Characterization of a thermostable β-glucuronidase from Thermotoga maritima expressed in Arabidopsis thaliana.

Jing Xu, Yong-Sheng Tian, Ri-He Peng, Bo Zhu, Jian-Jie Gao, Quan-Hong Yao

Agro-Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd., Shanghai, 201106, China.

Applied microbiology and biotechnology 2012 Sep

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TmGUSI, a gene identical to that encoding a thermostable β-glucuronidase in the hyperthermophilic anaerobe Thermotoga maritima, has been synthesized using a PCR-based two-step DNA synthesis and codon optimization for plants, and expressed in both Escherichia coli and Arabidopsis thaliana. TmGUSI expressed in transformed E. coli cells exhibited maximum hydrolytic activity at 65 °C and pH 6.5 and retained more than 80% activity after incubation at 85 °C for 30 min. TmGUSI activity in transgenic A. thaliana plants containing TmGUSI was also stable over the temperature range 65-80 °C. Our data suggest that β-glucuronidase from T. maritima can serve as a useful thermostable marker in higher plants.

Citation

Jing Xu, Yong-Sheng Tian, Ri-He Peng, Bo Zhu, Jian-Jie Gao, Quan-Hong Yao. Characterization of a thermostable β-glucuronidase from Thermotoga maritima expressed in Arabidopsis thaliana. Applied microbiology and biotechnology. 2012 Sep;95(5):1211-9