Metagenomic analysis revealing the metabolic role of microbial communities in the free amino acid biosynthesis of Monascus rice vinegar during fermentation

Free amino acid(FAA)is the important component of vinegar that infl uences quality perception and consumer acceptance.FAA is one of the major metabolites produced by microorganisms;however,the microbial metabolic network on FAA biosynthesis remains unclear.Through metagenomic analysis,this work aimed to elucidate the roles of microbes in FAA biosynthesis during Monascus rice vinegar fermentation.Taxonomic profiles from functional analyses showed 14 dominant genera with high contributions to the metabolism pathways.The metabolic network for FAA biosynthesis was then constructed,and the microbial distribution in different metabolic pathways was illuminated.The results revealed that 5 functional genera were closely involved in FAA biosynthesis.This study illuminated the metabolic roles of microorganisms in FAA biosynthesis and provided crucial insights into the functional attributes of microbiota in vinegar fermentation.

N^+离子注入选育色素产生菌Monascus的研究

应用 1 0keV的N+离子注入,对产红色素红曲菌进行了诱变选育。在确定最佳注入剂量后,经过两轮离子注入诱变筛选,选育出 1株高产突变株,红色素色价提高 3 8.1 %,且色调适宜。经传代培养,其高产性状能稳定遗传。对该菌株的发酵特性也进行了研究。

氨基酸对于Monascus ruber生物合成Monacolin K影响的研究

在Monacolin K发酵中添加氨基酸后发现较高质量浓度的氨基酸高度抑制了Monacolin K的产量。0.1 g.L-1的D-甲硫氨酸在发酵第4 d添加可以提高产量30%以上,而L-甲硫氨酸则没有此功能,D,L-甲硫氨酸因D-甲硫氨酸的关系也有一定的增产效果。以甲硫氨酸代替蛋白胨作为主要氮源,则抑制了polyketide途径的前期步骤,因此严重抑制了色素及Monacolin K的生产。另外,发现1 g.L-1的L-苯丙氨酸的添加时间越早越有利于Monacolin K的生产,在起始时添加发酵单位可达135.9 mg.L-1,可能是因为L-苯丙氨酸经过脱氨后,可以进入polyketide途径从而促进了Monacolin K的生产。

产红色素真菌Monascus sanguineus的分离及其色素提取条件研究

采用组织块分离法首次从药用植物地黄中分离出一株产红色素内生真菌RJL03,根据形态学特征和r DNA ITS1-5.8S-ITS2序列分析的手段鉴定为Monascus sanguineus。采用响应面法(RSM)对菌株RJL03中红色素提取工艺条件进行研究。在单因素实验的基础上,以提取液OD值为指标,利用Box-Behnken中心组合实验设计原理(BBD)优化菌株中红色素的有机溶剂超声波辅助浸提(UAE)工艺。结果显示,该色素提取液在515 nm处有最大吸收波长,最佳提取工艺为温度70℃、提取时间34 min、乙醇浓度50%。在此条件下提取菌株中的红色素,得到的提取液OD值为2.587,可比优化前提高约1.5倍。

产红色素真菌Monascus sanguineus的液态发酵条件研究

从药用植物地黄中分离出一株产红色素内生真菌RJL03(Monascus sanguineus),为进一步提高菌株RJL03的红色素产量,采用响应面法(RSM)对其液态摇瓶发酵条件进行研究。在单因素实验的基础上,以发酵液OD值为指标,利用Box-Behnken中心组合实验设计原理(BBD)优化菌株液态发酵产红色素的工艺条件。结果显示,该色素发酵液在474 nm处有最大吸收波长,最佳发酵工艺为温度30℃、装瓶量67%、初始p H自然(7)。在此条件下液态发酵菌株,显著提高了菌株RJL03的红色素产量。

Study on the Monascus Pigment of Monascus Strains JF-02 Fermentation

[ Objective] The aim was to study the fermentation technology of monascus pigment of monascus strains JF-02. [ Method] Single factor experiment was carried out to study the influence of temperature, initial pH, culture time, different agricultural byproduct, and nitrogen source on monascus pigment in fermentation solution. Meanwhile, orthogonal experiment was conducted to get the optimal culture medium and cultivation condition. [ Resultl The optimal gene in the pigment of monascus pigment was 200 g/L of rice, 30 g/L of sweet potato powder, 10 g/L of glucose, 15 g/L of monosodium glutamate, 0.1% of zinc sulfate, and 0.1% of magnesium sulfate. The optimal culture condition was 30 ℃ and initial pH was 6.0. Fermentation time was 72 h, but when 24-L fermentation pot was used, culture time can last to 84 h. [ Condusion] The study provided theoretical basis for the development and application of monascus strains.

