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1.
J Biotechnol ; 341: 137-145, 2021 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-34601020

RESUMO

Extractive fermentation has been proven to be efficient in enhancing the secretion and production of secondary metabolites in submerged fermentation by Monascus spp., owing to increased cell membrane permeability and resolved product inhibition. In this study, we investigated the regulation effect of Triton X-100 on cell growth and secondary metabolite biosynthesis in submerged fermentation of M. purpureus DK. The results show that the maximum monascus pigments (MPs), citrinin (CIT) production, and specific growth rate are 136.86 U/mL, 4.57 mg/L, and 0.04 h-1, respectively, when 3 g/L of Triton X-100 is supplemented after fermentation for 10 d, and the extracellular MPs and CIT increased by 127.48% and 288.57%, respectively. RT-qPCR shows that the expression levels of MPs and CIT biosynthesis gene clusters are significantly upregulated, whereas those of glycolysis, tricarboxylic acid cycle, respiratory chains, and ATP synthase are downregulated. This study provides a vital strategy for extractive fermentation under extreme environmental conditions for further enhancing MP production.


Assuntos
Citrinina , Monascus , Suplementos Nutricionais , Fermentação , Monascus/metabolismo , Octoxinol , Pigmentos Biológicos/metabolismo
2.
Appl Microbiol Biotechnol ; 105(18): 6691-6706, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34463799

RESUMO

Natural water-soluble Monascus pigments (WSMPs) have been in increasing demand but have not been able to achieve industrial production due to the low production rate. This study aimed to improve the biosynthesis and secretion of extracellular yellow pigments (EYPs) through submerged fermentation with Monascus ruber CGMCC 10,910 supplemented with sodium starch octenyl succinate (OSA-SNa). The results demonstrated that the yield was 69.68% and 48.89% higher than that without OSA-SNa in conventional fermentation (CF) and extractive fermentation (EF), respectively. The mainly increased EYP components were Y3 and Y4 in CF, but they were mainly Y1 and Y2 as well as secreted intracellular pigments, including Y5, Y6, O1, and O2, in EF. Scanning electron microscopy analysis revealed that the mycelium presented an uneven surface profile with obvious wrinkles and small fragments with OSA-SNa. It was found that a higher unsaturated/saturated fatty acids ratio in the cell membrane resulted in increased permeability and facilitated the export of intracellular yellow pigments into the broth with OSA-SNa treatment. In addition, a higher NAD+/NADH ratio and glucose-6-phosphate dehydrogenase activity provided a reducing condition for yellow pigment biosynthesis. Gene expression analysis showed that the expression levels of the key genes for yellow pigment biosynthesis were significantly upregulated by OSA-SNa. This study provides an effective strategy to promote the production of WSMPs by microparticle-enhanced cultivation using OSA-SNa. KEY POINTS: • OSA-SNa addition facilitated the production of Monascus yellow pigments. • Mycelial morphology and membrane permeability were affected by OSA-SNa. • The key gene expression of yellow pigments was upregulated.


Assuntos
Monascus , Fermentação , Monascus/metabolismo , Pigmentos Biológicos/metabolismo , Sódio , Amido , Succinatos , Água
3.
J Photochem Photobiol B ; 217: 112164, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33676287

RESUMO

Light or low frequency magnetic field (LF-MF) as one of the cultivation environments affects secondary metabolites (SMs) production of M. purpureus. Phytochrome (Phy) is a hybrid histidine kinase possessing dual properties of photoreceptor and kinase to sense red and far-red light. The interaction effects of LF-MF and light on SMs of M. purpureus was investigated by knocking out the Phy-like gene in M. purpureus (MpPhy) by homologous recombination. A MpPhy-deletion (ΔMpPhy) strain produced less Monascus pigments (MPs) and monacolin K (mon K) than the wild-type (WT) strain and reduced citrinin production by 78.3% on 10th day but didn't affect the biomass. These results indicated that the MpPhy gene is involved in SMs biosynthesis of M. purpureus. MPs production in WT was decreased significantly when the inoculum was exposed to white/blue/green/red light (500 Lux). But it in ΔMpPhy was no significant difference when exposed to white/red light. The colony size of ΔMpPhy was smaller on potato dextrose agar media containing 0.01% SDS. These results indicated that the deletion of MpPhy gene affected the aerial hyphae and increased sensitivity to cell membrane stress but decreased sensitivity to red light. The inoculum of both WT and ΔMpPhy was exposure to the LF-MF (50 Hz). The accumulation of WT secondary metabolites was not changed, while SMs production of ΔMpPhy was significantly enhanced under exposed to 2.0 mT LF-MF. This indicated that the decrease of SMs caused by the deletion of MpPhy gene was restored by LF-MF. It revealed that there is a crosstalk between magnetoreception and photosensitivity.


