CN102228823A - Modified beer waste yeast adsorbent, preparation method and application thereof - Google Patents
Modified beer waste yeast adsorbent, preparation method and application thereof Download PDFInfo
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- CN102228823A CN102228823A CN2011101059530A CN201110105953A CN102228823A CN 102228823 A CN102228823 A CN 102228823A CN 2011101059530 A CN2011101059530 A CN 2011101059530A CN 201110105953 A CN201110105953 A CN 201110105953A CN 102228823 A CN102228823 A CN 102228823A
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Abstract
The invention relates to the technical field of biosorbent preparation and application, and specifically discloses a modified beer waste yeast adsorbent, a preparation method and an application thereof. The modified beer waste yeast adsorbent provided by the present invention is a novel adsorbent which is prepared from a cross-linked beer waste yeast modified through citric acid, and provides good absorbability for lysozyme such that the modified beer waste yeast adsorbent can be applicable for extracting the lysozyme.
Description
Technical field
The present invention relates to the preparation and the applied technical field of biological adsorption agent, be specifically related to a kind of modification beer waste yeast adsorbent and its production and application.
Background technology
Beer waste yeast is the accessory substance of fermentation industry, be rich in-OH ,-NH
2,-functional groups such as COOH.As a kind of adsorbent cheaply, often be used to adsorb the harmful substance in the waste water.Through the various functional groups of its surface grafting that are everlasting, increase active sites and count to strengthen its adsorption capacity.Many researchs are reported, utilize the group on beer waste yeast surface, carry out derivative reaction, come the chelating adsorbing metal ions.But the research that the beer waste yeast through modification is used to adsorb lysozyme, Shang Weijian has report.
Lysozyme is a kind of enzyme that is in daily use, be present in mainly that egg is clear, in the body fluid of people's tears, saliva and other animals, it can destroy-acetylmuramic acid in the cell membrane and the β-1 between the N-n acetylglucosamine n, 4 glycosidic bonds, make the insoluble glutinous polysaccharide of cell membrane resolve into the solubility glycopeptide, cause the effusion of cell wall rupture Dissolve things inside and make bacterolysis.Because have this character, lysozyme is used (R.Islam as anticorrisive agent usually, J.Kite, A.S.Baker, A.Ching Jr., M.R.Islam, African J.Biotechnology.Affinity purification of hen egg lysozyme using sephadex.5 (2006) 1902.1515-1520).Usually extract lysozyme from egg is clear, wherein the content of lysozyme is 0.34% (G.Zhang et al.Tris (hydroxymethyl) aminomethane-modified magnetic microspheres for rapid affinity purificationof lysozyme.Talanta 83 (2011) 1515-1520).Because the foreign protein of coexistence was more during egg was clear, therefrom separate, Purification of Lysozyme is cumbersome.At present, the separation of relevant lysozyme, purifying mainly contains following these methods: film separates (R.Ghosh, S.S.Silva, Z.F.Cui, Lysozyme separation by hollow-fibre ultrafiltration, 19-24 Biochem.Eng.J.6 (2000) 19-24), ion-exchange (E.Lichan, S.Nakai, J.Sim, D.B.Bragg, K.V.Lo, Lysozyme Separation from Egg White by Cation Exchange Column Chromatography, FoodSci.51 (1986) 1032-1036.C.Guerin-Dubiard, M.Pasco, A.Hietanen, A.Q.del Bosque, F.Nau, T.Croguennec, Hen egg white fractionation by ion-exchange chromatography.J.Chromatogr.A 1090 (2005) 58-67), (Y.C.Cheng is separated out in crystallization, R.F.Lobo, S.I.Sandler, A.M.Lenhoff, Kinetics and equilibria of lysozyme precipitation and crystallization in concentrated ammonium sulfate solutions.Biotechnology and Bioengineering.
