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| 利用重组大肠杆菌合成几丁寡糖的研究 | |
| 引用本文： | 张大伟,楚杰,郝永任,李明华,王鹏. 利用重组大肠杆菌合成几丁寡糖的研究[J]. 生物工程学报, 2007, 23(3): 525-529 |
| 作者姓名： | 张大伟 楚杰 郝永任 李明华 王鹏 |
| 作者单位： | 1. 山东省应用微生物技术重点实验室,济南250014;山东大学微生物技术国家重点实验室,济南,250100 2. 山东省应用微生物技术重点实验室,济南250014 3. 山东大学微生物技术国家重点实验室,济南,250100 |
| 摘 要： | 提取根瘤菌Mesorhizobium.loti基因组,克隆编码N-乙酰氨基葡萄糖转移酶nodC基因,插入质粒pUC19的lac启动子的下游,构建并筛选出能够合成几丁寡糖的重组大肠杆菌DCL-3。利用优化的MMYNG培养基,重组大肠杆菌DCL-3在10L发酵罐中培养26h后,培养液菌体浓度测定OD560=10.8,几丁寡糖得率达到526mg/L。收集重组细菌的细胞并煮沸破碎,利用活性炭的吸附和P4凝胶层析对几丁寡糖产物进行分离纯化。纯化产物的液质分析(LC-ESI-MS)结果表明主要寡糖产物为几丁四糖(m/z,831[M H] )和几丁五糖(m/z,1034[M H] )。 |
| 关 键 词： | 几丁寡糖 N-乙酰氨基葡萄糖转移酶 重组大肠杆茵 液质联用技术 |
| 文章编号： | 1000-3061（2007）03-0525-05 |
| 修稿时间： | 2006-10-302006-11-28 |
Biosynthesis of Chitooligosaccharides by Recombinant Escherichia coli |
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| ZHANG Da-Wei,CHU Jie,HAO Yong-Ren,LI Ming-Hua and WANG Peng. Biosynthesis of Chitooligosaccharides by Recombinant Escherichia coli[J]. Chinese journal of biotechnology, 2007, 23(3): 525-529 | |
| Authors: | ZHANG Da-Wei CHU Jie HAO Yong-Ren LI Ming-Hua WANG Peng |
| Affiliation: | Shandong Key Laboratory of Microbial Technology, Shandong Academy of Sciences, Jinan 250014, China;State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China;Shandong Key Laboratory of Microbial Technology, Shandong Academy of Sciences, Jinan 250014, China;Shandong Key Laboratory of Microbial Technology, Shandong Academy of Sciences, Jinan 250014, China;Shandong Key Laboratory of Microbial Technology, Shandong Academy of Sciences, Jinan 250014, China;State Key Laboratory of Microbial Technology, Shandong University, Jinan 250100, China |
| Abstract: | Acetyl-N-glucosaminyltransferase gene (nodC) was successfully cloned to Escherichia coli from Mesorhizobium loti. The recombinant E.coli harboring nodC gene was able to synthesize chitooligosaccharides (COs) in MMYNG medium. In optimized condition, a yield of 526 mg/L was obtained after 26 h cultivation in 10 L bioreactor. COs concentration reached up to 4.5% of the cell dry weight. The COs products were purified by charcoal adsorption and Bio-gel P4 chromatography, penta-N-acetylchitopentaose (m/z, 1034[M H] )and tetra-N-acetylchitopentaose (m/z, 831[M H] ) were identified as the dominating COs product using the method of liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS). |
| Keywords: | chitooligosaccharides acetyl-N-glycosaminyltransferase recombinant E.coli LC-ESI-MS coupling technique |
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