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.

The Himalayan Mountains are placed among the globally recognized biodiversity hot spots. While the Indian Himalayan Region (IHR) has been subjected to extensive studies on plant and animal biodiversity, microbial diversity is now being studied for its bioprospection. The present paper deals with the evaluation of bacterial diversity in high-altitude soil samples from IHR following polyphasic approach including comparison between the MALDI-TOF mass spectrometry and 16S rRNA gene sequencing for species-level identification. Initially, a culture collection of large number of bacterial isolates was established in the laboratory. Performing morphological and biochemical screenings, sixty-one representative isolates were selected for mass spectrometry and gene sequencing. Both the methods emerged with bacterial identification showing maximum number of Bacillus followed by Pseudomonas species. The other frequently isolated strains belonged to the genera Alcaligenes, Carnobacterium, Lysinibacillus, Microbacterium, Paenarthrobacter, Rhodococcus, Serratia and Stenotrophomonas. Although the MALDI-TOF technique appeared to be advantageous as less time-consuming in comparison with 16S rRNA-based method, the discrepancies at species level indicated the limited database of MALDI Biotyper and species complexity in the genera. The remarkable characteristics of the bacterial isolates were their tolerance to wide range of pH and temperature. Their potential to produce industrially valuable enzymes indicated their importance in bioprospection. Accessioning of these bacterial isolates in microbial culture collections is a cautious effort for their availability to conduct advanced research on these cold-adapted bacteria in future.

Production of laccase by a cold and pH tolerant strain ofPenicillium pinophilumhas been investigated under different cultural conditions for up to 35 days of incubation. The fungus was originally isolated from a low temperature environment under mountain ecosystem of Indian Himalaya. The estimations were conducted at 3 temperatures (15, 25, and 35°C), a range of pH (3.5–11.5), and in presence of supplements including carbon and nitrogen sources, vitamins, and antibiotics. Optimum production of laccase was recorded at 25°C (optimum temperature for fungal growth) and 7.5 pH. The production of enzyme was recorded maximum on day 28 (11.6±0.52 U/L) following a slow decline at day 35 of incubation (10.6±0.80 U/L). Fructose and potassium nitrate (0.2%) among nutritional supplements, chloramphenicol (0.1%) among antibiotics, and folic acid (0.1%) among vitamins were found to be the best enhancers for production of laccase. Relatively lower but consistent production of laccase for a longer period is likely to be an ecologically important phenomenon under low temperature environment. Further, enhancement in production of enzyme using various supplements will be useful for its use in specific biotechnological applications.