Endophytes are microorganisms that live within plant tissues that are potential sources of novel bioactive compounds, including enzymes. We have identified endophytes of the Australian native plant Eremophilia longifolia which were screened for the production of industrially useful enzymes. Seventeen fungal endophytes were isolated from the leaves of E. longifolia and enzyme production was investigated within a range of pH (3.5, 5.5, 7 and 9) and temperatures (9, 25, 37 and 50 °C). Amylase was the most common enzyme encountered with numerous isolates showing production throughout the temperature and pH ranges. Protease production was also seen over the conditions tested but was more dominant at lower pH and temperature. Activity was not observed for other enzymes including ligninase, xylanase and cellobiohydrolase. Enzymes from isolates of Preussia minima, Alternaria sp. and an unclassified fungus, which showed highest activity in screening assays, were investigated further. Enzyme production was verified by zymography and the amylase activity of P. minima was found to be significantly greater than that of Aspergillus oryzae particularly in alkaline conditions and low temperature which are desirable properties for the detergent industry. This work shows that enzymes with potential use in industry can be readily identified in fungal endophytes.