The Structure Of Bioactive Components Of Red Kojic And Its Health-Care Function Evaluation

The main focuses of the paper are to extract bioactive components, i. e, Lovastatin and Monascus pigments from red kojic at a time. And the four fractions, Monascus yellow pigment, Lovastatin, alcohol-soluble Monascus red pigment and water-soluble Monascus red pigment, was obtained by silica gel column and identified their structures. The paper evaluated the anti-fatigue function and blood lipid regulation of alcohol-soluble Monascus red pigment (RK3) for the first time and probed into the effect of alcohol-soluble Monascus red pigment and Lovastatin on the gene expression of lipoprotein lipase (LPL) mRNA and low density lipoprotein receptor (LDLR) mRNA by adapting molecular-biological technology. The main results are as follows:1. Fermentation of functional red kojic with low-yielding Citrinin The red kojic was prepared by using mutagenic strain from Monascuspurpureus with the liquid-solid two-step fermentation technology. The content of Citrinin in fermentation product was lower than 0.1mg/kg ( the lowest quantity is 0. Img/kg) while the content of Lovastatin is from 0. 25 to 0.4 mg/kg by HPLC analysis.2. Extraction and Isolation of bioactive components from red kojic A new technology of extraction and purification for Lovastatin fromRed kojic was set up, i. e., extracted with ethyl acetate and purified by neutral aluminium oxide column. Collecting the light-color fraction eluted by petroleum ether- ethyl acetate (1: 1, v/v) as eluent , then concentrated, dried by vacuum at 40, N-hexane cleaned, dissolved in hot benzene again, cooled and crystallized. Then recrystallized in acetone contained water, the crystal of Lovastatin was obtained.A new isolated program of three-stage adverse current was used to extract Lovastatin and Monascus pigments from red kojic with 70% ethanol. And the functional mixture of Monascus pigments was prepared for the first time. In the mixture the content of Lovastatin is 1. 23mg/g and Citrinin lower than 0.1mg/kg, the production rate is higher than 16%.The four pure fractions , Monascus yellow pigment, Lovastatin, alcohol-soluble Monascus red pigment and water-soluble Monascus red pigment was prepared from the mixture pigments by silica gel column. No Citrinin was inspected out of the Monascus yellow pigment or Monascus red pigment obtained. The production rate of Monascus yellow pigment was 3. 56% and Monascus red pigment 6.64 %.3. The structure characteristics of bioactive substanceThe crystal of Lovastatin obtained is proved to be identify with literature by UV, IR, HPLC, HPLC-MS,HNMR and CNMR.It discovered that the Monascus yellow pigments mixture was made up of 4 yellow pigments by HPLC-MS, HPLC analysis. They are Monascine , Ankaflavine, and another two new Monascus yellow pigments. Their molecular weight is 332 and 360, formulas C19H24O5 and C21H28O5 respectively.The mixture of alcohol-soluble Monascus red pigment was composed of three main components, i.e., Monascorubramine, Rubropunctamine and a new Monascus red pigment by HPLC-MS, HPLC analysis. The molecular weight as new Monascus red pigment was 397, a possible formula was C23H27NO5.4. Effect of Lovastatin from Red Kojic on lipid metabolisn and its molecular mechanism in Hyperlipidemic MiceThe results showed that:(l) the Lovastatin test groups(15mg/kg. d, 30mg/kg. d) significantly decreased the serum TC, TG and LDL-C levels (p<0. 01) and the Lovastatin test groups (5mg/kg. d, 15mg/kg. d, 30mg/kg. d) significantly enhanced serum HDL-C level (p<0. 01) than high-fat control group; (2)the Lovastatin test groups(5mg/kg. d, 15mg/kg. d, 30mg/kg. d) significantly lowered serum AI (p<0. 01) than the high-fat control group; (3) TC, TG, MDA levels in the liver of test mice with Lovastatin test groups (5mg/kg. d, 15mg/kg. d, 30mg/kg. d) were significantly lowered(p<0.01) than in high-fat control group; (4) Lovastatin increased the activity of LPL mRNA with dose-dependency in the liver of test mice.The expression of LPL mRNA gene in the liver of testing mice of Lovastatin groups(5mg/kg. d, 15mg/kg. d and 30mg/kg.