Hello
A strain of Bacillus velezensis, producing β-glucanase and protease, was isolated from animal manure and we attempted to produce β-glucanase and protease after optimization of various process parameters with the solid-state fermentation.
How we can measure the following:
The growth and enzyme production profile over the fermentation period to show the stage at which enzyme production is started.
Any help will be appreciated.
You would get more answers if your question title indicated what the problem is. That you need help is already obvious from the fact that you post a question here.
You would get more appropriate answers, if you give more relevant details in your question: which organism are we talking about, and what growth medium? Which enzyme do you want to measure? Production of what is started?
Engelbert Buxbaum Thank you for your suggestion, I have edited the question.
For growth measure the OD of bacteria at different intervals of incubation at 600nm and for activities perform enzymes assays.
OD 600 is routinely used to assess cell density in liquid media, and it is convenient because it is non-destructive. On solid cultures, one could measure the diameter of the colonies, or one could take samples and measure protein or DNA. It may also be possible to use stains that become fluorescent when they bind to proteins or DNA (live stains) without interfering with growth. Molecular Probes (now part of Thermo) sells such things, check their handbook (https://www.thermofisher.com/de/de/home/global/forms/mp-handbook-download-request-form-2014.html)
As to the production of hydrolytic enzymes, one could, in theory, use a substrate labelled for FRET (two dyes in resonance close to each other on the same substrate molecule). As the enzyme hydrolyses the substrate, the distance between the fluorophores increases, and the fluorescence at the donor wavelength increases. That way, the activity of the enzyme could be monitored continuously. For proteases, such assays are commercially available, I don't know about glucanases.
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