enzyme extraction - (Dec/13/2007 )
Hi everyone,I am new here,panning El Dorado gold as in Zorro starring Antonio B. and A.Hopkins.I have polbags with oyster mushroom mycellium growing on sawdust,bran,lime medium. Trying unsuccessfully to get out some of the cellulase-water and 5% NaCl solution not showing enzyme after testing 2ml in tissue paper strip (~50mg) for 3-4h at room temperature at pH ~5, without glucose showing. The glucotest strip did not change colour.
I am hobbyist doing this in kitchen.My interest in cellulase stems from interest in bioethanol from wastewood which I can get free. Buying chemicals here in Malaysia is very costly and I do not have that much money as I am a pensioner. Someone here suggested laminar flow for $20k.Have to strike lottery for that!
But I will try out all suggestions within my means. With much appreciation.
I am hobbyist doing this in kitchen.My interest in cellulase stems from interest in bioethanol from wastewood which I can get free. Buying chemicals here in Malaysia is very costly and I do not have that much money as I am a pensioner. Someone here suggested laminar flow for $20k.Have to strike lottery for that!
But I will try out all suggestions within my means. With much appreciation.
Some suggestions: 1) try to break the materials down by using a blender, if you have not used one. Give it a couple of striks and see if that will help releasing the enzyme. Try to use 1% NaCl instead. 2) you may want to use more of the extract and/or let the reaction go for longer. 3) To prevent bacterial growth, you may want to use some sodium azide (NaN3), like a final concentration of 0.1% (weight/weight). It is not so expensive and you only need very little to do the job. Only that this chemical is slightly toxic, so handle with care and it is not very stable at acidic pH.
There maybe some publications on the enzyme from this mushroom. I will see if I can get my hand on it. not guaranty, though.
Found two papers on this subject, at least letting you know that there are several enzymes made by the fungus. I think you can either do glucose test, or by determining the weight loss of cellulose paper retrived, and dried to constant weight as rough estimation. good luck and have fun! Check your private message for the papers.
Thanks Genehunter-1. Ya-yes I'll try 1% NaCl and Na azide. Gross the author of DMSO paper simply withdraws 1ml samples at intervals from culture so DMSO might be good for cts. culture of enzymes.A friend here suggested Trichorderma mould as a good source.Many things to do now. Will keep you posted genehunter-1.