CN102174644A - Method for measuring beta-glucanase activity - Google Patents
Method for measuring beta-glucanase activity Download PDFInfo
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- CN102174644A CN102174644A CN2009100169555A CN200910016955A CN102174644A CN 102174644 A CN102174644 A CN 102174644A CN 2009100169555 A CN2009100169555 A CN 2009100169555A CN 200910016955 A CN200910016955 A CN 200910016955A CN 102174644 A CN102174644 A CN 102174644A
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- glucanase
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- 238000000034 method Methods 0.000 title claims abstract description 58
- 101710130006 Beta-glucanase Proteins 0.000 title claims abstract description 42
- 230000000694 effects Effects 0.000 title claims abstract description 28
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 50
- 239000008103 glucose Substances 0.000 claims abstract description 50
- 238000002835 absorbance Methods 0.000 claims abstract description 30
- PHOLIFLKGONSGY-CSKARUKUSA-N (e)-(3-methyl-1,3-benzothiazol-2-ylidene)hydrazine Chemical compound C1=CC=C2S\C(=N\N)N(C)C2=C1 PHOLIFLKGONSGY-CSKARUKUSA-N 0.000 claims abstract description 28
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 claims abstract description 26
- 229920002498 Beta-glucan Polymers 0.000 claims abstract description 26
- -1 potassium ferricyanide Chemical compound 0.000 claims abstract description 7
- 239000000243 solution Substances 0.000 claims description 42
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 24
- 239000003153 chemical reaction reagent Substances 0.000 claims description 23
- 102000004190 Enzymes Human genes 0.000 claims description 21
- 108090000790 Enzymes Proteins 0.000 claims description 21
- 238000006243 chemical reaction Methods 0.000 claims description 14
- FPFSGDXIBUDDKZ-UHFFFAOYSA-N 3-decyl-2-hydroxycyclopent-2-en-1-one Chemical compound CCCCCCCCCCC1=C(O)C(=O)CC1 FPFSGDXIBUDDKZ-UHFFFAOYSA-N 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000006460 hydrolysis reaction Methods 0.000 claims description 10
- 230000007062 hydrolysis Effects 0.000 claims description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 239000002253 acid Substances 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 2
- 239000001384 succinic acid Substances 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000005259 measurement Methods 0.000 abstract 1
- 239000000758 substrate Substances 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 description 3
- 238000001976 enzyme digestion Methods 0.000 description 3
- BYGOPQKDHGXNCD-UHFFFAOYSA-N tripotassium;iron(3+);hexacyanide Chemical compound [K+].[K+].[K+].[Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] BYGOPQKDHGXNCD-UHFFFAOYSA-N 0.000 description 3
- 210000002421 cell wall Anatomy 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 235000007319 Avena orientalis Nutrition 0.000 description 1
- 244000075850 Avena orientalis Species 0.000 description 1
- 235000007558 Avena sp Nutrition 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
- 235000007238 Secale cereale Nutrition 0.000 description 1
- 244000082988 Secale cereale Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 125000000218 acetic acid group Chemical class C(C)(=O)* 0.000 description 1
- 108010050181 aleurone Proteins 0.000 description 1
- 235000013405 beer Nutrition 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000012817 gel-diffusion technique Methods 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 239000010977 jade Substances 0.000 description 1
- 230000036632 reaction speed Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 238000004879 turbidimetry Methods 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 238000009941 weaving Methods 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for measuring beta-glucanase activity, which comprises the following steps: A, drawing a standard correspondence curve between the glucose absorbance value measured by an MBTH method and a corresponding concentration; and B, measuring the absorbance value equivalent to glucose generated after the beta-glucan is hydrolyzed by the beta-glucanase to be measured with the MBTH method; and determining the content of reducing sugar equivalent to glucose generated in unit time in the enzymolysis product of the beta-glucanase to be measured according to the standard correspondence curve between the glucose concentration and the measured corresponding absorbance value drawn in the previous step, and calculating the beta-glucanase activity based on the content. The detection limit and working concentration range in glucose measurement in the invention are obviously lower than those of a DNS method, a potassium ferricyanide method and Somogyi-Nelson, thus the sensitivity of the method is obviously higher than that of the three methods; and moreover, by using the method, the dosage of the substrate beta-glucan is greatly reduced, and the experiment cost can be remarkably lowered.
Description
Technical field
What the present invention relates to is a kind of method of measuring activity of beta-glucanase, is specifically related to relate in particular to the method for MBTH method mensuration activity of beta-glucanase the selection of key parameter in the mensuration process.
