|Table of Contents|

The transcriptional regulation characteristics of xylose-inducible promoter in Bacillus licheniformis(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

Issue:
2019 03
Page:
695-701
Research Field:
Articles
Publishing date:

Info

Title:
The transcriptional regulation characteristics of xylose-inducible promoter in Bacillus licheniformis
Author(s):
LIU Xiang1 2 LI Youran1 2 ZHANG Liang1 2 DING Zhongyang1 2 XU Sha1 2 GU Zhenghua1 2 & SHI Guiyang1 2**
1 National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, China 2 School of Biotechnology, Jiangnan University, Wuxi 214122, China
Keywords:
Bacillus licheniformis xylose operon RT-qPCR glucose stress fermentation process transcriptional regulation
CLC:
TQ92: Q78
PACS:
DOI:
10.19675/j.cnki.1006-687x.201807042
DocumentCode:

Abstract:
The xylose operon is a commonly used as a expression element in Bacillus spp., but our present understanding of its function remains only at the level of its static mechanism, whereas studies on its transcriptional regulation characteristics in the fermentation process have only been done rarely. Obtaining further understanding of this operon’s characteristics during the fermentation process should help to provide a scientific basis for the application of xylose-inducible expression systems in the fermentation process. In this study, the dinitrosalicylic acid (DNS) method was used to control the glucose content during the fermentation process, and then quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) technology and high-performance liquid chromatography (HPLC) were used to accurately analyze the residual sugar content and quantify the transcription levels of the operon gene, respectively, in samples collected during the fermentation process. As a result, the glucose concentration was controlled within a desired range so that the growth and metabolism of Bacillus licheniformis in the samples were stable. Meanwhile, a systematic method was established for detecting the expression levels of the xylose-inducible promoter gene during the fermentation process, which revealed the transcriptional regulation characteristics of the xylose-inducible promoter gene in the fermentation process. The transcription level of the xylose-inducible promoter gene increased until the stationary phase, with the highest transcription level attained at the end of the logarithmic growth or pre-stabilization phase, which was increased by about 14 times after 7 h, and then after this it began to decline. Further, glucose concentrations of 20?180 g/L inhibited the transcription of this promotor gene, and the degree of inhibition was consistent across this concentration range. When the glucose content was low or zero, and xylose was present, the transcription level of the promoter was extremely high. The results of this study indicated that the xylose-inducible system with Bacillus licheniformis as the host has the best induction effect at the end of the logarithmic growth phase of microbial fermentation, and when there is little or no glucose in the environment and xylose is present conditions are even more favorable for promoter expression. These findings are significant because they could be used to guide the optimization of the application of inducible fermentation by xylose-induced recombinant Bacillus licheniformis.

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Last Update: 2019-06-25