CN106893702B - Improve the active Low Temperature Plasma Treating method of 'beta '-mannase - Google Patents
Improve the active Low Temperature Plasma Treating method of 'beta '-mannase Download PDFInfo
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- CN106893702B CN106893702B CN201611219440.1A CN201611219440A CN106893702B CN 106893702 B CN106893702 B CN 106893702B CN 201611219440 A CN201611219440 A CN 201611219440A CN 106893702 B CN106893702 B CN 106893702B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N13/00—Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- 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
Abstract
The present invention relates to a kind of active Low Temperature Plasma Treating methods of raising 'beta '-mannase, the following steps are included: trestle table is arranged in the processing chamber housing of apparatus for low-temperature plasma treatment, trestle table is equipped with porous material, and 'beta '-mannase is put on the porous material；Processing chamber housing is emptied, working gas, and discharge treatment 2-12min are passed through, wherein gas pressure intensity is 20-95Pa, discharge power 50-300W.Method provided by the invention 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, it is environmentally protective, energy-saving and emission-reduction have boundless market prospects.
Description
Technical field
The present invention relates to enzyme modified working process technical field more particularly to a kind of raising beta-mannase enzymatic activitys
Low Temperature Plasma Treating method.
Background technique
'beta '-mannase is hydrolyzed using β -1,4-D- mannopyranose as the manna oligosacchride of main chain, mannocarolose (sweet dew
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 generating 'beta '-mannase.Bacterium
In bacillus, pseudomonad, vibrios, in the aspergillus of fungi, trichoderma, yeast, mould, bracket fungus, sclerotinite and actinomyces
Streptomycete all be generate 'beta '-mannase common monoid.
'beta '-mannase has efficient specificity, reaction condition mildly, to environment without dirt as a kind of biocatalyst
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 reduces cost, promotes growth of animal etc. for improving the digestion and absorption of feed.As a kind of preceding place in textile industry
Biological enzyme is managed, cotton fabric is refined, degumming process etc. is carried out to bast fiber fabrics, efficiently avoids traditional chemical degumming institute
The problems such as bring chemicals dosage is big, and energy consumption water consume is high, and environmental pollution is serious.However, as biological enzyme formulation, β-sweet dew
That there is also catalytic activity in use is big by process conditions such as extraneous pH value, temperature for dextranase, and Activity and stabill is inadequate
Height, it is low to degradation of substrates rate the problems such as.Thus, how to further increase, improve the catalytic activity of 'beta '-mannase and steady
It is qualitative, to its catalytic efficiency is enhanced, expands its application range and use condition, be of great significance.
Low temperature plasma is the 4th state of substance after solid-state, liquid, gaseous state, when applied voltage reach gas
When thermoelectricity is pressed, gas molecule is breakdown, and generating includes the mixtures such as electronics, ion, atom and free radical.Low temperature plasma category
In the upper state of excitation, ionization, the negative electrical charge of electronics and the positive charge sum of ion are equal, macroscopically externally not aobvious electric
Property, it is 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.
Summary of the invention
In order to solve the above technical problems, the object of the present invention is to provide active low temperature of a kind of raising 'beta '-mannase etc.
Gas ions processing method, method provided by the invention 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, environmentally protective, section
Energy emission reduction, has boundless market prospects.
A kind of active Low Temperature Plasma Treating method of raising 'beta '-mannase of the invention, comprising the following steps:
(1) a transparent horizontal bracket platform, bracket are set in the processing chamber housing of apparatus for low-temperature plasma treatment
Platform is equipped 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 treatment 2-12min, wherein gas pressure intensity is
20-95Pa, discharge power 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
For organic polymer fiber or inorganic polymer fiber.
Further, in step (1), 'beta '-mannase is solid-state 'beta '-mannase.
Further, in step (1), by mannosan, thinly, equably laying is 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, intake valve is opened, to
After steady air current, electric discharge device is opened.
Further, in step (2), working gas is one or more of oxygen, nitrogen and argon gas.
Further, in step (2), when working gas is oxygen, gas pressure intensity 65-95Pa, discharge power 50-
100W, discharge treatment time are 2-10min.
Further, in step (2), when working gas is nitrogen, gas pressure intensity 65-80Pa, discharge power 50-
250W, discharge treatment time are 8-10min.
Further, in step (2), when working gas is argon gas, gas pressure intensity 20-65Pa, discharge power 50-
300W, discharge treatment time are 8-12min.
Further, further comprising the steps of after step (2):
Collection step (2) treated 'beta '-mannase, it is sealed at 2-8 DEG C.