Assuntos
Luz , Monascus/metabolismo , Fitocromo/genética , Metabolismo Secundário/efeitos da radiação , Biomassa , Citrinina/biossíntese , Meios de Cultura/química , Lovastatina/biossíntese , Monascus/citologia , Monascus/crescimento & desenvolvimento , Mutagênese , Fitocromo/metabolismo , Pigmentos Biológicos/metabolismo
4.
Braz J Microbiol ; 52(2): 587-596, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33651332

RESUMO

Three culture media were studied for red pigment production by Monascus ruber in submerged cultivation: rice flour (20 g L-1), sugarcane molasses (30 g L-1), and, finally, molasses + rice flour (10 g L-1+10 g L-1); all culture media were added of 5 g L-1 glycine as nitrogen source. Rice flour showed pigment production of 7.05 UA510nm and molasses 5.08 UA510nm, and the mixture of rice flour and molasses showed the best result of 16.38 UA510nm. Molasses culture presented good results for cell biomass production of 11.09 g L-1. With these results, it was observed that one substrate presented good pigment production (rice flour) and another attained better results for cell biomass growth (molasses), and a third medium containing 10 g L-1 of rice flour + 10 g L-1 of molasses was formulated. The results for this mixture showed satisfactory results, with global pigment productivity of 0.097 UA510nm h-1 and maximum productivity rate of 0.17 UA510nm h-1. The high production and productivity obtained for the mixture of rice flour and molasses indicated that the production of red pigment by submerged fermentation, using the mixture of these low-cost culture media, may be promising in terms of commercial production.


Assuntos
Farinha/microbiologia , Melaço/microbiologia , Monascus/metabolismo , Oryza/microbiologia , Pigmentos Biológicos/biossíntese , Saccharum/microbiologia , Biotransformação , Fermentação , Farinha/análise , Melaço/análise , Monascus/crescimento & desenvolvimento , Oryza/metabolismo , Saccharum/química , Resíduos/análise
5.
J Sci Food Agric ; 101(13): 5652-5659, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33740266

RESUMO

BACKGROUND: Monacolin K, an important secondary metabolite of Monascus, possesses a cholesterol-lowering effect and is widely used in the manufacture of antihypertensive drugs. In the present study, we constructed an extractive fermentation system by adding non-ionic surfactant and acquired a high monacolin K yield. The mechanism was determined by examining both cell morphology and the transcription levels of the related mokA-I genes in the monacolin K biosynthetic gene cluster. RESULTS: The monacolin K yield was effectively increased to 539.59 mg L-1 during extraction, which was an increase of 386.16% compared to that in the control group fermentation. The non-ionic surfactant showed good biocompatibility with Monascus. Electron scanning microscopy revealed alterations in the morphology of Monascus. The loosened mycelial structure and increased number of cell surface wrinkles were found to be related to the increased cell-membrane permeability and extracellular accumulation of monacolin K. Gene expression levels were measured via a quantitative reverse transciptase-polymerase chain reaction. By contrast, in the control group, mokA, mokB, mokC, mokD and mokF showed higher-level and longer-term expression in the extractive fermentation group, whereas mokE and mokG did not present a similar trend. The expression levels of mokH and mokI, encoding a transcription factor and efflux pump, respectively, were also higher than the control levels. CONCLUSION: The addition of a non-ionic surfactant to Monascus fermentation effectively increases the yield of monacolin K by transforming the fungus morphology and promoting the expression of monacolin K biosynthesis genes. © 2021 Society of Chemical Industry.


Assuntos
Proteínas Fúngicas/genética , Lovastatina/biossíntese , Monascus/crescimento & desenvolvimento , Monascus/metabolismo , Vias Biossintéticas , Fermentação , Proteínas Fúngicas/metabolismo , Monascus/genética , Família Multigênica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
6.
J Food Sci ; 86(3): 969-976, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33527354

