Two new Monascus metabolites with strong blue fluorescence isolated from red yeast rice

Red yeast rice obtained as cultures of Monascus AS3.4444 on rice was extracted and analyzed by high-performance liquid chromatography (HPLC). Two new Monascus metabolites with similar fluorescence spectra (lambda ex = 396 nm, lambda em = 460 nm) and UV absorption spectra (lambda max = 386 nm) were detected. They were isolated by rechromatography on a silica gel column and semipreparative HPLC, and two strong blue fluorescent compounds were obtained. Their structures were elucidated by electrospray ionization mass spectrometry (ESI-MS), electrospray ionization tandem mass spectrometry (ESI-MS/MS), intensive ESI-MS, and nuclear magnetic resonance spectroscopy ( (1)H NMR, (13)C NMR, COSY, and HMBC) studies. High-resolution mass spectrometry indicated the molecular formulas C 21H 24O 5 and C 23H 28O 5. The two new compounds, named monasfluore A and monasfluore B, respectively, contain a alkyl side chain, gamma-lactone, and propenyl group, whereas the more lipophilic compound, monasfluore B, is a higher homologue of monasfluore A, with the more lipophilic octanoyl instead of the hexanoyl side chain.     

New monascus metabolite isolated from red yeast rice (angkak,red koji)

Red yeast rice (angkak, red koji) obtained as cultures of Monascus purpureus on rice was extracted and analyzed by HPLC. In addition to the known red, orange, and yellow pigments and the mycotoxin citrinin, a new Monascus metabolite was detected. It is present in the original red yeast rice and formed in higher amounts when red yeast rice is heated. High-resolution mass spectrometry indicated the molecular formula C(15)H(12)O(4). The chemical structure was elucidated by analysis of NMR data. The new compound, named monascodilone, is characterized by a propenyl group on a pyrone ring, an aromatic ring, and a gamma-lactone group.     

Constituents of red yeast rice, a traditional Chinese food and medicine

Detailed analyses were undertaken of the natural constituents of red yeast rice, a traditional Chinese medicine and food known for centuries to improve blood circulation. Preparation of red yeast rice following ancient methods by fermenting the fungal strain Monascus purpureus Went on moist and sterile rice indicated the presence of a group of metabolites belonging to the monacolin family of polyketides, together with fatty acids, and trace elements. The presence of these compounds may explain in part the cholesterol-lowering ability associated with this traditional Chinese food.     

玉米固态发酵产红曲红色素补加营养物的优化（简报）

为了优化玉米固态发酵生产红曲红色素的主要补加营养物,采用析因试验设计确定了紫红曲霉M144固态发酵红曲红色素的主要影响因子为葡萄糖、硫酸镁和酵母膏添加量,采用Box-Behnken试验设计和响应面优化分析确定了补加营养物的最佳添加量分别为葡萄糖0.26%(w/w),硫酸镁0.43%(w/w),酵母膏0.25%(w/w),在此条件下红曲红色素红色价为2335.17U/g,色调为1.69,比未补加营养物时分别提高了55.67%和64.08%。通过对玉米固态发酵过程中补加营养物质的系统研究,提高了红曲红色素的产量和质量,可为玉米固态发酵红曲红色素的生产提供技术支持。     

几种吡啶类化合物对土壤硝化的抑制作用比较

为了探明吡啶类化合物对土壤硝化作用的抑制效应,采用室内微宇宙试验,研究了2-氯-6(三氯甲基)硫酸盐、2-氯-6(三氯甲基)盐酸盐、吡啶混合物和吡啶X类化合物对潮土、红壤和水稻土中铵态氮硝化的抑制作用。结果表明,在35 d培养期内,吡啶类化合物处理土壤硝态氮含量明显低于对照(未添加吡啶类化合物),吡啶类化合物对土壤中铵态氮的硝化抑制率介于2.91%~91.92%,抑制强度先逐渐升高后降低,在培养21 d时抑制强度达到峰值。不同类型吡啶类化合物对土壤中铵态氮的硝化抑制效果存在差异,吡啶盐酸盐类化合物优于其他几类化合物;吡啶类化合物对土壤中铵态氮的硝化抑制作用与土壤类型有关,对3种土壤中铵态氮的硝化抑制作用表现为潮土>水稻土>红壤。就同一土壤而言,硝化抑制强度随着吡啶类化合物用量的增加而增加。     

