With globalisation and growing health risks of synthetic colourants, search for pigments from natural sources has increased owing to their non-toxic nature. The present study highlights the bioprospection of natural pigment from a cold adapted fungal strain of Penicillium sp. (GBPI_P155), isolated from soil of Indian Himalayan region. The fungus produced insoluble and orange-coloured pigment in liquid medium with maximum production recorded in potato dextrose (PD) broth at 15°C and 3 pH, while maximum biomass was produced at 25°C and pH 3. While examining the effect of different mineral salts, and carbon and nitrogen sources on pigment production, maximum accumulation of pigment was recorded in PD broth supplemented with 2% maltose. Following production, extraction of pigment was performed using chloroform and characterised partially by UV/vis (λ_max at 495 nm and a shoulder peak at 530 nm) and Fourier Transform Infrared (FT-IR) spectroscopy. Thin layer chromatography of chloroform extract resulted in separation of pigment in three fractions with Rf values 0.911, 0.852 and 0.808, which were further analysed using Liquid Chromatography Mass Spectrometry (LC/MS). The overall approach resulted in identification of pigment as a mixture of different derivatives of carotenoids. The extracted pigment also possessed antimicrobial activity against Gram-positive and Gram-negative bacteria and actinobacteria.