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23 pages, 4959 KiB

Open AccessArticle

Characterization of Key Metabolic Markers in Hongqujiu Across Different Aging Years Using Metabolomics

by Yiyang Cai, Sunan Yan, Simei Huang, Bin Yang, Wenlan Mo, Lishi Xiao, Xiangyou Li and Zhiwei Huang

J. Fungi 2025, 11(5), 353; https://doi.org/10.3390/jof11050353 - 2 May 2025

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Abstract

Hongqujiu, one of the three principal varieties of yellow wine, is a traditional fermented beverage originating from China that employs Hongqu as the fermentation agent. In this study, an untargeted metabolomics approach based on gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS) [...] Read more.

Hongqujiu, one of the three principal varieties of yellow wine, is a traditional fermented beverage originating from China that employs Hongqu as the fermentation agent. In this study, an untargeted metabolomics approach based on gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS) was applied to systematically analyze the volatile compounds (VOCs) and non-volatile compounds (NVCs) in Hongqujiu across different aging years for the first time. The analysis identified a total of 262 VOCs and 2564 NVCs in samples of Hongqujiu aged for six distinct years. Based on metabolic differences, the samples were categorized into two groups: the low-year group (5-year, 6-year) and the high-year group (8-year, 10-year, 15-year, 20-year). Nineteen VOCs (e.g., 4-amino-butyric acid and diethanolamine) and thirty NVCs (e.g., palmitoylethanolamide and erinacine D) were identified as key differential metabolites distinguishing the low-year group from the high-year group. The higher-year group is enriched with a variety of substances with different flavors or biological activities, such as sugar derivatives, amino acids and their complexes, organic acids and their intermediate metabolites, steroids and terpenoid compounds, lipids and their derivatives, nitrogen-containing heterocycles, and aromatic compounds. The accumulation of these substances not only shapes the unique and rich flavor characteristics of aged red rice wine (such as the caramel aroma and umami peptide flavor), but also endows red rice wine with potential health benefits due to the physiological regulatory functions of some active ingredients. This study contributes to a deeper understanding of the composition and dynamic variations in metabolites in Hongqujiu, offering a scientific foundation for identifying aged Hongqujiu and conducting further research to enhance its quality. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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14 pages, 2544 KiB

Open AccessArticle

Transcriptome Analysis Revealed the Molecular Mechanism of Acetic Acid Increasing Monascus Pigment Production in Monascus ruber CICC41233

by Yan Wang, Weiwei Wu, Xiaoshu Wu, Weiyu Li, Jingjing Cui and Chuannan Long

J. Fungi 2025, 11(1), 49; https://doi.org/10.3390/jof11010049 - 9 Jan 2025

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Abstract

The addition of acetic acid to Monascus ruber cultures is usually used to inhibit the growth of heterotrophic bacteria; however, we found that acetic acid also promotes the growth of M. ruber CICC41233, as well as the synthesis of Monascus pigments (MPs). Compared [...] Read more.

The addition of acetic acid to Monascus ruber cultures is usually used to inhibit the growth of heterotrophic bacteria; however, we found that acetic acid also promotes the growth of M. ruber CICC41233, as well as the synthesis of Monascus pigments (MPs). Compared with no acetic acid or HCl addition, the diameter of M. ruber CICC41233 colonies increased significantly under acetic acid conditions. On the sixth day of fermentation, the yield of total pigments in M. ruber increased significantly by 9.97 times (compared with no acetic acid) and 13.9 times (compared with hydrochloric acid). The transcriptomics data showed that the differentially expressed genes between M. ruber with acetic acid and without acetic acid were mainly involved in starch and sucrose metabolism, glycolysis/gluconeogenesis, pyruvate metabolism, TCA cycle, and oxidative phosphorylation, and that these differentially expressed genes were not involved in amino acid metabolism. Gene expression analysis showed that the relative expression levels of MP synthesis genes (MpPKS5, MppA, MpFasB, MppB, MppD, and MppR2) were significantly up-regulated under acetic acid conditions. This study clarified the metabolic mechanism of acetic acid promoting the growth of M. ruber and the synthesis of MPs, which provided some theoretical guidance for the large-scale production of MPs in the industry in future. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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10 pages, 2631 KiB