The present invention is using different experiments atmosphere, gas pressure intensity, the processing conditions such as time and discharge power, to beta-mannase
Enzyme carries out the processing of low temperature plasma dry state, using energetic ion serial in low temperature plasma, electronics isoreactivity substance and β-
Mannosan enzyme effect is, it can be achieved that macromolecular and its Lian Shang functional group to 'beta '-mannase are modified, or even can play sky
Between conformation etc. change, from can lead to its activity and stability is obviously improved, being conducive to further to develop β-sweet dew and gather
Application potential of the carbohydrase in fields such as weavings.
According to the above aspect of the present invention, the present invention has at least the following advantages:
1, the present invention provides it is a kind of improve 'beta '-mannase Activity and stabill Low Temperature Plasma Treating method,
By comparing the 'beta '-mannase enzymatic activity before and after the processing of low temperature plasma, β-sweet dew after Low Temperature Plasma Treating is found
Enzyme and stability significantly improve；
2, the method for the present invention is the modification working process to standard biologic enzyme, the corona treatment period is short, it is easy to operate,
Convenient and efficient, processing cost is low；
3, the method for the present invention belongs to dry state processing, and useless to use chemicals, no pollution to the environment efficiently avoids passing
The problems such as chemicals dosage brought by chemical Degumming of uniting is big, and energy consumption water consume is high, and environmental pollution is serious.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention and the accompanying drawings.
Detailed description of the invention
Fig. 1 is the standard curve of 'beta '-mannase active testing of the present invention；
Fig. 2 is 'beta '-mannase processing result figure in the embodiment of the present invention 1,2,3；
Fig. 3 is 'beta '-mannase processing result figure in the embodiment of the present invention 4,5,6；
Fig. 4 is 'beta '-mannase processing result figure in the embodiment of the present invention 7,8,9.
Specific embodiment
With reference to the accompanying drawings and examples, specific embodiments of the present invention will be described in further detail.Implement below
Example is not intended to limit the scope of the invention for illustrating the present invention.
The reagent and its preparation method that the present invention uses are as follows:
DNS reagent: it weighs 3,5- dinitrosalicylic acid 3.15g and is dissolved in 500mL water, stirring 5s is placed on 45 DEG C of water-bath
In.It is then gradually adding the sodium hydroxide solution of 100mL 0.2g/mL, is stirred continuously simultaneously, guarantees that temperature is not higher than 48 DEG C, directly
It is as clear as crystal to solution.It is gradually added Rochelle salt 91.0g, phenol 2.50g and anhydrous sodium sulfite 2.50g again.After
Continue 45 DEG C of heating water baths, while water supplement 300mL, is stirred continuously, until the substance of addition is completely dissolved.Stop heating, it is cooling
To room temperature, add water constant volume 1000mL.It is filtered with fritted glass filter.Filtrate is taken, storage in a brown bottle, is kept in dark place.
It can be used after storing 7 days at room temperature, validity period is 6 months.
Buffer (pH=4.8,0.1mol/L): 0.1mol/L citric acid solution 460mL and 0.1mol/L sodium citrate are taken
Solution 540mL is uniformly mixed, and adjusts pH value to 4.8 spare.
D-MANNOSE solution (5mg/mL): it weighs drying at 105 DEG C and is accurate to up to the mannose 0.25000g of constant weight
0.0001g, with citric acid-trisodium citrate buffer solution of pH=4.8, the constant volume in 50mL volumetric flask.
Konjaku sol solution (2mg/mL): weighing 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, the production 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 of pH=4.8-trisodium citrate buffer solution, make concentration be 0,0.10mg/ml,
0.15mg/ml, 0.20mg/ml, 0.25mg/ml, 0.30mg/ml and 0.35mg/mlD- mannose standard solution.
7 25ml color-comparison tube numbers 1~7 are taken, the D-MANNOSE standard solution for drawing above-mentioned concentration series is each
2.0ml is sequentially added into colorimetric cylinder, then is separately added into 3.0mlDNS reagent, shakes up, 5min is heated in boiling water bath.Then
It takes out colorimetric cylinder and is cooled to room temperature, add water and be settled to 25ml, stood after shaking up.Using No. 1 pipe solution as blank control sample,
In the absorbance of sequentially determining within the scope of 400~700nm and record each sample on TU-1810 type spectrophotometry instrument
Value.Then using D-MANNOSE concentration C as ordinate, the absorbance value A for corresponding to maximum characteristic wave strong point (λ max/485nm) is
Abscissa establishes standard working curve.
Embodiment 1
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using argon gas to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects argon gas as working gas, operating condition are as follows: gas pressure intensity 20Pa, discharge power
100W handles time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, unlatching is put
Electric switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum are closed
Valve is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collects that treated that β-is sweet
Reveal dextranase, is sealed at 2-8 DEG C.
