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首页> 外文期刊>Journal of Experimental Botany >Maize C4-form phosphoenolpyruvate carboxylase engineered to be functional in C3 plants: mutations for diminished sensitivity to feedback inhibitors and for increased substrate affinity
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Maize C4-form phosphoenolpyruvate carboxylase engineered to be functional in C3 plants: mutations for diminished sensitivity to feedback inhibitors and for increased substrate affinity

机译：在C 3 植物中起作用的玉米C 4 -型磷酸烯醇丙酮酸羧化酶：突变导致对反馈抑制剂的敏感性降低以及底物亲和力增加

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摘要

Introducing a C4-like pathway into C3 plants is one of the proposed strategies for the enhancement of photosynthetic productivity. For this purpose it is necessary to provide each component enzyme that exerts strong activity in the targeted C3 plants. Here, a maize C4-form phosphoenolpyruvate carboxylase (PEPC, EC 4.1.1.31) was engineered for its regulatory and catalytic properties so as to be functional in the cells of C3 plants. Firstly, amino acid residues Lys-835 and Arg-894 of maize PEPC, which correspond to Lys-773 and Arg-832 of Escherichia coli PEPC, respectively, were replaced by Gly, since they had been shown to be involved in the binding of allosteric inhibitors, malate or aspartate, by our X-ray crystallographic analysis of E. coli PEPC. The resulting mutant enzymes were active but their sensitivities to the inhibitors were greatly diminished. Secondly, a Ser residue (S780) characteristically conserved in all C4-form PEPC was replaced by Ala conserved in C3- and root-form PEPCs to decrease the half-maximal concentration (S0.5) of PEP. The double mutant enzyme (S780A/K835G) showed diminished sensitivity to malate and decreased S0.5(PEP) with equal maximal catalytic activity (Vm) to the wild-type PEPC, which will be quite useful as a component of the C4-like pathway to be introduced into C3 plants.
机译：将C 4 样途径引入C 3 植物是提高光合作用生产力的一种策略。为此，有必要提供在目标C 3 植物中发挥强活性的每种酶。在此，对玉米C 4 型磷酸烯醇丙酮酸羧化酶（PEPC，EC 4.1.1.31）进行了工程改造，使其具有调控和催化特性，从而可以在C 3 细胞中发挥作用。 >植物。首先，玉米PEPC的氨基酸残基Lys-835和Arg-894（分别对应于大肠杆菌PEPC的Lys-773和Arg-832）被Gly取代，因为已证明它们参与了与PEPE的结合。通过我们对大肠杆菌PEPC的X射线晶体学分析得出苹果酸或天冬氨酸等变构抑制剂。所得的突变酶是有活性的，但是它们对抑制剂的敏感性大大降低了。其次，将在所有C 4 型PEPC中特征性保守的Ser残基（S780）替换为在C 3 -和根型PEPC中保守的Ala，以减少半衰期。 PEP的最大浓度（S 0.5 ）。双突变酶（S780A / K835G）对苹果酸的敏感性降低，S 0.5 （PEP）降低，最大催化活性（V m ）与野生型PEPC相等，它将作为引入C 3 植物的C 4 样途径的组成部分非常有用。

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    《Journal of Experimental Botany》 |2008年第7期|p.1811-1818|共8页
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    Tatsuya Endo1*† Yuko Mihara2† Tsuyoshi Furumoto23 Hiroyoshi Matsumura4 Yasushi Kai4‡ and Katsura Izui125§;

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    1Laboratory of Plant Physiology, Faculty of Agriculture, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan 2Laboratory of Plant Physiology, Graduate School of Biostudies, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502, Japan 3Department of Biological Science, Graduate School of Science, Hiroshima University, 1-3-1 Kagamiyama, Higashihiroshima, Hiroshima 739-8526, Japan 4Department of Materials Chemistry, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan 5Department of Biotechnological Science, Kinki University, 930 Nishimitani, Kinokawa, Wakayama 649-6493, Japan;

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