Volume 94(2006) _ and 177-188.C.J.Coen, J.M.Prausnitz, H.W.Blanch, Protein salting-out:Phase equilibria in two-protein systems Biotechnology and Bioengineering
Volume 53, Issue6,Pages 567-574,20 March 1997.) and affinity precipitation (T.Goto, T.Ohkuri, S.Shioi, Y.Abe, T.Imoto, T.Ueda, Crystal structures of K33 mutant hen lysozymes with enhanced activities.Journal of Biochemistry Volume 144, Issue 5, November 2008, Pages 619-623.).At present, with the method separation of absorption and the research of protein purification report is arranged more.Mainly come selective absorption or specificity combining target albumen (T.Kawai by static, ion-exchange, hydrogen bond and affine absorption, K.Saito, W.Lee, Protein binding to polymer brush, based on ion-exchange, hydrophobic and affinity interactions, J.Chromatogr.B 790 (2003) 131-142.).The research of this respect also has much, for example: A.X.Lu etc. are by introducing Fe (III) absorption lysozyme (A.X.Lu et al.Preparation of Fe (III)-immobilized collagen fiber for lysozyme adsorption.Colloids and Surfaces A:Physicochem.Eng.Aspects 301 (2007) 85-93.) on collagenous fibres, selection dyestuff parts such as C.Garcia-Diego adsorb lysozyme (C.Garcia-Diego, J.Cuellar.Preparation and characterization of a dye-ligand adsorbent for lysozyme adsorption.Chemical Engineering Journal 143 (2008) 337-348.), Z.Lei etc. utilize carbosphere absorption lysozyme (Z.Lei et al.Adsorption of lysozyme on spherical mesoporous carbons (SMCs) replicated from colloidal silica arrays by chemical vapor deposition.Journal of Colloid and Interface Science 339 (2009) 439-445.); D.Aktas Uygun and G.Zhang etc. are by magnetic microsphere absorption lysozyme or the like (D.Aktas Uygun et al.Magnetic hydrophobic affinity nanobeads for lysozyme separation.Materials Science and Engineering C 29 (2009) 2165-2173.G.Zhang et.al.Tris (hydroxymethyl) aminomethane-modified magnetic microspheres for rapid.affinity purification of lysozyme.Talanta 83,2011.).Yet the equal more complicated of these methods, cost are higher.So, be necessary to study a kind of simply, separate fast, the method for Purification of Lysozyme.
Summary of the invention
At the deficiencies in the prior art, the object of the present invention is to provide a kind of modification beer waste yeast adsorbent and its production and application.
Modification beer waste yeast adsorbent provided by the present invention is a kind of new adsorbent that makes with the crosslinked beer waste yeast of citric acid modification (crosslinking agent is a glutaraldehyde), this adsorbent has good adsorption capacity to lysozyme, can be applied to extract lysozyme.
In order to realize above-mentioned technical purpose, the present invention has taked following technical measures:
A kind of modification beer waste yeast adsorbent, the step of its preparation method is as follows:
(1) preliminary treatment of beer waste yeast:
Take by weighing beer waste yeast and place conical flask, the water that adds 8-12 times of weight, take out behind the 20-30 ℃ of water-bath vibration 0.5-1.5h and leave standstill 20-40min, abandon the upper strata turbid solution, it is colourless that water continues beer waste yeast is washed till supernatant, use absolute ethanol washing 2-4 time again after, leave standstill, suction filtration makes Separation of Solid and Liquid then, and the gained solid places 60 ℃ of baking ovens dry, standby;
(2) preparation of crosslinked beer waste yeast:
Take by weighing through the pretreated beer waste yeast of step (1) and join in the glutaraldehyde solution of 2wt%, the consumption of glutaraldehyde solution is the every g beer waste yeast of 30-35mL, behind 30-40 ℃ of shaking table oscillating reactions 10-12h, collect product, product washes with water earlier three times, use absolute ethanol washing again three times,, place 60 ℃ of oven dryings at last to remove the unreacted glutaraldehyde in product surface, obtain yellow crosslinked beer waste yeast (also claiming the unmodified yeast), standby;
(3), modify the preparation of beer waste yeast:
Take by weighing the crosslinked beer waste yeast of step (2) preparation, amount according to the crosslinked beer waste yeast of the every g of 10-13mL adds N, dinethylformamide (DMF), the citric acid of quality such as adding and crosslinked beer waste yeast and the sodium dihydrogen phosphate (NaH of 0.5 times of crosslinked beer waste yeast quality again
2PO
4As catalyst), place three-neck flask 130-140 ℃ of reaction 1.5-2.5h, and cooling for reflux, utilize the water that produces in the water knockout drum branch dereaction process; After reaction finished, suction filtration carried out Separation of Solid and Liquid, and it is colourless washing solid product to filtrate with water, use absolute ethanol washing 2-4 time again, place 60 ℃ of oven dryings at last, promptly get and modify beer waste yeast (also claiming to modify yeast), modification beer waste yeast adsorbent promptly of the present invention.Reaction equation is as follows:
The advantage and the beneficial effect of the inventive method are as follows:
Prepare new adsorbent with the citric acid modification beer waste yeast, not only raw material beer waste yeast wide material sources, cheap and easy to get can make full use of a large amount of waste yeasts that brewing industry produces.And, it is carried out simple modification after, regeneration, and synthetic method is simple, needed reaction reagent kind, consumption are less, do not need harsh experiment condition, cost is relatively low.Adsorbent behind the modification, in the process to the lysozyme test for nominal samples, adsorption conditions is easier to reach, and the adsorbent consumption is few, energy of adsorption comparatively fast reaches balance, and adsorption capacity is than higher.The acidity and the temperature range that are suitable for are bigger, can use in the acidity of broad and temperature range.