Monascus pilosus-fermented black soybean inhibits lipid accumulation in adipocytes and in high-fat diet-induced obese mice

Objective:To explore the anti-obesity effects and the mechanism of action of Monascus pilosus(M.pilosus)-fermented black soybean(MFBS)extracts(MFBSE)and MFBS powders(MFBSP)in adipocytes and high-fat diet(HFD)-induced obese mice,respectively.Methods:Black soybean was fermented with M.pilosus,and the main constituents in MFBS were analyzed by HPLC analysis.In vitro,MFBSE were examined for anti-adipogenic effects using Oil-Red O staining.In vivo,mice were fed a normal-fat diet(NFD)control,HFD control or HFD containing 1 g/kg MFBSP for 12 weeks,and then body weight gain and tissues weight measured.Real-time PCR and western blot assay were used to determine the mechanism of anti-adipogenic effects.Results:MFBSE inhibited lipid accumulation in 3T3-L1 adipocytes without exerting cell cytotoxicity.MFBSP treatment in HFD-fed mice significantly decreased the body weight gain compared with the HFD control mice.MFBSE and MFBSP treatment resulted in significantly lower mRNA levels of adipogenesis-related genes,such as peroxisome proliferator-activated receptorγ(PPARγ),fatty acid-binding protein 4(FABP4),and fatty acid synthase(FAS),in adipocytes and in white adipose tissue(WAT)of HFD-induced obese mice.Conclusions:These results suggest that the anti-obesity effects of MFBS are elicited by regulating the expression of adipogenesis-related genes in adipocytes and WAT of HFDinduced obese mice.

Cloning and Sequence Analysis of the Full-length cDNA of a Novel yp05 Gene Associated With Citrinin Production in Monascus aurantiacus

Objective To obtain the full-length cDNA of a novel gene （named yp05） associated with citrinin production-related genes in Monascus aurantiacus. Methods Total RNA was extracted from mycelium, 3＇ and 5＇ cDNA end of yp05 gene was amplified using smartTM trace cDNA amplification kit, and the full-length cDNA of a novel gene （named yp05） was obtained from the electronic assembly of 3＇-RACE and 5＇- RACE products. Results This yp05 gene was 787 bp including a 597 bp open reading frame （ORF） and encoded a deduced protein with 199 amino acid residues, and the amino acid sequence of this protein was found similar with the sequences of many fungal manganese-superoxide dismutases in the GenBank with the aid of BLASTp. The transcription ofyp05 gene in Monascus strains was analyzed with the aid of Northern blotting. The transcription of yp05 gene was only detected in Monascus strains, provided that citrinin was produced. Conclusion The transcription of yp05 gene belongs to differential expression genes of citfinin yielded from Monascus and has no correlation with the biosynthesis pathway of red pigments.

Repeated-batch Cultivation of Encapsulated Monascus purpureus by Polyelectrolyte Complex for Natural Pigment Production

In general,productions of natural pigment in submerged microorganism culture were much less than that in solid-state fermentation,because the solid-state culture can provide a support carrier for the mycelium. To improve natural pigment production,the cultivation of Monascus purpureus in submerged encapsulated cell was investigated. Monascus purpureus immobilized in polyelectrolyte complex(PEC) microcapsules,which were pre-pared by sodium cellulose sulphate(NaCS) and poly-dimethyl-diallyl-ammonium chloride(PDMDAAC),was a good substitute for submerged cell culture because it mimicked the solid-state environment. The repeated-batch process with encapsulated cells was studied in flasks and a bubble column. The results indicated that the bubble column was more suitable for the encapsulation culture than the shaking flasks because of its good mass transfer performance and minor shear stress on cells. Owing to the protection of the microcapsule's membrane,Monascus purpureus in microcapsules increased approximately three times over that in free cell culture with negligible cell leakage to the medium. The pigment production in the bubble column finally reached 3.82(OD500) ,which was two times higher than in free cell culture. In addition,the duration of each batch was shortened to 15% of that in free cell culture.