Background technology
Beta-glucan is the non-starch based polysaccharide of a class, it is by β-1, and 3-glucoside bond and/or β-1, the chain polysaccharide that the 4-glucoside bond is formed, be the moiety of plant cell wall, be present in the aleurone layer and albuminous cell wall of cereal class (comprising barley, oat, rye and wheat etc.).Beta-glucanase has important hydrolytic action to the glycosidic link in the beta-glucan.At present, this enzyme has been widely used in industries such as beer, feed, weaving, papermaking, medicine.The detection method of this enzyme activity has viscosimetry, fluorescent method, chromogenic substrate method, gel diffusion method, reducing sugar test method etc., and wherein the method for normal employing is DNS method, Tripotassium iron hexacyanide method and a Somogyi-nelson method in the reducing sugar test method.Because these three kinds of method sensitivity are lower, be difficult to measure the initial velocity of enzyme digestion reaction, therefore, accuracy is relatively poor.
The present invention is the vigor that developer is surveyed beta-glucanase with MBTH (3-methyl-2-[4-morpholinodithio ketone hydrazone) reagent.This method is not subjected to the interference of low-concentration acetic acid salt and succinate damping fluid, and gained enzyme concn curve and enzyme digestion reaction speed are linear dependence in the enzyme concn scope of broad.
Summary of the invention
At the deficiency of prior art, the invention provides a kind of measuring method of activity of beta-glucanase, this method adopts 3-methyl-2-[4-morpholinodithio ketone hydrazone (MBTH) method to measure the vigor of beta-glucanase.
Below introduce the measuring method of a kind of activity of beta-glucanase of the present invention in detail, A makes the glucose absorbance that records with the MBTH method and the standard corresponding relation curve of its respective concentration; B detects the absorbance that is equivalent to glucose that produces behind the beta-glucan enzymic hydrolysis beta-glucan to be measured with the MBTH method; Determine the content of the reducing sugar that is equivalent to glucose that in the unit time, produced in the enzymolysis product of beta-glucanase to be measured according to the standard corresponding relation curve between rapid glucose concn of making of previous step and the corresponding absorbance that records, calculate the vigor of beta-glucanase with this.
Optimally; The concrete steps of above-mentioned steps A are; Glucose absorbance and its respective concentration standard corresponding relation curve that effect MBTH method records; Get the glucose standardized solution 0.6mL of 6-15 group different concns (0-30 μ g/mL) respectively, add 0.6mL 0.5 normal NaOH solution, mixing, add MBTH reagent 0.6mL again and add ferric ammonium sulfate reagent 1.2mL while hot behind 75-85 ℃ of heating in water bath 5-20min, the absorbance of glucose is measured in room temperature cooling back in wavelength 620-680nm place; Each concentration is made three parallel samples; Above dosage can increase and decrease in proportion; According to concerning drawing standard corresponding relation curve between every group of glucose concn and the corresponding absorbance that records;
Optimally; The concrete steps of above-mentioned steps B are; Detect the vigor of beta-glucanase with the MBTH method; With concentration is the beta-glucan solution of measuring temperature that is preheated to of 1-5mg/mL, be added to be preheated in the beta-glucan enzyme solution of measuring temperature and carry out enzyme digestion reaction, measuring temperature is 25-40 ℃, hydrolysis time is in the 60min, get hydrolyzed solution rapidly with the abundant mixing of isopyknic 0.5 normal NaOH solution with termination reaction (can in reaction process time segment sampling detect), getting 1.2mL joins in the test tube and (makees three parallel samples), add MBTH reagent 0.6mL again and behind 75-85 ℃ of heating in water bath 5-20min, add ferric ammonium sulfate reagent 1.2mL while hot, absorbance is measured in room temperature cooling back in wavelength 620-680nm place, above dosage can increase and decrease in proportion.Determine the vigor of beta-glucanase to be measured according to the standard corresponding relation curve between the concentration of the rapid glucose of making of previous step and the corresponding absorbance that records; The unit of activity of beta-glucanase can be defined as under these conditions, and in every milliliter of reaction system, it is an enzyme activity unit that per minute generation 1nmol is equivalent to the required enzyme amount of glucose.Wherein the method for making of MBTH reagent is that the 3-methyl-2-[4-morpholinodithio ketone hydrogencarbonate aqueous solution of 3mg/mL and the dithiothreitol (DTT) solution equal-volume of 1mg/mL mix promptly, and matching while using is effective in one day.
Optimally; The method for making of above-mentioned ferric ammonium sulfate reagent is 0.5% (FeNH
4(SO
4)
2) 12H
2O, 0.5% thionamic acid, 0.5 equivalent hydrochloric acid.
