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The anaerobic gut fungi (Neocallimastigomycota), first described almost 50 years ago, hold extraordinary potential for biotechnology. Anaerobic fungi could contribute to bioenergy and bio-based chemical production via their ability to degrade lignocellulose, may enhance animal health and production, and are now being revealed to have interesting biosynthetic potential for development. In this chapter, we briefly describe the biology of these species, discuss recent efforts to identify and exploit anaerobic fungal enzymes, and survey challenges and opportunities for their adoption in a variety of biotechnology applications.