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| 奶牛瘤胃微生物元基因组文库中脂肪酶的筛选与酶学性质 | |
| 引用本文： | 赵圣国,王加启,刘开朗,朱雅新,卜登攀,李旦,于萍.奶牛瘤胃微生物元基因组文库中脂肪酶的筛选与酶学性质[J].生物工程学报,2009,25(6):869-874. |
| 作者姓名： | 赵圣国 王加启 刘开朗 朱雅新 卜登攀 李旦 于萍 |
| 作者单位： | 1. 中国农业科学院北京畜牧兽医研究所动物营养学周家重点实验室,北京,100193;甘肃农业大学动物科学技术学院,兰州,730070 2. 中国农业科学院北京畜牧兽医研究所动物营养学周家重点实验室,北京,100193 3. 中国科学院微生物研究所微生物资源前期开发周家重点实验室,北京,100080 |
| 基金项目： | 国家科技支撑计划(No.2006BAD04A03)资助~~ |
| 摘 要： | 利用含有三油酸甘油酯的脂肪酶选择性筛选培养基,从奶牛瘤胃微生物元基因组文库15360个克隆中,筛选得到了18个脂肪酶阳性克隆,其插入片段大约为60kb,并且各个克隆的插入片段各不一样。利用p-NPP法对脂肪酶克隆的脂肪酶活性分析,表明均具有大小不等的脂肪酶活性。底物特异性分析表明Lipase6、Lipase7和Lipase8分别对C16底物(对硝基苯棕榈酸酯)、C12底物(对硝基苯月桂酸酯)和C16底物(对硝基苯棕榈酸酯)水解能力最强。Lipase6、Lipase7、Lipase8的脂肪酶最适pH为7.5;Lipase8的脂肪酶活性半衰期随反应温度的升高而缩短,70oC时能达到30min。本研究所筛选的脂肪酶具有不同的底物特异性和较好的热稳定性,这对于工业化生产具有一定的应用潜力。 |
| 关 键 词： | 瘤胃微生物 元基因组文库 脂肪酶 底物特异性 最适pH 热稳定性 |
| 收稿时间： | 2/9/2009 12:00:00 AM |
Screening and characterization of lipase from a metagenome library of dairy rumen microflora |
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| Shengguo Zhao,Jiaqi Wang,Kailang Liu,Yaxin Zhu,Dengpan Bu,Dan Li and Ping Yu.Screening and characterization of lipase from a metagenome library of dairy rumen microflora[J].Chinese Journal of Biotechnology,2009,25(6):869-874. | |
| Authors: | Shengguo Zhao Jiaqi Wang Kailang Liu Yaxin Zhu Dengpan Bu Dan Li and Ping Yu |
| Institution: | State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China; College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China;State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China; College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China |
| Abstract: | Using lipase segregation agar containing trioleoylglycerol,we obtained 18 lipase positive clones by screening from a metagenome library of dairy rumen microflora containing 15 360 clones.The average insert size of lipase positive clones was about 60 kb.Lipase enzyme activity analysis by p-NPP method indicated that Lipase6,Lipase7 and Lipase8 had higher lipolytic activities to substrates of p-nitrophenyl palmitate(C16),p-nitrophenyl alaurate(C12) and p-nitrophenyl palmitate(C16) respectively.The optimum pH o... |
| Keywords: | rumen bacterium metagenome library lipase substrate optimum pH thermo stability |
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