CN109810960A - A kind of method that spray drying process prepares alkalescent xylanase preparation - Google Patents
A kind of method that spray drying process prepares alkalescent xylanase preparation Download PDFInfo
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- CN109810960A CN109810960A CN201711152090.6A CN201711152090A CN109810960A CN 109810960 A CN109810960 A CN 109810960A CN 201711152090 A CN201711152090 A CN 201711152090A CN 109810960 A CN109810960 A CN 109810960A
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- alkalescent xylanase
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Abstract
The present invention relates to microbial fermentation and enzyme preparation technical fields, and in particular to a kind of method that wheat bran fermentation combines spray drying to prepare alkalescent xylanase preparation.It is fermented using wheat bran, and wheat bran ferments through bacillus pumilus produces alkalescent xylanase, and the producing enzyme vigor 2700IU/ml of fermentation liquid is greatly improved in terms of producing enzyme vigor；The alkaline xylose enzyme preparation of the spray-dried preparation of fermentation liquid, eliminates ammonium sulfate precipitation, greatly reduces discharging of waste liquid.So that the producing enzyme vigor in alkalescent xylanase preparation process is improved four times or more, greatly improves production efficiency, be conducive to technological industrialization.The processing step that the present invention uses is 1) fermentation；2) enzyme preparation.
Description
Technical field
The present invention relates to microbial fermentation technologies and enzyme preparation technical field, and in particular to spray drying preparation alkalinity wood is poly-
The method of carbohydrase preparation.
Background technique
Zytase (Xylanase) is the enzyme of single-minded degradation of xylan, belongs to hydrolase, including a variety of restriction endonucleases and
Excision enzyme.The alkalescent xylanase that bacillus pumilus generates has stability under alkaline condition and effect is most strong, can incite somebody to action
Xylan hydrolysis is xylo-oligosaccharide or xylose.Therefore it can be used for the celluloses manufacturing industry such as papermaking, ramie processing, alkalescent xylanase
It can make hemicellulose (predominantly xylan) selectively partial hydrolysis, without damaging fibre strength, reduce subsequent technique
The dosage of middle chemicals reduces pollution of the harmful chemicals object to environment.Spray drying prepares alkalescent xylanase preparation, Ke Yidai
Manufacture enzyme preparation technique is precipitated for traditional ammonium sulfate precipitation, the addition of chemicals ammonium sulfate and the discharge of waste liquid is saved, reduces enzyme
Preparation produces the pollution to environment.
Microbial fermentation produces alkalescent xylanase and starts late in China, is in phase of basic research mostly, domestic autonomous
Product is rare, and product enzyme activity is low.Chinese patent literature (ZL200510018574.2) discloses a kind of system of alkalescent xylanase
Preparation Method, what is utilized is saccharomycetes to make fermentation production zytase, and fermentation liquid producing enzyme vigor is 426IU/ml.Chinese patent literature
(ZL200610020049.9) it is poly- to produce alkalinity wood also with saccharomycetes to make fermentation for the preparation method of alkalescent xylanase disclosed in
Carbohydrase.Its fermentation liquid producing enzyme vigor is 650IU/ml.
Summary of the invention
The method that spray drying process provided by the invention prepares alkalescent xylanase preparation, is fermented using wheat bran, and wheat bran is through short
Bacillus pumilus fermentation produces alkalescent xylanase, and the producing enzyme vigor 2700IU/ml of fermentation liquid is obtained very big in terms of producing enzyme vigor
Improve；The alkaline xylose enzyme preparation of the spray-dried preparation of fermentation liquid, eliminates ammonium sulfate precipitation, greatly reduces discharging of waste liquid.
So that the producing enzyme vigor in alkali zytase preparation process is improved four times or more, greatly improves production efficiency, be conducive to technology
Industrialization.
To achieve the above object, the technical solution adopted by the present invention are as follows: spray drying process prepares alkalescent xylanase preparation
Method the step of are as follows:
1) it ferments: fluid nutrient medium A being packed into after fermentor and adjusts pH to 7.5-8.0, after conventional sterilant, according to volume ratio 2-5%
Sterile working accesses the good bacillus pumilus strain of activation culture, is 37 DEG C, speed of agitator 300-500r/min in temperature, leads to
Tolerance 25-35L/min, tank press 0.03-0.06MPa, fermentation period 24-36h；Producing enzyme vigor is surveyed with 3,5- dinitrosalicylic acid system
Stop fermentation when up to 2500IU/ml；
2) preparation of enzyme preparation: use high-efficiency plate frame formula filter, under 0.05-0.1MPa pressure remove fermentation liquid in thallus and
Insoluble impurities, acquisition brown supernatant liquid are enzyme solution；Dextrin is added according to the mass ratio of 8-12%, stirs evenly, in air inlet temperature
Be spray-dried under the conditions of 180-200 DEG C, 80-100 DEG C of leaving air temp of degree, obtain the alkalescent xylanase preparation of powdery solid at
Product, product enzyme activity 10000-20000IU/g, yield 1000000-1500000IU/L fermentation liquid.
