title: Characterization of a thermostable β-glucuronidase from Thermotoga maritima expressed in Arabidopsis thaliana.
abstract: 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.
authors.affiliation: Agro-Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Rd., Shanghai, 201106, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xu
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Tian
authors.forename: Ri-He
authors.initials: RH
authors.lastname: Peng
authors.forename: Bo
authors.initials: B
authors.lastname: Zhu
authors.forename: Jian-Jie
authors.initials: JJ
authors.lastname: Gao
authors.forename: Quan-Hong
authors.initials: QH
authors.lastname: Yao
mesh_terms: D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression; D005966:Glucuronidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D020124:Thermotoga maritima / Q000201:enzymology* / Q000235:genetics; D013997:Time Factors
chemical_list: D011994:Recombinant Proteins; D005966:Glucuronidase