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Sequence and transcriptional analysis of the genes responsible for curdlan biosynthesis in Agrobacterium sp. ATCC 31749 under simulated dissolved oxygen gradients conditions

  • Applied Microbial and Cell Physiology
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Abstract

Expression at the mRNA level of ten selected genes in Agrobacterium sp. ATCC 31749 under various dissolved oxygen (DO) levels during curdlan fermentation related to electron transfer chain (ETC), tricarboxylic acid (TCA) cycle, peptidoglycan/lipopolysaccharide biosynthesis, and uridine diphosphate (UDP)-glucose biosynthesis were determined by qRT-PCR. Experiments were performed at DO levels of 30%, 50%, and 75%, as well as under low-oxygen conditions. The effect of high cell density on transcriptional response of the above genes under low oxygen was also studied. Besides cytochrome d (cyd A), the transcription levels of all the other genes were increased at higher DO and reached maximum at 50% DO. Under 75% DO, the transcriptional levels of all the genes were repressed. In addition, transcription levels of icd, sdh, cyo A, and fix N genes did not exhibit significant fluctuation with high cell density culture under low oxygen. These results suggested a mechanism for DO regulation of curdlan synthesis through regulation of transcriptional levels of ETCs, TCA, and UDP-glucose synthesis genes during curdlan fermentation. To our knowledge, this is the first report that DO concentration apparently regulates curdlan biosynthesis in Agrobacterium sp. ATCC 31749 providing essential lead for the optimization of the fermentation at the industrial scale.

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Acknowledgments

The authors declare that they have no conflict of interest. We would like to thanks Dr. João Carlos Setubal (Virginia Bioinformatics Institute) for providing us with helpful advice on the manuscript preparation. This work was supported by the research grants from the National Natural Science Foundation of China (20676055, 60604028, and 20806034), and Ministry of Science and Technology of China (National Basic Research Program of China, 2007CB714303 and 2011BAD23B00). The work was also supported by the Program for Introducing Talents of Discipline to the Universities, no. 111-2-06. The authors would like to thank these organizations for their financial support.

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  1. Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Hong-Tao Zhang, Xiao-Bei Zhan, Zhi-Yong Zheng, Jian-Rong Wu, Xiao-Bin Yu, Yun Jiang & Chi-Chung Lin

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  1. Hong-Tao Zhang
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Correspondence to Xiao-Bei Zhan.

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S1

PCR detection of the ETC component genes in the genome Agrobacterium sp. ATCC 31749. Agarose gel analysis of products of PCR reactions is performed with the total DNA as template and the primers listed in “Materials and methods”. The abbreviations of gene names are indicated above each lane. “M” indicated DNA molecular size marker (100 bp DNA Ladder, Takara) (JPEG 84 kb)

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Zhang, HT., Zhan, XB., Zheng, ZY. et al. Sequence and transcriptional analysis of the genes responsible for curdlan biosynthesis in Agrobacterium sp. ATCC 31749 under simulated dissolved oxygen gradients conditions. Appl Microbiol Biotechnol 91, 163–175 (2011). https://doi.org/10.1007/s00253-011-3243-1

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