Open AccessArticle

Improvement of Monacolin K and Pigment Production in Monascus by 5-Azacytidine

by Chan Zhang, Haijiao Wang, Qing Sun, Arzugul Ablimit, Huijun Dong, Congcong Wang, Duchen Zhai, Bobo Zhang, Wenlin Hu, Chengjian Liu and Chengtao Wang

J. Fungi 2024, 10(12), 819; https://doi.org/10.3390/jof10120819 - 26 Nov 2024

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Abstract

Monascus species are known to produce various secondary metabolites with polyketide structures, including Monacolins, pigments, and citrinin. This study investigates the effects of 5-azacytidine on Monascus M1 and RP2. The dry weight, red, yellow, and orange pigment values, and Monacolin K yield of [...] Read more.

Monascus species are known to produce various secondary metabolites with polyketide structures, including Monacolins, pigments, and citrinin. This study investigates the effects of 5-azacytidine on Monascus M1 and RP2. The dry weight, red, yellow, and orange pigment values, and Monacolin K yield of both Monascus strains were measured, and their hyphae observed through electron microscopy. The experimental group showed higher dry weights and pigment values than the control group for both strains. However, Monacolin K production increased substantially only for Monascus M1. Electron micrographs revealed surface wrinkles and large protrusions in both strains after 5-azacytidine treatment. As a potent DNA methylation-promoting agent, 5-azacytidine is very useful for epigenetic and cancer biology studies and for studying secondary metabolism in fungi. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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14 pages, 4004 KiB

Open AccessArticle

Comparative Effects of Monascin and Monascinol Produced by Monascus pilosus SWM-008 on Pro-Inflammatory Factors and Histopathological Alterations in Liver and Kidney Tissues in a Streptozotocin–Nicotinamide-Induced Rat Model

by Pei-Xin Yang, Ya-Wen Hsu, Tzu-Ming Pan and Chun-Lin Lee

J. Fungi 2024, 10(12), 815; https://doi.org/10.3390/jof10120815 - 25 Nov 2024

Cited by 1 | Viewed by 1140

Abstract

Monascinol (Msol), an analog of monascin (MS) produced by Monascus pilosus, possesses potential anti-inflammatory properties. This study compares the effects of M. pilosus SWM-008 fermented red mold rice, which contains the functional components MS and Msol, on liver and kidney damage related [...] Read more.

Monascinol (Msol), an analog of monascin (MS) produced by Monascus pilosus, possesses potential anti-inflammatory properties. This study compares the effects of M. pilosus SWM-008 fermented red mold rice, which contains the functional components MS and Msol, on liver and kidney damage related to diabetic complications in rats. An animal model of liver and kidney injury was induced by an intraperitoneal injection of streptozotocin (STZ) at 65 mg/kg body weight combined with nicotinamide (NA) at 150 mg/kg body weight. Our findings indicate that Msol significantly reduces STZ-NA induced pro-inflammatory markers, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2) in both liver and kidney tissues. Significant improvements were noted in the histopathological assessments. Msol was more effective than MS in suppressing renal IL-1β and COX-2 expressions. In summary, the findings indicate that Msol shows potential as a novel therapeutic agent for treating liver and kidney injuries associated with diabetic complications. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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21 pages, 7014 KiB

Open AccessArticle

Molecular Mechanism of Mok I Gene Overexpression in Enhancing Monacolin K Production in Monascus pilosus

by Zhiwei Huang, Lishi Xiao, Wenlan Mo, Yaru Zhang, Yiyang Cai, Simei Huang, Zhiting Chen and Chuannan Long

J. Fungi 2024, 10(10), 721; https://doi.org/10.3390/jof10100721 - 16 Oct 2024

Cited by 4 | Viewed by 1295

Abstract

Monascus species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the mok I gene from the MK synthesis gene cluster in Monascus species can significantly increase MK production; however, the molecular [...] Read more.

