Dekkera bruxellensis (anomorph Brettanomyces) is a hemiascomycetes yeast with a genome size ranging from under 20 to over 30 Mb. The yeast is a promising candidate for fuel ethanol production from both traditional starch based feed stocks as well as lignocellulose based feed stocks. In fact in at least one large scale continuous fermentation D. bruxellensis replaced the inoculated Saccharomyces strain and the fermentation continued at high levels of production for over two years. It has a high ethanol tolerance, in the wine industry it does not generally start growing until completion of the primary Saccharomyces fermentation. It is resistant to inhibitors formed during pretreatment of lignocellulose feed stock and it is able to utilize a number of sugars created during processing which are not used by Saccharomyces: cellibiose and arabinose. Additionally, Dekkera may prove useful in replacing the petrochemical production of acetic acid. In this proposal we suggest completion of the D. bruxellensis CBS 2499 genome sequence. Over 40% of the genome has already been sequenced and a number of genetic techniques have been developed. We hope that this sequence will help in developing control measures for Dekkera in alcohol fermentations and enhance the genetic engineering of Dekkera and other yeasts for the production of industrially important chemicals. Finally, the sequence may provide an interesting study in parallel evolution with Saccharomyces, as both yeasts inhabit the same environments but have developed alternative metabolic strategies to allow efficient utilization of metabolic resources. It may be possible to exploit these differences in order to enhance the metabolic capabilities of Saccharomyces.