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| 地芽孢杆菌Y565-5分离鉴定及其木糖异构酶基因xylA的克隆表达和酶学性质 | |
| 引用本文： | 张洁,黄志勇,王钦宏,王永莉,王硕.地芽孢杆菌Y565-5分离鉴定及其木糖异构酶基因xylA的克隆表达和酶学性质[J].微生物学通报,2011,38(8):1147-1154. |
| 作者姓名： | 张洁 黄志勇 王钦宏 王永莉 王硕 |
| 作者单位： | 1. 天津科技大学,食品营养与安全重点实验室,天津,300457;中国科学院天津工业生物技术研究所,天津,300108 2. 中国科学院天津工业生物技术研究所,天津,300108 3. 中国科学院兰州地质所,甘肃,兰州,730000 4. 天津科技大学,食品营养与安全重点实验室,天津,300457 |
| 基金项目： | 纤维素乙醇高温发酵与生物炼制子课题(No. KSCX1-YW-11E); 中国科学院知识创新工程重要方向项目(KZCX2-YW-Q05-05) |
| 摘 要： | 从甘肃玉门油田地表土中分离到一株嗜热木糖利用菌,地芽孢杆菌Y565-5。利用PCR方法从该菌株中克隆得到一个木糖异构酶基因,xylA。该基因开放阅读框长1182 bp,编码394个氨基酸,XylA氨基酸序列与Geobacillus sp.Y412MC52相似性达到99%。将xylA基因克隆到原核表达载体pET-28a(+)上,得到重组质粒pET-28a(+)-xylA,然后将此重组质粒转化至BL21(DE3)中,经IPTG诱导后,通过SDS-PAGE电泳检测出明显的45 kD(相对分子质量)特异性蛋白质条带,并且通过半胱氨酸咔唑法检测出表达产物具有木糖异构酶的活性。对其酶学性质的研究发现,XylA最适温度为90°C,最适pH值为8.0。 |
| 关 键 词： | Geobacillus sp.Y565-5 木糖异构酶 克隆 表达 |
Cloning, expression and characterization of xylose isomerase, XylA from Geobacillus sp. Y565-5 |
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| ZHANG Jie,HUANG Zhi-Yong,WANG Qin-Hong,WANG Yong-Li and WANG Shuo.Cloning, expression and characterization of xylose isomerase, XylA from Geobacillus sp. Y565-5[J].Microbiology,2011,38(8):1147-1154. | |
| Authors: | ZHANG Jie HUANG Zhi-Yong WANG Qin-Hong WANG Yong-Li and WANG Shuo |
| Institution: | 1. Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology, Tianjin 300457, China; 2. Tian-jin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300108, China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300108, China;2. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300108, China;3. Lanzhou Institute of Geology, Chinese Academy of Sciences, Lanzhou, Gansu 730000, China;1. Key Laboratory of Food Nutrition and Safety, Tianjin University of Science & Technology, Tianjin 300457, China |
| Abstract: | Xylose-utilizing and thermophilic Geobacillus sp.Y565-5 was isolated from surface soil of an oilfield in Yumen Town,Gansu Province,China.A xylose isomerase(XylA) gene was cloned from the strain by PCR.The open reading frame of xylA(1 182 bp) encoded a protein of 394 amino acids,which showed high sequence homology(99% identity) with that of Geobacillus sp.Y412MC52.The in-tact coding region was subcloned into pET28a(+) vector and expressed in Escherichia coli BL21(DE3).The molecular weight of the recombinant ... |
| Keywords: | Geobacillus sp Y565-5 xylose isomerase cloning expression |
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