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Fig. 5 | Microbial Cell Factories

Fig. 5

From: Pichia pastoris MutS strains are prone to misincorporation of O-methyl-l-homoserine at methionine residues when methanol is used as the sole carbon source

Fig. 5

Methionine supplementation reduces the amount of the methoxine incorporation. Nanobody B was produced in the CBS7435MutS strain in shake flasks. Methionine was added at the indicated concentration when cells were switched to methanol containing medium. In one condition (4 × 400) a methionine concentration of 400 mg ml−1 was added four times (every 12 h after the cells were switched to methanol containing medium). Relative abundance of the Δ−16 Da variant was done via RP-HPLC analysis followed by total mass measurement by ESI-Q-TOF-MS. Expression was verified on SDS-PAGE and Coomassie staining

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