Optimally; Above-mentioned beta-glucan is a solvent with Succinic Acid buffered soln or the hac buffer of 50mM.
Optimally; The optimal wavelength of said determination absorbance is 650nm.
The quantity of the advantage of the present invention glucose end group that to be the present invention produced after to beta-glucan enzymic hydrolysis beta-glucan with the MBTH method detects the vigor of determining beta-glucanase with this; Glucose and oligomeric beta-glucan are the products of beta-glucan enzymic hydrolysis, and therefore the sensitivity that the glucose endgroup content is detected is directly determining the sensitivity to the activity of beta-glucanase detection.This method is easier, quick, accurate, inexpensive than turbidimetry.Detectability and working concentration scope all are starkly lower than DNS method and Tripotassium iron hexacyanide method, so the sensitivity of present method is apparently higher than DNS method and Tripotassium iron hexacyanide method.
Embodiment
Embodiment 1:
Key instrument: constant temperature water bath vibrator, SHZ-82 type, state China Electrical Appliances Co., Ltd; Electric-heated thermostatic water bath, HH-4 type, state China Electrical Appliances Co., Ltd; Digital ph, Mettler Toledo Inc.; The UV2100 ultraviolet spectrophotometer, Shanghai UNICO Instr Ltd.; Beta-glucanase, Ningxia jade of the He family Bioisystech Co., Ltd; Beta-glucan, hundred special pure macromole Science and Technology Ltd.s; Pipettor, Finnpipette Finland thunder is vigorous.
Solution allocation
MBTH develop the color the 3-methyl-2-[4-morpholinodithio ketone hydrogencarbonate aqueous solution of liquid: 3mg/mL and 1mg/mL dithiothreitol (DTT) solution balanced mix promptly, matching while using, in one day effectively.
Glucose standardized solution (30 μ g/mL): accurately take by weighing the glucose 0.3000g that is dried to permanent quality, with the dissolving of the hac buffer of 50mMpH5.5, shift and be settled to 100mL, be made into the glucose standardized solution of 3.00mg/mL; Therefrom get the 1mL glucose solution, shift and be settled to 100mL, promptly get the glucose standardized solution of 30 μ g/mL.
The method for making of ferric ammonium sulfate reagent is 0.5% (FeNH
4(SO
4)
2) 12H
2O, 0.5% thionamic acid, 0.5 equivalent hydrochloric acid.
Steps A is made the glucose absorbance that records with the MBTH method and the standard corresponding relation curve of its respective concentration;
The glucose standardized solution 0.5mL that adds 8 groups of different concns respectively, add 0.5mL 0.5 normal NaOH solution, mixing, add MBTH reagent 0.5mL again and add ferric ammonium sulfate reagent 1mL while hot behind 80 ℃ of heating in water bath 11-13min, the absorbance of glucose is measured in room temperature cooling back in wavelength 650nm place; Each concentration is made three parallel samples; According to concerning drawing standard corresponding relation curve between every group of glucose solution concentration and the corresponding absorbance that records;
B is with the activity of beta-glucanase of MBTH method detection;
With its concentration of 5mL beta-glucan solution is 2mg/mL, the pH value is 5.5, join in the Erlenmeyer flask preheating 10min in 30 ℃ shaking bath, add the beta-glucan enzyme solution 5mL to be measured reaction that is hydrolyzed, for determining that suitable beta-glucan enzyme concn to be measured can become beta-glucanase solution dilution to be measured the different concns gradient to detect one by one, hydrolysis time is in the 60min, get hydrolyzed solution 2mL (can in reaction process time segment sampling detect), add in the 2mL 0.5 normal NaOH solution (making three parallel samples) rapidly, mixing, getting 1mL joins in the test tube, add MBTH reagent 0.5mL again and behind 80 ℃ of heating in water bath 11-13min, add ferric ammonium sulfate reagent 1.2mL while hot, absorbance is measured in room temperature cooling back in wavelength 650nm place, determine the amount of the reducing sugar that is equivalent to glucose that contained in the enzymolysis product of beta-glucanase to be measured according to the standard corresponding relation curve between the glucose concn of the rapid making of previous step and the corresponding absorbance that records.The unit of activity of beta-glucanase can be defined as under these conditions, and to produce the enzyme amount that 1nmol is equivalent to the amount of glucose be an enzyme activity unit to per minute in every milliliter of reaction system, thereby define the vigor of calculating wooden beta-glucanase according to enzyme activity.
