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The complex was then concentrated using an Amicon Ultra Centrifugal Filter (MWCO, 100 kDa) and subjected to size-exclusion chromatography (SEC) on a Superdex 200 Increase 10/300 column (GE Healthcare) that was pre-equilibrated with 20mM HEPES pH 7.4, 100mM NaCl, 2mM MgCl 2 , 1M peptide, 0.01% (w/v) LMNG and 0.0006% (w/v) CHS to separate complex from contaminants

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This extended fermentation process produces various gases, including hydrogen sulfide (H2S) , which is the culprit behind the distinct rotten-egg smell

After the addition of DTT (15 l, 31 mg ml 1 stock in ddH 2 O) to quench excess IAA, samples were shaken at 37 C for further 30 min (850 rpm, Thermomixer, Eppendorf)