For a safe and sustainable environment, effective microbes as biocontrol agents are in high demand. We have isolated a new strain DTU001, investigated its antifungal spectrum, sequenced its genome, and uncovered the production of lipopeptides in HPLC-HRMS analysis. To test the antifungal efficacy, extracts of DTU001 was tested against a range of twenty human or plant pathogenic fungi. We demonstrate that inhibitory potential of DTU001 against selected fungi is superior in comparison to single lipopeptide, either iturin or fengycin. The isolate showed analogous biofilm formation to other closely related . To further support the biocontrol properties of the isolate, coculture with demonstrated that DTU001 exhibited excellent antiproliferation effect against . In summary, the described isolate is a potential antifungal agent with a broad antifungal spectrum that might assist our aims to avoid hazardous pathogenic fungi and provide alternative to toxicity caused by chemicals.