The treatment of azo dye effluent from the dyeing process, as well as the sustainable development of the environment, can all be achieved through the use of green mycoremediation. In this in vitro study, Trichothecium roseum was isolated by the sedimentation technique from the environment of an unplasticized polyvinyl chloride pipe manufacturing plant and used to decolourize and optimise the biosorption percentage of Azo Lanasyn Navy M-DNL dye. T. roseum was also used to test the decolourization percentage of the dyes Lanasyn Navy M-DNL, Isolan Black 2S-LDN, and Isolan Yellow 2S-GLN. The outcomes showed that Lanasyn Navy M-DNL dye has the highest decolourization percentage (94%). Lanasyn Navy M-DNL dye was also found to be adsorbed onto the surface of T. roseum using scan electron microscopy and Fourier-transform infrared spectroscopy investigations. Under ideal optimisation parameters, the biosorption percentage of Lanasyn Navy M-DNL dye by beads of immobilised T. roseum cells was 96%, 97%, 96%, 97%, and 96%, respectively, for beads number 60, fresh weight 1 g, incubation temperature 25 °C, pH 6, dye concentrations 10 mg/l, and sucrose concentrations 10 g/l. Langmuir and Freundlich adsorption isotherms show good agreement between the Freundlich adsorption isotherm model and the adsorption process. Also, the elution cycle was found to be effective enough to be employed for five cycles using sixty beads of immobilised T. roseum cells. The current study suggests that T. roseum is an influencer of Lanasyn Navy M-DNL dye biosorption to create a successful green strategy for handling Lanasyn Navy M-DNL dye-contaminated effluents.