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PubMed2004

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.

Citrinin↗
PubMed2003

Genetic transformation of Monascus purpureus DSM1379.

Monascus purpureus was transformed into hygromycin B resistance with hygromycin B phosphotransferase (hph) fused to Aspergillus nidulans trpC or a putative Monascus purpureus gpd1 promoter by electroporation. Among five strains, only M. purpureus DSM1397 was a competent recipient. Normal growth and sporulation on media containing up to 500 mg hygromycin B l(-1) occurred up to five generations. Upon transformation of the strain with the green fluorescent protein gene (sgfp) as a model gene and hph as a selection marker, characteristic green fluorescence was observed under fluoromicroscopy indicating successful transformation.

Drug Resistance, Fungal↗
PubMed2005

Cloning and sequence analysis of putative glyceraldehyde-3-phosphate dehydrogenase gene from Monascus purpureus KCCM11832.

Using a synthetic oligonucleotide probe, glyceraldehyde-3-phosphate dehydrogenase gene (gpd1) was cloned from Monascus purpureus KCCM11832. The 2834 bp EcoRV-HindIII region harbored 1183 bp 5'-UTR containing such regulatory elements as CT box, common in fungal gpd's, and gpd box previously found exclusively in Aspergillus gpd's. Full-length cDNA was cloned by PCR, and its sequence was determined. Transcription starting point was located 88 bp upstream from start codon. Polyadenylation signal sequence occurred 201 bp downstream from stop codon. Region from start codon ATG to stop codon TAA including introns showed 62 approximately 69% nucleotide sequence identity to those of Aspergillus gpd's. Significant bias in third position, with pyrimidines favored over purines, was observed in codon usage. The deduced amino acid sequence had 81 approximately 85% identity to Aspergillus gpd's. Monascus purpureus GPD was located at the same clade with Aspergillus GPD's.

Amino Acid Sequence↗
PubMed2005

Polyketide synthase gene responsible for citrinin biosynthesis in Monascus purpureus.

Citrinin produced by Aspergillus, Penicillium, and Monascus species is a polyketide compound that has nephrotoxic activity in mammals and is bactericidal toward gram-positive bacteria. To avoid the risk of citrinin contamination in other fermentation products produced by Monascus purpureus, knowledge of the citrinin biosynthetic genes is needed so that citrinin-nonproducing strains can be generated. We cloned a polyketide synthase (PKS) gene from M. purpureus with degenerate primers designed to amplify the conserved region of a ketosynthase domain of a fungal PKS. A 13-kb genomic DNA fragment was identified that contained a full-length PKS gene (pksCT) of 7,838 bp with a single 56-bp intron. pksCT encodes a 2,593-amino-acid protein that contains putative domains for ketosynthase, acyltransferase, acyl carrier protein (ACP), and a rare methyltransferase. There was no obvious thioesterase domain, which usually is downstream of the ACP domain in multi-aromatic-ring PKSs. pksCT transcription was correlated with citrinin production, suggesting that the pksCT gene product was involved in citrinin biosynthesis. Homologous recombination between the wild-type allele and a truncated disruption construct resulted in a pksCT-disrupted strain of M. purpureus. The disruptant did not produce citrinin, but a pksCT revertant generated by successive endogenous recombination events in the pksCT disruptant restored citrinin production, indicating that pksCT encoded the PKS responsible for citrinin biosynthesis in M. purpureus.

Amino Acid Sequence↗
PubMed2006

Lipid-lowering effect of monascus garlic fermented extract (MGFE) in hyperlipidemic subjects.

