CN102586207A - Method for preparing neutral bleaching xylanase - Google Patents
Method for preparing neutral bleaching xylanase Download PDFInfo
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- CN102586207A CN102586207A CN2012100520007A CN201210052000A CN102586207A CN 102586207 A CN102586207 A CN 102586207A CN 2012100520007 A CN2012100520007 A CN 2012100520007A CN 201210052000 A CN201210052000 A CN 201210052000A CN 102586207 A CN102586207 A CN 102586207A
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Abstract
The invention provides a method for preparing neutral bleaching xylanase, which comprises the following steps of: taking pichia pastoris as production strains, adopting a test-tube slant for activating, shake-flask amplification culturing, amplification culturing in a primary seeding tank, amplification culturing in a secondary seeding tank, and fermentation culturing in a fermentation tank. Due to the adoption of the method for preparing the neutral bleaching xylanase, the blank of application of the xylanase to a textile dyeing and bleaching process is filled, the filling power of cottonseed hulls can be enhanced so as to enable the removing effect of the cottonseed hulls to be good, the whiteness is obviously increased, the fiber cannot be damaged, the fiber strength cannot be affected, the weight loss of textile is reduced, the levelling property is good, the color yield is improved, the dyeing defective rate is reduced greatly, the dye can be saved, the hand feel and the softness can be improved simultaneously, the cloth cover is more bright, the textures are clearer, the processing time is shortened, and the production efficiency is improved.
Description
Technical field
The present invention relates to a kind of product preparation method that is used to bleach, relate in particular to a kind of preparation method of neutral bleaching zytase.
Background technology
The main alkali oxidation style (2g/L alkali and 3-5g/L ydrogen peroxide 50) that adopts is carried out refining bleaching in the printing and dyeing pre-treating process at present, not only damages fiber, also environment is caused serious harm, increases the weight of WWT burden and energy consumption cost.It is a kind of green bleaching method of brand-new gentleness that neutral bleaching zytase helps the technology of floating, and can use with neutral meta-alkalescence polygalacturonase compatibility, at utmost keeps the intensity of fiber; Weightless little; Fabrics feel soft is abundant, and high resilience, the most important thing is that bleaching effect is good.
Xylan is a hemicellulose components main in the vegetable cell, accounts for 35% of vegetable cell dry weight, is a kind of abundant biomass resource, is occurring in nature the abundantest polysaccharide of content except that Mierocrystalline cellulose.It has two types, and a kind of is cereal endosperm camber ramose araboxylan; A kind of is the less araboxylan of branch, has uronic acid and galactose residue at side chain, is present in usually in the highly lignified tissue, for example the stem of grass class, corn straw and big wheat husk etc.In pulp cooking process, xylan is partly dissolved, sex change also is deposited on the fiber surface again.If use zytase, just can remove the xylan that part deposits again in this process.Increase the paper pulp matrix pores like this, stranded solubility xylogen is discharged, also made the chemical bleaching agent more effectively to penetrate in the paper pulp simultaneously.Generally speaking, it can improve the bleachability of paper pulp, and therefore reduces the consumption of chemical bleaching agent.Zytase is more in the market is used in the middle of pulping bleaching, food and the feed, and zytase is used to weave, and still genus is blank in the biorefining bleaching.
In view of this, how with the zytase biorefining bleaching field that applies to weave, and it is not high to solve the cotton seed hulls loft; Whiteness increases not obvious; The easy damaged fiber influences fiber strength, and level-dyeing property, dye yield, dyeing fraction defective are not high; Problems such as the process time is long become an important research project.
Summary of the invention
The present invention provides a kind of preparation method of neutral bleaching zytase, is used to fill up zytase be used to the to weave blank of biorefining bleaching, enhances productivity.