The elution process of having adsorbed the adsorbent behind the lysozyme is simple, and eluant, eluent is exactly common NaCl solution, elution efficiency height.The adsorption capacity of the beer waste yeast after the citric acid modification is 10.6 times of unmodified, illustrates that modifying beer waste yeast has obviously improved its adsorption capacity to lysozyme.Adsorbent is reused four times, and adsorption rate still can keep 94.6% of first adsorption rate, illustrates that the regenerability of adsorbent of the present invention is good, can reuse.
Adsorbent of the present invention is applied to the actual sample egg when clear, extraction lysozyme from egg is clear fast and effeciently, adsorption capacity height.And good vigor recovery rate and very high purifying multiple are arranged.It is good that the biologically active of the lysozyme after the enrichment keeps.And The results in electrophoresis also confirms, the lysozyme in clear has purification preferably to egg to modify yeast.
Description of drawings
Fig. 1. be the unmodified yeast of embodiment 1 preparation and the infrared spectrogram of modifying yeast;
Fig. 1 shows that the unmodified yeast is at 1726cm
-1Do not have obvious peak, show that the unmodified yeast surface does not have carboxyl.Modify yeast at 1726cm
-1And 1167cm
-1For in the ester group-stretching vibration peak (Mao of C=O and C-O, J., Won, S.W., Vijayaraghavan, K., Yun, Y.-S., 2009.Surface modification of Corynebacterium glutamicum for enhanced Reactive Red 4 biosorption.Bioresour.Technol.100,1463-1466).3302cm
-1Be O-H and N-H stretching vibration peak, the peak of modification bacterium obviously broadens, show because the introducing of citric acid, make the surface of beer waste yeast contain more O-H (Deng, S., Ting, Y.P., 2005.Characterization of PEI-modified biomass and biosorption of Cu (II), Pb (II) and Ni (II) .Water Res.39,2167-2177.).These results show that citric acid has been grafted to the surface of beer waste yeast, have introduced a large amount of carboxyls on its surface.
Fig. 2. be the unmodified yeast of embodiment 1 preparation and the xps energy spectrum figure of modification yeast;
Fig. 3. be the unmodified yeast of embodiment 1 preparation and the O1s figure of modification yeast;
The result of Fig. 2 and Fig. 3 shows that bigger variation has taken place the ratio of modifying back each element of yeast surface, and especially the ratio of oxygen element improves very big.The ratio that table 1 has been listed the unmodified yeast and modified each element of yeast, the ratio of oxygen element brings up to 33.88% by original 24.36%.Reason is that the hydroxyl of the carboxyl of citric acid and yeast surface esterification has taken place has been connected more citric acid residue on the surface of beer waste yeast, make yeast surface-COOH content improves the also corresponding increase of the ratio of oxygen element greatly.Show that modification that citric acid passes through the esterification success arrived the surface of yeast, this result is consistent with infrared parsing.
Table 1 unmodified yeast and each element ratio of modification yeast are relatively
Fig. 4. be the unmodified yeast of embodiment 1 preparation and the acidity curve map of modification yeast absorption lysozyme;
Fig. 5. be the unmodified yeast of embodiment 1 preparation and the time plot of modification yeast absorption lysozyme;
Fig. 6. be the unmodified yeast of embodiment 1 preparation and the temperature profile of modification yeast absorption lysozyme;
Fig. 7. be the unmodified yeast of embodiment 1 preparation and the adsorption isotherm of modification yeast absorption lysozyme;
From figure, can know the unmodified yeast and modify yeast experiment adsorption capacity (maximal absorptive capacity) and be respectively 89.9mgg
-1, 954.7mgg
-1, the modification yeast is 10.6 times of unmodified yeast.