RESUMO

Additional moisture in preparing red koji, Monascus-fermented rice, is a characteristic production process. To determine how additional moisture affects red koji preparation as per quality, we compared the growth of Monascus purpureus, enzyme and pigment production, and related gene expressions using our findings. We considered two kinds of red koji: one prepared with additional moisture at the middle part of the preparation and the other prepared without additional moisture. Our results showed that additional moisture did not promote the growth of M. purpureus, but it was significantly increased the pigment (red and yellow) and tended to increase the α-amylase level and saccharification power. Although adding a high amount of moisture (approximately 60% moisture content) promoted pigment production, it slightly repressed enzyme production. In contrast, adding approximately 50% moisture content promoted enzyme production. These findings showed that the additional moisture can affect the quality of red koji on the purpose. The expression of 10 pigment biosynthetic gene clusters and two glycohydrolase genes in red koji after adding moisture was analyzed through real-time qPCR. Eight genes were upregulated within 1 hr after adding water, with mppR2 being the first upregulated gene within 30 min. The expression of genes as per pigment production quickly responded to additional moisture during solid-state fermentation. Moreover, acetyl-CoA, which is a starting substrate for pigment content in red koji was increased within 3 hr after adding water. This study first described the relationship between additional moisture and expression of pigment biosynthetic genes by Monascus spp. during red koji preparation.


Assuntos
Fermentação , Manipulação de Alimentos/métodos , Monascus/metabolismo , Oryza/microbiologia , Pigmentos Biológicos/genética , Água , Expressão Gênica , Monascus/enzimologia , Monascus/genética , Pigmentação , Pigmentos Biológicos/biossíntese , alfa-Amilases/metabolismo
7.
Appl Microbiol Biotechnol ; 105(5): 1905-1912, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33576885

RESUMO

Monascus pigments (MPs) are widely used natural colorants in Asian countries. The problems of low extracellular red pigment (ERP) and high citrinin remain to be solved in Monascus pigment production. The effect of lanthanum(III) ion (LaCl3) on Monascus purpureus fermentation was investigated in this study. The yields of ERP and biomass respectively reached maxima of 124.10 U/mL and 33.10 g/L by adding 0.4 g/L La3+ on the second day in the total 8-day fermentation; simultaneously, citrinin was decreased by 59.93% and 38.14% in the extracellular and intracellular fractions, respectively. Reactive oxygen species (ROS) levels were obviously improved by La3+ treatment, while the activities of catalase (CAT) and superoxide dismutase (SOD) were increased compared with the control. The ratio of unsaturated/saturated fatty acids in mycelia was increased from 2.94 to 3.49, indicating that the permeability and fluidity of the cell membrane were enhanced under La3+ treatment. Gene expression analysis showed that the relative expression levels of Monascus pigment synthesis genes (pksPT, mppB, mppD, MpFasB2, and MpPKS5) were significantly upregulated by La3+ treatment, and in contrast, the relative expression levels of citrinin synthesis genes (ctnA, pksCT and mppC) were markedly downregulated. This work confirmed that LaCl3 possesses the potential to induce red pigment biosynthesis and inhibit citrinin production in M. purpureus fermentation. KEY POINTS: • La3+ induced red pigment and inhibited citrinin production in Monascus fermentation. • La3+ regulated genes expression up for Monascus pigment and down for citrinin. • La3+ increased the UFAs in cell membrane to enhance the permeability and fluidity.


Assuntos
Citrinina , Monascus , Ásia , Fermentação , Lantânio , Monascus/metabolismo , Pigmentos Biológicos/metabolismo
8.
Food Chem ; 343: 128410, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33406573

RESUMO

Monascus, which is traditionally used in various Asian industries, produces several secondary metabolites during the fermentation process, including citrinin, a toxin whose impact limits the development of the Monascus industry. We have previously found that the addition of 2.0 g/L genistein to Monascus medium reduces citrinin production by approximately 80%. Here, we explored the molecular mechanisms whereby genistein affects citrinin production. We sequenced the Monascus genome and performed transcriptome analysis on genistein-treated and -untreated groups. Comparison between the two groups showed 378 downregulated and 564 upregulated genes. Among the latter, we further examined the genes related to citrinin biosynthesis and quantified them using quantitative real-time polymerase chain reaction (qRT-PCR). Genes orf5, pksCT, orf3, orf1, orf6, and ctnE were significantly downregulated, demonstrating that genistein addition indeed affects citrinin synthesis. Our results may lay the groundwork for substantial improvements in the Monascus fermentation industry.