According to the present invention, a kind of preparation method of neutral bleaching zytase is provided, may further comprise the steps:
1) is to produce bacterial classification with the pichia spp, adopts the test tube slant activation, shake the mode cultured strain of bottle amplification culture;
2) with bacterial classification inoculation to first class seed pot, tank pressure 0.07～0.08Mpa, air quantity 16～18m
3/ h, pH value nature was cultivated 15～20 hours for 29～31 ℃;
3), the bacterial classification weight in wet base that detects the first class seed pot amplification culture changes the secondary seed jar over to, tank pressure 0.07～0.08Mpa, air quantity 180～200m when reaching 30～40g/L
3/ h, pH value nature was cultivated 10～15 hours for 29～31 ℃;
4), the bacterial classification weight in wet base that detects secondary seed jar amplification culture changes fermentor tank over to when reaching 30～40g/L, tank pressure 0.07～0.08Mpa, and initial air quantity is 1000m
3/ h cultivates after 5 hours air quantity and opens to maximum, and initial mixing speed is 140r/min; Cultivate that mixing speed is adjusted to 180～220r/min after 8 hours, in culturing process, add ammoniacal liquor the pH value is maintained 5.5, the yeast culture temperature is 29～31 ℃; When reducing sugar is 0.1%～0.2%, add methyl alcohol, temperature is adjusted into 27～29 ℃; Inoculum size 5%～30%, ferment tank culture cycle are 120～130 hours;
5) stop adding methyl alcohol, blew sterile air 2 hours, begin jar, going out a jar pH value is 4.5～5.0.
Preferably, the used substratum of first class seed pot comprises neutral bleaching zytase preparing method's step 2): glucose 5Kg, peptone 5Kg, yeast powder 4Kg; Potassium primary phosphate 0.32Kg, ammonium sulfate 0.36Kg runs enemy 100ml; Add trace element after feeding intake, be settled to 150L, pH4.5～5.0;
Preferably, the used substratum of secondary seed jar in neutral bleaching zytase preparing method's the step 3) comprises: glucose 30Kg, peptone 15Kg; Potassium primary phosphate 9Kg, ammonium sulfate 9Kg runs enemy 1Kg; Add trace element after feeding intake, be settled to 1500L, pH4.8～5.3;
Preferably, the used substratum of fermentor tank comprises in neutral bleaching zytase preparing method's the step 4): peptone 22Kg, and potassium primary phosphate 160Kg, ammonium sulfate 180Kg runs enemy 10Kg, adds trace element after feeding intake, and is settled to 11m
3, pH5.5～6.0;
Preferably, go out jar real before discharging pipeline that disappears in neutrality bleaching zytase preparing method's the step 5), sterilized 30 minutes for 121～122 ℃;
Preferably, the neutrality the prepared bleaching zytase biorefining bleaching field that is used to weave;
Preferably, the prescription of the biorefining bleaching field that is used to weave comprises neutral bleaching zytase, permeate agent, laccase, Sodium Acid Pyrophosphate and trisodium phosphate;
Preferably; The weight percent of neutral bleaching zytase is 50%～80% of prescription in the prescription of neutral bleaching zytase; The weight percent of permeate agent is 5%～10% of prescription; The weight percent of laccase is 5%～15% of prescription, and the weight percent of Sodium Acid Pyrophosphate is 5%～10% of prescription, and the weight percent of trisodium phosphate is 5%～15% of prescription.
Beneficial effect of the present invention is:
Adopt neutral bleaching zytase of the present invention, filled up the blank that zytase is used on the textile printing and dyeing bleaching process, neutral bleaching zytase of the present invention can partly replace multiple auxiliary agents such as the caustic soda, refining agent, ydrogen peroxide 50, SYNTHETIC OPTICAL WHITNER, hydrogen peroxide bleaching stabilizer, removers in the traditional technology, simplifies original working method; No longer need transfer pH value, effectively shorten man-hour, significantly reduce TDS, COD, BOD index in the waste water, reduce environmental pollution; The removal cotton seed hulls is effective, and fabric whiteness obviously increases, and does not damage fiber, does not influence fiber strength; Fabric is weightless to be reduced, good level-dyeing property, and dye yield improves, and the dyeing fraction defective declines to a great extent; Can save dyestuff, improve feel and flexibility, make that cloth cover is more glossy, texture is more clear, handle combining with biopolishing; Fabric coloured light can not change, and shortens the process time, enhances productivity; The water loss of full technology can reduce more than 30%, and energy expenditure reduces simultaneously, practices thrift water power, steam.
