The yeast Kluyveromyces marxianus is a promising candidate for chemicals biosynthesis. It has rapid growth kinetics at temperatures upwards of 45 °C and can produce short and medium chain volatile esters and ethanol at high rates. Of particular interest is its capacity to produce ethyl acetate at rates upwards of 2 g/L/hr in aerated cultures. Both ethanol and ethyl acetate production rely on the activity of alcohol dehydrogenases to oxidize acetaldehyde to ethanol, which serves a substrate for ethyl acetate production. Little is known about the ester production pathways in K. marxianus. Prior findings suggest the presence of alcohol acetyltransferase (Atf) activity and the absence of significant reverse esterase activity. Our studies suggest the presence of one ATF and seven different alcohol dehydrogenase (ADH) genes in K. marxianus. In this work, we design a hybrid-synthetic RNA polymerase III promoter to create a CRISPR-Cas9 genome editing systems for K. marxianus. This system was used to construct a disruption library of ADH and ATF genes to study their function in volatile metabolite production. ATF disruption and overexpression analyses suggest a limited role of Atf in overall ethyl acetate biosynthesis in K. marxianus. ADH2 disruption resulted in reduced ethanol production along with accumulation of acetaldehyde suggesting an importance of Adh2 in ethanol production. The terminal step of ethyl acetate synthesis remains puzzling but overexpression of ADH7 shows promiscuous activity towards ethyl acetate production from hemiacetal. These findings serve as starting point for metabolic engineering approaches towards ethyl acetate and longer chain esters production.