<Cold water extract>
1. Pulverize 'freeze dried 겨우살이' and add 10 times volume(w/v) of PBS after that, 2~3 min homogenize using homogenizer , extract by agitating 8~16 hours on 4°C
2. Collect supernatant by centrifuge ( 8000 x g, 30 min, 4°C)
3. Eliminate impurities by using filter paper (Whatman, No. 2) and get filtrate.
4. Dialysis in same buffer 48h(2days) after that centrifuge (8000 x g, 30 min, 4°C) for eliminate precipitate and filter supernatant by using 0.45㎛ membrane filter.
5. Get the extract about 10% yield rate.
<Hot water extract>
1. Pulverize 150g 겨우살이and mix 1L water and heat 10 hours on 100°C after that get 750mL extract.
as a result, we get Conversion glucose 1% of solid content by using 'Adobe glucose saccharimeter' .
<Enzyme extract>
1. Fine grind freeze dried 겨우살이 to get under 200㎛ size.
2. Make suspension by mixing 150g 겨우살이 to 2L water.
3. Put 10% acetic acid solution to control suspension's pH to 5.2
4. Treat amylase(Econase, AB Enzyme Gmbn, Germany) 400㎕
and cellulose(Myconase, Girst Brocade Inc, Netherland) 40㎕ to 'pH controled suspension.'
5. Shake 'enzyme treated suspension' 10 hours in shaking incubator ( 53°C, 60Hz)
6. centrifuge (3500RPM) and separate 1.6L supernatant from shaked solution.
as a result, we get Conversion glucoes 2.5% of solid content by using 'Adobe glucose saccharimeter'.( 27% of theory yield rate)