|本期目录/Table of Contents|

[1]刘 翔 李由然 张 梁 丁重阳 徐 沙 顾正华 石贵阳**.地衣芽孢杆菌中木糖操纵子受葡萄糖胁迫的转录调控特性*[J].应用与环境生物学报,2019,25(02):1-10.[doi:10.19675/j.cnki.1006-687x.2018.07042]
 LIU Xiang,LI Youran,Zhang Liang,et al.The transcriptional regulation characteristics of xylose-inducible promoter in Bacillus licheniformis*[J].Chinese Journal of Applied & Environmental Biology,2019,25(02):1-10.[doi:10.19675/j.cnki.1006-687x.2018.07042]
点击复制

地衣芽孢杆菌中木糖操纵子受葡萄糖胁迫的转录调控特性*()
分享到:

《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年02期
页码:
1-10
栏目:
研究论文
出版日期:
2019-04-25

文章信息/Info

Title:
The transcriptional regulation characteristics of xylose-inducible promoter in Bacillus licheniformis*
文章编号:
201807042
作者:
刘 翔12 李由然12 张 梁12 丁重阳12 徐 沙12 顾正华12 石贵阳12**
1江南大学粮食发酵工艺与技术国家工程实验室 江苏 无锡 214122 2江南大学生物工程学院 江苏 无锡 214122
Author(s):
LIU Xiang12 LI Youran12 Zhang Liang12 Ding Zhongyang12 Xu Sha 12 Gu Zhenghua 12 SHI Guiyang12**
1 National Engineering Laboratory for Cereal Fermentation Technology, Wuxi, Jiangsu 214122, China 2 School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China
关键词:
地衣芽孢杆菌木糖操纵子实时荧光定量PCR葡萄糖胁迫发酵过程转录调控
Keywords:
Bacillus licheniformis xylose operon RT-qPCR glucose stress fermentation process transcriptional regulation
DOI:
10.19675/j.cnki.1006-687x.2018.07042
摘要:
木糖操纵子是芽孢杆菌中常用的表达元件,但目前对它的认识只停留在静态机理层面,关于它在发酵过程中转录调控特性的研究鲜见报道。进一步认识它在发酵过程中的特性,有助于对木糖诱导表达系统在地衣芽孢杆菌发酵过程中的应用给予科学的依据。本研究利用qPCR技术探究了在葡萄糖胁迫下地衣芽孢杆菌木糖诱导的木糖异构酶基因在发酵过程中的转录水平,考察了菌体的生长状态,并通过DNS法及高效液相色谱(High Performance Liquid Chromatography, HPLC)法测定了发酵过程中糖浓度变化。结果显示,在实验所设置的地衣芽孢杆菌代谢相对稳定的条件下,地衣芽孢杆菌木糖启动子转录强度在稳定期以前均呈增加的趋势,在对数生长末期或稳定前期转录强度最高,约是7 h时的14倍,随后呈下降趋势;进一步发现,20?180 g/L葡萄糖浓度均对其表现为抑制,且抑制程度一致,当葡萄糖含量极少或者没有,而木糖存在的情况下,启动子转录强度极高。本研究表明,以地衣芽孢杆菌为宿主的木糖诱导系统在菌体对数生长末期诱导效果最佳;当环境中葡萄糖含量极少或者没有,而木糖存在的情况下,更有利于启动子表达。这对利用木糖诱导型重组地衣芽孢杆菌诱导发酵有一定的启示与指导意义。
Abstract:
The xylose operon is a commonly used expression element in Bacillus spp, but the present understanding of it only stays at the static mechanism level, and studies on its transcriptional regulation characteristics in the fermentation process are rarely reported. Further understanding its characteristics in the fermentation process, is help ful to give scientific basis for the application of xylose-inducible expression system in the fermentation process. In this work, the DNS method is used to control the glucose content in the fermentation process, the RT-qPCR technology and the High Performance Liquid Chromatography (HPLC) are used to accurately analyzed residual sugar and quantify the transcriptional level in the samples collected during the fermentation process, respectively. As a result, the glucose content was controlled at the desired concentration range, the growth and metabolism of Bacillus licheniformis were stable. Meanwhile, a systematic methods was established for detecting the expression intensity of the xylose-inducible promoter gene in the fermentation process, which revealed the transcriptional regulation characteristics of the xylose-inducible promoter gene in the fermentation process. The transcriptional intensity of xylose-inducible promoter was increasing before the stationary phase, the highest transcriptional intensity was obtained at the end of logarithmic growth or pre-stabilization, which was about 14 times than 7 h, and then b egan to decline. Furtherly, 20?180 g/L glucose concentration showed inhibition and the degree of inhibition was consistent. When the glucose content was little or no, and the xylose was present , the transcriptional intensity of the promoter was extremely high. The results of this work indicated the xylose-inducible system with Bacillus licheniformis as the host has the best induction effect at the end of logarithmic growth; when the glucose content in the environment is little or no, and the presence of xylose is more favorable for promoter expression. This has certain inspirations and guiding significance for the inducible fermentation by the xylose-induced recombinant Bacillus licheniformis.? ??

相似文献/References:

[1]杨革,刘艳,李桂芝.地衣芽孢杆菌γ-谷氨酰转移酶的分离和纯化[J].应用与环境生物学报,2008,14(03):432.
 YANG Ge,et al..Purification and Characterization ofγ- Glutamyl Transferase from Bacillus licheniformis QBL-033[J].Chinese Journal of Applied & Environmental Biology,2008,14(02):432.
[2]平芮巾,孙 谧*刘均忠王跃军郝建华张胜军.响应面法优化海洋细菌MP2酯酶发酵条件[J].应用与环境生物学报,2008,14(04):544.
[3]訾楠,沈微,石贵阳,等.地衣芽孢杆菌生麦芽糖α-淀粉酶的基因克隆与鉴定[J].应用与环境生物学报,2009,15(01):130.[doi:10.3724/SP.J.1145.2009.00130]
 ZI Nan,SHEN Wei,SHI Guiyang,et al.Gene Cloning and Characterization of Bacillus licheniformis Maltogenic α-amylase[J].Chinese Journal of Applied & Environmental Biology,2009,15(02):130.[doi:10.3724/SP.J.1145.2009.00130]

备注/Memo

备注/Memo:
收稿日期: 2018-07-29 接受日期 Accepted: 2018-08-15*国家“十三五”重点研发计划(2016YFD0401400)、国家自然科学基金青年基金项目(31401674)和江苏省科技项目(BE2016628)资助 **通讯作者(E-mail: gyshi@jiangnan.edu.cn)点击摘要页题目后的“PDF”可下载阅读全文;本文为已录用的作者修定稿,尚未经编辑全面修改。引用本文请注明出处本刊;发表刊期和页码将以正式出版时的安排为准,但DOI确定不变。
更新日期/Last Update: 2018-11-14