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Applied Microbiology and Biotechnology Nov 2023Monascus sp. is an important food microbial resource with the production of cholesterol-lowering agent lovastatin and other healthy metabolites. However, the mycotoxin...
Monascus sp. is an important food microbial resource with the production of cholesterol-lowering agent lovastatin and other healthy metabolites. However, the mycotoxin citrinin naturally produced by Monascus sp. and the insufficient productivity of lovastatin limit its large-scale use in food industry. The aim of this paper is to modify a lovastatin-producing strain Monascus pilosus GN-01 through metabolic engineering to obtain a citrinin-free M. pilosus strain with higher yield of lovastatin. The citrinin synthesis regulator gene ctnR was firstly disrupted to obtain GN-02 without citrinin production. Based on that, the lovastatin biosynthesis genes (mokC, mokD, mokE, mokF, mokH, mokI, and LaeA) were, respectively, overexpressed, and pigment-regulatory gene (pigR) was knocked out to improve lovastatin production. The results indicated ctnR inactivation effectively disrupted the citrinin release by M. pilosus GN-01. The overexpression of lovastatin biosynthesis genes and pigR knockout could lead higher contents of lovastatin, of which pigR knockout strain achieved 76.60% increase in the yield of lovastatin compared to GN-02. These studies suggest that such multiplex metabolic pathway engineering in M. pilosus GN-01 is promising for high lovastatin production by a safe strain for application in Monascus-related food. KEY POINTS: • Disruption of the regulator gene ctnR inhibited citrinin production of M. pilosus. • Synchronous overexpression of biosynthesis gene enhanced lovastatin production. • pigR knockout enhanced lovastatin of ΔctnR strain of M. pilosus.
PubMed: 37672068
DOI: 10.1007/s00253-023-12747-2 -
Food Chemistry Jan 2024A co-fermentation strategy was explored to rapidly improve the characteristic components and quality of Pericarpium Citri Reticulatae (PCR) using Monascus anka and...
A co-fermentation strategy was explored to rapidly improve the characteristic components and quality of Pericarpium Citri Reticulatae (PCR) using Monascus anka and Saccharomyces cerevisiae, and the enzymatic mechanism was investigated. The results showed that the free flavonoid content of fermented PCR was 48.12% higher than that of unfermented PCR after 12 days of co-fermentation, resulting in stronger antioxidant activity. d-Limonene, γ-terpinene, proline (Pro), arginine (Arg), and serine (Ser) contributed the most to the flavors of citrus, herb, and sweet citrus based on odor and taste activity value analysis. Metabolomics and multivariate statistics showed that 55 components were differentially metabolized during co-fermentation, and ten metabolic pathways were closely related to metabolism. Furthermore, five hydrolases participated in the release and conversion of the active ingredients. This study provides an effective processing method for PCR and is conducive to the development of new PCR functional health foods.
Topics: Fermentation; Metabolomics; Arginine; Candy; Functional Food; Saccharomyces cerevisiae
PubMed: 37657346
DOI: 10.1016/j.foodchem.2023.137227 -
World Journal of Microbiology &... Aug 2023L-glutaminase is a hydrolytic enzyme with wide biotechnological applications. Mostly, these enzymes are employed in the feed industry for flavor enhancement and...
L-glutaminase is a hydrolytic enzyme with wide biotechnological applications. Mostly, these enzymes are employed in the feed industry for flavor enhancement and acrylamide mitigation. Also, L-glutaminase may have antiviral and antineoplastic effects making it a good choice for pharmaceutical applications. In this study, the strain Monascus ruber URM 8542 was identified through classical and molecular taxonomy using partial sequencing of β-tubulin and calmodulin genes. Subsequently, the optimal culture conditions were evaluated by submerged fermentation (L-glutamine 10 g.L) for L-glutaminase excretion. The isolate was identified as M. ruber URM 8542 which showed significant extracellular enzyme production with a yield of 11.4 times in relation to the specific activity of intracellular L-glutaminase. Regarding the optimization experiments, several factors such as L-glutamine concentration, temperature, and pH were compared using a full factorial design (2). The concentrations greater than 1% proved to be significantly better for glutaminase production (R = 0.9077). Additionally, the L-glutaminase was optimally active at pH 7.0 and 30 ºC. The L-glutaminase was remarkably stable across an alkaline pH range (7.0-8.0) and had a thermal stability ranging from 30 ºC to 60 ºC for 1 h. Taken together, these findings suggest that the L-glutaminase produced by M. ruber is a promising candidate for pharmacological application, although further studies need to be performed. To the best of our knowledge, this is the first report of L-glutaminase production by Monascus ruber.