Embodiment 2:
Key instrument: electric-heated thermostatic water bath, HH-4 type, state China Electrical Appliances Co., Ltd; Digital ph, Mettler Toledo Inc.; The UV2100 ultraviolet-visible spectrophotometer, UNICO(Shanghai) Instruments Co., Ltd.; Beta-glucanase, Xinhuayang Biological Co., Ltd., Wuhan; Beta-glucan, hundred special pure macromole Science and Technology Ltd.s; Pipettor, Finnpipette Finland Lei Bo group.
Solution allocation
MBTH develop the color the 3-methyl-2-[4-morpholinodithio ketone hydrogencarbonate aqueous solution of liquid: 3mg/mL and 1mg/mL dithiothreitol (DTT) solution balanced mix promptly, matching while using, in one day effectively.
Glucose standardized solution (30 μ g/mL): accurately take by weighing the glucose 0.3000g that is dried to permanent quality, with the dissolving of the hac buffer of 50mMpH4.0, shift and be settled to 100mL, be made into the glucose standardized solution of 3.00mg/mL; Therefrom get the 1mL glucose solution, as the operation of preceding method and be settled to 100mL, promptly get the glucose standardized solution of 30 μ g/mL.
The method for making of ferric ammonium sulfate reagent is 0.5% (FeNH
4(SO
4)
2) 12H
2O, 0.5% thionamic acid, 0.5 equivalent hydrochloric acid.
Steps A is made the glucose absorbance that records with the MBTH method and the standard corresponding relation curve of its respective concentration;
The glucose standardized solution 1mL that adds 8 groups of different concns respectively, add 1mL 0.5 normal NaOH solution, mixing, add MBTH reagent 1mL again and add ferric ammonium sulfate reagent 2mL while hot behind 80 ℃ of heating in water bath 11-13min, the absorbance of glucose is measured in room temperature cooling back in wavelength 650nm place; Each concentration is made three parallel samples; According to concerning drawing standard corresponding relation curve between every group of glucose solution concentration and the corresponding absorbance that records;
B is with the vigor of MBTH method detection beta-glucanase;
(its concentration is 2mg/mL with 0.25mL beta-glucan solution, the pH value is 5.5), add in the test tube preheating 10min in 30 ℃ water-bath, add the beta-glucan enzyme solution 0.25mL that the is preheated to 30 ℃ to be measured reaction that is hydrolyzed, for determining that suitable beta-glucan enzyme concn to be measured can become beta-glucanase solution dilution to be measured the different concns gradient to detect one by one, hydrolysis time is 30 minutes, add the abundant mixing of 0.5mL 0.5 normal NaOH solution rapidly with termination reaction, add 0.5mL MBTH reagent, abundant mixing, behind 80 ℃ of heating in water bath, add ferric ammonium sulfate reagent 1mL while hot, absorbance is measured in room temperature cooling back in wavelength 650nm place, make three parallel samples as method, determine the amount of the reducing sugar that is equivalent to glucose that contained in the enzymolysis product of beta-glucanase to be measured according to the standard corresponding relation curve between rapid glucose concn of making of previous step and the corresponding absorbance that records.The unit of activity of beta-glucanase can be defined as under these conditions, and per minute produces 1nmol to be equivalent to the required enzyme amount of glucose be an enzyme activity unit in every milliliter of reaction system, thereby defines the vigor of calculating wooden beta-glucanase according to enzyme activity.
Certainly, above-mentioned explanation is not to be limitation of the present invention, and the present invention also is not limited to aforesaid operations, and all variations of making in essential scope of the present invention, remodeling, interpolation or replacement all should belong to protection scope of the present invention.
Claims (6)
1. the measuring method of an activity of beta-glucanase is characterized in that;
A makes the absorbance of the glucose that records with the MBTH method and the standard corresponding relation curve of its respective concentration;
B detects the absorbance that is equivalent to glucose that produces behind the beta-glucan enzymic hydrolysis beta-glucan to be measured with the MBTH method; Determine the content of the reducing sugar that is equivalent to glucose that in the unit time, produced in the enzymolysis product of beta-glucanase to be measured according to the standard corresponding relation curve between rapid glucose concn of making of previous step and the corresponding absorbance that records, calculate the vigor of beta-glucanase with this.
