Improving the thermostability of Pseudoalteromonas Porphyrae κ-carrageenase by rational design and MD simulation.
Improving the thermostability of Pseudoalteromonas Porphyrae κ-carrageenase by rational design and MD simulation.
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DOI：
10.1186/s13568-024-01661-z
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发表时间：
2024-01-20
期刊：
影响因子：
3.7
通讯作者：
Zhu, Yanbing
中科院分区：
文献类型：
--
作者：
Sang, Yuyan;Huang, Xiaoyi;Li, Hebin;Hong, Tao;Zheng, Mingjing;Li, Zhipeng;Jiang, Zedong;Ni, Hui;Li, Qingbiao;Zhu, Yanbing
关键词：
κ-卡拉胶酶的工业应用因其较差的热稳定性而受到限制。在这项研究中，基于折叠自由能变化（ΔΔG）和使用分子动力学（MD）模拟对来自紫菜假交替单胞菌（WT）的碱性κ-卡拉胶酶KCgCD进行的灵活性分析，突变体S190R被鉴定为具有改善的热稳定性。 50°C孵育30分钟后，S190R的残留活性为63.7%，比WT高25.7%。通过差示扫描量热法测定的 S190R 和 WT 的 Tm 值分别为 66.2 °C 和 64.4 °C。 S190R 的最适温度比 WT 高 10 °C。与未经处理的κ-卡拉胶相比，S190R 产生的κ-卡拉胶水解产物显示出更高的黄嘌呤氧化酶抑制活性。 S190R的MD模拟分析表明，F5中的残基（V186-M194和P196-G197）和F3中的关键残基R150显示出柔性下降，而靠近催化中心的T169-N173残基显示出柔性增加。这些改变的灵活性可能是突变体S190R热稳定性提高的原因。本研究提供了一种结合ΔΔG计算和MD模拟的合理设计策略，以提高κ-卡拉胶酶的热稳定性，使其更好的工业应用。 基于ΔΔG和MD模拟，通过合理设计鉴定出突变体κ-卡拉胶酶S190R。 突变的 κ-卡拉胶酶 S190R 提高了热稳定性。 突变酶处理的 κ-卡拉胶表现出高黄嘌呤氧化酶抑制活性。
The industrial applications of the κ-carrageenases have been restricted by their poor thermostability. In this study, based on the folding free energy change (ΔΔG) and the flexibility analysis using molecular dynamics (MD) simulation for the alkaline κ-carrageenase KCgCD from Pseudoalteromonas porphyrae (WT), the mutant S190R was identified with improved thermostability. After incubation at 50 °C for 30 min, the residual activity of S190R was 63.7%, 25.7% higher than that of WT. The Tm values determined by differential scanning calorimetry were 66.2 °C and 64.4 °C for S190R and WT, respectively. The optimal temperature of S190R was 10 °C higher than that of WT. The κ-carrageenan hydrolysates produced by S190R showed higher xanthine oxidase inhibitory activity compared with the untreated κ-carrageenan. MD simulation analysis of S190R showed that the residues (V186–M194 and P196–G197) in F5 and the key residue R150 in F3 displayed the decreased flexibility, and residues of T169–N173 near the catalytic center displayed the increased flexibility. These changed flexibilities might be the reasons for the improved thermostability of mutant S190R. This study provides a useful rational design strategy of combination of ΔΔG calculation and MD simulation to improve the κ-carrageenase’s thermostability for its better industrial applications. Mutant κ-carrageenase S190R is identified by rational design based on ΔΔG and MD simulation. Mutant κ-carrageenase S190R increases the thermostability. Mutant enzyme-treated κ-carrageenan exhibits high xanthine oxidase inhibitory activity.
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影响因子：
2.9
作者：
Zhu J;Fan H;Periole X;Honig B;Mark AE
通讯作者：
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影响因子：
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作者：
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源自红色海藻的寡糖：生产，特性以及潜在的健康和化妆品应用。
DOI：
10.3390/molecules23102451
发表时间：
2018-09-25
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
Cheong KL;Qiu HM;Du H;Liu Y;Khan BM
通讯作者：
Khan BM
影响因子：
6
作者：
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通讯作者：
Ochsenreither K
DOI：
10.3923/pjbs.2008.1779.1784
发表时间：
2008-07-15
期刊：
Pakistan journal of biological sciences : PJBS
影响因子：
--
作者：
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通讯作者：
Shahi, Majid Mohamad