Research Status and Application Prospects of Monascus sp.

Monascus sp. has been widely applied for a long time in China. In recent years, the effective physiological active substances produced by Monascus sp. have been extensively investigated. The classification, cultivation characteristics, physiological and biochemical properties, primary metabolites, secondary metabolites and application prospects of Monascus sp. were summarized and discussed in this paper.

Serial Analysis of Gene Expression in Monascus aurantiacus Producing Citrinin

To construct a tag expression library of Monascus aurantiacus that could produce citrinin maximally on the thirteenth (0.966 mg/mL) day in the submerged culture. Methods Total RNA was extracted from the mycelium, cDNA was synthesized using the SuperScript choice system, and then, a SAGE library was successfully constructed according to the MicroSAGE method. Results Five hundred and ninety eight clones were obtained in SAGE library, and 120 clones were picked out randomly for identification and sequencing purpose. Eighty nine clones had positive inserts, 26 clones had no inserts and the remaining 5 clones had no site of NlaⅢ enzyme in inserts. There were seven repeated clones. Conclusion With the aid of SAGE2000 software, 901 tags were obtained from 89 clones, representing 686 unique transcripts. Six unique tags of them belong to highly expressed genes (Number of tags≥10) and 143 unique tags to moderately expressed genes (repeat tags≥2).

Monascus vinegar alleviates high-fat-diet-induced inflammation in rats by regulating the NF-κB and PI3K/AKT/mTOR pathways

Monascus vinegar(MV) is a typical fermented food with various health-promoting effects. This study aimed to evaluate the role of MV in alleviating high-fat-diet-induced inflammation in rats with hyperlipidemia and elucidate the possible regulatory mechanisms. In the study, serum lipid profiles, liver pathology and liver inflammatory cytokines were analyzed in hyperlipidemia rats with MV(0.5 mL/kg mb, 2 mL/kg mb). Results showed that the administration of MV alleviated dyslipidemia by decreasing the serum and liver levels of triglyceride and total cholesterol. Increase in hepatic lipase and carnitine palmitoyl transferase 1(CPT-1)levels and decrease in hepatocyte steatosis, nephritis, and intestinal tissue injury in the HD group showed that high-dose MV can significantly suppress hepatic lipid accumulation and steatosis. In addition, compared with the model(MOD) group, the HD group showed significantly down-regulated the level of serum or hepatic alanine aminotransferase(ALT), aspartate aminotransferase(AST), CPT-1, interleukin(IL)-2, IL-6, IL-12,and tumor necrosis factor α(TNF-α). Moreover, the HD group showed repressed hepatic nuclear factor κB(NF-κB) pathway and inactivated phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway mitigated liver inflammation. Similar results were obtained from cell experiments. Collectively, these findings revealed that MV might attenuate high-fat-diet-induced inflammation by inhibiting the NF-κB and PI3K/Akt/mTOR pathways.

Extracts of Monascusus Purpureus beyond Statins——Profile of Efficacy and Safety of the Use of Extracts of Monascus Purpureus

Extracts of Monascus purpureus have always been considered a natural source of Iovastatin, the precursor of the world＇s largest selling class of drugs. In actual fact, the fungus contains many other substances （flavonoids, polyunsaturated fats, pyrrolinic compounds etc.） with a wide variety of other actions. The most recent studies have shown that it has an action on the glycemic metabolism, and on the mechanisms of adipogenesis, also an effects on the endothelium and on postprandial vasodilation. These effects are more extensive and complex than those of statins alone. And new strains of Monascus purpureus have recently been patented where the presence of statins is only one of the therapeutic components of the fungus. In particular, the increase in secondary components, such as flavonoids, which coincides with a more complex therapeutic action, probably making the new extracts of Monascus purpureus, the ideal candidate for the treatment of the metabolic syndrome.

