I thing for this experiment you can use commercially available cellulase and other cell wall degrading enzymes. without disturbing the cell contents, you can degrade the cell wall alone. the secondary metabolites will be released into the outer liquid medium and after proper filtration and removal protein  and other contaminants you can measure the secondary metabolite through spectrophotometry. These enzymes will specifically act on cell wall only. 

Hi, there are many methods for that, one of them you can use ultrasonic to break cell wall, or by freezing the fungal mycelia on liquid nitrogen and grind with mortar 

Sonication is a good and simple method. 

prepare a suspension of fungi, break the cell wall using sonicator device, centrifuge the suspension and use the supernatant. cell density of your suspension should be adjusted to a specific value.

Thanks for your answers.

I agree with ultrasonic or sonication method....because it is simple and available to me. but i didn't find the protocol detail such as device power, suspension volume, process time, pause and start time for it....!!

are there the other simple ways for the breaking? or can anyone help me to use the ultrasonic?

Liquid nitrogen is one of the most used methods for fungal protein and DNA extractions and for this reason could be used to break the fungal walls, however the right method depends on what you want to do afterwards or what you want to recover.

If  sonication is better for you, you could try the cycles and times that are reported in the attached paper.

Good luck,

Diana

Dear Diana

Thank you for the paper and your answer. the paper is very useful for me :) 

I try to use the liquid nitrogen method,either.

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Yes, I think, the used liquid nitrogen is better

oh! I'm confused.

my desired metabolite has polyketide structure. according to this structure, is the liquid nitrogen method the best to extract it?

so, when is  the ultrasonic method used for the breaking?

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Yes, you can used the ultrasonic method, around ten min is enough

I used 10-15 cycles of 30 sec sonication with 10-15 sec intervals in my work.

It depends on the fungi you use for your study.

You can also use glass beads to break down the fungal cell wall. But both methods (i.e liquid nitrogen and sonication) can work ok. It all depends after the lysis what you want to do: enzyme or metabolite extraction. I hope it helps