烟曲霉素5 L罐发酵条件优化及中试
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摘要
以烟曲霉(Aspergillus fumigatus)CEA-1701为生产菌株,通过高效液相色谱(HPLC)法测定发酵液中烟曲霉素的含量,考察碳源、pH值、搅拌速度、发酵温度和接种量对5 L发酵罐中烟曲霉素产量的影响,优化了5 L罐的发酵工艺条件。结果表明,甘油作为唯一的碳源,发酵温度30℃,搅拌速度800 r/min,接种量为3%,自然pH条件下,通气量0.5 m~3/h,烟曲霉CEA-1701产烟曲霉素效果理想,发酵168 h烟曲霉素产量达到97.76 mg/L。将优化后的工艺进行20 L罐的放大试验,0.5%接种量发酵252 h时烟曲霉素获得最高产量达到130.57 mg/L,中试放大效果良好。
Using Aspergillus fumigatus CEA-1701 as the production strain, the content of fumagillin in the fermentation broth was determined by HPLC, and the effect of the carbon source, pH value, stirring speed, fermentation temperature and inoculum on the fumagillin production in the 5 L fermenter were investigated, and the fermentation process in 5 L fermenter was optimized. The results showed that wsing glycerol as the sole carbon source, fermentation temperature 30 ℃, stirring speed 800 r/min, inoculum 3%, natural pH, ventilatory volume 0.5 m~3/h, and the fumagillin production reached 97.76 mg/L at 168 h by A. fumigatus CEA-1701. The optimized process was subjected to a 20 L fermenter pilot-scale test. The highest fumagillin production was 130.57 mg/L with 0.5% inoculum and fermentation time 252 h, and the effect of scale-up experiment was good.
Using Aspergillus fumigatus CEA-1701 as the production strain, the content of fumagillin in the fermentation broth was determined by HPLC, and the effect of the carbon source, pH value, stirring speed, fermentation temperature and inoculum on the fumagillin production in the 5 L fermenter were investigated, and the fermentation process in 5 L fermenter was optimized. The results showed that wsing glycerol as the sole carbon source, fermentation temperature 30 ℃, stirring speed 800 r/min, inoculum 3%, natural pH, ventilatory volume 0.5 m~3/h, and the fumagillin production reached 97.76 mg/L at 168 h by A. fumigatus CEA-1701. The optimized process was subjected to a 20 L fermenter pilot-scale test. The highest fumagillin production was 130.57 mg/L with 0.5% inoculum and fermentation time 252 h, and the effect of scale-up experiment was good.
引文
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[22]吴升山.黑曲霉DB056产柚苷酶发酵工艺的研究[D].厦门:集美大学,2010.
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[2]BAILEY L.Effect of fumagillin upon Nosema apis(Zander)[J].Nature,1953,171(4344):212-213.
[3]HIGES M,NOZAL M J,ALVARO A,et al.The stability and effectiveness of fumagillin in controlling Nosema ceranae(Microsporidia)infection in honey bees(Apis mellifera)under laboratory and field conditions[J].Apidologie,2011,42(3):364-377.
[4]SALEH M,KUMAR G,ABDEELBAKI A A,et al.Development of a novel in vitro method for drug development for fish;application to test efficacy of antimicrosporidian compounds[J].Veterinary Record,2014,175(22):561.
[5]KENT M L,SHAW R W,SANDERS J L.Microsporidia in Fish[M].Microsporidia.Wiley-Blackwell,2014.
[6]INGBER D,FUJITA T,KISHIMOTO S,et al.Synthetic analogues of fumagillin that inhibit angiogenesis and suppress tumour growth[J].Nature,1990,348(6301):555-557.
[7]RAVINDRAN B,WONG J W,SELVAM A,et al.Microbial biodegradation of proteinaceous tannery solid waste and production of a novel value added product-metalloprotease[J].Bioresource Technol,2016,217:150-156.
[8]MOLINA J M,GOGUEL J,SARFATI C,et al.Trial of oral fumagillin for the treatment of intestinal microsporidiosis in patients with HIV infection[J].Aids,2000,14(10):1341.
[9]VAN DER HEEVER J P,THOMPSON T S,CURTIS J M,et al.Fumagillin:an overview of recent scientific advances and their significance for apiculture[J].J Agric Food Chem,2014,62(13):2728-2737.
[10]BRA KENHIELM E,CAO R,GAO B,et al.Angiogenesis inhibitor,TNP-470,prevents diet-induced and genetic obesity in mice[J].Circul Res,2004,94(12):1579-1588.
[11]LIJNEN H R,FREDERIX L,VAN HOEF B.Fumagillin reduces adipose tissue formation in murine models of nutritionally induced obesity[J].Obesity,2010,18(12):2241.
[12]AN J,WANG L,PATNODE M L,et al.Physiological mechanisms of sustained fumagillin-induced weight loss[J].Jci Insight,2018,3(5).
[13]KIM E J,SHIN W H.General pharmacology of CKD-732,a new anticancer agent:effects on central nervous,cardiovascular,and respiratory system[J].Biol Pharmaceut Bull,2005,28(2):217-223.
[14]HUANG W F,BOCQUET M,LEE K C,et al.The comparison of r DNAspacer regions of Nosema ceranae isolates from different hosts and locations[J].J Invert Pathol,2008,97(1):9-13.
[15]SANG J S,AHN J B,PARK K S,et al.A Phase Ib pharmacokinetic study of the anti-angiogenic agent CKD-732 used in combination with capecitabine and oxaliplatin(XELOX)in metastatic colorectal cancer patients who progressed on irinotecan-based chemotherapy[J].Investigat New Drug,2012,30(2):672-680.
[16]曹葳蕤,薛晓锋,吴黎明.烟曲霉素及其防治蜜蜂微孢子虫病的研究进展[J].中国农业科技导报,2016,18(2):52-58.
[17]杨思佳.家蚕微孢子虫Met-AP2基因验证与烟曲霉素治疗效果检测[D].广州:华南农业大学,2016.
[18]余龙江.发酵工程原理与技术应用[M].北京:化学工业出版社,2006.
[19]肖安风,倪辉,吴升山,等.黑曲霉产柚苷酶的发酵条件优化[J].中国食品学报,2011,11(6):89-97.
[20]唐诗潮,李晓凤,袁琨.不同培养条件对黑曲霉细胞生长及水解特性的影响研究[J].现代食品科技,2017(8):195-200.
[21]陈杰.海洋芽孢杆菌B-9987发酵工艺优化及初步放大[D].上海:华东理工大学,2013.
[22]吴升山.黑曲霉DB056产柚苷酶发酵工艺的研究[D].厦门:集美大学,2010.
[23]PETERSON M H,GOLDSTEIN A W,DENISON F W.p H control in production of fumagillin:US 2803586 A[P].1957-08-20.