Modified mutation method for screening low citrinin-producing strains of Monascus purpureus on rice culture

Monascus purpureus NTU 601 is a strain that produces monacolin K, gamma-aminobutyric acid (GABA), and citrinin under solid culture conditions. Because citrinin is a mycotoxin and possesses nephrotoxic and hepatoxic effects, it has a negative impact on the acceptance of red mold rice by people. In this research, a simple and quick selection method for mutant strains with low citrinin production was designed based on the fact that citrinin possesses antibacterial activity for Bacillus subtilis and will form an inhibition zone around the colony of the Monascus strain. The mutant strain M. purpureus N 301 only produced 0.23 +/- 0.01 ppm citrinin, which was 50% less than that of the parent strain, and the monacolin K production was 481.29 +/- 7.98 ppm and maintained 91% productivity. M. purpureus N 310, the other mutant strain, produced 0.27 +/- 0.01 ppm citrinin, which was 41% less than that of the parent strain, and the monacolin K production was 526.29 +/- 5.54 ppm, which showed no significant changes when compared with the parent strain. The GABA content of the two strains was 5000 ppm, which is similar to that of the parent strain. The results showed that the method could be used to select red mold rice with low citrinin production.     

利用返回式飞船选育优良红曲霉菌

利用“神舟 3号”返回式飞船搭载红曲霉菌 0 70 8。对搭载后的菌株进行复壮、分离和筛选 ,以高产洛伐他汀和耐硒为选择条件 ,筛选到 8株耐硒又高产洛伐他汀的菌株。经连续发酵试验证明变异菌株的优良特性可以保持稳定。     

Improving the ratio of monacolin K to citrinin production of Monascus purpureus NTU 568 under dioscorea medium through the mediation of pH value and ethanol addition

Using dioscorea root as substrate of Monascus species was found to stimulate monacolin K (cholesterol-lowering agent) formation in our previous study, but the mycotoxin-citrinin has never been studied. This study used dioscorea root as the liquid medium to culture Monascus purpureus NTU 568 using a 6.6 L jar fermentor. Culture pH value, dioscorea concentration, and ethanol concentration were used as the factors of response surface methodology (RSM) to investigate the optimal culture condition for high monacolin K production and low citrinin formation. Monacolin K and citrinin formation of M. purpureus NTU 568 under submerged dioscorea medium were respectively found to be significantly formed by 148% and 147%, as compared to that under submerged rice medium. The reason is due to the pH value (3.5) of dioscorea medium involved in the formation of Monascus cell amount and secondary metabolite. RSM results further indicated that lowering the pH value to 2.5 would result in high monacolin K and citrinin concentrations as well as high biomass in fixed dioscorea amount, implying that pH value may stimulate the formation of monacolin K and citrinin through increasing Monascus cell amount. Lowering dioscorea and ethanol concentration was able to increase the ratio of monacolin K level to citrinin level. The optimal culture condition (pH 5.7, 1% dioscorea concentration, and 0.5% ethanol concentration) would increase monacolin K levels to 27.9 mg/g (by 47%) and decrease citrinin level to 2.15 microg/g (by 54%), as compared to control conditions (pH 3.5, 5% dioscorea, and ethanol free).     