Topics: Glutaminase; Glutamine; Ice Cream; Monascus
PubMed: 37632594
DOI: 10.1007/s11274-023-03733-x -
Journal of Agricultural and Food... Sep 2023Exopolysaccharides from genistein-stimulated (G-EMP) exhibited immunomodulatory potential in vitro, but whether it had immune-enhancing effects in vivo and its...
Exopolysaccharides from genistein-stimulated (G-EMP) exhibited immunomodulatory potential in vitro, but whether it had immune-enhancing effects in vivo and its potential mechanism are not yet known. Here, the immunomodulatory effects of G-EMP were investigated by establishing an immunosuppressed mouse model treated with cyclophosphamide (Cy). The results suggested that G-EMP effectively alleviated the signs of weight reduction and diet reduction caused by Cy, increased fecal water content and splenic index, and decreased the oxidative stress of the liver. Simultaneously, G-EMP improved Cy-induced intestinal injury by restoring villus length, increasing the number of cupped cells, upregulating the expression of mucin and tight junction proteins, and downregulating the ratio of apoptotic proteins (Bax/Bcl-2). It also boosted the levels of mouse colonic cytokines, CD4 and CD8 T cells. Additionally, G-EMP markedly enhanced immunomodulation via the activation of PI3K/AKT-MAPKs/NF-κB signal pathways. Furthermore, G-EMP intervention displayed a positive association with most immunological indexes by elevating the levels of short-chain fatty acids, varying gut microbiota composition, and enhancing beneficial bacteria (, and ). These findings demonstrated that G-EMP can strengthen immunity, repair intestinal mucosal damage, regulate gut microbiota, and be a potential source of prebiotics.
Topics: Animals; Mice; NF-kappa B; Genistein; Monascus; Phosphatidylinositol 3-Kinases; Proto-Oncogene Proteins c-akt; Gastrointestinal Microbiome; CD8-Positive T-Lymphocytes; Cyclophosphamide
PubMed: 37611142
DOI: 10.1021/acs.jafc.3c03186 -
Molecules (Basel, Switzerland) Jul 2023Fish bone fermented using (FBF) has total phenols and functional amino acids that contribute to its anti-oxidant and anti-inflammatory properties. Colorectal cancer,...
Fish bone fermented using (FBF) has total phenols and functional amino acids that contribute to its anti-oxidant and anti-inflammatory properties. Colorectal cancer, one of the most prevalent cancers and the third largest cause of death worldwide, has become a serious threat to global health. This study investigates the anti-cancer effects of FBF (1, 2.5 or 5 mg/mL) on the cell growth and molecular mechanism of HCT-116 cells. The HCT-116 cell treatment with 2.5 or 5 mg/mL of FBF for 24 h significantly decreased cell viability ( < 0.05). The S and G2/M phases significantly increased by 88-105% and 25-43%, respectively ( < 0.05). Additionally, FBF increased the mRNA expression of caspase 8 (38-77%), protein expression of caspase 3 (34-94%), poly (ADP-ribose) polymerase (PARP) (31-34%) and induced apoptosis (236-773%) of HCT-116 cells ( < 0.05). FBF also increased microtubule-associated protein 1B light chain 3 (LC3) (38-48%) and phosphoinositide 3 kinase class III (PI3K III) (32-53%) protein expression, thereby inducing autophagy (26-52%) of HCT-116 cells ( < 0.05). These results showed that FBF could inhibit HCT-116 cell growth by inducing S and G2/M phase arrest of the cell cycle, apoptosis and autophagy. Thus, FBF has the potential to treat colorectal cancer.