Assuntos
Citrinina/biossíntese , Genisteína/farmacologia , Monascus/química , Transcriptoma/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Ontologia Genética , Genes Fúngicos , Monascus/genética , Monascus/metabolismo , Família Multigênica , Reação em Cadeia da Polimerase em Tempo Real , Regulação para Cima/efeitos dos fármacos
9.
Food Microbiol ; 95: 103689, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33397619

RESUMO

This study investigated the effects of cofactor metabolism on secondary metabolite production in M. purpureus through the application of different cofactor engineering strategies. Total pigment production dramatically increased by 39.08% and 40.89%, and yellow pigment production increased by 74.62% and 114.06% after the addition of 1.0 mg/L of the exogenous cofactor reagents methyl viologen and rotenone, respectively, in submerged batch-fermentation. The extracellular red pigment tone changed to yellow with the application of electrolytic stimulation at 800 mV/cm2, but almost no citrinin production was detected. In addition, the total pigment, yellow pigment and citrinin production increased by 35.46%, 54.89% and 6.27% after disruption of the nuoⅠ gene that encodes NADH-quinone oxidoreductase, respectively. Thus, cofactor metabolic engineering strategies could be extended to the industrial production of Monascus pigment or high yellow pigment with free citrinin production.


Assuntos
Meios de Cultura/metabolismo , Monascus/genética , Monascus/metabolismo , Paraquat/metabolismo , Pigmentos Biológicos/biossíntese , Rotenona/metabolismo , Citrinina/biossíntese , Engenharia Metabólica , Metabolismo Secundário
10.
Chem Biol Drug Des ; 97(1): 77-86, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32666679

RESUMO

A high-throughput screening assay was developed and applied to a large library of natural product extract samples, in order to identify compounds which preferentially inhibited the in vitro 2D growth of a highly metastatic osteosarcoma cell line (MG63.3) compared to a cognate parental cell line (MG63) with low metastatic potential. Evaluation of differentially active natural product extracts with bioassay-guided fractionation led to the identification of lovastatin (IC50  = 11 µm) and the limonoid toosendanin (IC50  = 26 nm). Other statins and limonoids were then tested, and cerivastatin was identified as a particularly potent (IC50  < 0.1 µm) and selective agent. These compounds potently and selectively induced apoptosis in MG63.3 cells, but not MG63. Assays with other cell pairs were used to examine the generality of these results. Statins and limonoids may represent unexplored opportunities for development of modulators of osteosarcoma metastasis. As cerivastatin was previously approved for clinical use, it could be considered for repurposing in osteosarcoma, pending validation in further models.


Assuntos
Produtos Biológicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Ensaios de Triagem em Larga Escala/métodos , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Neoplasias Ósseas/metabolismo , Neoplasias Ósseas/patologia , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Humanos , Lovastatina/química , Lovastatina/isolamento & purificação , Lovastatina/farmacologia , Melia/química , Melia/metabolismo , Monascus/química , Monascus/metabolismo , Osteossarcoma/metabolismo , Osteossarcoma/patologia , Extratos Vegetais/química , Piridinas/química , Piridinas/isolamento & purificação , Piridinas/farmacologia , Sementes/química , Sementes/metabolismo
11.
J Sci Food Agric ; 101(6): 2561-2569, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33063356

RESUMO

BACKGROUND: Hyperlipidemia is one of the metabolic disorders that poses a great threat to human health. This study is aimed at investigating the potential hypolipidemic properties of extract from peanut meal fermented with Bacillus natto and Monascus in mice fed with a high-fat diet. Herein, 60 male C57BL/6J mice were randomly divided into six groups: four control groups, comprised of a normal group, a model (M) group, a positive control group (atorvastatin 10 mg kg-1 ), and a nonfermented peanut meal extract group (150 mg kg-1 ), and two experimental groups, comprised of a fermented peanut meal extract low-dose group (50 mg kg-1 ) and a fermented peanut meal extract high-dose group (FH, 150 mg kg-1 ). RESULTS: Body weight (P = 0.001) and levels of serum total cholesterol (P = 0.007), triacylglycerol (P = 0.040), low-density lipoprotein cholesterol (P < 0.001), and leptin (P < 0.001) were remarkably decreased in the FH group, whereas the serum high-density lipoprotein cholesterol levels were increased (P < 0.001) by 78.3% compared with the M group. Ileum tissue stained with hematoxylin and eosin showed that the ileal villus detachments in mice were improved, and the villus height was increased by supplementation with extract from fermented peanut meal. Moreover, the expressions of intestinal ZO-1 (P = 0.003) and occludin (P = 0.013) were elevated in the FH group, compared with the M group. CONCLUSION: Extract of peanut meal fermented by B. natto and Monascus can effectively improve hyperlipidemia caused by a high-fat diet in mice, via regulating leptin and blood lipid levels, and protect the intestinal mucosal barrier, which provides evidence for its anti-hyperlipidemia effects and is a research basis for potential industrial development. © 2020 Society of Chemical Industry.