New bioactive orange pigments with yellow fluorescence from Monascus-fermented dioscorea

Red mold dioscorea (RMD) is a fermented product of Monascus purpureus NTU 568 using dioscorea as culture substrate. To investigate the bioactive components of RMD, six orange pigments including four new azaphilones with yellow fluorescence, monapilol A-D (1-4), and known monascorubrin (5) and rubropunctatin (6) were isolated and characterized. Structural elucidation of new isolates was based on nuclear magnetic resonance ((1)H NMR, (13)C NMR, COSY, HMQC, and HMBC) and other spectroscopic analyses. The structures of monapilols (1-4) were similar to those of monascorubrin (5) and rubropunctatin (6); however, the hydroxyl group (8-OH) in compounds 1-4 substituted for the C-8 carbonyl in compounds 5 and 6. Biological evaluation indicated that compounds 1-4 inhibited nitric oxide (NO) production on lipopolysaccharide-stimulated RAW 264.7 cells. Compounds 1-4 also exhibited antiproliferative activities against human laryngeal carcinoma (HEp-2) and human colon adenocarinoma (WiDr).     

Evaluation of citrinin occurrence and cytotoxicity in Monascus fermentation products

Monascus purpureus and its fermentation products have been used in food coloring and meat preservation in Asia for centuries and have also been recently used as dietary supplements because of their cholesterol-lowering ability. However, the presence of the mycotoxin citrinin (CTN), a secondary metabolite of Monascus species, in fermentation products is a potential threat to public health. In the present study, HPLC was used to analyze CTN levels in lipid and aqueous extracts of commercialized Monascus products. CTN was detected in lipid extracts of all examined samples at concentrations varying between 0.28 and 6.29 microg/g, but was not found in aqueous extracts. When human embryonic kidney cells (HEK293) were incubated for 72 h with Monascus extracts, the concentrations causing 50% cell death by all lipid extracts were in the range of 1.8-4.7 mg/mL, whereas aqueous extracts showed a lower cytotoxicity. Incubation of HEK293 cells with 60 microM pure CTN for 72 h caused cell viability to fall to 50% of control levels. In addition, coadministration of pure CTN and lipid extracts from Monascus samples significantly enhanced CTN cytotoxicity for HEK293 cells using the MTT assay. These results provide the first information about the cytotoxic effects of various Monascus samples and CTN-Monascus mixtures on a human cell line.     

Purification and characterization of an antimicrobial chitinase extracellularly produced by Monascus purpureus CCRC31499 in a shrimp and crab shell powder medium

Monascus purpureus CCRC31499 produced an antimicrobial chitinase when it was grown in a medium containing shrimp and crab shell powder (SCSP) of marine wastes. An extracellular antimicrobial chitinase was purified from the culture supernatant to homology. The chitinase had a molecular weight of approximately 81,000 and a pI of 5.4. The optimal pH, optimum temperature, and pH stability of the chitinase were pH 7, 40 degrees C, and pH 6-8, respectively. The activity of the chitinase was activated by Fe(2+) and strongly inhibited by Hg(2+). The unique characteristics of the purified chitinase include high molecular weight, nearly neutral optimum pH, protease activity, and antimicrobial activity with bacteria and fungal phytopathogens. This is also the first report of isolation of a chitinase from a Monascus species.     

Synthesis and herbicidal activity of 2-cyano-3-substituted-pyridinemethylaminoacrylates

Two series of 2-cyano-3-substituted-pyridinemethylaminoacrylates, namely 12 new (Z)-2-cyano-3-methylthio-3-substituted-pyridinemethaneaminoacrylates and 10 new (Z)-2-cyano-3-alkyl-3-substituted-pyridinemethaneaminoacrylates, were synthesized as herbicidal inhibitors of photosystem II (PSII) electron transport. All of these compounds were confirmed by (1)H NMR, elemental, IR, and mass spectrum analyses. Their herbicidal activities were evaluated. Some compounds exhibited excellent herbicidal activities, even at a dose of 75 g/ha. A suitable substituent at the 2-position of the pyridine ring and the well-fit group at the 3-position of acrylate were essential for high herbicidal activity. 2-Cyanoacrylates containing a substituted pyridine ring provide higher herbicidal activities than parent compounds containing phenyl. These PSII inhibitor herbicides are safe to corn, which is a major crop in China.     

