Monascus pigments are promising sources of natural food colorants, and their productivity can be improved by a novel extractive fermentation technology. In this study, we investigated the variations in pigment characteristics and biosynthetic gene expression levels in resting cell culture systems combined with extractive fermentation in Monascus anka GIM 3.592. Although the biomass was low at about 6 g/L DCW, high pigment titer of approximately 130 AU470 was obtained in the resting culture with cells from extractive fermentation, illustrating that it had a good biocatalytic activity for pigment synthesis. The oxidation–reduction potential value correlated with the rate of relative content of the intracellular orange pigments to the yellow pigments (O/Y, r > 0.90, p < 0.05), indicating that the change in pigment characteristics may be responsible for the cellular redox activity. The up- or down-regulation of the pigment biosynthetic genes (MpFasA2, MpFasB2, MpPKS5, mppD, mppB, mppR1, and mppR2) in the resting culture with extractive culture cells was demonstrated by real-time quantitative polymerase chain reaction analysis. Moreover, the mppE gene associated with the yellow pigment biosynthesis was significantly (p < 0.05) down-regulated by about 18.6%, whereas the mppC gene corresponding to orange pigment biosynthesis was significantly (p < 0.05) up-regulated by approximately 21.0%. These findings indicated that extractive fermentation was beneficial for the biosynthesis of the intracellular orange pigment. The mechanism described in this study proposes a potential method for the highly efficient production of Monascus pigments.
This is a preview of subscription content, log in to check access.
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no competing interests.
The authors gratefully acknowledge the financial support of the National Natural Science Foundation of China (No. 31271925) and the Special Project on the Integration of Industry, Education and Research of Guangdong Province, China (No. 2013B090600015), as well as the Science and Technology Program of Guangzhou, China (No. 2014 J4100192).
Balakrishnan B, Karki S, Chiu S, Kim H, Suh J, Nam B, Yoo Y, Chen C, Kwon H (2013) Genetic localization and in vivo characterization of a Monascus azaphilone pigment biosynthetic gene cluster. Appl Microbiol Biot 97:6337–6345CrossRefGoogle Scholar
Balakrishnan B, Kim H, Suh J, Chen C, Liu K, Park S, Kwon H (2014) Monascus azaphilone pigment biosynthesis employs a dedicated fatty acid synthase for short chain fatty acyl moieties. J Korean Soc Appl Bi 57:191–196CrossRefGoogle Scholar
Balakrishnan B, Park S, Kwon H (2017) A reductase gene mppE controls yellow component production in azaphilone polyketide pathway of Monascus. Biotechnol Lett 39:163–169CrossRefPubMedGoogle Scholar
Carels M, Shepherd D (1977) The effect of different nitrogen sources on pigment production and sporulation of Monascus species in submerged, shaken culture. Can J Microbiol 23:1360–1372CrossRefPubMedGoogle Scholar
Chen D, Xue C, Chen M, Wu S, Li Z, Wang C (2016) Effects of blue light on pigment biosynthesis of Monascus. J Microbiol 54:305–310CrossRefPubMedGoogle Scholar
Chen G, Shi K, Song D, Quan L, Wu Z (2015) The pigment characteristics and productivity shifting in high cell density culture of Monascus anka mycelia. BMC Biotechnol 15(1):72CrossRefPubMedPubMedCentralGoogle Scholar
Chen G, Wu Z (2016) Production and biological activities of yellow pigments from Monascus fungi. World J Microb Biot 32(8):1–8CrossRefGoogle Scholar
Chen G, Huang T, Bei Q, Tian X, Wu Z (2017a) Correlation of pigment production with mycelium morphology in extractive fermentation of Monascus anka GIM 3.592. Process Biochem 58:42–50CrossRefGoogle Scholar
Chen G, Tang R, Tian X, Qin P, Wu Z (2017b) Change of Monascus pigment metabolism and secretion in different extractive fermentation process. Bioprocess Biosyst Eng 40(6):857–866CrossRefPubMedGoogle Scholar
Chen G, Bei Q, Shi K, Tian X, Wu Z (2017c) Saturation effect and transmembrane conversion of Monascus pigment in nonionic surfactant aqueous solution. AMB Express 7(1):24CrossRefPubMedPubMedCentralGoogle Scholar
Du C, Zhang Y, Li Y, Cao Z (2007) Novel redox potential-based screening strategy for rapid isolation of Klebsiella pneumoniae mutants with enhanced 1,3-propanediol-producing capability. Appl Environ Microb 73(14):4515–4521CrossRefGoogle Scholar
