The NEMO project provides novel efficient enzymes and microbes for 2nd generation bioethanol production. It generates through metabolic engineering and mutagenesis & screening approaches robust yeast strains that have a broad substrate range and can (co-)ferment C6 and C5 sugars to ethanol with high productivity (rate and yield), and that are significantly more stress tolerant, i.e. inhibitor, ethanol and thermotolerant than the current S.cerevisiae strains used in ethanol production. The NEMO project also identifies and improves enzymes for hydrolysis of biomasses relevant for Europe. Novel enzymes are identified and improved through various approaches, based on screening, broad comparative genomics analyses, and protein engineering. These efforts will generate more thermostable enzymes for high temperature hydrolysis, more efficient enzymes for hydrolysis of the resistant structures in lignocellulose such as crystalline cellulose and lignin-hemicellulose complexes, enzymes with reduced affinity on lignin, and efficient thermo and mesophilic enzyme mixtures that are optimised and tailor-made for the relevant biomasses for Europe and European industry. These novel biocatalysts are tested in an iterative manner in process relevant conditions, including also pilot-scale operations, which ensure that the novel enzymes and microbes will be superior in real process conditions. Furthermore, optimal enzyme, microbe and process regime combinations are identified, providing basis for the development of the most economic and ecoefficient overall processes. The impact of the NEMO project on 2nd generation bioethanol production is significant because it provides most realistic but widely applicable technologies that could be exploited broadly by European industry. Its impact goes also much beyond bioethanol because NEMO provides technology improvements that are directly applicable and crucial for efficient and economic production of also other biofuels and bulk chemicals.