Cloning, Expression and Characterization Studies on Xylanase Gene, xynA15, from Alicyclobacillus sp.A15

doi:10.3969/j.issn.1008-0864.2010.06.20

›› 2010, Vol. 12 ›› Issue (6): 114-119.DOI: 10.3969/j.issn.1008-0864.2010.06.20

Previous Articles Next Articles

Cloning, Expression and Characterization Studies on Xylanase Gene, xynA15, from Alicyclobacillus sp.A15

WANG Jian-she1,2, BAI Ying-guo2, YIN Jun1, YAO Bin2

(1.College of Life Sciences, Inner Mongolia Agricultural University, Hohhot|2.Key Laboratory of Feed Biotechnology,
Ministry of Agriculture, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China)

Received:2010-03-22 Revised:2010-06-22 Online:2010-12-15 Published:2010-09-15

脂环酸芽孢杆菌Alicyclobacillus sp. A15木聚糖酶基因xynA15的克隆表达及性质研究

王建设1,2,柏映国2,尹俊1,姚斌2

(1.内蒙古农业大学生命科学学院, 呼和浩特 010018|2.中国农业科学院饲料研究所, 农业部饲料生物技术重点开放实验室, 北京 100081)

通讯作者: 姚斌,研究员,博士,博士生导师,从事微生物工程研究。Tel:010-82106053; E-mail: yaobin@caas-bio.net.cn。尹俊,教授,博士,硕士生导师,从事动物分子遗传学研究。E-mail: yinjunparis@163.com
作者简介:王建设,硕士研究生,从事微生物遗传学研究。E-mail: wongjaction@163.com。
基金资助:
国家转基因生物新品种培育重大专项(2008ZX003-002);国家肉鸡产业技术体系(nycytx-42-G2-05)资助。

Abstract

Abstract:

Alicyclobacillus sp.A15 was isolated from the outflow of a hot spring in Baoshan City, Yunnan Province, China. xynA15 was cloned from the strain through homogolous cloning and TAIL-PCR, which consists of 990 bp and encodes 329 amino acids and one stop codon. Then it was inserted into the prokaryotic expression vector pET-22b (+) and transformed into Escherichia coli BL21 (DE3). Xylanase activity was detected in the cultured supernantant after induced by Isopropyl-β-d-1-thiogalactopyranoside (IPTG). The optimal temperature and pH was 50℃ and 7.0, respectively for XynA15 activity. XynA15 contains higher activity at 35~60℃ and better thermal stability at 50℃.

Key words: Alicyclobacillus sp.A15, xylanase, enzymology character, prokaryotic expression

摘要：

从云南保山市的一眼温泉中分离到一株嗜热嗜酸菌,脂环酸芽孢杆菌Alicyclobacillus sp.A15。通过同源克隆和TAIL-PCR方法,从该菌株中克隆得到一个木聚糖酶基因,xynA15,全长990 bp,编码329个氨基酸和一个终止密码子。将xynA15基因克隆到原核表达载体pET-22b(＋)上,并转化至BL21(DE3),经过IPTG诱导,其表达产物具有木聚糖酶的活性。对其酶学性质研究发现,XynA15的最适温度为50℃,最适pH值为70,不仅在较广的温度范围（35℃~60℃）内具有较高的酶活,而且在50℃具有较好的热稳定性。

关键词: Alicyclobacillus sp.A15;木聚糖酶;酶学性质;原核表达

CLC Number:

Q786

WANG Jian-she1,2, BAI Ying-guo2, YIN Jun1, YAO Bin2. Cloning, Expression and Characterization Studies on Xylanase Gene, xynA15, from Alicyclobacillus sp.A15[J]. , 2010, 12(6): 114-119.

王建设1,2,柏映国2,尹俊1,姚斌2. 脂环酸芽孢杆菌Alicyclobacillus sp. A15木聚糖酶基因xynA15的克隆表达及性质研究[J]. , 2010, 12(6): 114-119.