The formula of fluid nutrient medium A in the step 1) is as follows: wheat bran 5.0-8.0%, peptone 0.5-2.0%, ferment
Female cream 0.2-0.5%, ammonium chloride 0.5-1.5%, sodium chloride 0.01-0.025%, dipotassium hydrogen phosphate 0.2-0.5%, calcium chloride 0.05-
0.1%, magnesium sulfate 0.05-0.1%, excess water.
Compared with prior art, the invention has the advantages that and effect: the present invention is using microbial fermentation and spraying dry
Dry production alkalescent xylanase has the advantage that, thermal stability is strong, optimal reactive temperature 50 compared with common zytase
DEG C, it is stable under alkaline condition, the celluloses such as papermaking processing industry using when do not have to be adjusted pH value, reduce technique stream
Journey has apparent economic benefit, and the use that the use of the alkalescent xylanase of efficient enzyme activity can reduce bleaching agent is conducive to protect
Environment.Simultaneous spray drying is used for the production of alkalescent xylanase, can replace ammonium sulfate precipitation technique, avoids chemicals sulphur
The addition of sour ammonium and the discharge of waste liquid, can not only cut down the consumption of raw materials, but also can mitigate environmental protection of enterprise load.
Four, specific embodiment
Embodiment 1:
Spray drying prepares the step of method of alkalescent xylanase preparation are as follows:
Step 1: fermentation:
Fluid nutrient medium A is packed into after fermentor and adjusts pH to 8.0, after conventional sterilant, is accessed according to 5% sterile working of volume ratio
The good bacillus pumilus strain of activation culture is 37 DEG C, speed of agitator 500r/min, ventilatory capacity 28L/min in temperature, tank pressure
0.05MPa, fermentation period is for 24 hours；It is surveyed when producing enzyme vigor reaches 2500IU/ml with 3,5- dinitrosalicylic acid system and stops fermentation；
Step 2: the preparation of enzyme preparation:
With high-efficiency plate frame formula filter, thallus and insoluble impurities in fermentation liquid are removed under 0.1MPa pressure, obtains brown
Supernatant liquid is enzyme solution；Dextrin is added according to 10% mass ratio, stirs evenly, at 190 DEG C of inlet air temperature, 90 DEG C of leaving air temp
Under the conditions of be spray-dried, obtain the alkalescent xylanase finished dosage form of powdery solid, product enzyme activity 20000IU/g, yield
1500000IU/L fermentation liquid.
That mentions in step 1 has fluid nutrient medium A formula as follows:
Wheat bran 6.0%, peptone 1.0%, yeast extract 0.3%, ammonium chloride 0.9%, sodium chloride 0.018%, dipotassium hydrogen phosphate 0.4%, chlorine
Change calcium 0.08%, magnesium sulfate 0.07%, excess water.
Wheat Straw Pulp Bleaching test:
The alkalescent xylanase of above-mentioned preparation is for non-bleached wheat straw pulp bleaching in paper technology, Straw Pulp concentration 5.0%,
PH9.6,50 DEG C, enzyme preparation dosage 1g/kg over dry Straw Pulp acts on 1.0h.Improve wheat straw pulp brightness 6%.
Embodiment 2:
Spray drying prepares the step of method of alkalescent xylanase preparation are as follows:
Step 1: fermentation:
Fluid nutrient medium A is packed into after fermentor and adjusts pH to 7.8, after conventional sterilant, is connect according to 3.5% sterile working of volume ratio
Enter the good bacillus pumilus strain of activation culture, is 37 DEG C, speed of agitator 400r/min, ventilatory capacity 35L/min in temperature, tank
Press 0.04MPa, fermentation period 30h；It is surveyed when producing enzyme vigor reaches 2500IU/ml with 3,5- dinitrosalicylic acid system and stops fermentation；
Step 2: the preparation of enzyme preparation:
With high-efficiency plate frame formula filter, thallus and insoluble impurities in fermentation liquid are removed under 0.08MPa pressure, obtains brown
Supernatant liquid is enzyme solution；Dextrin is added according to 8% mass ratio, stirs evenly, at 180 DEG C of inlet air temperature, 80 DEG C of items of leaving air temp
It is spray-dried under part, obtains the alkalescent xylanase finished dosage form of powdery solid, product enzyme activity 15000IU/g, yield
1300000IU/L fermentation liquid.
That mentions in step 1 has fluid nutrient medium A formula as follows:
Wheat bran 8.0%, peptone 2.0%, yeast extract 0.5%, ammonium chloride 1.5%, sodium chloride 0.025%, dipotassium hydrogen phosphate 0.5%, chlorine
Change calcium 0.1%, magnesium sulfate 0.1%, excess water.
Wheat Straw Pulp Bleaching test:
The alkalescent xylanase of above-mentioned preparation is for non-bleached wheat straw pulp bleaching in paper technology, Straw Pulp concentration 5.0%,
PH9.6,50 DEG C, enzyme preparation dosage 1.5g/kg over dry Straw Pulp acts on 1.0h.Improve wheat straw pulp brightness 6%.
