Fig. 4

Ethyl acetate biosynthesis in K. marxianus by alcohol-O-acetyltransferase. a RT-qPCR analysis of wild-type K. marxianus ATF transcript levels at lag (5 h), log (10 h), and stationary (18 h) phases. mRNA copy number represents total number of transcripts for 5 ng of isolated RNA. b KmATF disruption reduces ethyl acetate production in both aerobic and anaerobic conditions. Cultures were grown at 37 °C for 10 h (aerobic) and 14 h (anaerobic). c Lysate Atf activity from S. cerevisiae with overexpressed KmATF and ScATF1. Rates are reported per mg of lysate protein. Enzyme expression was confirmed by Western blot analysis using anti-c-MYC and anti-GAPDH antibodies. Bars and error bars represent the arithmetic mean and standard deviation of triplicate biological samples. Statistical significance (P < 0.05) is indicated by asterisk and was determined using a T test