Confirmation of E1 expression and secretion in C. bescii using Western blot analysis. Concentrated extracellular proteins (10 µg) were electrophoresed in a 15 % gradient Mini-Protean TGX gel (BIO-RAD) and electro-transferred to a PVDF membrane (ImmobilonTM-P; Millipore). The membrane was then probed with an E1 monoclonal antibody. Truncated version of E1 produced in Streptomyces lividans (lane 1); parent strain JWCB018 ΔpyrFA
ldh::ISCbe4 Δcbe1 grown at 75 °C (lane 2); E1 expression strain JWCB052 ΔpyrFA
acel0614 (E1) grown at 75 °C (lane 3), 70 °C (lane 4), and 65 °C (lane 5); MW, MagicMark™ molecular weight marker (Invitrogen).