2. the measuring method of activity of beta-glucanase according to claim 1 is characterized in that; The concrete steps of above-mentioned steps A are; Get the glucose standardized solution 0.6mL that the 6-15 group has different concns respectively, concentration range is between the 0-30 mcg/ml, add 0.6mL 0.5 normal NaOH solution, mixing, add MBTH reagent 0.6mL again and add ferric ammonium sulfate reagent 1.2mL while hot behind 75-85 ℃ of heating in water bath 8-20min, absorbance is measured in room temperature cooling back in wavelength 620-680nm place; Each concentration is made three parallel samples; Above dosage can increase and decrease in proportion; According to concerning drawing standard corresponding relation curve between every group of glucose concn and the absorbance that records;
3. the measuring method of activity of beta-glucanase according to claim 1 and 2 is characterized in that; The concrete steps of above-mentioned steps B are; With final concentration is the beta-glucan solution of measuring temperature that is preheated to of 1-5mg/mL, join in the beta-glucan enzyme solution that is preheated to relevant temperature and begin hydrolysis reaction, hydrolysis temperature is 25-40 ℃, hydrolysis time is in the 60min, this enzymolysis solution is mixed with termination reaction rapidly with isopyknic 0.5 normal NaOH solution, can in reaction process, take a sample stage by stage, therefrom getting 1.2mL joins and makees three parallel samples in the test tube, add MBTH reagent 0.6mL again and behind 75-85 ℃ of heating in water bath 5-20min, add ferric ammonium sulfate reagent 1.2mL while hot, absorbance is measured in room temperature cooling back in wavelength 620-680nm place, the concentration of reduced sugar that is equivalent to glucose that standard corresponding relation curve between glucose concn of making according to steps A and the corresponding absorbance that records is determined in the enzymolysis product to be contained is calculated the vigor of beta-glucanase with this; The unit of activity of beta-glucanase can be defined as under these conditions, and it is an enzyme activity unit that every milliliter of reaction system per minute produces the required enzyme amount of amount that 1nmol is equivalent to glucose; Wherein the method for making of MBTH reagent is that the 3-methyl-2-[4-morpholinodithio ketone hydrogencarbonate aqueous solution of 3mg/mL and the dithiothreitol (DTT) solution equal-volume of 1mg/mL mix promptly, and matching while using is effective in one day.
4. the measuring method of activity of beta-glucanase according to claim 3 is characterized in that; The method for making of above-mentioned ferric ammonium sulfate reagent is 0.5%FeNH
4(SO
4)
212H
2O, 0.5% thionamic acid, 0.5 equivalent hydrochloric acid.
5. the measuring method of activity of beta-glucanase according to claim 3 is characterized in that; Above-mentioned beta-glucan enzyme solution is a solvent with Succinic Acid or the acetum of 50mM.
6. the measuring method of activity of beta-glucanase according to claim 3 is characterized in that; The optimal wavelength of said determination absorbance is 650nm.
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| CN2009100169555A CN102174644A (en) | 2009-06-30 | 2009-06-30 | Method for measuring beta-glucanase activity |
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|---|---|---|---|
| CN2009100169555A CN102174644A (en) | 2009-06-30 | 2009-06-30 | Method for measuring beta-glucanase activity |
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| CN102174644A true CN102174644A (en) | 2011-09-07 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103243148A (en) * | 2013-05-06 | 2013-08-14 | 上海市农业科学院 | Method for detecting enzyme activity of beta-dextranase |
| CN105506059A (en) * | 2016-01-18 | 2016-04-20 | 黑龙江大学 | Method for determining activity of beta-mannase |
| CN108690867A (en) * | 2017-04-11 | 2018-10-23 | 南京农业大学 | A kind of enzyme activity determination method of n-glycosylase |
| CN110514655A (en) * | 2019-08-29 | 2019-11-29 | 云南大学 | The measuring method of endoglucanase enzyme activity in a kind of wizened bacterium |
-
2009
- 2009-06-30 CN CN2009100169555A patent/CN102174644A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| GORDON E. ANTHON ET AL.: "Determination of Reducing Sugars with 3-Methyl-2-benzothiazolinonehydrazone", 《ANALYTICAL BIOCHEMISTRY》 * |
| 张永勤等: "β-葡聚糖酶活力测定方法的研究进展", 《食品科学》 * |
| 张永勤等: "还原糖的可见风光光度法研究进展", 《食品与发酵工业》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103243148A (en) * | 2013-05-06 | 2013-08-14 | 上海市农业科学院 | Method for detecting enzyme activity of beta-dextranase |
| CN105506059A (en) * | 2016-01-18 | 2016-04-20 | 黑龙江大学 | Method for determining activity of beta-mannase |
| CN108690867A (en) * | 2017-04-11 | 2018-10-23 | 南京农业大学 | A kind of enzyme activity determination method of n-glycosylase |
| CN110514655A (en) * | 2019-08-29 | 2019-11-29 | 云南大学 | The measuring method of endoglucanase enzyme activity in a kind of wizened bacterium |
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Application publication date: 20110907 |