Separation and Purification of Monascus Pigments from Red Yeast Rice

It is difficult to extract beneficial components from Monascus pigments due to their complex composition. The low purity of Monascus pigment preparations limits their further application in research and production. Therefore, this study was conducted to optimize the conditions for extraction and isolation of Monascus pigments from red yeast rice, to improve the purity of Monascus pigment preparations. Three fractions were isolated using column chromatography from red yeast rice, they were red, orange and yellow respectively. Then, the three fractions were analyzed for composition and purity using thin layer chromatography and liquid chromatography-mass spectrometry(LC-MS). The chromatograms revealed that the yellow and orange fractions had complex compositions that were difficult to be separated, while the red fraction consisted of fewer components. Finally, a single component was isolated from the red fraction using a thin layer chromatography plate, identified as monascorubramine according to its molecular mass.

Effect of <i>Monascus</i>Fermentation on Aroma Patterns of Semi-Dried Grass Carp

The aim of the present study is to investigate the effects of Monascus fermentation on the aroma pattern of semi-dried grass carp. Semi-dried fish was fermented using Monascus purpureus GDMCC3.439. The volatile flavor substances present in fresh fish, semi-dry fish and Monascus fermented semi-dried fish were compared by simultaneous distillation and extraction combined with gas chromatograph-mass spectrometer (GC-MS). The results showed that alcohols, aldehydes and ketones were the main components of the flavor of the unfermented and fermented semi-dried grass crap. Monascus fermentation could significantly affect the volatile flavor substances of semi-dried grass carp. Moreover, the processing of semi-dried fish fermented by Monascus could not only effectively improve the fishy smell of fresh fish, but also make up for the defect of the pickled flavor of semi-dried fish. Eighteen of the main volatile components in semi-dried fish fermented by Monascus were identified by relative odor activity value (ROAV), as follows: 1-octene-3-ol, phenylethanol, hexanal, heptanal, nonanal, (E)-2-octenal, 3-Methyl-1-butanal, benzaldehyde, (E)-2-nonenal, (E,E)-2,4-heptadienal, (E)-2-Decenal, phenylacetaldehyde, (E,E)-2.4-decadienal, tetradecanal, 2,3-butanone, 2,3-octanedione, alpha-pinene, 2-pentane furan.

Ecological Dyeing of Polyamide 6, 6(PA66) Fabrics with Monascus Pigments in Decamethylcyclopentasiloxane(D5) Solvent

Synthetic dyes in the aqueous media have been commonly used for textile dyeing, resulting in resource and environmental pressure arising from consumption of water and environmentally unfriendly chemicals. In this study, an eco-friendly process of dyeing polyamide 6, 6(PA66) fabrics with natural Monascus pigments in decamethylcyclopentasiloxane(D5) solvent has been developed to minimize water consumption and effluent generation. The influence of processing parameters including dyeing temperature, dyebath pH and dyeing time on dyeing effects was explored. It was shown that color strength and color fastness of the samples dyed in D5 media were higher than those of the samples dyed in the aqueous media. Moreover, PA66 fabrics exhibited the highest color strength, good color fastness and a bacteriostatic rate of 53.6% against Staphylococcus aureus when it was dyed at pH of 3.5 and temperature of 80 ℃ for 30 min.

Process Optimization on Monascus Pigment Produced by Monascus

[Objective] This study was conducted to optimize rice making process and improve the content of Monascus pigment. [Method]Selecting water addition( A),sterilization temperature( B),sterilization time( C),soaking time( D) as experimental factors,L_9( 3~4) orthogonal test was carried out to optimize rice making process. [Result] The four factors all had very significant effect on the yield of Monascus pigment,the optimal combination was A_1B_2C_2D_2,namely water addition of 30 ml,sterilization temperature at 121 ℃,sterilization time of 23 min,and soaking time of 24 h. The level of Monascus reached 2 884 U/g under this condition. [Conclusion] The study has great practical significance to Monascus rice production enterprises.

