Hayashi N R, Arai H, Kodama T, Igarashi Y
Department of Biotechnology, University of Tokyo, Japan.
Biochem Biophys Res Commun. 1997 Dec 18;241(2):565-9. doi: 10.1006/bbrc.1997.7853.
The cbbQ and cbbO genes are located downstream from the RubisCO genes (cbbLS) in the thermophilic hydrogen-oxidizing bacterium, Pseudomonas hydrogenothermophila. Recombinant RubisCO enzymes were purified from E. coli cells which were transformed with plasmids expressing cbbLS, cbbLSQ, cbbLSO, or cbbLSQO. Co-expression of cbbQ and/or cbbO with cbbLS made the maximal rates of carboxylation (Vmax) of the recombinant RubisCOs about two-fold higher than that of the enzyme derived from only cbbLS. The RubisCOs with high Vmax also had a high stability when undergoing ultrasonic treatment. The results of the circular dichroism spectra and the 8-anilino-1-naphthalenesulfonate binding assay indicated that these recombinant RubisCOs were conformationally different to each other.
在嗜热氢氧化细菌嗜热氢假单胞菌中,cbbQ和cbbO基因位于核酮糖-1,5-二磷酸羧化酶/加氧酶基因(cbbLS)的下游。从用表达cbbLS、cbbLSQ、cbbLSO或cbbLSQO的质粒转化的大肠杆菌细胞中纯化重组核酮糖-1,5-二磷酸羧化酶。cbbQ和/或cbbO与cbbLS共表达使重组核酮糖-1,5-二磷酸羧化酶的最大羧化速率(Vmax)比仅由cbbLS产生的酶高出约两倍。具有高Vmax的核酮糖-1,5-二磷酸羧化酶在进行超声处理时也具有高稳定性。圆二色光谱和8-苯胺基-1-萘磺酸盐结合测定的结果表明,这些重组核酮糖-1,5-二磷酸羧化酶在构象上彼此不同。