Protective effect of Monascus-fermented red mold rice against alcoholic liver disease by attenuating oxidative stress and inflammatory response

Monascus purpureus NTU 568 fermented rice is reported to exhibit a wide variety of biological effects, including antitumor, antihypertriglyceridemia, antioxidant, and anti-inflammatory activities. However, its role in the pathogenesis of alcoholic liver disease remains obscure. In this study, the hepatoprotective effects of Monascus-fermented red mold rice (RMR) was evaluated in vivo using chronic alcohol-induced mice as an experimental model. The alcohol-induced mice were orally treated with RMR at 307.5 mg/kg (1-fold), 615 mg/kg (2-fold), and 1537.5 mg/kg (5-fold) for 5 weeks, whereas controls received vehicle only. Treatment with RMR significantly attenuated the increased level of serum transaminases (aspartate aminotransferase and alanine aminotransferase) and hepatic triglyceride and total cholesterol accumulation. Furthermore, RMR elevates hepatic antioxidant ability that reduced hepatic cell damage (steatosis) and decreased tissue inflammatory cytokine levels. These findings suggest that Monascus-fermented RMR may represent a novel, protective strategy against alcoholic liver disease by attenuating oxidative stress, inflammatory response, and steatosis.     

Anti-tumor and anti-inflammatory properties of ankaflavin and monaphilone A from monascus purpureus NTU 568

An azaphilonidal derivative monaphilone A (MA) was recently isolated from the fermented products of Monascus purpureus NTU 568 by our laboratory. We report here the exploration of apoptosis-related and anti-inflammatory properties of MA and ankaflavin (AK) by some experiments about inducing death of human laryngeal carcinoma cell line HEp-2 and reducing inflammatory responses on murine macrophage RAW 264.7 cells. We employed a ssDNA enzyme-linked immunosorbent assay (ELISA) kit to investigate the nuclear changes of early apoptosis induced by AK and MA on HEp-2 cells and used a western blot and an enzyme activity assay to demonstrate the activation of caspase-3, caspase-8, and caspase-9 by MA and AK. Our studies revealed that AK and MA may decrease lipopolysaccharide (LPS)-induced inflammatory responses, including nitrite productions and expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) in RAW 264.7 cells. All evidence support that azaphilonidal derivatives from M. purpureus NTU 568, such as AK and MA, are suitable for the development of chemotherapy or chemopreventive agents.     

Protective Effect of Deferricoprogen Isolated from Monascus purpureus NTU 568 on Citrinin-Induced Apoptosis in HEK-293 Cells

Monascus species have traditionally been used in Asian food, with rice as their fermentation substrate. Red mold rice (RMR) contains citrinin, a nephrotoxic agent capable of exerting oxidative stress and cellular apoptosis. We investigated the components in RMR that could minimize the adverse effects of citrinin. Combining chemical separations and bioactivity assays, we identified an antioxidative component called deferricoprogen (DFC) in the fermented rice of Monascus purpureus NTU 568. The DFC structure was confirmed by nuclear magnetic resonance (NMR) and mass spectra analysis. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging activity of DFC was similar to that of vitamin E. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and flow cytometric analysis showed the effect of DFC and citrinin on cell viability and cell cycle. DFC was found to be protective against the cytotoxicity and cell death induced by citrinin on human embryonic kidney (HEK-293) cells. DFC also demonstrated anti-apoptotic property in preventing citrinin-induced apoptosis.     

Odor-active headspace components in fermented red rice in the presence of a monascus species

Headspace components from rice and agar with (experimental) and without (control) inoculation with a Monascus spp. were investigated. Kinetics studies were carried out. Using rice as a substrate, 10 and 19 compounds were found for the control and the experimental groups, respectively, at day 14. Experimental group compounds were composed mainly of alcohols, ketones, and esters, whereas control group compounds were composed of aldehydes and ketones. With agar as a substrate, only five and three components were found in the control and experimental groups, respectively. Five alcohols, four esters, two ketones, and one furan with odor activity values (OAV) >1 dominated the overall flavor of the product. With liquid inoculation, the first six components with high OAVs were in the following order: 3-methyl-1-butanol (17) > ethanol (14) > ethyl acetate (10) > 2-methyl-1-propanol (15) > ethyl butanoate (11) > 3-methylbutyl acetate (13). Kinetic studies showed that most compounds reached their maximum concentrations at 10-12 days. Many compounds identified in the model red rice were reported in commercial red sufus, and several appeared to contribute solely by red rice.     