Description of drawings
The reader with reference to advantages after the embodiment of the present invention, will become apparent all respects of the present invention.Wherein,
Fig. 1 is according to the present invention, preparing method's schema of neutral bleaching zytase.
Embodiment
With reference to the accompanying drawings, embodiment of the present invention is done further to describe in detail.
Fig. 1 is according to the present invention, preparing method's schema of neutral bleaching zytase.As shown in Figure 1, the preparation method of neutral bleaching zytase of the present invention comprises the slant strains cultivation, shakes a bottle amplification culture, first class seed pot amplification culture, secondary seed jar amplification culture, fermentor tank mixed fermentation.Wherein, the raw material that the preparation method of neutral bleaching zytase needs comprises the first class seed pot substratum: glucose 5Kg, and peptone 5Kg, yeast powder 4Kg, potassium primary phosphate 0.32Kg, ammonium sulfate 0.36Kg runs enemy 100ml, trace element; Secondary seed jar substratum: glucose 30Kg, peptone 15Kg, potassium primary phosphate 9Kg, ammonium sulfate 9Kg runs enemy 1Kg, trace element; Fermentation tank culture medium: peptone 22Kg, potassium primary phosphate 160Kg, ammonium sulfate 180Kg runs enemy 10Kg, trace element; Sugared in batches: glucose 800Kg, trace element; Glycerine: glycerine 600Kg, trace element, the trace element in the raw material comprises sal epsom, ferrous sulfate, calcium sulfate, vitriolate of tartar.
Embodiment 1
1, slant strains is cultivated
Adopt pichia spp as producing bacterial classification, used BMGY slant medium comprises: yeast powder 1%, and peptone 2%, glycerine 1%, YNB 1.34%, transfers pH to 6.0, adds 2.5% agar then, and 30 ℃ of static cultivations 48 hours are put into 4 ℃ of refrigerators and are preserved.
2, shake a bottle amplification culture
The single colony inoculation of picking in the triangular flask that contains 500ml BMGY, 30 ℃, the 250r/min overnight cultures, OD reaches at 2～6 o'clock, in culture is added jar.
3, first class seed pot amplification culture
According to first class seed pot prescription batching, add trace elements such as magnesium, iron, sal epsom after feeding intake, raw material is settled to 150L in jar; Disappear in fact and adopt direct air inlet, guarantee the real back volume that disappears at 160L, pH is 4.5; Be seeded to first class seed pot after disappearing in fact and carry out amplification culture, tank pressure 0.07Mpa, air quantity 16m
3/ h does not open stirring, and pH value nature was cultivated 15 hours for 29 ℃.Change kind of back 0 hour and begin to measure, per hour measure once subsequently, pH is remained on about 4.5, change kind of back 0 hour and observe by fermentation beginning microscopy by fermentation; Per hour observe once, decide initial value by middle detection before reducing sugar changes kind after sterilization, after 12 hours cycles the second time mensuration, per subsequently 3 hours mensuration is once; The weight in wet base of bacterial classification is begun to measure by middle control changeing kind of back 0 hour, and 12 hours cycles were measured for the second time, measured once in per subsequently 3 hours; Reach 30g/L in weight in wet base, comprehensively judge according to mash viscosity, smell, the microscopy thalli growth is good; Growing way is vigorous, and no microbiological contamination phenomenon changes the secondary jar over to.
