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Trichoderma is a well-known soil-borne fungus, highly efficient producer of extracellular enzymes including chitinases. The aim of this study was to recover a chitinase from fermentation waste after harvesting Trichoderma koningiopsis Th003 conidia and assess its potential as an enhancer of Beauveria bassiana insecticidal activity against Diatraea saccharalis. T. koningiopsis was produced by solid fermentation, conidia were harvested, and a crude extract (CE) was recovered by washing the residual substrate (rice:wheat bran). The partially purified chitinase (PPC) (75 kDa product) with N-acetyl-β-glucosaminidase activity was obtained by chromatography to 29.3-fold with optimal activity at pH 5 and 55°C. Both the CE and the PPC were mixed with B. bassiana Bv062 conidia and assessed in a bioassay against D. saccharalis larvae. The CE and PPC from T. koningiopsis Th003 did not affect the germination or viability of B. bassiana conidia and enhanced its insecticidal activity when used at 0.06 U/ml enzymatic activity with a 24.5% reduction in B. bassiana lethal time (LT 90 ). This study demonstrated the potential of chitinases produced by T. koningiopsis in solid fermentation to be recovered from the waste substrate and used as an additive to enhance B. bassiana, adding value to the main waste from the Trichoderma biopesticide/biofertilizer industries., (© 2021 Wiley-VCH GmbH.)