Structure-antifeedant activity relationship of clerodane diterpenoids. Comparative study with withanolides and azadirachtin

A structure-antifeedant activity relationship (SAR) study of clerodane diterpenoids was carried out. Attention was focused on the feeding-deterrent activities exhibited toward Tenebrio molitor by clerodane diterpenoids and withanolides. Azadirachtin was chosen as a reference compound. SAR studies on the clerodane compounds indicate that the stereoelectronic factors are more important than the hydrophobic aspects as determinants of antifeedant activity. A furan ring in the side chain and a carbonyl alpha,beta-unsaturated (or spiro-epoxide) group appear to be indispensable for the biological response. A conformational study indicate that the optimum interatomic distance between these moieties is a range between 9.5 and 10.5 A. In addition, a similar stereoelectronic response was found among withanolides and azadirachtin. On the basis of these results it is reasonable to imagine a closely related chemical mechanism for these compounds.     

Lipophilic extracts from banana fruit residues: a source of valuable phytosterols

The chemical composition of the lipophilic extracts of unripe pulp and peel of banana fruit 'Dwarf Cavendish' was studied by gas chromatography-mass spectrometry. Fatty acids, sterols, and steryl esters are the major families of lipophilic components present in banana tissues, followed by diacylglycerols, steryl glucosides, long chain fatty alcohols, and aromatic compounds. Fatty acids are more abundant in the banana pulp (29-90% of the total amount of lipophilic extract), with linoleic, linolenic, and oleic acids as the major compounds of this family. In banana peel, sterols represent about 49-71% of the lipophilic extract with two triterpenic ketones (31-norcyclolaudenone and cycloeucalenone) as the major components. The detection of high amounts of steryl esters (469-24405 mg/kg) and diacylglycerols (119-878 mg/kg), mainly present in the banana peel extract, explains the increase in the abundance of fatty acids and sterols after alkaline hydrolysis. Several steryl glucosides were also found in significative amounts (273-888 mg/kg), particularly in banana pulp (888 mg/kg). The high content of sterols (and their derivatives) in the 'Dwarf Cavendish' fruit can open new strategies for the valorization of the banana residues as a potential source of high-value phytochemicals with nutraceutical and functional food additive applications.     

New lipophilic tyrosyl esters. Comparative antioxidant evaluation with hydroxytyrosyl esters

New lipophilic esters of tyrosol, a naturally occurring phenol with interesting biological properties, have been synthesized in good yields by a chemoselective procedure, using lipase from Candida antarctica or p-toluenesulfonic acid as catalysts. Their antioxidant activities have been evaluated by the Rancimat test in lipophilic food matrices, as well as by FRAP and ABTS assays in methanolic solutions, and compared with those of previously synthesized hydroxytyrosyl esters. Free tyrosol, hydroxytyrosol, butylhydroxytoluene, and alpha-tocopherol were used as standards. All methods used for the antioxidant activity evaluation emphasized the high influence of the ortho-diphenolic structure on the antioxidant capacity, tyrosol and its derivatives being less active than hydroxytyrosol and its analogues and even less than BHT and alpha-tocopherol. In addition, the Rancimat test revealed a lower activity for ester derivatives than for their respective reference compounds (HTy or Ty), in agreement with the polar paradox. On the other hand, FRAP and ABTS methods reported an opposite behavior between the synthetic esters and their respective references. Thus, hydroxytyrosyl esters were more active than HTy, whereas tyrosyl esters were less active than Ty. The length and nature of the acyl side chain did not seem to play an important role in the antioxidant activity of either the hydroxytyrosyl or tyrosyl ester series, since no significant differences were observed among them.