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首页> 外文期刊>Journal of Applied Glycoscience >Insertion of a Loop Structure into the “Loopless” GH19 Chitinase from Bryum coronatum
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Insertion of a Loop Structure into the “Loopless” GH19 Chitinase from Bryum coronatum

机译:将环状结构插入冠状番茄的“无环” GH19几丁质酶中

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Chitinases belonging to the GH19 family have diverse loop structure arrangements. A GH19 chitinase from rye seeds (RSC-c) has a full set of (six) loop structures that form an extended binding cleft from -4 to +4 (“loopful”), while that from moss (BcChi-A) lacks several loops and forms a shortened binding cleft from -2 to +2 (“loopless”). We herein inserted a loop involved in sugar residue binding at subsites +3 and +4 of RSC-c (Loop-II) into BcChi-A (BcChi-A+L-II), and the thermal stability and enzymatic activity of BcChi-A+L-II were then characterized and compared with those of BcChi-A. The transition temperature of thermal unfolding decreased from 77.2 ?C (BcChi-A) to 63.3 ?C (BcChi-A+L-II) by insertion of Loop-II. Enzymatic activities toward the chitin tetramer (GlcNAc)4 and the polymeric substrate glycol chitin were also suppressed by the Loop-II insertion to 12 and 9 %, respectively. The Loop-II inserted into BcChi-A was found to be markedly flexible and disadvantageous for protein stability and enzymatic activity.
机译:属于GH19家族的几丁质酶具有不同的环结构安排。来自黑麦种子(RSC-c)的GH19几丁质酶具有完整的(六个)环状结构,形成从-4到+4的扩展结合裂隙(“环状”),而来自苔藓(BcChi-A)的几丁质缺乏循环并形成从-2到+2的缩短的结合裂隙(“无环”)。我们在本文中将一个涉及糖残基结合的环插入RSc-c(Loop-II)的亚位+3和+4(BcChi-A + L-II),以及BcChi-A的热稳定性和酶活性。然后鉴定A + L-II并与BcChi-A进行比较。通过插入Loop-II,热解折叠的转变温度从77.2°C(BcChi-A)降至63.3°C(BcChi-A + L-II)。通过Loop-II插入分别将甲壳素四聚体(GlcNAc)4和聚合底物二醇甲壳质的酶活性抑制到12%和9%。发现插入BcChi-A的Loop-II具有明显的柔韧性,并且对蛋白质稳定性和酶活性不利。

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