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Method of heat-stabilizing α-glucan phosphorylase(GP)

机译:热稳定α-葡聚糖磷酸化酶(GP)的方法

摘要

An α-glucan phosphorylase having improved thermostability, which obtained by modifying natural α-glucan phosphorylase, and a method for producing this α-glucan phosphorylase having improved thermostability are provided. The natural α-glucan phosphorylase is derived from a plant, this α-glucan phosphorylase having improved thermostability has an amino acid residue which is different from that of the natural α-glucan phosphorylase in at least one position selected from the group consisting of a position corresponding to position 4 in a motif sequence 1L or 1H, a position corresponding to position 4 in a motif sequence 2, and a position corresponding to position 7 in a motif sequence 3L or 3H, and wherein the enzyme activity of α-glucan phosphorylase having improved thermostability at 37° C., after heating in a 20 mM citrate buffer (pH 6.7) at 60° C. for 10 minutes, is 20% or more of the enzyme activity of the α-glucan phosphorylase having improved thermostability at 37° C., before heating.
机译:本发明提供了通过对天然α-葡聚糖磷酸化酶进行修饰而得到的热稳定性提高的α-葡聚糖磷酸化酶以及该热稳定性提高的α-葡聚糖磷酸化酶的制造方法。天然α-葡聚糖磷酸化酶来自植物,该热稳定性提高的α-葡聚糖磷酸化酶在选自由以下组成的组中的至少一个位置具有与天然α-葡聚糖磷酸化酶不同的氨基酸残基。对应于基序序列 1 L或 1 H中位置 4 的位置,对应于A中位置 4 的位置基序序列 2 ,以及基序序列 3 L或 3 H中与位置 7 相对应的位置,以及其中,在20 mM柠檬酸盐缓冲液(pH 6.7)中于60°C加热10分钟后,在37°C下具有改善的热稳定性的α-葡聚糖磷酸化酶的酶活性为该酶的20%或更高。加热前在37℃下具有改善的热稳定性的α-葡聚糖磷酸化酶。

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