产紫杉醇内生真菌枝状枝孢霉MD2的发酵条件优化  

Optimization of fermentation conditions of Cladosporium cladosporioides MD2 for 10-DAB and taxol production

作　　者：苗莉云[1] 张鹏[2,3] 周蓬蓬 余龙江[2] 

机构地区：[1]运城学院生命科学系,山西运城044099 [2]华中科技大学生命科学与技术学院资源生物与生物技术研究所,湖北武汉430074 [3]湖北省农业科学研究院经济作物研究所,湖北武汉430064

出　　处：《微生物学通报》2013年第6期1033-1040,共8页Microbiology China

基　　金：国家自然科学基金项目(No.20776058;20906036);中国博士后科学基金项目(No.20100471183);运城学院自然科学基金项目(No.CY2012004)

摘　　要：【目的】通过优化内生真菌枝状枝孢霉MD2的发酵条件,提高10-去乙酰巴卡亭Ⅲ(10-DAB)和紫杉醇(Taxol)的产量。【方法】采用单因素试验分析不同的培养基初始pH值、培养温度、摇床转速和培养时间对10-DAB和紫杉醇产量的影响,优化枝状枝孢霉MD2的培养条件;以YES为基本培养基,采用单因素试验和正交试验分析添加苯甲酸钠、苯丙氨酸、丝氨酸和甘氨酸4种前体物对10-DAB和紫杉醇产量的影响,优化枝状枝孢霉MD2的培养基组分。【结果】优化后发酵条件为:在初始pH为5.0的300 mL YES培养基中,添加15 mg/L苯甲酸钠、25 mg/L苯丙氨酸、5 mg/L丝氨酸、15 mg/L甘氨酸,接种1 mL枝状枝孢霉MD2的孢子悬液(107 108个孢子/mL),28.0°C、220 r/min发酵培养12 d。在此条件下,枝状枝孢霉MD2的生物量、10-DAB和紫杉醇的产量分别为15.5 g/L、471.5μg/L和569.5μg/L,与初始发酵条件相比,分别提高了1.3、3.6和3.4倍。【结论】首次获得了枝状枝孢霉MD2生产10-DAB和紫杉醇的较适摇瓶发酵条件,可为进一步放大发酵培养提供参考。[Objective] The objective of this study was to increase the yield of 10-deacetylbaccatin III （10-DAB） and taxol in Cladosporium cladosporioides MD2 through optimizing fermentation conditions. [Methods] The culture conditions of C. cladosporioides MD2 were optimized by single factor experiments to investigate the effect of different initial pH values of medium, culture temperatures, rotational speeds, and culture time on the yield of 10-DAB and taxol. The fermentation medium components were optimized by single factor and orthogonal experiments to investigate the effect of four precursors on the yield of 10-DAB and taxol. [Resultsl The optimization results were that 1 mL spore suspension of C. cladosporioides MD2 （107-108 spores） was inoculated into 300 mL YES medium with the ini- tial pH of 5.0, which was supplemented with 15 mg/L sodium benzoate, 25 mg/L L-phenylalanine, 5 mg/L serine and 15 mg/L glycine, and then cultured at 28.0 ℃ and 220 r/min for 12 days. Under the optimized fermentation conditions, the biomass, 10-DAB yield and taxol yield of C. cladosporioides MD2 were respectively 15.5 g/L, 471.5 μg/L and 569.5 μg/L, about 1.3 times, 3.6 times and 3.4 times of that under the initial fermentation con- ditions. [Conclusion] A suitable fermentation condition of C. cladosporioides MD2 for 10-DAB and taxol production in flask culture was obtained for the first time, and these results provided a reference for its scale-up fermentation.

关 键 词：内生真菌 枝状枝孢霉MD2 10-去乙酰巴卡亭Ⅲ 紫杉醇 发酵条件 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]