Assuntos
Arachis/metabolismo , Bacillus/metabolismo , Hiperlipidemias/dietoterapia , Hipolipemiantes/metabolismo , Metabolismo dos Lipídeos , Monascus/metabolismo , Extratos Vegetais/metabolismo , Animais , Arachis/microbiologia , HDL-Colesterol , LDL-Colesterol , Dieta Hiperlipídica/efeitos adversos , Fermentação , Humanos , Hiperlipidemias/etiologia , Hiperlipidemias/metabolismo , Mucosa Intestinal/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Alimentos de Soja/análise , Alimentos de Soja/microbiologia , Triglicerídeos/metabolismo
12.
Food Chem ; 338: 128025, 2021 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-32927200

RESUMO

Short-chain fatty acid esters are important flavor chemicals in Chinese traditional fermented Baijiu. Monascus purpureus was recognized as an important microorganism contributing to ester synthesis. However, the molecular basis for ester synthesis was still lacking. The present work combined genome sequencing, transcriptome sequencing, gene library construction, and enzyme engineering to discover a novel catalyst from M. purpureus (isolated from Baijiu fermentation starter). Enzyme LIP05, belonging to the α/ß hydrolase family, was identified to synthesize short-chain fatty acid esters under aqueous phase. After deleting the lid domain of LIP05, the synthesis of ethyl pentanoate, ethyl hexanoate, ethyl octanoate, or ethyl decanoate was achieved. Ethyl octanoate with the highest conversion ratio of 93.7% was obtained with the assistance of ultrasound. The study reveals the molecular basis for synthesizing short-chain fatty acid esters by M. purpureus and will promote the application of the species or the enzyme in food industry.


Assuntos
Hidrolases/metabolismo , Monascus/genética , Água/química , Vinho/análise , Biocatálise , Caproatos/química , Caproatos/metabolismo , Ésteres/química , Ésteres/metabolismo , Evolução Molecular , Concentração de Íons de Hidrogênio , Hidrolases/genética , Monascus/metabolismo , Sonicação , Temperatura
13.
J Sci Food Agric ; 101(1): 185-193, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-32623720

RESUMO

BACKGROUND: The rapid development of the rice wine industry has increased the demand for raw materials worldwide. A fungal strain with good adaptability to rice wine brewing conditions, which can also enhance the utilization rate of raw materials (URRM), thus increasing the production efficiency, was sought in the present research. RESULTS: The strain FJMR24 was successfully isolated and screened from 35 fermentation starters and exhibited high amylase activity (2200.9 ± 18.5 U g-1 ) and high glucoamylase activity (2330.4 ± 31.9 U g-1 ). Based on a morphological examination and a sequence analysis of the internal transcribed spacer (ITS) gene and ß-tubulin gene, FJMR24 was identified as Monascus purpureus, which is an edible and versatile fungus that plays a dominant role in the processing of Hong Qu. A moderate pH of 5-6 under incubation at 35 °C for 5-6 days was favorable for the growth and enzyme production of FJMR24. The strain could also tolerate the extreme conditions of 15-45 °C, 18% ethanol (v/v), and an acidity of pH 2. The excellent fermentation adaptability of FJMR24 might enable it to retain high enzyme activity during rice wine brewing. As a result of the action of FJMR24, the URRM of the base liquor increased by around 26% due to increased starch hydrolysis efficiency, which was mainly due to the high unit enzyme activity of FJMR24. CONCLUSION: This study provides perspectives for the application of a M. purpureus strain with high starch hydrolysis activity for enhancing the URRM in traditional rice wine brewing. © 2020 Society of Chemical Industry.


Assuntos
Monascus/isolamento & purificação , Monascus/metabolismo , Oryza/microbiologia , Vinho/análise , Amilases/genética , Amilases/metabolismo , Fermentação , Microbiologia de Alimentos , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Glucana 1,4-alfa-Glucosidase/genética , Glucana 1,4-alfa-Glucosidase/metabolismo , Monascus/enzimologia , Monascus/genética , Oryza/metabolismo , Amido/metabolismo , Vinho/microbiologia
14.
Prep Biochem Biotechnol ; 51(2): 120-126, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33342390

RESUMO

Ginseng (Panax ginseng C. A. Meyer) is a famous Traditional Chinese Medicine, which is widely used to treat cardiovascular disease. Monascus ruber (M. ruber) is a fungus used in food and medicine fermentation, and lovastatin, its metabolite, is used extensively in the treatment of dyslipidemia. In this study, ginseng has been fermented by M. ruber, and the response surface methodology (RSM) was applied to optimize fermentation parameters to obtain optimal fermentation system, with further exploring to lipid-lowering activity of P. ginseng C. A. Meyer-M. ruber fermentation products (PM). The concentration of ginseng, temperature, and rotating speed were set as variables and the lovastatin yield was optimized by a Box-Behnken design (BBD) analyzed by RSM. The binding capacity of PM for sodium taurocholate and sodium cholate was assayed by UV spectrophotometry. The highest content of lovastatin production (85.53 µg g-1) was obtained at a ginseng concentration of 1.96%, temperature of 30.11 °C, and a rotating speed of 160.47 rpm. PM exhibited bile acid binding capacity, which was stronger than unfermented ginseng. The RSM can be used to optimize the fermentation system to obtain the best fermentation process. In addition, the fermentation of ginseng by M. ruber can enhance the lipid-lowering effect.