Monascus Garlic Fermented Extract (MGFE) is a unique material produced from garlic fermented using Monascus pilosus, which contains characteristic compounds such as dimerumic acid and monacolin K. In this study, we examined the effect of MGFE on hyperlipidemic subjects. Fifteen subjects aged 33-59 years (11 men and 4 women) participated. All the subjects had either hypercholesterolemia (> or = 220 mg/dl) or hyper-low-density lipoprotein cholesterolemia (> or = 140 mg/dl), but only 4 of the 15 (27%) were hypertriglycemic (> or = 150 mg/dl). All subjects received two capsules of MGFE (225 mg/capsule) after breakfast and dinner for 4 weeks. After an overnight fast, blood was taken 0, 2 and 4 weeks after the start of MGFE intake, and 2 weeks after MGFE withdrawal. MGFE significantly reduced serum total cholesterol (TC) and low-density lipoprotein cholesterol levels 2 and 4 weeks after the start of MGFE intake as compared with the baseline. Although the level of high-density lipoprotein cholesterol (HDL-C) was unaffected at any time, the atherogenic index calculated from the value of TC and HDL-C was significantly reduced 2 and 4 weeks after the start of MGFE intake. These effects of MGFE tended to disappear within 2 weeks after withdrawal. Triglyceride (TG) and lipid peroxide levels were not reduced dramatically, but TG levels in hypertriglycemic subjects tended to reduce as compared with the baseline value. No abnormal changes in blood biochemical parameters or adverse effects were observed in any of the subjects. Our present results indicate that MGFE attenuates hyperlipidemia, suggesting that MGFE is a potent agent for preventing arteriosclerotic diseases.

Adult↗
PubMed2000

Suspended rice particles for cultivation of Monascus purpureus in a tower-type bioreactor.

Cultivation of Monascus purpureus (CCRC 31615) for the production of natural pigments was investigated. Traditionally, Monascus species were grown on rice by solid-state culture. For large-scale cultivation, solid-state cultures were associated with some problems such as contamination and scale-up. By using submerged cultures with rice particles, a stirred-tank fermentor was not suitable for submerged cultures as the impeller tended to break the particles into small pieces. A conventional bubble column was also unsuitable as its mixing capability was poor. In the present study, a modified bubble column with wire-mesh draft tubes was employed for the cultivation of M. purpureus. The proposed column had a shorter mixing time and a higher oxygen transfer rate relative to the conventional bubble column. The production of pigments using the proposed column was up to 80% higher than that achieved using the conventional bubble column.

Ascomycota↗
PubMed2000

Nitric oxide-mediated endothelium-dependent relaxation of rat thoracic aorta induced by aqueous extract of red rice fermented with Monascus ruber.

Vasodilatory effects of aqueous extract of red rice fermented with Monascus ruber IFO32318 were examined on the isolated rat aorta. The water phase of fermented rice with Monascus (WP/FRM, 0.1-10 mg/ml) caused a transient relaxation of the endothelium-intact rat aorta precontracted with norepinephrine (NE, 300 nM). The WP/FRM-induced relaxation was abolished by removal of endothelium or in the presence of N(G)-nitro-L-arginine (L-NNA, 10 microM), a nitric oxide (NO) synthase inhibitor. Neither atropine, a muscarinic receptor antagonist (10 microM), nor indomethacin, a cyclooxygenase inhibitor (10 microM), altered the WP/FRM-induced endothelium-dependent relaxation. gamma-Aminobutyric acid (GABA), one of the principle components of the extract, did not affect the muscle tension of the aorta with intact endothelium. In addition, WP/FRM increased the production of NO in primary cultured endothelial cells from human umbilical vein. The enhanced production of NO by WP/FRM was diminished by pretreatment with L-NNA (10 microM). In conclusion, WP/FRM induces relaxation of rat aorta by releasing NO from endothelium. There seem to be some unknown factor(s) other than acetylcholine (Ach) and GABA, in the aqueous extract of red rice, which stimulate vascular endothelial cells to produce and/or release NO leading to endothelium-dependent relaxation by WP/FRM.

Acetylcholine↗
PubMed2002

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.

Animals↗
PubMed2002

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.