Monascus species are capable of producing various active metabolites, including monacolin K (MK) and pigments. Studies have shown that the overexpression of the mok I gene from the MK synthesis gene cluster in Monascus species can significantly increase MK production; however, the molecular mechanism has not yet been fully elucidated. Therefore, this study focused on the mok I gene of Monascus pilosus to construct overexpression strains of the mok I gene, resulting in high-yield MK production. Sixteen positive transformants were obtained, seven of which produced 9.63% to 41.39% more MK than the original strain, with no citrinin detected in any of the transformants. The qRT-PCR results revealed that the expression levels of mok I in the transformed strains TI-13, TI-24, and TI-25 increased by more than 50% compared to the original strain at various fermentation times, with the highest increase being 10.9-fold. Furthermore, multi-omics techniques were used to analyze the molecular mechanisms underlying enhanced MK production in transformed strains. The results indicated that mok I overexpression may enhance MK synthesis in M. pilosus by regulating the expression of key genes (such as MAO, HPD, ACX, and PLC) and the synthesis levels of key metabolites (such as delta-tocopherol and alpha-linolenic acid) in pathways linked to the biosynthesis of cofactors, the biosynthesis of unsaturated fatty acids, tyrosine metabolism, ubiquinone and other terpenoid-quinone biosynthesis, alpha-linolenic acid metabolism, and glycerophospholipid metabolism. These findings provide a theoretical basis for further study of the metabolic regulation of MK in Monascus species and for effectively enhancing their MK production. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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15 pages, 4166 KiB

Open AccessArticle

Mutation Breeding of Monascus to Produce a High Yield of Orange Pigment and Low Citrinin Content Using the ARTP Method

by Chan Zhang, Qing Sun, Le Yang, Arzugul Ablimit, Huijun Dong, Haijiao Wang, Congcong Wang and Chengtao Wang

J. Fungi 2024, 10(8), 553; https://doi.org/10.3390/jof10080553 - 5 Aug 2024

Cited by 1 | Viewed by 1757

Abstract

Monascus is a filamentous fungus with a long history of application in China, which can produce a variety of secondary metabolites, including Monascus red pigments, Monascus orange pigments, Monascus yellow pigments, and citrinin. There is widespread attention being paid to natural pigments because [...] Read more.

Monascus is a filamentous fungus with a long history of application in China, which can produce a variety of secondary metabolites, including Monascus red pigments, Monascus orange pigments, Monascus yellow pigments, and citrinin. There is widespread attention being paid to natural pigments because of their safety. Among the many natural pigments, orange pigment has a wide range of applications because of its unique color, but current production levels in the orange pigment industry are limited to a certain extent due to the insufficiently wide range of sources and low production. In this study, the ARTP mutation was used to obtain a strain with high-yield orange pigment and low citrinin. The strain RS7 was obtained through two-step mutagenesis, and all three pigments were improved to different degrees. The color value of orange pigment was elevated from the original 108 U/mL to 180 U/mL, an increase of 66.7% compared to the original strain, and the citrinin content was reduced by 69%. The result of microscopic morphology showed that RS7 has more wrinkles and is more convex than the R1 strain, but there was little change between the two strains. Therefore, the ARTP mutation influenced the growth and the biosynthesis of pigments in Monascus. In addition, the conditions of ultrasonic extraction of Monascus pigments were optimized using the response surface, and the separation of pigments was achieved with the method of thin-layer chromatography. Pigment stability results showed that the temperature had no significant effect on orange pigment, while tea polyphenol could improve its stability. This study generated a strain with high-yielding orange pigment and could lay a foundation for the future application of Monascus orange pigment in the food industry. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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18 pages, 3170 KiB

