caa a22 4500 605500371 CHVBK 20210128100555.0 cr unu---uuuuu 210128e20150401xx s 000 0 eng 10.1007/s00253-014-6362-7 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6362-7 Two UDP-glucuronic acid decarboxylases involved in the biosynthesis of a bacterial exopolysaccharide in Paenibacillus elgii [Elektronische Daten] [Ou Li, Chao-Dong Qian, Dao-qiong Zheng, Pin-Mei Wang, Yu Liu, Xin-Hang Jiang, Xue-Chang Wu] Xylose is described as a component of bacterial exopolysaccharides in only a limited number of bacterial strains. A bacterial strain, Paenibacillus elgii, B69 was shown to be efficient in producing a xylose-containing exopolysaccharide. Sequence analysis was performed to identify the genes encoding the uridine diphosphate (UDP)-glucuronic acid decarboxylase required for the synthesis of UDP-xylose, the precursor of the exopolysaccharide. Two sequences, designated as Peuxs1 and Peuxs2, were found as the candidate genes for such enzymes. The activities of the UDP-glucuronic acid decarboxylases were proven by heterologous expression and real-time nuclear magnetic resonance analysis. The intracellular activity and effect of these genes on the synthesis of exopolysaccharide were further investigated by developing a thymidylate synthase based knockout system. This system was used to substitute the conventional antibiotic resistance gene system in P. elgii, a natural multi-antibiotic resistant strain. Results of intracellular nucleotide sugar analysis showed that the intracellular UDP-xylose and UDP-glucuronic acid levels were affected in Peuxs1 or Peuxs2 knockout strains. The knockout of either Peuxs1 or Peuxs2 reduced the polysaccharide production and changed the monosaccharide ratio. No polysaccharide was found in the Peuxs1/Peuxs2 double knockout strain. Our results show that P. elgii can be efficient in forming UDP-xylose, which is then used for the synthesis of xylose-containing exopolysaccharide. Springer-Verlag Berlin Heidelberg, 2015 UDP-glucuronic acid decarboxylases nationallicence Bacterial exopolysaccharide nationallicence Paenibacillus elgii nationallicence Gene knockout nationallicence Autotrophic mutant strain nationallicence Li Ou College of Life Sciences, Zhejiang Sci-Tech University, 310018, Hangzhou, People's Republic of China aut Qian Chao-Dong College of Life Sciences, Zhejiang Chinese Medical University, 310053, Hangzhou, People's Republic of China aut Zheng Dao-qiong Ocean College, Zhejiang University, 310058, Hangzhou, People's Republic of China aut Wang Pin-Mei Ocean College, Zhejiang University, 310058, Hangzhou, People's Republic of China aut Liu Yu Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut Jiang Xin-Hang Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut Wu Xue-Chang Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 3127-3139 0175-7598 99:7<3127 2015 99 253 https://doi.org/10.1007/s00253-014-6362-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6362-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Li Ou College of Life Sciences, Zhejiang Sci-Tech University, 310018, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Qian Chao-Dong College of Life Sciences, Zhejiang Chinese Medical University, 310053, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Zheng Dao-qiong Ocean College, Zhejiang University, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Wang Pin-Mei Ocean College, Zhejiang University, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Liu Yu Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Jiang Xin-Hang Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Wu Xue-Chang Institute of Microbiology, College of Life Sciences, Zhejiang University, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/7(2015-04-01), 3127-3139 0175-7598 99:7<3127 2015 99 253