基于2-酮基-D-葡萄糖酸高产的发酵条件优化研究
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摘要
为了实现异维生素C前体2-酮基-D-葡萄糖酸在沙雷氏菌Serratia sp. FMME043中的高效生产,在30 L发酵罐中对补料策略和发酵条件进行了优化,建立了补料分批发酵工艺。最终确定发酵策略为:溶氧控制在40%,在发酵16、22 h及28 h时分别补加64、80 g和96 g硫酸铵,并且当残糖浓度降至15～25 g/L时,分五次等量补加801 g葡萄糖,优化后,发酵44 h,2-KGA的产量和转化率分别达到268.5 g/L和1.04 g/g,分别比优化前提高了58.4%和9.9%。本研究结果为目前报道的产量和转化率在国内外属领先水平,为2-酮基-D-葡萄糖酸及异维生素C工业化生产打下了坚实的基础。
In order to achieve the high production of 2-keto-D-gluconic acid(2-KGA) by Serratia sp. FMME043,feeding strategy and fermentation conditions are optimized in 30 L fermentor,and fed batch fermentation is established. The optimal fermentation conditions are as follows:the dissolved oxygen is 40%,the quality of 64,80 g,and 96 g ammonium sulfate is added at 16,22 h,and 28 h,respectively,and the quality of 801 g glucose is supplemented with five times at the residual sugar concentration between 15～25 g/L. Under these conditions,at 44 h,the production and the yield of2-KGA reached 268.5 g/L and 1.04 g/g,which are 58.4% and 9.9% higher than that not optimized,respectively.
In order to achieve the high production of 2-keto-D-gluconic acid(2-KGA) by Serratia sp. FMME043,feeding strategy and fermentation conditions are optimized in 30 L fermentor,and fed batch fermentation is established. The optimal fermentation conditions are as follows:the dissolved oxygen is 40%,the quality of 64,80 g,and 96 g ammonium sulfate is added at 16,22 h,and 28 h,respectively,and the quality of 801 g glucose is supplemented with five times at the residual sugar concentration between 15～25 g/L. Under these conditions,at 44 h,the production and the yield of2-KGA reached 268.5 g/L and 1.04 g/g,which are 58.4% and 9.9% higher than that not optimized,respectively.
引文
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[3]SUN Wenjing,ZHAO Fenmei,ZHAO Shenwang,et al. Effects of the Bacteriophages on 2-keto-D-gluconic acid fermentation in industrial process[J]. Food Science,2005,26(8):36-41.(in Chinese)
[4]ZHAO Fenmei,SUN Wenjing,WANG Muhua,et al. Selection of phage-resistant strains from 2-keto-D-gluconic acid producing strain[J]. Industial Microbiology,2000,30(4):45-49.(in Chinese)
[5]DANIEL Y N B,MARK D. Enhanced product formation in continuous fermentations with microbial cell recycle[J].Biotechnology and Bioengineering,1981,23(2):373-389.
[6]SUN Wenjing,ZHAO Fenmei,YANG Qingwen,et al. Studies on the fermentative production of 2-keto-D-gluconic acid by Pseudomonas fluorescens in fed-batch culture[J]. Food Science,2004,23(11):115-117.(in Chinese)
[7]ZHENG Zhiyong,ZHAO Zhanjun. Immobilization of Pseudomonzs cell for production of 2-keto-D-gluconie acid[J]. Journal of Shanxi Agricultural University,2003,23(2):158-160.(in Chinese)
[8]NIU Panqing,YANG Aihua,YANG Songxin,et al. Screening and identification of 2-keto-D-gluconic acid-producing strain[J].The Chinese Journal of Process Engineering,2012,12(6):1008-1013.(in Chinese)
[9]TENG W H,SUN W J,YU B,et al. Continuous conversion of rice starch hydrolysate to 2-keto-D-gluconic acid by Arthrobacter globiformis C224[J]. Biotechnology and Bioprocess Engineering,2013,18(4):709-714.
[10]冯晓燕. 2-酮基-D-葡萄糖酸补料分批发酵工艺的优化及动力学研究[D].石家庄:河北师范大学,2009.
[11]LI Jiawei,LIU Long,LI Jianghua,et al. Strategy of two-stage agitation rate for citric acid Production[J]. Journal of Food Science and Biotechnology,2014,33(2):131-136.(in Chinese)
[12]WEI Gongyuan,WANG Dahui,CHEN Jiian. Analysis of batch fermentation process of glutathione under different control modes of dissolved oxygen[J]. CIESC Jorunal,2007,9:2330-2332.(in Chinese)
[13]GAO Nianfa,ZHANG Xiianqing,WU Di. The effects of dissolve oxygen on pyruvate fermentation[J]. Journal of Tianjing University of Science&Technology,2004,19(3):5-7.(in Chinese)
[14]LIN Yaohui,BAI Zengmei,WANG Yongshun,et al. D-isoascorbic acid sodium indirect fermentation medium test[J]. Food and Fermentation Industries,1985,11(4):18-24.(in Chinese)
[15]SUN Wenjing,YANG Qingwen,HUANG Huiying,et al. Studies on the fermentation conditions of Pseudomonas fluorescens AR4Producing 2-Keto-D-gluconic Acid[J]. Food Science,2004,25(10):46-49.(in Chinese)
[16]JIANG Mingzhu,BAI Zhaoxi,ZHANG Junxian,et al. Fermentation test of 2-keto-D-gluconic acid by Arthrobacter globiformis K1022[J]. The Industrial Microbial,1989,19(1):3-7.(in Chinese)
[17]CAO Guifang,LIANG Gaiqin,YI Guanglin. Fermentation conditions of food antioxidant-D-isoascorbic acid[J]. Microbiology,1988,15(1):7-11.(in Chinese)
[18]MISENHEIMER T J ANDERSON R F,LAGODA A A,et al. Production of 2-Keto-gluconic acid by Serratia macrcescens[J].Applied Microbiology and Biotechnology,1965,13(3):393-396.
[19]ZHANG Wei,XIE Zhipeng,LUO Wei,et al. Fermentation process optimization and kinetics studies of 2-keto-D-gluconic acid production by Serratia sp. BK-98[J]. CIESC Jorunal,2011,62(5):1371-1376.(in Chinese)