Embodiment 3:
Spray drying prepares the step of method of alkalescent xylanase preparation are as follows:
Step 1: fermentation:
Fluid nutrient medium A is packed into after fermentor and adjusts pH to 7.5, after conventional sterilant, is connect according to 2.0% sterile working of volume ratio
Enter the good bacillus pumilus strain of activation culture, is 37 DEG C, speed of agitator 300r/min, ventilatory capacity 25L/min in temperature, tank
Press 0.03MPa, fermentation period 36h；It is surveyed when producing enzyme vigor reaches 2500IU/ml with 3,5- dinitrosalicylic acid system and stops fermentation；
Step 2: the preparation of enzyme preparation:
With high-efficiency plate frame formula filter, thallus and insoluble impurities in fermentation liquid are removed under 0.05MPa pressure, obtains brown
Supernatant liquid is enzyme solution；Dextrin is added according to 12% mass ratio, stirs evenly, at 200 DEG C of inlet air temperature, 90 DEG C of leaving air temp
Under the conditions of be spray-dried, obtain the alkalescent xylanase finished dosage form of powdery solid, product enzyme activity 10000IU/g, yield
1000000IU/L fermentation liquid.
That mentions in step 1 has fluid nutrient medium A formula as follows:
Wheat bran 5.0%, peptone 0.5%, yeast extract 0.2%, ammonium chloride 0.5%, sodium chloride 0.01%, dipotassium hydrogen phosphate 0.2%, chlorination
Calcium 0.05%, magnesium sulfate 0.05%, excess water.
Wheat Straw Pulp Bleaching test:
The alkalescent xylanase of above-mentioned preparation is for non-bleached wheat straw pulp bleaching in paper technology, Straw Pulp concentration 5.0%,
PH9.6,50 DEG C, enzyme preparation dosage 2g/kg over dry Straw Pulp acts on 1.0h.Improve wheat straw pulp brightness 6%.
Bacillus pumilus number CGMCC1.2160 in embodiment is purchased and is entrusted in Chinese microorganism strain preservation management
Member's meeting common micro-organisms center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, postcode: 100101.Signature:
Bacillus, epitheton: pumilus, Chinese scientific name: bacillus pumilus.
Step 1) -3 in embodiment) in, the enzyme activity (IU) mentioned is defined as: with 1% xylan solution, temperature 55
DEG C, pH8.0, under the conditions of reacting 10min, enzyme amount required for the 1 μm of ol reduced sugar (in terms of xylose) of release in 1 minute is defined as one
Enzyme activity unit (IU).
Claims (2)
1. the method that spray drying prepares alkalescent xylanase preparation, it is characterised in that: the processing step are as follows:
1) it ferments:
Fluid nutrient medium A is packed into after fermentor and adjusts pH to 7.5-8.0, after conventional sterilant, according to the sterile behaviour of volume ratio 2-5%
Make the good bacillus pumilus strain of access activation culture, is 37 DEG C, speed of agitator 300-500r/min in temperature, ventilatory capacity
25-35L/min, tank press 0.03-0.06MPa, fermentation period 24-36h；Producing enzyme vigor is surveyed with 3,5- dinitrosalicylic acid system to reach
Stop fermentation when 2500IU/ml；
2) preparation of enzyme preparation:
With high-efficiency plate frame formula filter, thallus and insoluble impurities in fermentation liquid are removed under 0.05-0.1MPa pressure, is obtained
Brown supernatant liquid is enzyme solution；Dextrin is added according to the mass ratio of 8-12%, stirs evenly, at 180-200 DEG C of inlet air temperature, out
It is spray-dried under the conditions of 80-100 DEG C of air temperature, obtains the alkalescent xylanase finished dosage form of powdery solid, product enzyme activity
10000-20000IU/g, yield 1000000-1500000IU/L fermentation liquid.
2. the method that the spray drying according to claim 1 prepares alkalescent xylanase preparation, it is characterised in that:
The processing step 1) in fluid nutrient medium A formula it is as follows:
Wheat bran 5.0-8.0%, peptone 0.5-2.0%, yeast extract 0.2-0.5%, ammonium chloride 0.5-1.5%, sodium chloride 0.01-
0.025%, dipotassium hydrogen phosphate 0.2-0.5%, calcium chloride 0.05-0.1%, magnesium sulfate 0.05-0.1%, excess water.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114891769A (en) * | 2022-06-12 | 2022-08-12 | 内蒙古溢多利生物科技有限公司 | Preparation method of xylanase |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114891769A (en) * | 2022-06-12 | 2022-08-12 | 内蒙古溢多利生物科技有限公司 | Preparation method of xylanase |
| CN114891769B (en) * | 2022-06-12 | 2023-11-28 | 内蒙古溢多利生物科技有限公司 | Preparation method of xylanase |
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Application publication date: 20190528 |