Tracking of pigment accumulation and secretion in extractive fermentation of Monascus anka GIM 3.592

BACKGROUND Monascus pigments are promising sources for food and medicine due to their natural food-coloring functions and pharmaceutical values. The innovative technology of extractive fermentation is used to promote pigment productivity, but reports of pigment trans-membrane secretion mechanism are rare. In this study, tracking of pigment accumulation and secretion in extractive fermentation of Monascus anka GIM 3.592 was investigated. RESULTS The increased vacuole size in mycelia correlated with fluorescence intensity (r > 0.85, p < 0.05), which indicates that intracellular pigments with strong fluorescence accumulated in the cytoplasmic vacuole. After adding nonionic surfactant Triton X-100, the uptake of rhodamine123 (Rh123) and 1-N-phenylnaphthylamine (NPN) and the release of K+ and Na+ rapidly increased, demonstrating that the physiological performances of the cell membrane varied upon damaging the integrity, increasing the permeability, and changing the potential. Simultaneously, the fatty acid composition also varied, which caused a weak fluidity in the membrane lipids. Therefore, the intracellular pigments embedded in Triton X-100 were secreted through the ion channels of the cell membrane. Dense, spherical pigment-surfactant micelles with an average size of 21 nm were distributed uniformly in the extraction broth. Based on the different pigment components between extractive fermentation and batch fermentation, a threefold decrease in the NAD+/NADH ratio in mycelia and a more than 200-fold increase in glucose-6-phosphate dehydrogenase (G6PDH) activity in extracellular broth occurred, further suggesting that a reduction reaction for pigment conversion from orange pigments to yellow pigments occurred in non-aqueous phase solution. CONCLUSIONS A putative model was established to track the localization of Monascus pigment accumulation and its trans-membrane secretion in extractive fermentation. This finding provides a theoretical explanation for microbial extractive fermentation of Monascus pigments, as well as other non-water-soluble products.