Assuntos
Ácidos e Sais Biliares/química , Fermentação , Lovastatina/química , Monascus/metabolismo , Reatores Biológicos , Biotecnologia/métodos , Química Farmacêutica/métodos , Técnicas In Vitro , Lipídeos/química , Medicina Tradicional Chinesa , Oryza , Panax , Ligação Proteica , Colato de Sódio/química , Espectrofotometria Ultravioleta , Ácido Taurocólico/química , Temperatura
15.
J Sci Food Agric ; 101(9): 3836-3842, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33336368

RESUMO

BACKGROUND: Monascus fermentation byproduct (MFB) is a biowaste generated after food colorants are extracted. Using MFB to produce probiotics (Bacillus subtilis) is a sustainable way for the entire production to be used as food or animal feed additives. However, due to the rigidity of the Monascus mycelium cell wall, B. subtilis cannot sufficiently utilize the nutrients in MFB, leading to low biomass production efficiency. We studied the effects of ultrasonic treatment, papain, ß-glucanase, and chitosanase, and their combinations on improving the levels of soluble components from MFB. The effects of these treatments on mycelium cell walls were visualized using scanning electron microscopy, and their influence on B. subtilis production was analyzed. RESULTS: Ultrasonic treatment increased the soluble components by 210 g kg-1 , including 50 g kg-1 protein and 120 g kg-1 carbohydrates. An enzyme mixture increased the soluble components by 160 g kg-1 , including 30 g kg-1 protein and 90 g kg-1 carbohydrates. The combination of the two methods achieved the highest increase of soluble components (up to 400 g kg-1 ) leading to a maximum B. subtilis production of 1 × 1011 colony-forming unit mL-1 . This yield was about 20 times greater than that using untreated MFB and about eight times greater than treatments using only ultrasonic or enzymatic methods. CONCLUSION: The productivity of B. subtilis production using MFB as the sole medium can be greatly improved by ultrasound or enzymes, which cause the release of intercellular components or cell wall components. © 2020 Society of Chemical Industry.


Assuntos
Bacillus subtilis/crescimento & desenvolvimento , Monascus/química , Resíduos/análise , Bacillus subtilis/metabolismo , Biocatálise , Parede Celular/química , Meios de Cultura/análise , Meios de Cultura/metabolismo , Fermentação , Glicosídeo Hidrolases/química , Monascus/metabolismo , Micélio/química , Micélio/metabolismo , Papaína/química , Probióticos/análise , Probióticos/metabolismo , Ultrassom
16.
Microb Cell Fact ; 19(1): 224, 2020 Dec 07.
Artigo em Inglês | MEDLINE | ID: mdl-33287814

RESUMO

BACKGROUND: Monascus azaphilone pigments (MonAzPs), which were produced by Monascus species, have been used as important food colorant and food supplements for more than one billion people during their daily life. Moreover, MonAzPs recently have received more attention because of their diverse physiological activities. However, the high microbial production of MonAzPs is still not always guaranteed. Herein, the aim of this study was to develop an efficient biotechnological process for MonAzPs production. RESULTS: In this study, exogenous cyclic adenosine monophosphate (cAMP) treatment not only induced MonAzPs production, but also stimulated the expression of a cAMP phosphodiesterase gene, named as mrPDE, in M. purpureus HJ11. Subsequently, MrPDE was identified as a cAMP phosphodiesterase by in vitro enzymatic reaction with purified enzyme. Further, a gene knockout mutant of mrPDE was constructed to verify the activation of cAMP signalling pathway. Deletion of mrPDE in M. purpureus HJ11 improved cAMP concentration by 378% and enhanced PKA kinase activity 1.5-fold, indicating that activation of cAMP signalling pathway was achieved. The ΔmrPDE strain produced MonAzPs at 8563 U/g, with a 2.3-fold increase compared with the WT strain. Moreover, the NAPDH/NADP+ ratio of the ΔmrPDE strain was obviously higher than that of the wild type strain, which led to a higher proportion of yellow MonAzPs. With fed-batch fermentation of the ΔmrPDE strain, the production and yield of MonAzPs achieved 332.1 U/mL and 8739 U/g. CONCLUSIONS: A engineered M. purpureus strain for high MonAzPs production was successfully developed by activating the cAMP signalling pathway. This study not only describes a novel strategy for development of MonAzPs-producing strain, but also provides a roadmap for engineering efforts towards the production of secondary metabolism in other filamentous fungi.


