Microflex system

Growth of type strain 901379 (the first isolate, from 2013) was assessed using 7H10 agar plates + 10% OADC growth supplement, after incubation for 14 days at either 30°C or 37°C. A standard series of biochemical tests and morphological examination were performed as described in (Pfyffer, 2015 ). MALDI-TOF MS profiles were acquired on a Bruker microflex system and interpreted using the Bruker mycobacterial database (version 3.0) (Detailed methods are in Supplementary File 1).

Purification of all (glyco)peptides and their corresponding analyses were performed on a 1,260 Agilent Infinity system. The analytical RP-HPLC method uses a Phenomenex Aeris Peptide C18 column (150 mm × 4.6 mm, 3.6 μm, 100 Å) at 0.8 ml/min flow rate or a Vydac Denali C18 column (250 mm × 4.6 mm, 5 μm, 120Å) at 1 ml/min flow rate, with 0.1% TFA in water (A) and 0.1% TFA in acetonitrile (B) as the eluents. The elution gradient for analytical RP-HPLC purification was 0–60% B over 30 min. The preparative RP-HPLC method uses the Grace Vydac monomeric C18 column (250 mm × 22 mm, 15–20 μm, 300Å) at 10 ml/min flow rate, with 0.1% TFA in water (A) and 0.1% TFA in acetonitrile (B) as the eluents. The elution gradient for preparative RP-HPLC purification was 0–50% over 110 min. The (glyco)peptides were detected at 214 nm by using a UV-Vis detector (Agilent 1,260 Infinity DAD). Purified (glyco)peptides were characterized by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) with a Voyager-DE STR system or a Bruker Microflex system, using α-cyano-4-hydroxycinnamic acid as matrix.