Fig. 8. (the langmuir isotherm equations is the unmodified yeast for preparing for embodiment 1: C with langmuir adsorption isotherm simulation curve with modifying yeast absorption lysozyme
e/ q
e=1/bq
m+ C
e/ q
m, q
eAdsorbance during for molecular balance, q
mBe maximal absorptive capacity, C
eThe concentration of lysozyme in the solution during for adsorption equilibrium, b is a constant);
Fig. 9. (the Freundlich isotherm equations is the unmodified yeast for preparing for embodiment 1: lnq with Freundlich adsorption isotherm simulation curve with modifying yeast absorption lysozyme
e=lnK+lnC
e/ n, q
eAdsorbance during for molecular balance, q
mBe maximal absorptive capacity, C
eThe concentration of lysozyme in the solution during for adsorption equilibrium, K and n are empiricals);
Table 2 is Langmuir and the Freundlich adsorption isotherm analog parameter of Fig. 8 and Fig. 9
The gained linearly dependent coefficient as can be seen from table, the process of unmodified yeast and modification yeast absorption lysozyme is accords with Langmuir adsorption isotherm pattern more, the absorption that shows lysozyme is based on the monolayer chemisorbed, and unmodified yeast and modification yeast are respectively 82.17mgg to the theoretical maximum adsorption capacity of lysozyme
-1And 909.09mgg
-1, more identical with the experiment value among Fig. 7.
Figure 10 is electrophoretogram (a. standard protein (Marker) of modification yeast Purification of Lysozyme from egg is clear of embodiment 1 preparation; B. pure lysozyme standard specimen; C. original egg white; D. adsorbed solution; E. the lysozyme that elutes).As can be seen from the figure, the supernatant behind the absorption lysozyme is the 4th swimming lane d, does not almost have lysozyme, illustrates that the beer waste yeast of modifying is more complete to the absorption of lysozyme.The 5th swimming lane e is the lysozyme that elutes after the adsorbents adsorb, and purity is higher, and the content of foreign protein is less.Illustrate and modify yeast separation, enrichment lysozyme from egg is clear quickly and efficiently.
The specific embodiment
The beer waste yeast that the present invention relates to is the discarded object that general beer process industry produces, and the actual beer waste yeast that uses is taken from Hubei brewery among the present invention.
Embodiment 1:
A kind of modification beer waste yeast adsorbent, the step of its preparation method is as follows:
1. the preliminary treatment of beer waste yeast: take by weighing the 15.0g beer waste yeast in the 250mL conical flask, add 150mL distilled water, take out behind 25 ℃ of water-bath vibration 1h and leave standstill 30min, abandon the upper strata turbid solution, it is colourless that water continues beer waste yeast is washed till supernatant, use absolute ethanol washing 3 times again after, leave standstill, suction filtration makes Separation of Solid and Liquid then, and the gained solid places 60 ℃ of baking ovens dry, standby;
2. the preparation of crosslinked beer waste yeast: 1.5g is joined in the glutaraldehyde solution of 50mL 2%wt through the pretreated beer waste yeast of step 1, behind 35 ℃ of shaking table oscillating reactions 12h, collect product, product washes with water earlier three times, use absolute ethanol washing again three times, to remove the unreacted glutaraldehyde in product surface, place 60 ℃ of oven dryings at last, obtain yellow crosslinked beer waste yeast (also claiming the unmodified yeast), standby;
3. modify the preparation of beer waste yeast: the crosslinked beer waste yeast that takes by weighing the preparation of 2g step 2 joins 25mL N, in the dinethylformamide (DMF), adds 2g citric acid and 1g sodium dihydrogen phosphate (NaH again
2PO
4), place 140 ℃ of three-neck flasks reaction 2h, and cooling for reflux, utilize the water that produces in the water knockout drum branch dereaction process; After reaction finished, suction filtration carried out Separation of Solid and Liquid, and it is colourless washing solid product to filtrate with water, use absolute ethanol washing again 3 times, place 60 ℃ of oven dryings at last, promptly get and modify beer waste yeast (also claiming to modify yeast), modification beer waste yeast adsorbent promptly of the present invention.
It is prepared that the adsorbent of using in following examples is embodiment 1.