Hajjaj H, Klaebe A, Loret MO, Tzedakis T, Goma G, Blanc PJ (1997) Production and identification of N-glucosylrubropunctamine and N-glucosylmonascorubramine from Monascus ruber and occurrence of electron donor–acceptor complexes in these red pigments. Appl Environ Microb 63:2671–2678Google Scholar
Hajjaj H, Klaebe A, Goma G, Blanc PJ, Barbier E, Francois J (2000) Medium-chain fatty acids affect citrinin production in the filamentous fungus Monascus ruber. Appl Environ Microb 66(3):1120–1125CrossRefGoogle Scholar
Hu Z, Zhang X, Wu Z, Qi H, Wang Z (2012a) Perstraction of intracellular pigments by submerged cultivation of Monascus in nonionic surfactant micelle aqueous solution. Appl Microbiol Biot 94(1):81–89CrossRefGoogle Scholar
Hu Z, Zhang X, Wu Z, Qi H, Wang Z (2012b) Export of intracellular Monascus pigments by two-stage microbial fermentation in nonionic surfactant micelle aqueous solution. J Biotechnol 162(2):202–209CrossRefPubMedGoogle Scholar
Juzlova P, Martinkova L, Kren V (1996) Secondary metabolites of the fungus Monascus: a review. J Ind Microbiol Biot 16(3):163–170CrossRefGoogle Scholar
Julsing MK, Kuhn D, Schmid D, Buhler B (2012) Resting cells of recombinant E. coli show high epoxidation yields on energy source and high sensitivity to product inhibition. Biotechnol Bioeng 109(5):1109–1119Google Scholar
Kang B, Zhang X, Wu Z, Qi H, Wang Z (2013) Solubilization capacity of nonionic surfactant micelles exhibiting strong influence on export of intracellular pigments in Monascus fermentation. Microb Biotechnol 6(5):540–550CrossRefPubMedPubMedCentralGoogle Scholar
Kitamoto D, Fuzishiro T, Yanagishita H, Nakanc T, Nakahara T (1992) Production of mannosylerythritol lipids as biosurfactants by resting cells of Candida antarctica. Biotechnol Lett 14(4):305–310Google Scholar
Lin TF, Demain AL (1993) Resting cell studies on formation of water soluble red pigments by Monascus sp. J Ind Microbiol Biot 12:361–367Google Scholar
Liu JZ, Yang HY, Weng LP, Ji LN (1999) Synthesis of glucose oxidase and catalase by Aspergillus niger in resting cell culture system. Lett Appl Microbiol 29(5):337–341Google Scholar
Lin Y, Wang T, Lee M, Su N (2008) Biologically active components and nutraceuticals in the Monascus-fermented rice: a review. Appl Microbiol Biot 77(5):965–973CrossRefGoogle Scholar
Liu Q, Xie N, He Y, Wang L, Shao Y, Zhao H, Chen F (2014) MpigE, a gene involved in pigment biosynthesis in Monascus ruber M7. Appl Microbiol Biot 98:285–296CrossRefGoogle Scholar
Liu S (2012) Study on S-poly-L-lysine biosynthesis and metabolic regulation. South China University of Technology, China, DissertationGoogle Scholar
Lu F, Huang Y, Zhang X, Wang Z (2017) Biocatalytic activity of Monascus mycelia depending on physiology and high sensitivity to product concentration. AMB Express 7(1):88CrossRefPubMedPubMedCentralGoogle Scholar
Patakova P (2013) Monascus secondary metabolites: production and biological activity. J Ind Microbiol Biot 40(2):169–181CrossRefGoogle Scholar
Shao Y, Lei M, Mao Z, Zhou Y, Chen F (2014) Insights into Monascus biology at the genetic level. Appl Microbiol Biot 98(9):3911–3922CrossRefGoogle Scholar
Shi K, Song D, Chen G, Pistolozzi M, Wu Z, Quan L (2015) Controlling composition and color characteristics of Monascus pigments by pH and nitrogen sources in submerged fermentation. J Biosci Bioeng 120(2):145–154CrossRefPubMedGoogle Scholar
Sun W, Xiao F, Wei Z, Cui F, Yu L, Yu S, Zhou Q (2015) Non-sterile and buffer-free bioconversion of glucose to 2-keto-gluconic acid by using Pseudomonas fluorescens AR4 free resting cells. Process Biochem 50:493–499Google Scholar
Tsuge Y, Kawaguchi H, Sasaki K, Tanaka T, Kondo A (2014) Two-step production of D-lactate from mixed sugars by growing and resting cells of metabolically engineered Lactobacillus plantarum. Appl Microbiol Biot 98:4911–4918CrossRefGoogle Scholar
Wang B, Zhang X, Wu Z, Wang Z (2015) Investigation of relationship between lipid and Monascus pigment accumulation by extractive fermentation. J Biotechnol 212:167–173CrossRefPubMedGoogle Scholar
Wang B, Zhang X, Wu Z, Wang Z (2016) Biosynthesis of Monascus pigments by resting cell submerged culture in nonionic surfactant micelle aqueous solution. Appl Microbiol Biot 100:7083–7089CrossRefGoogle Scholar
Wang M, Huang T, Chen G, Wu Z (2017) Production of water-soluble yellow pigments via high glucose stress fermentation of Monascus ruber CGMCC 10910. Appl Microbiol Biot 101(8):3121–3130CrossRefGoogle Scholar
Willrodt C, Hoschek A, Buhler B, Schmid A, Julsing MK (2015) Decoupling production from growth by magnesium sulfate limitation boosts de novo limonene production. Biotechnol Bioeng. https://doi.org/10.1002/bit.25883