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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短杆菌的“无环” GH19几丁质酶中

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摘要

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℃ (BcChi-A) to 63.3℃ (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多种环结构安排。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（“ loopful”），而来自苔藓（BcChi-A）的缺乏循环并形成从-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℃（BcChi-A）降至63.3℃（BcChi-A + L-II）。通过Loop-II插入分别抑制了对几丁质四聚体（GlcNAc）_4和聚合底物二醇几丁质的酶活性，分别为12％和9％。发现插入到BcChi-A中的Loop-II具有明显的柔性，并且不利于蛋白质的稳定性和酶活性。

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