在Komagataella phaffii中对Cu/Zn超氧化物脱酶 (SOD1) 的Codon优化生产:功能性表征和高产发酵策略
Abdulqader Al-Adeeb1, Sahibzada Muhammad Aqeel1, Qiuya Gu1
1Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214000, Jiangsu, China.
International journal of biological macromolecules
|October 29, 2025
概括
在Komagataella phaffii中增强了复合铜/超氧化脱酶 (SOD1) 生产,实现了抗氧化剂应用的高产量和稳定性.
科学领域:
- 生物技术是生物技术.
- 酶工程是什么? 酶工程是什么?
- 分子生物学分子生物学
背景情况:
- 铜/超氧化物脱酶 (SOD1) 对于中和有害的超氧化物激素至关重要.
- 目前生产复合真核SOD1的方法在产量和稳定性方面存在局限性.
研究的目的:
- 开发一种高效且可扩展的方法来生产重组SOD1.1.
- 描述产生的SOD1.1的酶性质和稳定性.
主要方法:
- 将Saccharomyces bayanus SOD1基因的克隆和编码优化转化为Komagataella phaffii表达载体pPIC9K.
- 在摇瓶和5L发酵器中表达和优化SOD1.
- 酶分析,稳定性测试 (pH,温度) 和分子对接用于表征.
主要成果:
- 在K. phaffii.中成功表达和分泌复合SOD1.
- 达到2074.9个单位/毫克的最大震动瓶活性和15120个单位/毫克的发酵器活性.
- 在pH 6.0和40°C时表现出最佳活性,具有显著的热稳定性 (在60°C时2小时后70%的活性).
结论:
- 使用K. phaffii. 开发了一种高效,可扩展的战略,用于使用K. phaffii. 进行重组SOD1生产.
- 生产的SOD1具有有利的酶特性和稳定性,可用于工业和治疗用途.
- 这种系统促进了SOD1在抗氧化剂配方中的更广泛应用.
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