Monascus purpureus M-32 fermented soybean meal improves the growth,immunity parameters,intestinal morphology,disease resistance,intestinal microbiota and metabolome in Pacific white shrimp(Litopenaeus vannamei)

This study was conducted to evaluate the effects of Monascus purpureus M-32 fermented soybean meal(MFSM)on growth,immunity,intestinal morphology,intestinal microbiota,and intestinal metabolome of Pacific white shrimp(Litopenaeus vannamei).Four groups of diets were formulated,including control group(30%fish meal and 30%soybean meal[SBM]included in the basal diet)and three experimental groups which MFSM replaced 20%(MFSM20),40%(MFSM40),and 60%(MFSM60)of SBM in control group,respectively.Results showed that the soluble proteins larger than 49 kDa in MFSM were almost completely degraded.Meanwhile,the crude protein,acid-soluble protein,and amino acid in MFSM were increased.The results of shrimp culture experiment showed that the replacement of SBM with MFSM decreased FCR(P<0.001)and content of malondialdehyde(P=0.007)in the experimental groups,and increased weight gain rate(P=0.006),specific growth rate(P=0.002),survival rate(P=0.005),in-testinal villus height(P<0.001),myenteric thickness(P=0.002),the activities of superoxide dismutase(P=0.002),and lysozyme(P=0.006)in experimental groups,as well as increased content of calcium(Ca2+)and phosphorus(PO43-)in blood and muscle,and enhanced resistance to Vibrio parahaemolyticus infection.The gut microbiota of MFSM groups was significantly different from that of the control group,and the abundance of Actinobacteria and Verrucomicrobia increased significantly in the MFSM60 group,whereas Proteobacteria and Firmicutes decreased.Compared with the control group,there were sig-nificant changes in the levels of several intestinal metabolites in the MFSM60 group,including leuko-triene C5,prostaglandin A1,taurochenodeoxycholic acid,carnosine,and itaconic acid.The fermentation of SBM by the strain M.purpureus M-32 has the potential to enhance the nutritional quality of SBM,promote the growth of L.vannamei,boost immune response,improve intestinal morphology and microbiota composition,as well as influence intestinal metabolites.

Monascus pigment-protected bone marrow-derived stem cells for heart failure treatment

Mesenchymal stem cells(MSCs)have demonstrated significant therapeutic potential in heart failure(HF)treatment.However,their clinical application is impeded by low retention rate and low cellular activity of MSCs caused by high inflammatory and reactive oxygen species(ROS)microenvironment.In this study,monascus pigment(MP)nanoparticle(PPM)was proposed for improving adverse microenvironment and assisting in transplantation of bone marrow-derived MSCs(BMSCs).Meanwhile,in order to load PPM and reduce the mechanical damage of BMSCs,injectable hydrogels based on Schiff base cross-linking were prepared.The PPM displays ROS-scavenging and macrophage phenotype-regulating capabilities,significantly enhancing BMSCs survival and activity in HF microenvironment.This hydrogel demonstrates superior biocompatibility,injectability,and tissue adhesion.With the synergistic effects of injectable,adhesive hydrogel and the microenvironment-modulating properties of MP,cardiac function was effectively improved in the pericardial sac of rats.Our results offer insights into advancing BMSCs-based HF therapies and their clinical applications.

Mphy gene reveals the involvement of light source in the regulation of pigment metabolism in Monascus ruber

The growth,development and pigment metabolism of Monascus are regulated by light.However,the function of the photoreceptor gene in the Monascus genome has not yet been clearly verified.In this paper,the gene sequence Mphy,which is homologous to plant phytochrome,was firstly found from Monascus ruber C100 by comparing it with the genome.Then,taking M.ruber C100 as the main research object,by knocking out and overexpressing the Mphy gene,the involvement of Mphy in the regulation of light on the growth and development of Monascus and the metabolism of Monascus pigments(MPs)were preliminarily explored.Compared with the original strain C100,the Mphy knockout strain(ΔMphy)and the overexpression strain(OE-Mphy)grew slowly and produced low spores.At the same time,under different light conditions,red light can promote the growth and sporulation of Monascus,while blue light can inhibit it.MPs are positively regulated by the Mphy gene,that is,the deletion of the Mphy gene reduces the production of MPs,while the overexpression of the gene increases the accumulation of the pigment,especially under red light conditions.These results indicate that the homologous genes of phytochromes are involved in the process of light-regulated growth,development and pigment metabolism in Monascus.Based on the above results,we found that under red light conditions,Mphy controls the process of pigment metabolism by affecting key enzymes in TCA metabolism and key signaling pathways such as the MAPK signaling pathway.