Topics: Animals; Humans; Monascus; Phosphatidylinositol 3-Kinases; Cell Line, Tumor; Apoptosis; Colorectal Neoplasms; Autophagy
PubMed: 37570647
DOI: 10.3390/molecules28155679 -
Foods (Basel, Switzerland) Jul 2023In order to study the effects and mechanism of Monascus on the quality of hairtail surimi, high-throughput sequencing technology, headspace solid-phase...
In order to study the effects and mechanism of Monascus on the quality of hairtail surimi, high-throughput sequencing technology, headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC/MS), and electronic nose techniques were used to investigate the changes in the quality, microbial diversity, and volatile flavor compounds of Monascus-fermented hairtail surimi (MFHS) during fermentation. The results showed that the total volatile basic nitrogen (TVB-N) index of hairtail surimi fermented by Monascus for 0-5 h met the requirements of the national standard. Among them, the 1 h group showed the best gel quality, which detected a total of 138 volatile substances, including 20 alcohols, 7 aldehydes, 12 olefins, 4 phenols, 12 alkanes, 8 ketones, 15 esters, 6 acids, 16 benzenes, 4 ethers, and 8 amines, as well as 26 other compounds. In addition, the dominant fungal microorganisms in the fermentation process of MFHS were identified, and a Spearman correlation analysis showed that 16 fungal microorganisms were significantly correlated with the decrease in fishy odor substances in the fermented fish and that 8 fungal microorganisms were significantly correlated with the increase in aromatic substances after fermentation. In short, Monascus fermentation can eliminate and reduce the fishy odor substances in hairtail fish, increase and improve the aromatic flavor, and improve the quality of hairtail surimi gel. These findings are helpful for revealing the mechanism of the quality formation of fermented surimi and provide guidance for the screening of starter culture in the future.
PubMed: 37569159
DOI: 10.3390/foods12152891 -
Heliyon Aug 2023are widely used around the world, mainly in purging and removal of endogenous active substances, such as anthraquinone and its derivatives. However, the potential...
are widely used around the world, mainly in purging and removal of endogenous active substances, such as anthraquinone and its derivatives. However, the potential toxicity of anthraquinones to the liver, kidney, and intestinal limits the application of . In this study, we aimed at safe regulation of to degrade anthraquinones, boosting medicinal properties and reducing toxicity and potency with fermentation strains H1102 for fermentation were selected as the initial strain which was capable of producing high yields of functional pigment and low yields of hazardous citrinin. The anthraquinone degradation rate reached 41.2%, with 212.2 U mL of the pigment and approximately 0.038 mg L of the citrinin under optimal fermentation conditions followed by response surface streamlining, which met the requirements of reducing toxicity, increasing efficiency of fermented . Furthermore, the culture had no observable toxic effect on HK-2 and L-02 cells and further inhibited cell apoptosis and necrosis. Overall, our results showed that fermentation could provide an alternative strategy for toxicity reduction of herbal medicines as well as efficacy enhancement.
PubMed: 37560635
DOI: 10.1016/j.heliyon.2023.e18735 -
Food Chemistry Jan 2024This study aims to explore the core microbiota of kombucha and to discover potential correlations between microbiota and volatile flavor compounds. The total acidity and...
