Hi Mohamad!

what we ussually do, is to stirr 2 g of solid medium in 10 ml water for 20 minuts. Sometimes it helps to add twin20 or another detergent, but some people say it can interfere with the quantification. You can also grind the medium before stirring if it is agglomerated.

best

Francisco

I appreciate it. Thanks for sharing!

Hi Mohamad, I recommend a protocol that I use with good results, you can use 4g of substrate and suspend it in 50 mL of citrate buffer (50mM, pH 5.0) in a shaker for 30 minutes. Separate the solids by filtration, and the resulting liquid centrifuge (4 ◦C, 5000 rpm, 3 min) to obtain the supernatant (crude extract) that you can store in falcon tubes at −20 ◦C for further determination. I hope this procedure can help you, although I would suggest that if you want to isolate lignin-modifying enzymes, use a solid submerged culture of your substrate because it allows you to obtain greater enzymatic activity and also allows the purification of these proteins in a simpler way and with greater yield and purity.

greetings and good luck !!!