4, secondary seed jar amplification culture
According to secondary seed jar prescription batching, add trace elements such as magnesium, iron, sal epsom after feeding intake, raw material is settled to 1500L in jar; Disappear in fact and adopt direct air inlet, guarantee the real back volume that disappears at 1500L, pH is 4.8; Be seeded to the secondary seed jar after disappearing in fact and carry out amplification culture, tank pressure 0.07Mpa, air quantity 180m
3/ h does not open stirring, and pH value nature was cultivated 10 hours for 29 ℃.Change kind of back 0 hour and begin to measure, per hour measure once subsequently, pH is remained on about 4.8, change kind of back 0 hour and observe by fermentation beginning microscopy by fermentation; Per hour observe once, decide initial value by middle detection before reducing sugar changes kind after sterilization, cultivate after 4 hours per 4 hours mensuration once; The weight in wet base of bacterial classification is begun to measure by middle control changeing kind of back 0 hour, measures once in per subsequently 4 hours, cultivates after 8 hours per 2 hours mensuration once; Reach 50g/L in weight in wet base, comprehensively judge according to mash viscosity, smell, the microscopy thalli growth is good; Growing way is vigorous, and no microbiological contamination phenomenon changes fermentor tank over to.
5, fermentor tank mixed fermentation
Each composition of fermentation tank culture medium is mixed in rotary spherical digester,, sterilized 60 minutes through 121 ℃ in three normal atmosphere; Fermentor tank is dropped in the cooling back; Change bacterial classification over to fermentor tank simultaneously, the back that feeds intake adds trace elements such as magnesium, iron, sal epsom, and raw material in the jar is settled to 11m
3, disappear in fact and adopt direct air inlet, guarantee that the real back volume that disappears is at 12.5m
3, accent pH to 5.5, the back that disappears is in fact prepared to change and is planted.
The glucose 500Kg that in the secondary jar, disappears adds trace element after feeding intake, and is settled to 1000L; The back disappear in fact as in the fermentor tank of twice adding of sugar just, and the glucose 800Kg that in the feed supplement jar, disappears adds trace element after feeding intake; Be settled to 1500L, sugaring adds in the fermentor tank as stream in the back that disappears in fact, in addition at feed supplement jar 30% glycerine that disappears: 600Kg glycerine; Add trace element after feeding intake, be settled to 1.8m
3, the back that disappears in fact added in the fermentor tank in the mixed stage of raising.
Fermentor tank tank pressure 0.07Mpa, initial air quantity is 1000m
3/ h cultivates air quantity 1300m after 5 hours
3More than/the h, open to maximum, dissolved oxygen requires greater than 20%; Initial mixing speed is 140r/min; Cultivate that mixing speed is adjusted to 180r/min after 8 hours, in culturing process, add ammoniacal liquor the pH value is maintained 5.5, the yeast culture temperature is 29 ℃; Inoculum size 5%, ferment tank culture cycle are 120 hours.Change and plant the back, per hour measure the pH value one time, the pH value is maintained about 5.5, carry out microscopy simultaneously and detect observation, per hour once by the fermentation beginning.Reducing sugar is begun to measure by middle control changeing kind of back 0 hour; Per subsequently 4 hours once, and weight in wet base is begun to measure by middle control changeing kind of back 0 hour, and per subsequently 4 hours once; Amino nitrogen is begun to measure by middle control changeing kind of back 0 hour; Measured in per subsequently about 24 hours once, cultivate and begin to measure enzyme by middle control after 48 hours and live, per subsequently about 12 hours mensuration once.Basically consumed at a big jar reducing sugar, reducing sugar is 0.1%, and weight in wet base 90g/L stops to flow sugaring; Blew sterile air 1 hour, and obviously dropped to characteristic with pH value nothing, begin to flow glycerol adding, initial flow is 60L/h; Stream adds methanol induction simultaneously, and temperature is adjusted into 27 ℃, to improve yield of enzyme.
In the culture cycle ending phase, if the microscopy thalline is aging, enzyme is lived in increasing and is lower than 2% and can considers to put jar, receive specifically go out jar time after, stop methyl alcohol, blow sterile air about 2 hours, the contact extraction personnel discharging pipeline that disappears begins jar.Stop air compressor machine according to producing a upward operation jar number contact compressed air station, go out a jar pH and be adjusted into 4.5.Culture transferring and stream add pipeline and must disappear in fact thoroughly.Disappear line temperature at 121 ℃ in fact, and disinfecting time is about 30 minutes.