Assuntos
AMP Cíclico/metabolismo , Engenharia Metabólica , Monascus/metabolismo , Pigmentos Biológicos/biossíntese , 3',5'-AMP Cíclico Fosfodiesterases/genética , 3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Benzopiranos , Fermentação , Técnicas de Inativação de Genes , Genes Fúngicos , Monascus/genética , NADP/metabolismo , Metabolismo Secundário , Transdução de Sinais
17.
Acta Sci Pol Technol Aliment ; 19(4): 445-456, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33179484

RESUMO

BACKGROUND: The addition of fatty acids and other molecules to culture media may intensify the production of biomolecules, such as monascus pigments, however, few studies of this have been developed. Thus, the objective of the present study was to investigate the effects of adding sodium octanoate to the culture medium, with a view to increasing the synthesis and production of the pigments produced by Monascus ruber CCT 3802 on solid and submerged cultivations. METHODS: Monacus ruber CCT 3802 was cultivated on solid and submerged media supplemented with different concentrations of sodium octanoate. The radial growth rate of the colonies was obtained from the declivity of the linear regression of the radius of the colonies as a function of cultivation time and the kinetics of submerged cultivations were performed. The filtrate obtained was submitted to scanning spectrophotometry at a range from 350 to 550 nm and the color parameters were determined by using the CIELAB color system. The data were submitted to a univariate analysis of variance (ANOVA) and the means obtained for each treatment submitted to Tukey's test using Statistica version 5.0 software at a 5% level of significance. RESULTS: Sodium octanoate exerted a strong influence on growth and pigment production in solid and submerged cultivations. The values for L*, a* and b* were positive for pigments produced, with regards to colors close to red and yellow. In the media supplemented with 1.0 mM and 1.5 mM of sodium octanoate, the production of red pigments became expressive from 48 hours-cultivation, increasing considerably from the second to the fourth days. This shows that supplementation with sodium octanoate provides a greater production of pigments in a shorter time interval than the control culture, which required 144 hours of cultivation to present a higher value for AU510nm, which directly influenced pigment productivity. CONCLUSIONS: The addition of sodium octanoate exerted a significant influence on both microbial growth and pigment production in both solid and submerged cultivations. The supplementation of the submerged cultures with sodium octanoate was responsible for an expressive production of pigments in just 48 hours, whereas 144 hours were necessary in the absence of sodium octanoate. These results are promising for increasing the productivity of pigment production, including possibilities for application on an industrial scale.


Assuntos
Caprilatos , Cor , Meios de Cultura/química , Monascus/efeitos dos fármacos , Pigmentação , Pigmentos Biológicos/biossíntese , Cinética , Monascus/crescimento & desenvolvimento , Monascus/metabolismo
18.
Appl Microbiol Biotechnol ; 104(24): 10451-10463, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33165660

RESUMO

Monascus purpureus, a pigment-producing ascomycetous fungus, has been traditionally used for red rice preparation using solid-state fermentation. The objective of this study was to develop an improved pigment-producing strain of M. purpureus MTCC 1090 through genome shuffling followed by detailed analytical estimations of pigments and other bioactive compounds produced by the fusant. Protoplast formation was optimum with 12 h-old mycelia incubated at 30 °C with cellulase, lyticase, and chitinase (40:1:1) for 5 h. Four UV-induced mutants that produced 13.1-39.5% higher amount of yellow, orange, and red pigments in fermented low-grade (cheap) broken rice were used as parents for genome shuffling. After the first round of fusion, four fusants with 35.9-60.52% higher pigment production capabilities were fused again, and finally the fusant F2-19 with distinct culture characteristic was selected under multi-selection pressure. It consistently produced 67%, 70%, and 76% higher content of yellow, orange, and red pigments respectively as compared to the wild-type. High-performance liquid chromatography (HPLC) analysis also reveals clear variation in pigment productions between wild-type and the fusant. Furthermore, HPLC analysis of F2-19 fermented rice extract confirms the production of 186 ± 8.71 and 3810 ± 29.81 mg/kg mevinolin and gamma-aminobutyric acid respectively. Citrinin was not detected. F2-19 fermented rice also has high antioxidant activity (7.92 ± 0.32 mg/g trolox equivalent), with good amount of phenolics (18.0 ± 0.95 mg/g gallic acid equivalent) and flavonoids (2.7 ± 0.26 mg/g quercetin equivalent). Thus, genome shuffling was successfully implemented on M. purpureus for the first time to develop a citrinin-free, better-performing fusant that holds future biotechnological potential. KEY POINTS: • Genome shuffling was performed by recursive protoplast fusion in Monascus purpureus. • The selected fusant, F2-19, was used in solid-state fermentation using low-grade rice. • It produced 67-76% higher content of yellow, orange, and red pigments than the wild-type. • HPLC detected 186 mg/kg mevinolin and 3810 mg/kg γ-aminobutyric acid, but no citrinin. • F2-19 shows high antioxidant activity with good amount of phenolics and flavonoids. Graphical abstract.


