Detection of immobilized yeast cells and sucrase

related topics . First, the experimental purpose
1. Learn the method of
of yeast
cells.
2. To master the detection method of sucrosease vitality.
II, experimental principle
solid phase enzyme, also known as fixed enzyme, is a combination of water-soluble enzyme and solid water insoluble support (or carrier) by physical and chemical treatment methods. Enzyme fixation can transform the enzyme into an insoluble substance while retaining the vitality of the enzyme, which has many advantages that water-soluble enzymes do not have in the catalytic reaction.
methods of fixing enzymes commonly used in the united States are:
. 1. Physical adsorption method;
2. carrier association method (key binding method);
3. cross-linking method;
4. encoursing method;
in terms of its application technology, it is divided into fixed enzymes and fixed cells.
compared with water-soluble enzymes, the advantages of fixed enzymes are mainly increased mechanical strength, improved stability, recycling and repeated use.
, instruments and
reagents
instruments test tubes
, 1 ml
straws
4, water bath pot.
reagents
. 1. Several grams of sodium alginate;
2. K yeast liquid
3. 10% sucrose liquid: said to take 10 grams of sucrose water content to 100 ml;
4. Filin A, B liquid.
4, operating step
l, called sodium seaweed acid 1 gram added to 100 ml of water, micro-fire
heating
dissolved and cooled to about 30 degrees C, the pre-prepared 3 grams of karst yeast (or K yeast
cultured
30 ml liquid) suspension mixed. Then pour into the
funnel
with hose and hemostamine clip at the bottom and let it slowly drip into a 4% calcium chloride solution to make a spherically fixed yeast with a diameter of 2-3 mm.
the stationary yeast into the column and add 10% sucrose from the top of the column to control a certain flow rate (10-17 drops per minute). The hydrolysate that flows first flows back for 10 minutes. The resulting outflow is a glycolyte hydrolyzed by the fixed K yeast, which is made up of a mixture of glucose and fructose.
2, sucrose enzyme detection
absorption of ferrine A, B liquid each 1 ml in
drying
test tube, add hydrolyte 1 ml, boiling water insulation, observe the color reaction. The precipitation of copper oxide indicates that sucrose has been hydrolyzed and sucrose enzymes are present in the tubes.
the blanks were compared with 10% sucrose, and the others were the same.
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