Embodiment 2
1, slant strains is cultivated
Adopt pichia spp as producing bacterial classification, used BMGY slant medium comprises: yeast powder 1%, and peptone 2%, glycerine 1%, YNB 1.34%, transfers pH to 6.0, adds 2.5% agar then, and 30 ℃ of static cultivations 48 hours are put into 4 ℃ of refrigerators and are preserved.
2, shake a bottle amplification culture
The single colony inoculation of picking in the triangular flask that contains 500ml BMGY, 30 ℃, the 250r/min overnight cultures, OD reaches at 2～6 o'clock, in culture is added jar.
3, first class seed pot amplification culture
According to first class seed pot prescription batching, add trace elements such as magnesium, iron, sal epsom after feeding intake, raw material in the jar is settled to 150L; Disappear in fact and adopt direct air inlet, guarantee the real back volume that disappears at 180L, pH is 5.0; Be seeded to first class seed pot after disappearing in fact and carry out amplification culture, tank pressure 0.08Mpa, air quantity 18m
3/ h does not open stirring, and pH value nature was cultivated 20 hours for 31 ℃.Change kind of back 0 hour and begin to measure, measured once in per subsequently 7 hours, pH is remained on about 5.0, change kind of back 0 hour and observe by fermentation beginning microscopy by fermentation; Per 7 h observation once decide initial value by middle detection before reducing sugar changes kind after sterilization, mensuration for the second time after 12 hours cycles, and per subsequently 5 hours mensuration is once; The weight in wet base of bacterial classification is begun to measure by middle control changeing kind of back 0 hour, and 12 hours cycles were measured for the second time, measured once in per subsequently 5 hours; Reach 40g/L in weight in wet base, comprehensively judge according to mash viscosity, smell, the microscopy thalli growth is good; Growing way is vigorous, and no microbiological contamination phenomenon changes the secondary jar over to.
4, secondary seed jar amplification culture
According to secondary seed jar prescription batching, add trace elements such as magnesium, iron, sal epsom after feeding intake, raw material is settled to 1500L in jar; Disappear in fact and adopt direct air inlet, guarantee the real back volume that disappears at 1700L, pH is 5.3; Be seeded to the secondary seed jar after disappearing in fact and carry out amplification culture, tank pressure 0.08Mpa, air quantity 200m
3/ h does not open stirring, and pH value nature was cultivated 15 hours for 31 ℃.Change kind of back 0 hour and begin to measure, measured once in per subsequently 3 hours, pH is remained on about 5.3, change kind of back 0 hour and observe by fermentation beginning microscopy by fermentation; Per 5 h observation are once decided initial value by middle detection before reducing sugar changes kind after sterilization, cultivate after 4 hours per 4 hours mensuration once; The weight in wet base of bacterial classification is begun to measure by middle control changeing kind of back 0 hour, measures once in per subsequently 4 hours, cultivates after 8 hours per 2 hours mensuration once; Reach 50g/L in weight in wet base, comprehensively judge according to mash viscosity, smell, the microscopy thalli growth is good; Growing way is vigorous, and no microbiological contamination phenomenon changes fermentor tank over to.
5, fermentor tank mixed fermentation
Each composition of fermentation tank culture medium is mixed in rotary spherical digester,, sterilized 60 minutes through 121 ℃ in three normal atmosphere; Fermentor tank is dropped in the cooling back; Change bacterial classification over to fermentor tank simultaneously, the back that feeds intake adds trace elements such as magnesium, iron, sal epsom, and raw material in the jar is settled to 11m
3, disappear in fact and adopt direct air inlet, guarantee that the real back volume that disappears is at 13.5m
3, accent pH to 6.0, the back that disappears is in fact prepared to change and is planted.
The glucose 500Kg that in the secondary jar, disappears adds trace element after feeding intake, and is settled to 1000L; The back disappear in fact as in the fermentor tank of twice adding of sugar just, and the glucose 800Kg that in the feed supplement jar, disappears adds trace element after feeding intake; Be settled to 1500L, sugaring adds in the fermentor tank as stream in the back that disappears in fact, in addition at feed supplement jar 30% glycerine that disappears: 600Kg glycerine; Add trace element after feeding intake, be settled to 1.8m
3, the back that disappears in fact added in the fermentor tank in the mixed stage of raising.