Ascomycota↗
PubMed1982

Production of Extracellular Pigment by a Mutant of Monascus kaoliang sp. nov.

A hyperpigment-producing mutant, R-10847, was derived from Monascus kaoliang F-2 (ATCC 26264) through a series of mutagenesis steps. The mutant produced a large quantity of Monascus pigment when grown in mantou (steamed bread) by solid culture. The mutant produced pigments extracellularly by extruding the pigments outside the cell in a lump together with some viscous substances. The productivity of pigment was about 100-fold greater than that of the wild type. The mutant lost the capability of spore formation, the growth rate decreased, and both the size and quantity of conidia were reduced. The color of the pigment produced by the mutant changed from orange to deep red.

Journal Article↗
PubMed1975

Sexal reproductive cycle of Monascus in submerged shaken culture.

A strain of Monascus was grown in submerged, shaken culture using a glucose-salts mediumn. The formation of ascospores was studied by microscopic examination of samples taken at regular intervals for 6 days. All the stages of sexual reproduction previously described for Monascus were observed. The unusual feature of the strain studied was that it produced sexual structures and cliestothecia of greatly varying sizes.

Ascomycota↗
PubMed2000

[Monascus purpureus: a new fungus of allergenic relevance].

Anaphylactic reactions to food containing allergens in the consumption or preparation of food are well known. However, allergy in the preparation of sausages have rarely been described. In the present study a 26-year-old butcher was investigated who had a severe anaphylactic reaction developing sneezing, rhinitis, conjunctivitis, generalised pruritus, followed by widespread urticaria, Quincke's oedema and dyspnoe after starting to prepare sausages containing red yield rice. Red yield rice is produced from polished and washed rice by means of the fungus Monascus purpureus. It was the first time that Monascus purpureus could be shown as allergic agent by means of prick-to-prick test, Cellular Antigen Stimulation Test (CAST) and different other immunoblots.

Adult↗
PubMed2002

Case report. Monascus purpureus--a new fungus of allergologic relevance.

Anaphylactic reactions to food containing allergens in the consumption or preparation of food are well known. However, allergy in the preparation of sausages has rarely been described. In the present study a 26-year-old-butcher was investigated who had a severe anaphylactic reaction developing sneezing, rhinitis, conjunctivitis, generalised pruritus, followed by widespread urticaria, Quincke's oedema and dyspnoe after starting to prepare sausages containing red yield rice. Red yield rice is produced by the fungus Monascus purpureus. It was the first time that Monascus purpureus could be shown as allergic agent by means of prick-to-prick test, Cellular Antigen Stimulation Test (CAST) and different immunoblots.

Adult↗
PubMed1975

Studies on the Monascus isolated from the startor of Kaoliang Brandy.

Seven strains of Monascus spp. were isolated from the startors of Kaoliang Brandy in Taiwan and Kinmen. Their morphological, physiological, and biochemical characteristics were examined. Shape of colony, length of conidial chain, production of pigment and tolerance to NaCl were considered to be suitable keys to the classification of these isolates. It was found that there were at least 6 species of Monascus in the startors of Kaoliang Brandy, 2 from Kinmen and 4 from Taiwan.

Ascomycota↗
PubMed2005

Anti-tumor-initiating effects of monascin, an azaphilonoid pigment from the extract of Monascus pilosus fermented rice (red-mold rice).

Monascin (1) constitutes one of the azaphilonoid pigments in the extracts of Monascus pilosus-fermented rice (red-mold rice). Compound 1 was evaluated for its anti-tumor-initiating activity via oral administration on the two-stage carcinogenesis of mouse skin tumor induced by peroxynitrite (ONOO-; PN) or by ultraviolet light B (UVB) as an initiator and 12-O-tetradecanoylphorbol-13-acetate (TPA) as a promoter. Compound 1 exhibited marked inhibitory activity on both PN- and UVB-induced mouse skin carcinogenesis tests. These findings suggest that compound 1 may be valuable as potential cancer chemopreventive agent in chemical and environmental carcinogenesis.

