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Screening and Fermentation Optimization of Corynebacterium acetoacidophilum Mutant for High-level Production of L-proline |
LU Wei-ning1, LIAO Yu-ling1,2 |
1. Star Lake Bioscience Corporation Incorporated, Zhaoqing 526040,China;
2. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006,China |
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Abstract Corynebacterium acetoacidophilum was used as the original strain for stepwise mutation by whole genome fragmentation mutation (UV and NaNO2) and subsequent sulfaguanidine resistance screening. A L-proline over-producing mutant was obtained. The optimum contents of glucose, biotin and thiamin in shake-flask fermentation were 16%, 300μg/L and 400μg/L respectively, and pH is 6.8~7.0, where 25ml medium was cultured in 500ml shake-flask. After 72h by flask-shaking batch fermentation, the production of L-proline reached 75.6g/L, 5% higher than with the control. To investigate the effect of cell growth on the L-proline production, 50L fed-batch culture was performed at the specific growth rate (μ) of 0.06/h,0.08/h and 0.1/h.The results showed that the specific L-proline production rate reached 0.091g/(g.h) and the highest production was 82.1g/L, increased by 14% compared with the control when the specific growth rate (μ) was 0.08/h.
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Received: 15 July 2010
Published: 19 November 2010
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[1] 张伟国. L-脯氨酸产生菌的选育. 无锡轻工大学学报, 1996, 15 (1): 44-48. Zhang W G.Journal of Wuxi University of Light Industry,1996,15(1):44-48.
[2] 郑素慧, 李文军, 娄凯. L-脯氨酸发酵生产的研究进展. 新疆农业科学, 2007, 44 (S2): 6-10. Zheng S H,Li W J,Lou K. Xinjiang Agricultural Science,2007,44(S2):6-10.
[3] 郑素慧, 李文军, 娄凯. L-脯氨酸摇瓶发酵条件的优化. 食品研究与开发, 2008, 29 (6): 61-66. Zheng S H,Li W J,Lou K.Food Research and Development,2008,29(6):61-66.
[4] 张伟国, 顾正华, 唐鸿. L-脯氨酸发酵条件的研究. 江苏食品与发酵, 2000, 4 (103): 32-36. Zhang W G,Gu Z H,Tang H.Jiangsu Food and Fermentation,2000,4(103):32-36.
[5] Yoshinaga F. Method of producting L-proline by Fermentation. US3819483, 1974.
[6] Tsuchida T,Kubota K,Yoshinage F,et al. Improvement of L-proline production by sulfaguanidine resistant mutants derived from L-glutamic acid-producing bacteria. Agric Biol Chem, 1986, 50(9): 2201-2207.
[7] 谢畅丰, 陈超菊, 林远声. 发酵后处理方法研究 (Ⅺ)-L-脯氨酸高产酸研究. 中山大学学报论丛, 1998, (4): 22-26. Xie C F,Chen C J,Lin Y S.Supplement to the Journal of Sun Yatsen University,1998,4:22-26.
[8] 郑素慧. L-脯氨酸发酵条件的优化. 新疆:新疆农业大学食品科学学院, 2008. Zheng S H.Optimization of L-proline Fermentation Conditions.Xinjiang:Xinjig Agricultural University,Food Science,2008.
[9] 方佩静, 毛维颖, 陈琦. L-脯氨酸发酵研究. 微生物学报, 1982, 22 (4): 353-360. Fang P J,Mao W Y,Chen Q.Acta Microbiologica Sinica,1982,22(4):353-360.
[10] 褚志义, 史济平, 胡英杰. L-脯氨酸生物合成条件的研究.上海第一医学院学报, 1984, 11(4): 261-266. Zhu Z Y,Shi J P,Hu Y J.Acta Academiae Medicinae Primae Shanghai,1984,11(4):261-266.
[11] Jungoh Ahn. Effect of Growth Rate on the production of L-Proline in the Fed-batch Culture of Corynebacterium acetoacidophilum. Biotechnology and Bioprocess Engineering, 2004, 9: 326-329.
[12] Toshide Nakanishi. Process for Producing L-proline by Fermentation. US4444885, 1983.
[13] Toshide Nakanishi. A process for producing L-proline by Fermentation. GB2077262A, 1981. |
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