Fermentor tank tank pressure 0.08Mpa, initial air quantity is 1000m
3/ h cultivates air quantity 1300m after 5 hours
3More than/the h, open to maximum, dissolved oxygen requires greater than 20%; Initial mixing speed is 140r/min; Cultivate that mixing speed is adjusted to 220r/min after 8 hours, in culturing process, add ammoniacal liquor the pH value is maintained 5.5, the yeast culture temperature is 31 ℃; Inoculum size 30%, ferment tank culture cycle are 130 hours.Change and plant the back by the fermentation beginning, measured a pH value in per 3 hours, the pH value is maintained about 5.5, carry out the microscopy detection simultaneously and observe, per 5 hours once.Reducing sugar is begun to measure by middle control changeing kind of back 0 hour; Per subsequently 4 hours once, and weight in wet base is begun to measure by middle control changeing kind of back 0 hour, and per subsequently 4 hours once; Amino nitrogen is begun to measure by middle control changeing kind of back 0 hour; Measured in per subsequently about 24 hours once, cultivate and begin to measure enzyme by middle control after 48 hours and live, per subsequently about 12 hours mensuration once.Basically consumed at a big jar reducing sugar, reducing sugar is 0.2%, and weight in wet base 110g/L stops to flow sugaring; Blew sterile air 1 hour, and obviously dropped to characteristic with pH value nothing, begin to flow glycerol adding, initial flow is 80L/h; Stream adds methanol induction simultaneously, and temperature is adjusted into 29 ℃, to improve yield of enzyme.
In the culture cycle ending phase, if the microscopy thalline is aging, enzyme is lived in increasing and is lower than 2% and can considers to put jar, receive specifically go out jar time after, stop methyl alcohol, blow sterile air about 2 hours, the contact extraction personnel discharging pipeline that disappears begins jar.Stop air compressor machine according to producing a upward operation jar number contact compressed air station, go out a jar pH and be adjusted into 5.0.Culture transferring and stream add pipeline and must disappear in fact thoroughly.Disappear line temperature at 122 ℃ in fact, and disinfecting time is about 30 minutes.
Neutrality bleaching zytase with the neutral preparation method who bleaches zytase of the present invention prepares can be as a kind of system component of neutral bleaching zytase; This prescription comprises neutral bleaching zytase, laccase, Sodium Acid Pyrophosphate and trisodium phosphate; Permeate agent; In the present invention's one preferred embodiment, that permeate agent adopts is permeate agent AT-80, also can adopt AEO-9 or other model among other embodiment.Wherein, The weight percent of neutral bleaching zytase is 50%～80% of prescription; The weight percent of permeate agent AT-80 is 5%～10% of prescription; The weight percent of laccase is 5%～15% of prescription, and the weight percent of Sodium Acid Pyrophosphate is 5%～10% of prescription, and the weight percent of trisodium phosphate is 5%～15% of prescription.Neutral bleaching zytase of the present invention is added assistance bleaching in the neutral low-temperature refining enzyme.
Therefore, the preparation method by neutral bleaching zytase of the present invention has filled up the blank that zytase is used on the textile printing and dyeing bleaching method, not only can strengthen the cotton seed hulls loft and then make the cotton seed hulls removal effect good; And whiteness obviously increases, and does not damage fiber, does not influence fiber strength; Make the weightless reduction of fabric, good level-dyeing property, dye yield improves; The dyeing fraction defective declines to a great extent, and can save dyestuff, improves feel and flexibility simultaneously; Make that cloth cover is more glossy, texture is more clear, shorten the process time, enhance productivity.
In the preceding text, illustrate and describe embodiment of the present invention.But those of ordinary skill in the art can be understood, and under the situation without departing from the spirit and scope of the present invention, can also specific embodiments of the invention do various changes and replacement.These changes and replacement all drop in claims restricted portion of the present invention.