Animals↗
PubMed2006

New Monascus metabolites: structure elucidation and toxicological properties studied with immortalized human kidney epithelial cells.

Monascus sp. are used for the production of red yeast rice, which has been applied in Asian food and medicine for centuries. The fungi form several secondary metabolites, among others the monascopyridines A and B. We have now detected two new monascopyridines (named C and D) with similar UV absorption spectra and maxima at 306-307 nm in red rice fermented with M. purpureus. The new monascopyridines have the same chromophores as the already known and differ in a missing gamma-lactone ring only. Monascopyridine C has a hexanoyl side chain, whereas monascopyridine D is the higher homologue with an octanoyl side chain. The toxicological properties of monascopyridines were studied using immortalized human kidney epithelial cells, displaying cytotoxic effects in micromolar concentrations with median effective concentration values between 20.7 and 43.2 micromol/L, depending on the compound and method used. The monascopyridines C and D did not induce apoptosis. However, they caused a rise of the mitotic index from 3.21 +/- 0.27% (control) to maximum 8.14 +/- 0.89% (monascopyridine D) by an accumulation of cells in the metaphase with a simultaneous decrease of cells in the ana- and telophase. Monascopyridine-induced metaphase arrest is caused by a partial or complete loss of the spindle apparatus as indicated by the occurrence of abnormal metaphases and immunological staining of alpha-tubulin. These effects are indicating an aneuploidic potential and therefore the monascopyridines C and D might contribute to tumor formations.

Caspase 3↗
PubMed2006

Cytotoxic and antimitotic effects of N-containing Monascus metabolites studied using immortalized human kidney epithelial cells.

Recently the first Monascus metabolites with a pyridine ring were detected, the monascopyridines A and B. They are formally dehydrogenated derivatives of the red rice pigments rubropunctamine and monascorubramine. Because of their structural similarity, the toxicological effects of these secondary metabolites were studied using immortalized human kidney epithelial cells. The cytotoxicity was determined with the following different endpoint detection methods: metabolic activity, trypan blue exclusion, and electronic cell counting. The compounds led to EC(50) values between 11 and 31 micromol/L but the pigments caused a stronger reduction of the cell viability. Also, the apoptotic potential was examined by measuring caspase 3 activity and detecting apoptotic bodies, but none of the tested compounds induced apoptosis. All four substances caused a rise of the mitotic index to about 9% (100 micromol/L monascopyridine A and B) and 20% (25 micromol/L rubropunctamine and monascorubramine). The significant decrease of the ratio of cells in the ana- and telophase to cells in the prometa- and metaphase proved a stop of the mitosis at the meta- to anaphase control point. The compounds caused mitotic arrest and the formation of structural damages like c-mitosis through interaction with the mitotic spindle. These effects point to an aneuploidy inducing potential, which is linked to cancer formation.

4-Butyrolactone↗
PubMed2002

Effect of rice--glycerol complex medium on the production of Lovastatin by Monascus ruber.

Response surface methodology (RSM) was employed to study the effect of the composition of the rice-glycerol complex medium on the production of lovastatin (Lvs) by the ascomycete Monascus ruber in mixed solid-liquid (or submerged) cultures at 25 degrees C. Four components (rice powder, peptone, glycerol, glucose) were studied to evaluate the approximate polynomial for all dependent variables, explaining their effects on the production of Lvs. The best composition derived from RSM regression was (in g/L) rice powder 34.4, peptone 10.8,, glucose 129, KNO3 8.0, MgSO4.7H2O 4.0 and glycerol 36.4 mL/L. With this composition, the Lvs production was 157 mg/L after 10 d of cultivation. In comparison with glycerol and glucose, the rice powder becomes a more suitable carbon source and represents a great potential for the production of Lvs.

Glucose↗