This study aims to explore the core microbiota of kombucha and to discover potential correlations between microbiota and volatile flavor compounds. The total acidity and microbial colony numbers changed dramatically in different fermentation periods of kombucha. Microbial analysis based on high throughput sequencing technology showed that the bacteria of Komagataeibacter, Pseudomonas, Burkholderia, Ralstonia, Halomonas, Sphingomonas and fungi of Dekkera, Saccharomyces cerevisiae, Botryotrichum, Monascus, Pichia were the dominant genera. In addition, the correlation coefficients between the bacteria and fungi were different. The volatile flavor compounds of alcohols, acids, esters, aldehydes, ketones, phenolics, and terpenes were identified using headspace solid-phase microextraction combined with gas chromatography coupled with mass spectrometry. Typically, the concentrations of ethanol, acetic acid, and ethyl acetate was 71.59-248.23 μg/L, 97.73-849.00 μg/L, and 44.52-181.59 μg/L, respectively, during fermentation. This study is helpful to understand the dynamic changes of microbial communities and volatile flavor compounds during the fermentation of kombucha.
Topics: Fermentation; Gas Chromatography-Mass Spectrometry; Microbiota; Bacteria; Ethanol; Microbial Interactions; Volatile Organic Compounds
PubMed: 37544149
DOI: 10.1016/j.foodchem.2023.137060 -
Foods (Basel, Switzerland) Jul 2023To study the diversity of microbial flora in Hongqu sufu and analyze the characteristics of special flavor compounds, this study took self-made Hongqu sufu as the...
To study the diversity of microbial flora in Hongqu sufu and analyze the characteristics of special flavor compounds, this study took self-made Hongqu sufu as the research object. Dynamic changes in sufu during fermentation were studied. High-throughput sequencing (HTS) was used to analyze changes in the diversity of fungal and bacterial communities during fermentation. The results showed that at the phylum level, the dominant fungal phyla were identified, Mucormyces and Ascomycetes. The dominant bacterial phyla were Proteobacteria and Firmicutes. At the genus level, the dominant fungal genera were identified as , and . The dominant bacterial genera were , , , and . Headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) combined with technology that can dynamically change flavor compounds was explored to investigate the correlation between microbiota and flavor compounds. In different stages of fermentation, 75 main volatile organic compounds were identified, including seven alcohols, four acids, 16 alkanes, 14 olefins, seven kinds of aldehydes, two kinds of ketones, 10 kinds of esters, one kind of phenol, one kind of sulfur-containing compound, one benzene, and 12 other compounds. The correlation analysis between flora and flavor compounds showed that the fungi genera and were significantly correlated with most flavor compounds. Bacteria genera including , , and had a strong positive correlation with ethyl oleate.
PubMed: 37509892
DOI: 10.3390/foods12142800 -
Foods (Basel, Switzerland) Oct 2022pigments (MPs) are natural edible pigments with high safety and strong function, which have been widely used in food and health products. In this study, different types...
pigments (MPs) are natural edible pigments with high safety and strong function, which have been widely used in food and health products. In this study, different types of tea extracts (rich in polyphenols) were used to regulate the biosynthesis of MPs. The results showed that 15% ethanol extract of pu-erh tea (T11) could significantly increase MPs production in liquid fermentation of M3. Comparative transcriptomic and metabolomic analyses combined with reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to further explore the regulatory mechanism of T11 on the biosynthesis of MPs. Comparative transcriptomic analysis showed that there were 1503 differentially expressed genes (DEGs) between the Con group and the T11 group, which were mainly distributed in carbohydrate metabolism, amino acid metabolism, energy metabolism, lipid metabolism, metabolism of terpenoids and polyketides, etc. A total of 115 differential metabolites (DMs) identified by metabolomics between the Con and T11 groups were mainly enriched in glutathione metabolism, starch and sucrose metabolism, alanine, aspartic acid and glutamate metabolism and glycine, serine and threonine metabolism, etc. The results of metabolomics were basically consistent with those of gene transcriptomics, indicating that the regulatory effect of T11 on the biosynthesis of MPs is mainly achieved through affecting the primary metabolic pathway, providing sufficient energy and more biosynthetic precursors for secondary metabolism. In this study, tea extracts with low economic value and easy access were used as promoters of MPs biosynthesis, which may be conducive to the application of MPs in large-scale industrial production. At the same time, a more systematic understanding of the molecular regulatory mechanism of metabolism was obtained through multi-omics analysis.
PubMed: 37430908
DOI: 10.3390/foods11203159