利用变温提高汉逊酵母发酵生产谷胱甘肽的研究
详细信息Study on temperature-shift strategy on the overproduction of glutathione by Hansenula polymorpha
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摘要: 依据温度对细胞代谢过程的影响,建立了一种提高谷胱甘肽（Glutathione,GSH）产量的方法。通过单因素实验确定了多形汉逊酵母DL-1的最适生长温度、GSH合成的最佳温度和培养基初始pH分别为30、37℃和pH6.5;接着,通过变温调控分批发酵,确定最佳变温条件为:45℃培养16h后将温度降低为37℃继续培养32h,采用该方法摇瓶发酵后GSH总量及GSH含量分别为420.76mg/L和105.19mg/g,比恒温培养（37℃）分别提高了2.34倍和1.27倍,即胞内GSH积累幅度显著升高;最后,通过5L发酵罐扩大培养,生物量和GSH总量分别达42.35g/L和927.68mg/L。实验结果表明该方法可用于提高利用多形汉逊酵母的GSH产量。Abstract: An optimal two-stage temperature control strategy was established to improve the yield and the productivity of glutathione in the batch fermentation, according to the effects of temperature on cell metabolism. Through single-factor experiments, the optimum growth temperature of Hansenula polymorpha DL-1, the optimum biosynthesis temperature of GSH and initial pH value of medium were obtained. The result showed that the effect of temperature on glutathione production was different from that on cell growth and maximum glutathione was obtained at 37℃, but the optimum growth temperature was at 30℃. When the temperature shifted from 45℃to 37℃for further cultivation of 16h, a glutathione yield of 420.16mg/L and an intracellular glutathione content of 105.19mg/g were achieved, which was 2.34 and 1.27 times than that of the constant temperature fermentation, respectively. Subsequently, scale-up studies were conducted in a 5L fermenter, a cell dry weight of 42.35g/L and a glutathione content of 927.68mg/L were achieved. Therefore, the temperature-shift strategy would be a valuable alternative to improve glutathione production.
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[1] 庞德钦.高产谷胱甘肽 (GSH) 酵母菌种的选育及发酵条件研究[D].武汉:华中科技大学, 2007. [2] 陈俭梅.高产谷胱甘肽酵母菌株的选育及发酵条件研究[D].济南:山东师范大学, 2009. [3] 梁国斌.产朊假丝酵母生产谷胱甘肽过程控制与优化[D].无锡:江南大学, 2008. [4] 卫功元.发酵法生产谷胱甘肽及其高产策略研究[D].苏州:苏州大学, 2010. [5] 张文燕, 堵国城, 陈坚.流加发酵及添加L-半胱氨酸对产软假丝酵母高密度培养合成谷胱甘肽的影响[J].应用与环境微生物学报, 2007, 13 (2) :261-264. [6] 陈珊.酿酒酵母发酵生产谷胱甘肽的研究[J].现代食品科技, 2011, 27 (1) :72-76. [7] 陈凤菊, 卢善发, 胡顿孝, 等.多形汉逊酵母外源基因表达系统[J].生物工程学报, 2001, 17 (3) :247-249. [8] Wen S W, Zhang T, Tan T W.Utilization of amino acids to enhance glutathione production in Saccharomyces Cerevisiae[J].Enzyme and Microbial Technology, 2004, 35 (2) :501-507.
[9] 刘娟.高产谷胱甘肽优良酵母菌种的选育及发酵条件研究[D].北京:北京化工大学, 2003. [10] Tietze F.Enzymic method for quantitive determination of nanogram amounts of total and oxidized glutathione[J].Anal Biochem, 1969, 27 (3) :502-522.
[11] 陈娜, 乔长晟, 胡玉霞, 等.面包酵母中谷胱甘肽抽提方法的研究[J].现代食品科技, 2008, 24 (2) :157-160. [12] 卫功元, 李寅, 堵国城, 等.温度对谷胱甘肽的影响及动力学模型[J].生物工程学报, 2003, 19 (3) :258-362. [13] 王丽娜, 陈水钫, 张兵, 等.3, 5-二硝基水杨酸比色法测定多糖含量的研究进展[J].吉林医药学院学报, 2009, 8 (30) :299-301. [14] Vira M U, Viadimir M A, Alexander Y M, et al.Optimization of glutathione production in batch and fed-batch cultures by the wild-type and recombinant strains of the methylotrophic yeast Hansenula polymorpha DL-1[J].Biotechnology, 2011, 8 (11) :1472-6750.
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