Methaneontitan

The history of protoplast isolation says that the first isolation of protoplasts was done by von Klercker in 1892, by first plasmolysing and then slicing up plant tissues. But what were the specifications of the plasmolysis, e.g. salinity level and time in the solution?

Are there simple methods for extraction of chitinase from food such as bananas, avocados, kiwis, chestnut, beans and so on? That is, the extracted chitinase need not be entirely pure, just in significantly higher concentrations than in the food material the process starts with. It does, however, need to avoid degradation that permanently compromises chitinase activity. Temporary inactivation during the extraction process is acceptable as long as it is easily reactivated afterwards, but degraded homologs derived from chitinase that merely retain other attributes such as chemical similarity and allergenicity won't do it if their chitinase activity is permanently lost. Since the question is about simple methods, specialist reagents or chromatographs are not the answers the question seeks. Are there simple methods based on simple "reagents" such as water, ethanol, common household chemicals, easily extracted DIY substances and other easily obtainable chemicals? That is, the product need not be pure but it should increase chitinase levels compared to untreated food.

Do you know how much chitinase activity in units per milligram there is in different types of chitinase-rich food in their "natural" state (i.e. not extracted) before and after boiling? If it differs a lot depending on ethylene effects, I would like to know numbers for specific maturity stages too, ethylene causing some plants to mature. That is, I am asking about retention of functional chitinase, not chitinase-homologs that have lost their ability to degrade chitin. Do you know what pH values chitinase can be treated with without being permanently damaged? That is, how acidic or alkaline solutions chitinase can be placed in and still regain its full chitin-degrading ability when returned to moderate pH? I an not intending to mix chitinase into the agar, I am intending to treat the cells with chitinase before I place them on the agar. Just wanting to avoid contamination. By the way, my name means methane on Titan. CH4 on the Saturnian moon with a substantial atmosphere.

By what methods is it possible to kill the microbes in foods rich in chitinase without destroying the chitinase? I am thinking of killing the kinds of microbes that can grow on regular agar plates at room temperature.