The development of alternate sources for the production of natural pigments has been targeted to overcome the utilization of artificial coloring, which is dangerous to human health and the environment. Dyes extracted from microbial sources are more important for beneficial food industry use, especially Monascus spp. produces several critical secondary metabolites such as lovastatin, g-amino butyric acids, monascodilone, monascorubramine, monascin, ankaflavin, rubropunctatin, and citrinin. Lovastatin is a fungal polyketide that inhibits the rate-limiting enzymes HMG-CoA reductase, an essential precursor in cholesterol biosynthesis. The light source regulates fungi’ growth, metabolism, and reproduction and is necessary for fungi’ existence and distribution. The impact of different color lights (red, green, blue, yellow, and white, darkness) and different temperatures (27°C and 37°C) on extracellular and intracellular pigment yield, lovastatin production, and biomass of Monascus ruber was studied, and appropriate incubation temperature and time enhance the intracellular, extracellular pigment, and biomass production. However, when exposed to other color lights, fungus growth and pigment yield are significantly reduced in Monascus ruber. Then, fungi and pigment yield development is decreased when exposed to other color lights. It can be concluded that darkness influenced pigment production and biomass yield at both temperatures (27°C and 37°C). Similarly, the production of lovastatin and its concentration were analyzed by HPLC. The highest concentration of lovastatin was obtained at 27°C when exposed to red color light (302.6 mg/ml for extracellular fermentation broth) and (86.7 mg/ml for intracellular fermentation broth). At 37°C, the highest concentration of lovastatin was obtained from (571.5 mg/ml extracellular fermentation broth) when exposed to darkness and (170.4 mg/ml intracellular fermentation broth) exposed to red color light. Thus, the result provides the knowledge to enable us to explore the pigments and lovastatin yield for functional foods and large-scale industrial applications.
Protoplast fusion is a crucial technique for strain enhancement that brings genetic recombination to filamentous fungus and creates hybrid strains. In this present study, the intergenic fusion between M. ruber and P. ostreatus has been carried out to enhance pigments, secondary metabolites, and proximate values. Protoplasts from M. ruber and P. ostreatus were isolated and an equal amount of protoplast was taken and performed protoplast fusion. The regenerated colonies were plated in potato dextrose agar with selected biological markers (nystatin and fluconazole) for the confirmation of the hybrid. In a standardized condition, the protoplast yield of M. ruber and P. ostreatus was 2.31×107 cells/ml and 2.33×107 cells/ml, respectively. Acidic pH yielded high amounts of protoplasts than neutral and alkaline pH. KCl showed a high yield of protoplasts 2.36×106 cells /ml from M. ruber and 2.46×107cells/ml from P. ostreatus. The colony resistance to nystatin and fluconazole was confirmed that the obtained regenerated colonies were fusant. The fusant has no septation in its transverse walls with lipid droplets. Fusion of protoplast from M. ruber and P. ostreatus resulted in genetic inference at the metabolic levels, explaining the elevation of pigment in fusant colonies. The proximate composition of fusant strains was higher. In conclusion, we have developed a new strain with genetic variance capable of producing an increased level of pigment with nutritional value through protoplast fusion that allows the generation of fusant with the characteristics of parent strains and elucidates the effectiveness of fusant for commercial applications.
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