[1]	SUI Fu1,2, LIU Xiaolin1,2, XIE Zhihong1,3*. Sensing Mechanism of Receptor TlpA1 to Succinic Acid in Azorhizobium caulinodans ORS571 [J]. Journal of Agricultural Science and Technology, 2020, 22(10): 77-84.
[2]	YANG Wenhan, GUO Xiaojing, CHEN Yiqun, LV Junnan, XIE Fei, CAO Yunhe*. Studies on Secreted Expression of an Acidic Xylanase in Pichia pastoris and Evaluation of its in vitro Activity [J]. , 2013, 15(5): 59-66.
[3]	LI Yanan1, YU Lihong1, LI Yunkai1, MA Wenkang, ZHANG Wei2*. Gene Cloning and Analysis of Enzymology Property of an Thermophilic Xylanase from Humicola insolens Y1 [J]. , 2013, 15(4): 121-128.
[4]	CHENG Fei-fei1,2, ZHAO Jun-qi2, SHI Peng-jun2, LI Jiang1, YAO Bin2. Cloning, Enzymology Characteristics Analysis of a Xylanase Gene from Phialophora sp. P13 [J]. , 2012, 14(1): 85-90.
[5]	CUI Yan-fang, SUN Pei-long, LIU Xin-qi. Effect Comparison of EIAV gp45 and HIV gp120 Genes Using Different Expressional Systems [J]. , 2011, 13(4): 72-78.
[6]	ZHANG Fan1,2, SHI Peng-jun2, MIAO Li-hong1, YAO Bin2. Studies on Cloning of Acid Xylanase from Phialophora sp. G5 and its Characteristics [J]. , 2011, 13(3): 60-66.
[7]	KUI Hong1, LUO Hui-ying2, DONG Shou-liang1, YAO Bin2. Gene Cloning, Expression and Characterization of \|an Acidic Xylanase from Bispora betulina [J]. , 2010, 12(5): 109-115.
[8]	LUO Jian-jie1, WANG Ya-ru1, YUAN Tie-zheng1, BAI Ying-guo1, HUANG Huo-qing1, LUO. High-efficiency Expression of Xylanase XYN-W with High Specific Activity in Pichia pastoris [J]. , 2010, 12(5): 80-85.
[9]	LIU Yu1,2, YAN Cai-xia1, ZHANG Ting-ting1, LI Chun-juan1, . Cloning and Prokaryotic Expression of Peanut NBS-LRR Resistant Gene [J]. , 2010, 12(3): 73-78.
[10]	LIU Xin, SHI Peng-jun, YANG Pei-long, YAO Bin. Research Progress on Bifunctional Xylanases [J]. , 2010, 12(2): 50-56.
[11]	LI Rui-qin1, FENG Shu-dan1, YU Ying2, LV Zhao-zhi1, LI Li1, XU Ming-hua1, YIN Yu. Cloning, Bioinformatics Analysis and Prokaryotic Expression of Chitinase ClassⅡ Gene in Leymus chinensis [J]. , 2010, 12(2): 103-110.
[12]	LIU Xiao-dan1,2, YANG Pei-long2, LIU Yong-chao3, XU Xiu-hong2, . Studies on the Gene Cloning and its Enzymology Characteristics of a Neutral β-Mannanase from Paenibacillus sp.A1 [J]. , 2009, 11(5): 60-65.
[13]	YAO Guo-yu, LI Ning, SHI Peng-jun, CHEN Qiang, YAO Bin. Cloning, Expression and Characterization of a Xylanase Gene, xynBS9, from Streptomyces sp. S9 [J]. , 2009, 11(4): 64-70.
[14]	LIU Quan, LI Guang-yue, ZENG Hong-mei, YANG Xiu-fen, QIU De-wen. Acquisition of Microbial Protein Elicitor PeaT1 and Preliminary Research on Inducing Drought Resistance of Rice [J]. , 2009, 11(3): 51-55.
[15]	DUAN Wen-juan, YANG Jiao-yan, LI Zhuo-fu, ZHANG Wei\| ZHANG Zhi-fang . Expression of β-Galactosidase from Pyrococcus furiosus \|in E. coli and Analysis of Lactase Properties [J]. , 2008, 10(2): 76-81.

Cloning, Expression and Characterization Studies on Xylanase Gene, xynA15, from Alicyclobacillus sp.A15

脂环酸芽孢杆菌Alicyclobacillus sp. A15木聚糖酶基因xynA15的克隆表达及性质研究

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics