Determination of cellulase vitality.

. Objective To learnand master the principles and methods of determining cellulose enzyme vitality by 3,5-denitrification salific acid (DNS) method, and to understand the properties of cellulase.
, principle, . Cellulase is a multi-member enzyme consisting of C
1
ase, C
X
ase, and ¦-glucosinease. The role of C1 enzyme is to hydrolysate natural cellulose into amoeba cellulose, C
X
enzyme is to continue to hydrolysis amoeba cellulose into fibrooligosaccharides, and the role of ¦-glucosinase is to hydrolyzed fiber oligosaccharides into glucose. Cellulose enzyme hydrolyzed cellulose produced fiber dissaccharides, glucose and other reduced sugars can reduce alkaline conditions of 3,5-denitrification salivary acid (DNS), the production of brown-red amino
compounds
, at a wavelength of 540nm has the maximum light absorption, in a certain range of reduced sugar and the color intensity of the reaction fluid proportional relationship, the use of color method to determine the amount of reduced sugar production can determine the vitality of cellulose enzymes.
, experimental materials, major instruments and
reagents1. Experimental material
(1) cellulase preparation 500mg
(2) Xinhua quantitative filter paper 50mg / parts × 4
(3) skimmed cotton 50 mg / × 4
(4) sodium methyl cellulose (CMC) 510mg
(5) salgon glycoside 500mg
2. Main instrument
(1)722 type or other models of visible hydrometer
(2)
ringing temperature
water bath 2
(3) boiling water bath
(4) electric stove
(5) scissors
(6) Analysis of one-tenth of a million
balance(7) constant temperature
dyring
box
(8) refrigerator
(9)
test tube

(10) glue head dropper
(11) plug-and-cut Test tube 20mL×24
(12) pipe or adder 0.5 mL×3; 2mL×7
(13)
bottle
100 mL×6; 1 000 mL×3
(14) drum 50 mL×2; 100 mL×1; 500 mL×1
(15)
berred
100 mL ×. 6;500mL×3;1,000 mL×1
3. Reagents (all pure analysis)
(1) concentration of 1mg/mL of glucose standard liquid
Store in a refrigerator at 4 degrees C (available for 12 to 15 days).
(2)3,5-dinitrosic acid (DNS) solution
accurately named DNS 6.3g in a 500mL berries, dissolved with a small amount of distilled water, added 2mol/L NaOH solution2 62mL, added to 500mL containing 185g potassium sodium
4
H
4
O
6
KNa
2
O In a hot water solution of MW,282.22, add 5g crystalline phenol (C
6
H
5
OH, MW s94.11) and 5g sodium aqueous sulphate (Na Na Na)
2
SO
3
, MW - 126.04), stir dissolved, cooled and moved into a 1,000mL capacity bottle with distilled water to 1,000mL, fully mixed. Store in a brown bottle and use at room temperature for one week.
(3) 0.05 mol/L pH4.5 citric acid buffer A solution (0.1 mol/L citric acid solution): accurately named C
6
H
8
O
7
. H
2
O (MW-210.14) 21.014g dissolved in a 500mL begup, dissolved with a small amount of distilled water, moved into a 1,000mL capacity bottle and mixed with distilled water to 1,000mL. Keep the spare in the refrigerator at 4 degrees C. .