Induction of hypocrellin production by Triton X-100 under submerged fermentation with Shiraia sp. SUPER-H168.
ABSTRACT Hypocrellins are important photodynamic therapy compounds for cancer disease. The effect of surfactants on hypocrellin production of Shiraia sp. SUPER-H168 was evaluated under submerged fermentation condition. The production of hypocrellins could reach 780.6 mg/l with the addition of Triton X-100, confirmed by color reaction, high performance liquid chromatography, electrospray ionization mass spectrometry and nuclear magnetic resonance experiments. According to our observation, treatment of the culture at the beginning of the fermentation was most effective, and the yield of hypocrellins was much lower with the addition of Triton X-100 during the log phase and stationary phase. Shiraia sp. SUPER-H168 could not produce hypocrellin with the addition of other tested surfactants, such as Tween 40, Triton X-114 and SDS. The experimental results indicated that Shiraia sp. SUPER-H168 could not produce hypocrellins without Triton X-100 under submerged fermentation condition.
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ABSTRACT: Bamboos, regarded as therapeutic agents in ethnomedicine, have been used to inhibit inflammation and enhance natural immunity for a long time in Asia, and there are many bamboo associated fungi with medical and edible value. In the present study, a total of 350 fungal strains were isolated from the uncommon moso bamboo (Phyllostachys edulis) seeds for the first time. The molecular diversity of these endophytic fungi was investigated and bioactive compound producers were screened for the first time. All the fungal endophytes were categorized into 69 morphotypes according to culturable characteristics and their internal transcriber spacer (ITS) regions were analyzed by BLAST search with the NCBI database. The fungal isolates showed high diversity and were divided in Ascomycota (98.0%) and Basidiomycota (2.0%), including at least 19 genera in nine orders. Four particular genera were considered to be newly recorded bambusicolous fungi, including Leptosphaerulina, Simplicillium, Sebacina and an unknown genus in Basidiomycetes. Furthermore, inhibitory effects against clinical pathogens and phytopathogens were screened preliminarily and strains B09 (Cladosporium sp.), B34 (Curvularia sp.), B35 (undefined genus 1), B38 (Penicillium sp.) and zzz816 (Shiraia sp.) displayed broad-spectrum activity against clinical bacteria and yeasts by the agar diffusion method. The crude extracts of isolates B09, B34, B35, B38 and zzz816 under submerged fermentation, also demonstrated various levels of bioactivities against bambusicolous pathogenic fungi. This study is the first report on the antimicrobial activity of endophytic fungi associated with moso bamboo seeds, and the results show that they could be exploited as a potential source of bioactive compounds and plant defense activators. In addition, it is the first time that strains of Shiraia sp. have been isolated and cultured from moso bamboo seeds, and one of them (zzz816) could produce hypocrellin A at high yield, which is significantly different from the other strains published.PLoS ONE 04/2014; 9(4):e95838. · 3.53 Impact Factor
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ABSTRACT: Some endophyte isolates were isolated in a bamboo pole sample parasitized the fungus Shiraia bambusicola from Zhejiang Province. After screening through hypocrellin bacteriostatic effect and fermentation test, we got the isolate TX4 of bacterial elicitor and GZUIFR-TT1 of fungal elicitor which had certain effect to promote S. bambusicola to produce hypocrellin. The Plackett-Burman design was introduced to evaluate the effects of nine factors based on single-factor test. Yeast extract, glucose, and isolate GZUIFR-TT1 elicitor were found to be the critical activity factors for increasing the total hypocrellin production. So we identified the isolate GZUIFR-TT1 as Trametes sp. Through response surface methodology, we obtained the optimum production conditions as follows: yeast extract, 2.99 g/L; glucose, 32.45 g/L; and Trametes sp. elicitor, 81.40 μg/mL. Under the above conditions, the experimental value of hypocrellin production was 102.60 mg/L, compared with the control it increased about 7.90 times.Folia Microbiologica 11/2012; · 1.15 Impact Factor
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ABSTRACT: A rapid and sensitive analytical method based on reverse-phase high-performance liquid chromatography was first developed to simultaneously determine elsinochrome C (EC) and hypocrellin A (HA) in the submerged fermentation. The mobile phase consisted of acetonitrile-water 60:40 (v/v) with a flow-rate of 1 mL/min. The calibration curves were as follows: y = 37,625x + 249,775 for EC, y = 30,813x + 556,409 for HA and linear at the investigated concentration. The correlation coefficients (R(2) ) were 0.9989 and 0.9998 respectively for EC and HA. The limits of detection and quantification were 175 and 585 µg/L for EC and 205 and 610 µg/L for HA. The precisions of concentration and retention times were less than 2.5 and 0.3%. The recovery of the method was greater than 95.0%. The methodology was applied to analyze simultaneously EC and HA concentrations in a submerged fermentation, and was adequate for analysis of biosynthesis of perylenequinones. The method was also amplified to separate and purify EC and HA using a semi-preparative C(18) column. In addition, elsinochrome C was first identified in the submerged fermentation broth of Shiraia sp. SUPER-H168.Biomedical Chromatography 10/2011; 26(6):737-42. · 1.95 Impact Factor