Assuntos
Citrinina , Monascus , Embaralhamento de DNA , Fermentação , Lovastatina , Monascus/genética , Monascus/metabolismo , Pigmentos Biológicos/metabolismo
19.
BMC Genomics ; 21(1): 679, 2020 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-32998685

RESUMO

BACKGROUND: Species of the genus Monascus are considered to be economically important and have been widely used in the production of yellow and red food colorants. In particular, three Monascus species, namely, M. pilosus, M. purpureus, and M. ruber, are used for food fermentation in the cuisine of East Asian countries such as China, Japan, and Korea. These species have also been utilized in the production of various kinds of natural pigments. However, there is a paucity of information on the genomes and secondary metabolites of these strains. Here, we report the genomic analysis and secondary metabolites produced by M. pilosus NBRC4520, M. purpureus NBRC4478 and M. ruber NBRC4483, which are NBRC standard strains. We believe that this report will lead to a better understanding of red yeast rice food. RESULTS: We examined the diversity of secondary metabolite production in three Monascus species (M. pilosus, M. purpureus, and M. ruber) at both the metabolome level by LCMS analysis and at the genome level. Specifically, M. pilosus NBRC4520, M. purpureus NBRC4478 and M. ruber NBRC4483 strains were used in this study. Illumina MiSeq 300 bp paired-end sequencing generated 17 million high-quality short reads in each species, corresponding to 200 times the genome size. We measured the pigments and their related metabolites using LCMS analysis. The colors in the liquid media corresponding to the pigments and their related metabolites produced by the three species were very different from each other. The gene clusters for secondary metabolite biosynthesis of the three Monascus species also diverged, confirming that M. pilosus and M. purpureus are chemotaxonomically different. M. ruber has similar biosynthetic and secondary metabolite gene clusters to M. pilosus. The comparison of secondary metabolites produced also revealed divergence in the three species. CONCLUSIONS: Our findings are important for improving the utilization of Monascus species in the food industry and industrial field. However, in view of food safety, we need to determine if the toxins produced by some Monascus strains exist in the genome or in the metabolome.


Assuntos
Genes de Plantas , Especiação Genética , Monascus/genética , Pigmentos Biológicos/genética , Metabolismo Secundário , Monascus/classificação , Monascus/metabolismo , Família Multigênica , Filogenia , Pigmentos Biológicos/biossíntese
20.
J Agric Food Chem ; 68(42): 11839-11845, 2020 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-33035423

RESUMO

Food authenticity in the field of food dyes can be interpreted as the correctness of the coloring ingredients indicated. The Rapid UV/vis Spectroscopic Dye Authentication Assay (RaSDAY) presented in this work was used to verify the authenticity of water-soluble reddish colorings for food use. RaSDAY includes the processing of samples under different experimental conditions with pH variations and heat exposure. The absorbances measured are analyzed by principal component analysis and a k-nearest neighbors algorithm. As a result, classification of anthocyanins, betalains, and carmine and the detection of Monascus pigments, undeclared artificial food dyes, and reactive textile azo dyes can be performed by utilizing a rapid screening method. In 17 out of 20 samples of coloring food additives that were included in this work, reactive dyes, unpermitted Monascus pigments, and artificial food dyes were detected using the developed method. "Reactive Red 120", "Reactive Red 195", and "Reactive Red 198" were identified by subsequent 1H NMR spectroscopy in eight of those samples.


Assuntos
Compostos Azo/química , Corantes de Alimentos/química , Contaminação de Alimentos/análise , Monascus/metabolismo , Naftalenossulfonatos/química , Pigmentos Biológicos/química , Espectrofotometria Ultravioleta/métodos , Triazinas/química , Monascus/química , Pigmentos Biológicos/metabolismo
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