Abstract

The endophytic fungus Berkleasmium sp. Dzf12, isolated from Dioscorea zingiberensis, was found to produce palmarumycins C12 and C13 which possess a great variety of biological activities. Seven biocompatible water-immiscible organic solvents including n-dodecane, n-hexadecane, 1-hexadecene, liquid paraffin, dibutyl phthalate, butyl oleate and oleic acid were evaluated to improve palmarumycins C12 and C13 production in suspension culture of Berkleasmium sp. Dzf12. Among the chosen solvents both butyl oleate and liquid paraffin were the most effective to improve palmarumycins C12 and C13 production. The addition of dibutyl phthalate, butyl oleate and oleic acid to the cultures of Berkleasmium sp. Dzf12 significantly enhanced palmarumycin C12 production by adsorbing palmarumycin C12 into the organic phase. When butyl oleate was fed at 5% (v/v) in medium at the beginning of fermentation (day 0), the highest palmarumycin C12 yield (191.6 mg/L) was achieved, about a 34.87-fold increase in comparison with the control (5.3 mg/L). n-Dodecane, 1-hexadecene and liquid paraffin had a great influence on the production of palmarumycin C13. When liquid paraffin was added at 10% (v/v) in medium on day 3 of fermentation, the palmarumycin C13 yield reached a maximum value (134.1 mg/L), which was 4.35-fold that of the control (30.8 mg/L). Application of the aqueous-organic solvent system should be a simple and efficient process strategy for enhancing palmarumycin C12 and C13 production in liquid cultures of the endophytic fungus Berkleasmium sp. Dzf12.

Highlights

  • Plant endophytic fungi are micro-organisms which colonize the interior parts of host plants, typically causing no apparent symptoms of pathogenesis and being well recognized as a promising source of bioactive compounds [1,2,3]

  • Seven organic solvents including n-dodecane, n-hexadecane, 1-hexadecene, liquid paraffin, dibutyl phthalate (DBP), butyl oleate and oleic acid were respectively added to the culture broth of Berkleasmium sp

  • We report for the first time the enhancement of palmarumycin C12 and C13 production by the endophytic fungus Berkleasmium sp

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Summary

Introduction

Plant endophytic fungi are micro-organisms which colonize the interior parts of host plants, typically causing no apparent symptoms of pathogenesis and being well recognized as a promising source of bioactive compounds [1,2,3]. The palmarumycins C12 and C13, the two main spirobisnaphthalenes, were found to be released from the intracellular to the extracellular medium Some attempts such as medium optimization as well as the application of metal ions, macroporous resins and elicitors have been made to effectively improve palmarumycin C12 and C13 production [18,41,42,43,44]. Theaim of the present work was to explore the effects of seven biocompatible organic solvents, including n-dodecane, n-hexadecane, 1-hexadecene, liquid paraffin, dibutyl phthalate (DBP), butyl oleate and oleic acid on the production of palmarumycin C12 and C13 in liquid cultures of the endophytic fungus Berkleasmium sp. The most effective solvent with its addition time and concentration could be selected for application in the aqueous-organic two-phase culture process for maximizing palmarumycin C12 and C13 production

Results and Discussion
Endophytic Fungus and Culture Conditions
Organic Solvents and Two-Phase Culture
Measurement of Mycelia Biomass and Palmarumycins C12 and C13 Yield
Statistical Analysis
Conclusions
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