溶氧浓度对rhG-CSF工程菌高密度发酵的影响
Effect of dissolved oxygen on high density fermentation of E.coli in producing rhG-CSF
-
摘要: 目的 确定rhG-CSF工程菌高密度发酵的最佳溶氧浓度。 方法 通过通入纯氧,高密度发酵的溶氧值可以精确控制。考察不同溶氧浓度对工程菌的生长、质粒丢失率、乙酸积累情况以及rhG-CSF表达的影响,来确定溶氧的最佳控制范围。 结果 工程菌生长阶段溶氧控制在25%,诱导后控制在35%,最有利于菌体生长和外源蛋白表达,最终菌体密度为(79.6±2.1)g/L,表达量为(40.2±1.2)%。 结论 溶氧对工程菌rhG-CSF高密度发酵有显著影响。Abstract: Objective To study the density of the dissolved oxygen(DO) on fermentation of E.coli in producing rhG-CSF. Methods Pure oxygen was used to control DO precisely and to observe the growth of engineered bacteria, the lost of plasmids ,the accumulation of acetic acid from different DO density. The optimum density of DO for fermentation was determined. Results 25% in growth phase and 30% after induction was optimum density of DO for rhG-CSF expression and engineered bacteria growth. Under these conditions, the wet cell weight reached(79.6±2.1) g/L and the expression of rhG-CSF was(40.2±1.2)%. Conclusions The growth of engineered bacteria and the expression of rhG-CSF were influenced by the density of dissolved oxygen significantly.
-
Key words:
- rhG-CSF /
- high density fermentation /
- dissolved oxygen
-
[1] 张 力,李龙芸,林宏英,等,重组人粒细胞集落刺激因子对肿瘤患者白细胞的影响[J].中国临床药学杂志,2001,10(4):207. [2] 乌 垠,杨 红,陆 军,等,重组人粒细胞集落刺激因子在Escherichia coli中的表达研究[J].东北师大学报2000,32(4):46. [3] 李 寅,高海军,陈 坚.高细胞密度发酵技术[M].北京:化学工业出版社,2006: 4. [4] 卢圣栋,主编.现代分子生物学实验技术[M],北京:高等教育出版社,1993:590. [5] Yee L, Blanch HW. Recombinant trypsin production in high cell density fed-batch cultures in Escherichia coli[J]. Biotechnol Bioeng, 1993, 41(8): 781. [6] 俞俊棠,唐孝宣.生物工艺学[M].上海:华东化工学院出版社,1994. [7] Suare DC, Kilikian BV. Acetic acid accumulation in aerobic growth of recombinant Escherichia coli[J]. Process Biochemistry, 2000, 35: 1051. [8] 常海燕,范代娣,骆艳娥. 重组大肠杆菌高密度发酵生产类人胶原蛋白Ⅱ条件优化[J].微生物学通报,2009, 36(6): 870. [9] Meyer HP, Iechter AF. Production of cloned human leukocyt interferon by Bacillus subtilis: Optimal production is connected with rest rained growth[J]. Appl Environ Mi-crobiol, 1985, 50: 503. [10] Phue J.N, Shiloach J. Impact of dissolved oxygen concentration on acetate accumulation and physiology of E. coli BL21, evaluating transcription levels of key genes at different dissolved oxygen conditions[J]. Metabolic Engi-neering, 2005, 7: 353. [11] Yee L, Blanch HW. Recombinant trypsin production in high cell density fed-batch cultures in Escherichia coli[J]. Biotechnol Bioeng, 1993, 41(8): 781.
计量
- 文章访问数: 2784
- HTML全文浏览量: 205
- PDF下载量: 163
- 被引次数: 0