Embodiment 2: the adsorbent of the inventive method preparation is applied to adsorb lysozyme
Determining of adsorption conditions:
1. determining of absorption acidity: take by weighing the crosslinked beer waste yeast of 0.0200g and modification beer waste yeast adsorbent (all parallel take by weighing many parts) respectively, place the 50mL conical flask respectively, add 10.00mL 0.2mgmL
-1The lysozyme mark liquid of different acidity (pH=3.5,4.0,4.5,5.0,5.5,6.0,6.5,7.0,7.4,8.0), after adsorbing 12h, shaking table under the 120rpm takes out supernatant, mensuration under 25 ℃, calculate the adsorption rate of adsorbent to lysozyme, its adsorber acid line of writing music is seen Fig. 4;
For the modification beer waste yeast, along with the increase of pH, the adsorption rate of lysozyme increases gradually, and when pH4.0, adsorption rate reaches maximum, increases pH again, and adsorption rate is almost constant; In order to make lysozyme keep higher activity, select the experiment pH of pH7.4 as following examples.For crosslinked beer waste yeast, pH＜4 o'clock, lysozyme does not adsorb, and along with the increase of pH, adsorption rate increases.Under same pH, to compare with the modification beer waste yeast, the adsorption rate of crosslinked beer waste yeast is lower than the modification beer waste yeast all the time.
2. determining of adsorption time: take by weighing the crosslinked beer waste yeast of 0.0200g and modification beer waste yeast adsorbent (all parallel take by weighing many parts) respectively, place the 50mL conical flask respectively, all add the 0.2mgmL of 10.00mL with buffer solution (the 0.05M Tris-hydrochloric acid system) configuration of pH7.4
-1Lysozyme mark liquid, shaking table absorption under 120rpm under 25 ℃ is taken out supernatant and is measured behind the different time, calculate the adsorption rate of adsorbent to lysozyme, draws its adsorption time curve, sees Fig. 5;
Reach balance behind the modification beer waste yeast absorption 40min, absorption fully.Crosslinked beer waste yeast is along with the increase of adsorption time, and adsorption rate has slowly to be increased, but changes little.The adsorption process that this modification beer waste yeast is described can reach balance fast.The adsorption time of following examples is decided to be 40min.
3. determining of adsorption temp: take by weighing the crosslinked beer waste yeast of 0.0200g and modification beer waste yeast adsorbent (all parallel take by weighing many parts) respectively, place the 50mL conical flask respectively, all add the 0.2mgmL of 10.00mL with buffer solution (the 0.05M Tris-hydrochloric acid system) configuration of pH7.4
-1Lysozyme is marked liquid, puts into the shaking bath of different temperatures, takes out supernatant mensuration behind the absorption 40min down in 120rpm, and the calculating adsorbent is drawn its temperature curve to the adsorption rate of lysozyme, sees Fig. 6;
In the time of 20～45 ℃, temperature is little to the influence of adsorption rate, and adsorption rate has a maximum all more than 80% at 25 ℃, therefore selects 25 ℃ for use as the optimal adsorption temperature.
4. the lysozyme initial concentration is to the influence of adsorbance: take by weighing the crosslinked beer waste yeast of 0.0200g and modification beer waste yeast adsorbent (all parallel take by weighing many parts) respectively, place the 50mL conical flask respectively, add 10.00mL respectively with 0.0 of the buffer solution (0.05M Tris-hydrochloric acid system) of pH7.4 configuration, 0.2,0.4,0.5,0.7,1.0,2.0,3.0mgmL
-1Lysozyme mark liquid takes out supernatant behind the 120rpm absorption 40min under 25 ℃ and measures in shaking bath, calculate the adsorbance of adsorbent to lysozyme, draws its adsorption isotherm, sees Fig. 7;
Embodiment 3: the regenerability test of modification beer waste yeast adsorbent
1. get 0.0200g and modify yeast in the 50.0mL conical flask, add the 0.2mgmL of 10.00mL with buffer solution (the 0.05M Tris-hydrochloric acid system) configuration of pH7.4
-1Lysozyme mark liquid is put into 25 ℃ of shaking baths, and shaking table absorption 40min after leaving standstill, takes out supernatant under 120rpm, measures the adsorption rate (be the adsorption rate of using for the first time this moment) of lysozyme.Behind the adsorbent natural air drying to be left, add 10.00mL 1.0MNaCl solution, put into 25 ℃ of shaking baths, shaking table wash-out 40min under 120rpm is after leaving standstill, take out supernatant, repeat above-mentioned absorption-wash-out behind the adsorbent natural air drying to be left, adsorbent uses altogether four times, and the adsorption capacity when using for the 4th time still keeps 94.6% (the seeing Table 4) of using first, the regenerability that this adsorbent is described is good, can reuse.
