Experiment 73 Determination of amylase activity from the germination of the seed of things

Experiment 73 The determination of amylase activity from the seed of thingsprincipleseed storage material starch, in the germination process, mainly under the action of amylase hydrolyzing into a simple
organic
compound
, these substances are the materials that make up the new organ. Quantifying the duration of the starch glycation process under certain conditions can indicate the size of enzyme vitality.。Instrument medicinebalance ringing temperature
water bath pot research white magnetic platefunnel
funnel holderculture
dishes filter paper 1% starch solution 0.2mol/L phosphoric acid buffer, pH6(see Schedule 2) I-KI solution: 1. Raw iodine:
2
11g, KI22g, set capacity to 500ml.2. Standard thin iodine: take 15 ml of raw iodine liquid, plus K18g to 500 ml.3. Color thin iodine liquid: 2 ml of raw iodine liquid plus KI20g, fixed capacity to 500 ml.standard destection: said to take 0.12g dedexion, suspended in a small amount of water, and then moved to boiling water, cooling capacity to 200ml, take 1 ml plus 3 ml of standard iodine as a comparative color.。Step 1. 1. Preparation of experimental materials6 days before the experiment, hydroponics was used to grow several different environmental conditions of the growth of crop seedlings (rice, wheat)2. Preparation of amylase solution will be different seedlings washed with water, each said to take seedling endosperm part 0.5g, respectively placed in the research, plus 5 ml pH6 phosphoric acid buffer, careful grinding, filter paper
filtration
(or centrifugation), with the appropriate amount of buffer rinse, and finally fixed to 10 ml. 3. Insulation glycation Take 20 ml of 1% starch and 5 ml pH6 phosphate buffer in a triangle
sam
, balance in a 35c water bath for 15 min, add 1 ml of prepared enzyme solution, stir well, and immediately remember the time. 4. Color, observation and determination suction of the above mixture 1 drop on the white porcelain reaction board, plus 1 drop of thin iodine liquid, observe the color, compared with the standard paste, the same is the reaction to reach the end point, record the glycation time t. 。 f is a dilution multiply (20). 。 The U
a
is an enzyme vitality unit (which means 1 unit of enzyme activity per g of material digested per hour for 1 g of starch). 。