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提高Pyrococcus horikoshii OT3糖原分枝酶的热稳定性和分枝效率

Jing Zhu¹, Jie Long¹, Xingfei Li¹

¹The State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China; School of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi 214122, China; Collaborative Innovation Center of Food Safety and Quality Control in Jiangsu Province, Jiangnan University, Wuxi 214122, China.

International journal of biological macromolecules

|November 18, 2023

概括

工程化糖原分支酶 (GBEs) 显示了增强的耐热性,使粉在高温下能够有效地进行修改. 这一突破改善了GBE在食品工业中的应用.

科学领域:

生物化学生化学
酵素工程是什么? 酶工程是什么
食品科学食品科学食品科学

背景情况:

粉修饰需要具有高热稳定的酶,因为工业凝化温度很高.
目前的葡萄糖分枝酶 (GBEs) 具有有限的耐热性,限制了它们在粉修饰中的使用.
Pyrococcus horikoshii OT3 GBE 是一种高热性酶,但实际应用需要进一步改善热量.

研究的目的:

使用蛋白质工程策略,提高Pyrococcus horikoshii OT3 GBE的热稳定性.
为了研究突变对GBE活动,稳定性和粉修饰能力在高温下的影响.

主要方法:

利用PROSS策略和PDBePISA分析来识别和设计GBE二极管接口中的关键残留物.
创造了特定的突变 (例如,T508K,H415W,V140I) 来提高耐热性.
评估了酶活性,化温度,热处理后的残留活性和粉修饰效率 (α-1.6 糖化键含量).

主要成果:

突变T508K的化温度增加了3.1°C. 突变T508K的化温度增加了3.1°C.
对大多数突变物来说,最佳反应温度增加了10°C.
组合突变在95°C60小时后保持了40%以上的活性,与野生型GBE不同.
突变变异的玉米粉呈现出显著更高的α-1,6甘氨酸键含量和增加的短葡萄糖链 (DP < 13).

结论:

PROSS策略有效地提高了Pyrococcus horikoshii OT3 GBE的热稳定性.
工程 GBEs 证明了在工业食品应用中高效粉分支的潜力.
这种方法为开发用于生物技术过程的热稳定酶提供了一种可行的方法.

关键词:

酶工程是什么?酶工程是什么?糖原分支酶酶是如何分支的普洛斯斯 (Prossus) 是一个生长的植物.热稳定性热稳定性

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