Enzymatic activity and the test of enzymatic activity retention rate are carried out using following methods:
Beta-mannase enzyme solutions (1.0mg/mL): weighing the above method treated 'beta '-mannase 0.05000g,
It is accurate to 0.0001g, with citric acid-trisodium citrate buffer solution of pH=4.8, is settled to 50mL.
2mg/mL konjaku sol solution 1mL is added in color-comparison tube, is put into 45 DEG C of thermostat water baths and balances 10min,
Equilibrated 1.0mg/mL beta-mannase enzyme solutions 1mL equally in 45 DEG C of water-baths is added, is shaken up, while stopwatch is at once
Start timing.Colorimetric cylinder is taken out after accurate reaction 10min, 3mLDNS reagent is rapidly joined, is put into boiling water and boils at once after shaking up
Boil 5min.It is cooling with cold water after taking-up, add deionized water constant volume 25mL, shakes up.'beta '-mannase in the above process is replaced
On behalf of buffer, blank sample is prepared using same method.After 2mL buffer and 3mL DNS boiling are boiled 5min constant volume 25mL
Solution is baseline, measures the absorbance value that sample and blank sample are tested at 485nm.
Enzymatic activity calculates as follows:
Enzymatic activity definition: under the conditions of 45 DEG C, pH are 4.8,1g 'beta '-mannase 1min hydrolyzes konjac glucomannan and generates 1ug
D-MANNOSE is 1 catalysis activity unit, is indicated with U/g.Fitting side according to standard working curve obtained by above-mentioned experiment simultaneously
The calculation formula of journey, catalytic activity is as follows:
In formula, X indicates the 'beta '-mannase catalysis activity in sample, unit U/g；AEIt is test specimen through β-sweet dew
The absorbance of glycan enzyme catalyzed hydrolysis liquid；ABFor the absorbance of corresponding blank sample；K and b is the slope of standard curve respectively
And intercept；V is the volume of enzyme digestion reaction liquid, units/ml；T is beta-mannase enzyme catalyzed hydrolysis time, unit min；m
For the 'beta '-mannase quality being added in reaction, unit g.
After Low Temperature Plasma Treating, beta-mannase enzymatic activity retention rate R can be calculated according to following formula
It arrives:
In formula, R is enzymatic activity retention rate, X0And X1Respectively refer to the work of Low Temperature Plasma Treating front and back 'beta '-mannase
Property, unit U/g.
Beta-mannase enzymatic activity is 33451.52U/g before using the above method to measure Low Temperature Plasma Treating, and sheet
The processed beta-mannase enzymatic activity of embodiment is 35224.25U/g, and enzymatic activity retention rate is 105.30%.As a result such as Fig. 2
Shown in middle a.It can be seen that enzymatic activity retention rate is greater than 100% under 1 treatment conditions of embodiment, illustrate the enzyme of 'beta '-mannase
Activity is improved better than enzymatic activity before handling, enzymatic activity.
Embodiment 2
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using nitrogen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects nitrogen as working gas, operating condition are as follows: gas pressure intensity 65Pa, discharge power
100W handles time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, unlatching is put
Electric switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum are closed
Valve is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collects that treated that β-is sweet
Reveal dextranase, is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 2
Shown in b.The 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%.It can be seen that the enzymatic activity and enzymatic activity of the 'beta '-mannase under the treatment conditions of the present embodiment
Retention rate is all significantly improved.
Embodiment 3
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using oxygen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects oxygen as working gas, operating condition are as follows: gas pressure intensity 95Pa, discharge power
100W handles time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, unlatching is put
Electric switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum are closed
Valve is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collects that treated that β-is sweet
Reveal dextranase, is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 2
Shown in c.The 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 and enzymatic activity of 'beta '-mannase are protected under the treatment conditions of the present embodiment
Rate is stayed all to be improved.
Embodiment 4
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using oxygen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects oxygen as working gas, operating condition are as follows: gas pressure intensity 65Pa, discharge power 50W,
Handle time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, electric discharge is opened and opens
It puts row glow discharge into, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum valve are closed, so
After close intake valve, after reaction cavity air pressure is restored to atmospheric pressure, open cavity door, and collect treated beta-mannase
Enzyme is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 3
Shown in a.The 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%.It can be seen that the enzymatic activity and enzymatic activity of the 'beta '-mannase under the treatment conditions of the present embodiment
Retention rate is all improved.
