Pepper Leaf "golden Flower" Bacteria Fermented Product And Active Research

Pepper leaves are the leaves of the Solanaceae Capsicum Capsicum annuum L. which are annual herb. At present, China is the largest country of pepper cultivation area all over the world. Pepper leaves resources are very rich as its by-product. However, it have not been effectively utilized. Eurotium Cristatum can secrete a variety of extracellular enzymes, such as polyphenol oxidase, pectinase, cellulase, protease, etc. So it can use tannin, starch and other substances as its carbon source. Therefore, we want to test using the process of fennentation by Eurotium Cristatum to transform pepper leaves’s ingredients. The research was supported by the following results:After separation and purification, we got the strains JH which in Fu brick. And it was identified as Eurotium Cristatum. Strains JH and standard strains of3.0448were contrasted by colony and individual morphological. The results show that they were no significant difference in morphology.Pepper leaves were liquid fermented by Eurotium Gristatum. By HPLC analysis we found that there are some differences between the fermentation products. Then, fermented pepper leaves with the strain of3.0448. Pre-and post-fermentation products were divided into four polar fractions (Petroleum ether, ethyl acetate, n-butanol and water extracts) by liquid-liquid extraction method. Total extract and4parts analyzed by HPLC. Results show that Pre-and post-fermentation products had great changes especially petroleum ether and ethyl acetate component.Pre-and post-fermentation products and its each component were tested the antioxidant and antibacterial activity. Anti-oxidation test results show:after fermentation the product’s ABTS (2,2’-Azinobis-(3-ethylbenzthiazoline-6-sulphonate))scavenging capacity increased20%. Pre-and post-fermentation extract showed no obvious DPPH(1,1-Diphenyl-2-picrylhydrazyl radical2,2-Diphenyl-l-(2,4,6-trinitrophenyl)hydrazyl) radical scavenging activity. Ethyl acetate component of post-fermentation ABTS, DPPH scavenging capacity are the most prominent. Its IC50is0.093mg/mL and0.082mg/mL. Post-fermentation than pre-fennentation increased300%and140%. Antibacterial test results show:N-butanol part of post-fennentation and water parts of pre-and post-fermentation has some inhibitory activity of E. coli. Ethyl acetate part of post-fermentation and the water parts of pre-and post-fermentation has the activity of a certain inhibition of Bacillus subtilis. Only the ethyl acetate part of post-fennentation can inhibit Bacillus cereus. Ethyl acetate part of post-fennentation and water parts of pre-and post-fermentation has some inhibitory activity of Staphylococcus aureus.During the phytochemical investigation, compounds were isolated from the leaves of pepper by different means, such as column chromatography(CC) with different solid-phase including silica gel and Sephadex LH-20.All the compounds were identified on the basic of NMR spectral data. These compounds are (1) apigenin,(2)4’,7,8-Trihydroxyflavone,(3)5-methoxy-4’,7-dihydroxyflavone,(4)(Z)-6-((2-(3-(hydroxymethyl)pent-l-en-3-yl)-1H-indol-3-yl)methylene)-3-methox-yl-3-methylpiperazine-2,5-dione,(5) asperflavin,(6) apigenin-7-O-β-D-glucopyrano-side apiofuranosyl (1â†'2)-β-D-glucopyranoside. Petroleum ether component was analyzed by LC/MS and identified six compounds. They are (1) Dihydroactinidiolide,(2)Methyl tetradecanoate(3)1,3,5-Trimethyl-4-n-octadecyl cyclohexane,(4) Hexadecanoic acid, methylester (5)9-Octadecenoic acid (Z)-, methyl ester,(6) Methyl stearate.For antioxidant activity, medium was optimized. The result shows:When the medium formulation is8%pepper leaves,2%sucrose, the fennentation product has the highest ABTS and DPPH scavenging rate.