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Fig. 3 | Biotechnology for Biofuels

Fig. 3

From: Single mutation at a highly conserved region of chloramphenicol acetyltransferase enables isobutyl acetate production directly from cellulose by Clostridium thermocellum at elevated temperatures

Fig. 3

In vitro characterization of the wild-type CATSa and CATSa F97W variant. a SDS-PAGE of the purified CATSa and CATSa F97W. The black arrow indicates the expected size of expressed target proteins, including CATSa and CATSa F97W. Notations: column 1, crude cell extract of IPTG induced E. coli BL21(DE3) harboring pET_CATSa; column 2, His-tag purified CATSa; column 3, crude extract of IPTG induced E. coli BL21(DE3) harboring pET_ CATSa F97W; column 4, His-tag purified CATSa F97W; and M, protein ladder. b Melting curve of CATSa and CATSa F97W. The intensity was normalized by each maximum value. c Michaelis–Menten plots of CATSa and CATSa F97W for various isobutanol concentrations at 50 °C. The co-substrate, acetyl-CoA, was supplemented at the saturated concentration of 2 mM. The error bars represent standard deviation of three biological replicates

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Biotechnology for Biofuels and Bioproducts

ISSN: 2731-3654

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