The result of table 4 adsorbent reactivation performance test
Embodiment 4:
Adsorbent of the present invention is used for actual sample handles, step is as follows:
1. be (0.05M) PBS of 6.50 dilution egg white to 5 times volume with the pH value, fully the egg white solution behind the agitation and dilution makes the abundant mixing of egg white and buffer solution.Transfer the pH value to 4.60 of dilution then with 0.05M phosphoric acid, water-bath is heated to 100 ℃, and insulation 20min allows ovalbumin fully precipitate.The centrifugal 10min of cooling back 10000r/min, it is stand-by to get supernatant;
2. take by weighing 0.0200g modification beer waste yeast adsorbent in the 50mL conical flask, add the supernatant that 10.00mL step 1 obtains, put into 25 ℃ of shaking baths, react 40min down in 120rpm.Take out, leave standstill, get supernatant then.Survey the vigor of lysozyme.
3. in above-mentioned steps 1, got in the modification beer waste yeast adsorbent of supernatant, add 10.00mL1.0M NaCl solution, put into 25 ℃ of shaking baths, shaking table wash-out 40min under 120rpm, measure the enzyme activity of eluent, table 3 is the separation and purification result of chicken lysozyme from egg white.
Table 3
From egg is clear, extract and separate lysozyme (parallel three times), the results are shown in Table 3.The clear ratio vigor of egg is 168Umg before purifying
-1, after the extraction, the ratio vigor of lysozyme is 4605Umg in the adsorbent eluent
-1, the single step purification multiple is 27.5, the recovery rate of lysozyme total activity is 71.3%.
Ratio vigor before ratio vigor/purifying behind purifying multiple=purifying
Total activity before total activity/purifying behind enzyme activity recovery rate=purifying
Claims (2)
1. modification beer waste yeast adsorbent, the step of its preparation method is as follows:
(1) preliminary treatment of beer waste yeast:
Take by weighing beer waste yeast and place conical flask, the water that adds 8-12 times of weight, take out behind the 20-30 ℃ of water-bath vibration 0.5-1.5h and leave standstill 20-40min, abandon the upper strata turbid solution, it is colourless that water continues beer waste yeast is washed till supernatant, use absolute ethanol washing 2-4 time again after, leave standstill, suction filtration makes Separation of Solid and Liquid then, and the gained solid places 60 ℃ of baking ovens dry, standby;
(2) preparation of crosslinked beer waste yeast:
Take by weighing through the pretreated beer waste yeast of step (1) and join in the glutaraldehyde solution of 2wt%, the consumption of glutaraldehyde solution is the every g beer waste yeast of 30-35mL, behind 30-40 ℃ of shaking table oscillating reactions 10-12h, collect product, product washes with water earlier three times, use absolute ethanol washing again three times,, place 60 ℃ of oven dryings at last to remove the unreacted glutaraldehyde in product surface, obtain yellow crosslinked beer waste yeast, standby;
(3), modify the preparation of beer waste yeast:
Take by weighing the crosslinked beer waste yeast of step (2) preparation, amount according to the crosslinked beer waste yeast of the every g of 10-13mL adds N, dinethylformamide, add again and the citric acid of quality such as crosslinked beer waste yeast and the sodium dihydrogen phosphate of 0.5 times of crosslinked beer waste yeast quality, place three-neck flask 130-140 ℃ of reaction 1.5-2.5h, and cooling for reflux, utilize the water that produces in the water knockout drum branch dereaction process; After reaction finished, suction filtration carried out Separation of Solid and Liquid, and it is colourless washing solid product to filtrate with water, uses absolute ethanol washing 2-4 time again, places 60 ℃ of oven dryings at last, promptly.
2. the application of the described modification beer waste yeast of claim 1 adsorbent in the absorption lysozyme.
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| CN108328746A (en) * | 2018-01-29 | 2018-07-27 | 华中科技大学 | A kind of carboxymethyl modified chitosan/yeast adsorbent, it is prepared and application |
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| CN105561943B (en) * | 2016-03-14 | 2017-12-15 | 武汉轻工大学 | A kind of preparation method and application of magnetic rape stalk sorbing material |
| CN108102939A (en) * | 2018-01-25 | 2018-06-01 | 浙江树人学院 | A kind of modified waste beer yeast bacterium and preparation method thereof |
| CN108328746A (en) * | 2018-01-29 | 2018-07-27 | 华中科技大学 | A kind of carboxymethyl modified chitosan/yeast adsorbent, it is prepared and application |
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