A new Developer Note on characterizing the limited operational lifetime of Cytosol reactions is now online. Please share your comments and feedback below.
@surendra link please?
This is really fun and the data particularly in Figure 1 is quite rich! The effect has been observed in extract before; I myself have found that loss of the energy carrier (NTPs and their secondary energy source) is what causes this loss in activity. It reminded me as well of Anne Doerr’s paper Radware Bot Manager Captcha, where they found ribosome deactivation to be the cause of reaction termination in PURE. They also found deactivation of PURE over time independently of TXTL activity but I don’t think they further investigated this much.
Would be interesting to check what exact component of the SMix is the first limiting factor.
Also, from Figure 1, it looks like the depletion of whatever resource it is, is proceeding nearly linearly at first, which would also suggest an enzymatic reaction, rather than spontaneous decay.