Open AccessArticle

Optimization Co-Culture of Monascus purpureus and Saccharomyces cerevisiae on Selenium-Enriched Lentinus edodes for Increased Monacolin K Production

by Yi He, Huafa Lai, Jinxiao Liang, Lu Cheng, Lixia He, Haolin Wang, Qingqing Teng, Wenjing Cai, Rui Wang, Lisha Zhu, Zhengbin Pang, Dafu Zhang, Xingxing Dong and Chao Gao

J. Fungi 2024, 10(7), 503; https://doi.org/10.3390/jof10070503 - 20 Jul 2024

Cited by 2 | Viewed by 1384

Abstract

Selenium-enriched Lentinus edodes (SL) is a kind of edible fungi rich in organic selenium and nutrients. Monascus purpureus with high monacolin K (MK) production and Saccharomyces cerevisiae were selected as the fermentation strains. A single-factor experiment and response surface methodology were conducted to [...] Read more.

Selenium-enriched Lentinus edodes (SL) is a kind of edible fungi rich in organic selenium and nutrients. Monascus purpureus with high monacolin K (MK) production and Saccharomyces cerevisiae were selected as the fermentation strains. A single-factor experiment and response surface methodology were conducted to optimize the production conditions for MK with higher contents from selenium-enriched Lentinus edodes fermentation (SLF). Furthermore, we investigated the nutritional components, antioxidant capacities, and volatile organic compounds (VOCs) of SLF. The MK content in the fermentation was 2.42 mg/g under optimal fermentation conditions. The organic selenium content of SLF was 7.22 mg/kg, accounting for 98% of the total selenium content. Moreover, the contents of total sugars, proteins, amino acids, reducing sugars, crude fiber, fat, and ash in SLF were increased by 9%, 23%, 23%, 94%, 38%, 44%, and 25%, respectively. The antioxidant test results demonstrated that 1.0 mg/mL of SLF exhibited scavenging capacities of 40%, 70%, and 79% for DPPH, ABTS, and hydroxyl radicals, respectively. Using gas chromatography–ion mobility spectrometry technology, 34 unique VOCs were identified in SLF, with esters, alcohols, and ketones being the main components of its aroma. This study showed that fungal fermentation provides a theoretical reference for enhancing the nutritional value of SL. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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Review

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10 pages, 2047 KiB

Open AccessReview

From Random Perturbation to Precise Targeting: A Comprehensive Review of Methods for Studying Gene Function in Monascus Species

by Yunxia Gong, Shengfa Li, Deqing Zhao, Xi Yuan, Yin Zhou, Fusheng Chen and Yanchun Shao

J. Fungi 2024, 10(12), 892; https://doi.org/10.3390/jof10120892 - 23 Dec 2024

Viewed by 722

Abstract

Monascus, a genus of fungi known for its fermentation capability and production of bioactive compounds, such as Monascus azaphilone pigments and Monacolin K, have received considerable attention because of their potential in biotechnological applications. Understanding the genetic basis of these metabolic pathways [...] Read more.

Monascus, a genus of fungi known for its fermentation capability and production of bioactive compounds, such as Monascus azaphilone pigments and Monacolin K, have received considerable attention because of their potential in biotechnological applications. Understanding the genetic basis of these metabolic pathways is crucial for optimizing the fermentation and enhancing the yield and quality of these products. However, Monascus spp. are not model fungi, and knowledge of their genetics is limited, which is a great challenge in understanding physiological and biochemical phenomena at the genetic level. Since the first application of particle bombardment to explore gene function, it has become feasible to link the phenotypic variation and genomic information on Monascus strains. In recent decades, accurate gene editing assisted by genomic information has provided a solution to analyze the functions of genes involved in the metabolism and development of Monascus spp. at the molecular level. This review summarizes most of the genetic manipulation tools used in Monascus spp. and emphasizes Agrobacterium tumefaciens-mediated transformation and nuclease-guided gene editing, providing comprehensive references for scholars to select suitable genetic manipulation tools to investigate the functions of genes of interest in Monascus spp. Full article

(This article belongs to the Special Issue Monascus spp. and Their Relative Products)

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