Embodiment 5
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using nitrogen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects nitrogen as working gas, operating condition are as follows: gas pressure intensity 65Pa, discharge power
250W handles time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, unlatching is put
Electric switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum are closed
Valve is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collects that treated that β-is sweet
Reveal dextranase, is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 3
Shown in b.The 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, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using argon gas to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects argon gas as working gas, operating condition are as follows: gas pressure intensity 35Pa, discharge power
300W handles time 4min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, unlatching is put
Electric switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum are closed
Valve is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collects that treated that β-is sweet
Reveal dextranase, is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 3
Shown in c.The 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, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using oxygen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects oxygen as working gas, operating condition are as follows: gas pressure intensity 65Pa, discharge power 50W,
Handle time 2min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, electric discharge is opened and opens
It puts row glow discharge into, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum valve are closed, so
After close intake valve, after reaction cavity air pressure is restored to atmospheric pressure, open cavity door, and collect treated beta-mannase
Enzyme is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 4
Shown in a.The 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, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using nitrogen to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects nitrogen as working gas, operating condition are as follows: gas pressure intensity 65Pa, discharge power 50W,
Handle time 10min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, electric discharge is opened and opens
It puts row glow discharge into, starts countdown while opening discharge power.After the completion of processing, discharge switch, vacuum valve are closed, so
After close intake valve, after reaction cavity air pressure is restored to atmospheric pressure, open cavity door, and collect treated beta-mannase
Enzyme is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 4
Shown in a.The 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 and enzymatic activity of 'beta '-mannase retain under the present embodiment treatment conditions
Rate is all improved.
Embodiment 9
'beta '-mannase 0.200g is weighed, a transparent horizontal bracket platform is set in low-temperature plasma cavity,
The thin, planar perforated web of one expansion of setting 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 vacuum pump of system configuration, and using argon gas to reaction cavity carry out gas washing or
Gas displacement 2-3 times.This implementation selects argon gas as working gas, operating condition are as follows: gas pressure intensity 35Pa, discharge power
200W handles time 12min.After gas scrubbing, intake valve is opened, after atmosphere pressure and steady air current to be processed, is opened
Discharge switch carries out glow discharge, starts countdown while opening discharge power.After the completion of processing, discharge switch, true is closed
Empty valve, is then shut off intake valve, after reaction cavity air pressure is restored to atmospheric pressure, opens cavity door, and collect treated β-
Mannase is sealed at 2-8 DEG C.
Using the enzymatic activity and the test of enzymatic activity retention rate before and after the method test processes in embodiment 1, as a result as in Fig. 4
Shown in a.The 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 a preferred embodiment of the present invention, it is not intended to restrict the invention, it is noted that for this skill
For the those of ordinary skill in art field, without departing from the technical principles of the invention, can also make it is several improvement and
Modification, these improvements and modifications also should be regarded as protection scope of the present invention.
Claims (4)
1. a kind of active Low Temperature Plasma Treating method of raising 'beta '-mannase, which comprises the following steps:
(1) trestle table is set in the processing chamber housing of apparatus for low-temperature plasma treatment, the trestle table is equipped with porous material,
'beta '-mannase is placed on the porous material；The 'beta '-mannase is solid-state 'beta '-mannase；
(2) processing chamber housing is emptied, is passed through working gas, the working gas is in oxygen, nitrogen and argon gas
It is one or more of；When the working gas is oxygen, gas pressure intensity 65-95Pa, discharge power 50-100W, discharge treatment
Time is 2-10min；When the working gas is nitrogen, gas pressure intensity 65-80Pa, discharge power 50-250W, at electric discharge
The reason time is 8-10min；When the working gas is argon gas, gas pressure intensity 20-65Pa, discharge power 50-300W, electric discharge
The processing time is 8-12min.
2. the active Low Temperature Plasma Treating method of raising 'beta '-mannase according to claim 1, feature exist
In: in step (1), the porous material is perforated web or porous membrane.
3. the active Low Temperature Plasma Treating method of raising 'beta '-mannase according to claim 1 or 2, feature
Be: in step (1), the material of the porous material is macromolecular fibre or metallic fiber.
4. the active Low Temperature Plasma Treating method of raising 'beta '-mannase according to claim 1, feature exist
In: in step (2), discharge type is glow discharge.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201611219440.1A CN106893702B (en) | 2016-12-26 | 2016-12-26 | Improve the active Low Temperature Plasma Treating method of 'beta '-mannase |
Applications Claiming Priority (1)
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| CN103255118A (en) * | 2013-05-21 | 2013-08-21 | 中南大学 | Beta-mannase, coding gene as well as producing strain and application thereof |
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| 低温等离子体技术及其在作物种子中的应用;张波等;《江苏农业科学》;20140102;第41卷(第9期);第58-60页 * |
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