment method for improving activity of beta-mannase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611219440.1A CN106893702B (en) | 2016-12-26 | 2016-12-26 | Low-temperature plasma treatment method for improving activity of beta-mannase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106893702A true CN106893702A (en) | 2017-06-27 |
| CN106893702B CN106893702B (en) | 2019-10-01 |
Family
ID=59198960
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611219440.1A Active CN106893702B (en) | 2016-12-26 | 2016-12-26 | Low-temperature plasma treatment method for improving activity of beta-mannase |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106893702B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115633718A (en) * | 2022-10-19 | 2023-01-24 | 中国农业科学院油料作物研究所 | Method for improving plant milk enzymolysis efficiency through low-consumption physical double-field cooperation |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101701212A (en) * | 2009-11-13 | 2010-05-05 | 清华大学 | Method for improving enzyme activity by utilizing atmospheric pressure discharge plasma |
| CN103255118A (en) * | 2013-05-21 | 2013-08-21 | 中南大学 | Beta-mannase, coding gene as well as producing strain and application thereof |
-
2016
- 2016-12-26 CN CN201611219440.1A patent/CN106893702B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101701212A (en) * | 2009-11-13 | 2010-05-05 | 清华大学 | Method for improving enzyme activity by utilizing atmospheric pressure discharge plasma |
| CN103255118A (en) * | 2013-05-21 | 2013-08-21 | 中南大学 | Beta-mannase, coding gene as well as producing strain and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| S.R.法内斯托克等: "《生物高分子 第7卷 聚酰胺和蛋白质材料1》", 31 January 2005, 化学工业出版社 * |
| 张波等: "低温等离子体技术及其在作物种子中的应用", 《江苏农业科学》 * |
| 陈杰瑢: "《等离子体清洁技术在纺织印染中的应用》", 30 September 2005, 中国纺织出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115633718A (en) * | 2022-10-19 | 2023-01-24 | 中国农业科学院油料作物研究所 | Method for improving plant milk enzymolysis efficiency through low-consumption physical double-field cooperation |
| CN115633718B (en) * | 2022-10-19 | 2024-03-15 | 中国农业科学院油料作物研究所 | Method for improving plant lactosis enzymolysis efficiency through low-consumption physical double-field cooperation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106893702B (en) | 2019-10-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103111265B (en) | Preparation method of adsorbent for removing antibiotics in water, prepared adsorbent and application | |
| CN103835002B (en) | A kind of bluish dogbane low-temperature plasma and Bio-enzyme Combined Pre-treatment Degumming method | |
| CN104313891A (en) | Starch slurry desizing method adopting supercritical carbon dioxide as medium | |
| CN104328648A (en) | Desizing treatment method in supercritical carbon dioxide medium | |
| CN104372598A (en) | Supercritical carbon dioxide fluid desizing method by use of combined enzyme | |
| CN106893702B (en) | Low-temperature plasma treatment method for improving activity of beta-mannase | |
| CN108609823A (en) | Method for enhancing sludge dewatering performance | |
| CN105838608A (en) | Method for segment pretreatment on straws through ammonia water and hydrogen peroxide | |
| CN111072154A (en) | Preparation method of sewage treatment agent by using microorganisms | |
| CN100542660C (en) | A kind of raising infiltrating and vaporizing membrane to moisture from surface modifying method optionally | |
| CN105671091A (en) | Method for pretreating cotton straw with ionic liquid [Bmim]Cl | |
| CN206300424U (en) | A kind of improved sample drying case | |
| CN206176937U (en) | A microwave drying apparatus for piece granular materials | |
| CN205586995U (en) | Regeneration treatment device of active carbon | |
| CN106929503A (en) | Method for Improving Pectinase Activity | |
| CN106119295A (en) | Utilize NH4hCO3the method of pretreatment corn straw and the anaerobic fermentation method of corn straw | |
| CN208493798U (en) | Ammonia nitrogen exhaust gas automatic water sealing arrangement | |
| CN111154811A (en) | Pretreatment system for improving biodegradation efficiency of lignocellulose and application thereof | |
| CN106754863B (en) | Dry state treatment method for improving xylanase activity | |
| CN110628581A (en) | Portable temporary operation platform for bacterial culture | |
| CN110615696A (en) | Kitchen waste synergistic reaction agent and composting method thereof | |
| CN106982808B (en) | Termite bait, termite trapping and killing device and manufacturing method thereof | |
| CN206538423U (en) | A kind of medical test incubation water heating incubator | |
| CN103039269A (en) | Simple liquid strain inoculation device | |
| CN208497713U (en) | A kind of capacitance coupling plasma macromolecule surface processing unit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |