酿酒酵母工程菌发酵产青蒿酸的工艺优化
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摘要
通过酿酒酵母工程菌(Saccharomyces cerevisiae 1211)发酵制备青蒿酸,并通过单因素实验和响应面优化,考察了发酵温度、pH、半乳糖质量浓度、发酵碳源及发酵氮源等对青蒿酸发酵产量的影响。结果表明:在发酵温度30℃,发酵培养基初始pH=5.5,发酵培养基中蔗糖质量浓度91.8 g/L,半乳糖质量浓度10.1 g/L,硫酸铵质量浓度10.3 g/L,磷酸二氢钾质量浓度8.7 g/L的条件下,青蒿酸发酵产量可达(1529.7±12.6)mg/L,与未优化时的发酵产量相比,提升了67.1%。
The fermentation medium and conditions of artemisinic acid produced by engineered S.cerevisiae 1211, were optimized in this paper. The yield of artemisinic acid was used as evaluation index to investigate the effects of temperature, pH, carbon source, nitrogen source and galactose concentration on the fermentation process by using single factor experiments and response surface methodology.The results showed that the production of artemisinic acid could reach(1529.7±12.6) mg/L under the conditions of temperature 30 ℃, p H=5.5, sucrose 91.8 g/L, galactose 10.1 g/L, ammonium sulfate 10.3 g/L and monopotassium phosphate 8.7 g/L, which was increased by 67.1% compared with that in the initial medium.This study provided some reference for the further development of industrial production.
The fermentation medium and conditions of artemisinic acid produced by engineered S.cerevisiae 1211, were optimized in this paper. The yield of artemisinic acid was used as evaluation index to investigate the effects of temperature, pH, carbon source, nitrogen source and galactose concentration on the fermentation process by using single factor experiments and response surface methodology.The results showed that the production of artemisinic acid could reach(1529.7±12.6) mg/L under the conditions of temperature 30 ℃, p H=5.5, sucrose 91.8 g/L, galactose 10.1 g/L, ammonium sulfate 10.3 g/L and monopotassium phosphate 8.7 g/L, which was increased by 67.1% compared with that in the initial medium.This study provided some reference for the further development of industrial production.
引文
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[3]Fr?hlich T,Ndreshkjana B,Muenzner J K,et al.Synthesis of novel hybrids of thymoquinone and artemisinin with high activity and selectivity against colon cancer[J].ChemMedChem,2017,12(3):226-234.
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[8]Kong J Q,Wang W,Cheng K D,et al.Research progresses in synthetic biology of artemisinin[J].Acta pharmaceutica Sinica,2013,48(2):193-205.
[9]Paddon C J,Westfall P J,Pitera D J,et al.High-level semi-synthetic production of the potent antimalarial artemisinin[J].Nature,2013,496(7446):528-532.
[10]Ro D K,Paradise E M,Ouellet M,et al.Production of the antimalarial drug precursor artemisinic acid in engineered yeast[J].Nature,2006,440(7086):940-943.
[11]Kung S H,Lund S,Murarka A,et al.Approaches and recent developments for the commercial production of semi-synthetic artemisinin[J].Frontiers in Plant Science,2018,9:87.
[12]Lenihan J R,Tsuruta H,Diola D,et al.Developing an industrial artemisinic acid fermentation process to support the cost-effective production of antimalarial artemisinin-based combination therapies[J].Biotechnology Progress,2008,24(5):1026-1032.
[13]Paddon C J,Keasling J D.Semi-synthetic artemisinin:A model for the use of synthetic biology in pharmaceutical development[J].Nature Reviews Microbiology,2014,12(5):355.
[14]Martin V J,Pitera D J,Withers S T,et al.Engineering a mevalonate pathway in Escherichia coli for production of terpenoids[J].Nature biotechnology,2003,21(7):796.
[15]Wang Dong(王冬),Dai Zhubo(戴住波),Zhang Xueli(张学礼).Production of plant-derived natural products in yeast cells-Areview[J].Acta Microbiologica Sinica(微生物学报),2016,56(3):516-529.
[16]Guo Rui(郭睿),Ding Mingzhu(丁明珠),Yuan Yingjin(元英进).Construction of artificial yeast cell for producing amorphadiene,andoptimization of fermentation[J].Journal of Chemical Industry and Engineering(China)(化工学报),2015,66(1):378-385.
[17]Wu Tao(吴涛),Wu Shengming(吴胜明),Yin Qing(殷晴),et al.Biosynthesis of amorpha-4,11-diene,a precursor of the antimalarial agent artemisinin,in Escherichia coli through introducing mevalonate pathway[J].Chinese Journal of Biotechnology(生物工程学报),2011,27(7):1040-1048.
[18]Chen W,Ma L,Zhou P,et al.A novel feedstock for biodiesel production:The application of palmitic acid from schizochytrium[J].Energy,2015,86:128-138.
[19]Xu Su(徐速),Sun Libin(孙立斌),Zhao Qingxia(赵清霞),et al.Optimization of solid-state fermentation process on antibiotic alternative bacillus subtilis probiotics[J].Journal of Chinese Institute of Food Science and Technology(中国食品学报),2016,16(8):132-139.
[20]Wang Dahong(王大红),Wei Lanlan(韦兰兰),Zhang Shuaiying(张帅滢),et al.Optimization of fermentation medium for producing strain of natamycin using response surface methodology[J].Fine Chemicals(精细化工),2017,34(11):1233-1238,1245.
[21]Zhang Qianwen(张倩雯),Wang Chunxia(王春霞),Yang Liyun(杨丽芸),et al.Research on the factors affecting Saccharomyces cerevisiae fermentation[J].China Brewing(中国酿造),2015,34(12):14-19.
[22]Li Hua(李华),Shan Zuhua(单祖华),Wang hua(王华).Inhibition effect of carbon dioxide on the growth and metabolism of Saccharomyces cerevisiae[J].Science&Technology Review(科技导报),2008,26(5):31-34.