Claims (8)
1. the preparation method of a neutral bleaching zytase is characterized in that, may further comprise the steps:
1) is to produce bacterial classification with the pichia spp, adopts the test tube slant activation, shake the mode cultured strain of bottle amplification culture;
2) with said bacterial classification inoculation to first class seed pot, tank pressure 0.07～0.08Mpa, air quantity 16～18m
3/ h, pH value nature was cultivated 15～20 hours for 29～31 ℃;
3), the bacterial classification weight in wet base that detects said first class seed pot amplification culture changes the secondary seed jar over to, tank pressure 0.07～0.08Mpa, air quantity 180～200m when reaching 30～40g/L
3/ h, pH value nature was cultivated 10～15 hours for 29～31 ℃;
4), the bacterial classification weight in wet base that detects said secondary seed jar amplification culture changes fermentor tank over to when reaching 30～40g/L, tank pressure 0.07～0.08Mpa, and initial air quantity is 1000m
3/ h cultivates after 5 hours air quantity and opens to maximum, and initial mixing speed is 140r/min; Cultivate that mixing speed is adjusted to 180～220r/min after 8 hours, in culturing process, add ammoniacal liquor the pH value is maintained 5.5, the yeast culture temperature is 29～31 ℃; When reducing sugar is 0.1%～0.2%, add methyl alcohol, temperature is adjusted into 27～29 ℃; Inoculum size 5%～30%, ferment tank culture cycle are 120～130 hours;
5) stop adding said methyl alcohol, blew sterile air 2 hours, begin jar, going out a jar pH value is 4.5～5.0.
2. the preparation method of neutral bleaching zytase as claimed in claim 1 is characterized in that step 2) described in the used substratum of first class seed pot comprise: glucose 5Kg; Peptone 5Kg, yeast powder 4Kg, potassium primary phosphate 0.32Kg; Ammonium sulfate 0.36Kg runs enemy 100ml, adds trace element after feeding intake; Be settled to 150L, pH4.5～5.0.
3. the preparation method of neutral bleaching zytase as claimed in claim 1 is characterized in that the used substratum of secondary seed jar described in the step 3) comprises: glucose 30Kg; Peptone 15Kg, potassium primary phosphate 9Kg, ammonium sulfate 9Kg; Run enemy 1Kg; Add trace element after feeding intake, be settled to 1500L, pH4.8～5.3.
4. the preparation method of neutral bleaching zytase as claimed in claim 1 is characterized in that the used substratum of the fermentor tank described in the step 4) comprises: peptone 22Kg; Potassium primary phosphate 160Kg, ammonium sulfate 180Kg runs enemy 10Kg; Add trace element after feeding intake, be settled to 11m
3, pH5.5～6.0.
5. the preparation method of neutrality bleaching zytase as claimed in claim 1 is characterized in that, goes out jar real before discharging pipeline that disappears in the step 5), sterilizes 30 minutes for 121～122 ℃.
6. the preparation method of neutral bleaching zytase as claimed in claim 1 is characterized in that, the neutrality of preparing with said preparation method is bleached the zytase biorefining bleaching field that is used to weave.
7. the preparation method of neutral bleaching zytase as claimed in claim 6 is characterized in that, the prescription of the said biorefining bleaching field that is used to weave comprises neutral bleaching zytase, permeate agent, laccase, Sodium Acid Pyrophosphate and trisodium phosphate.
8. the preparation method of neutral bleaching zytase as claimed in claim 6; It is characterized in that; The weight percent of said neutral bleaching zytase is 50%～80% of a said prescription, and the weight percent of said permeate agent is 5%～10% of a said prescription, and the weight percent of said laccase is 5%～15% of a said prescription; The weight percent of said Sodium Acid Pyrophosphate is 5%～10% of a said prescription, and the weight percent of said trisodium phosphate is 5%～15% of a said prescription.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103923846A (en) * | 2013-12-23 | 2014-07-16 | 华北制药集团新药研究开发有限责任公司 | Pichia pastoris culturing medium |
| CN104928271A (en) * | 2015-07-06 | 2015-09-23 | 唐亚军 | Preparation method for xylanase |
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