在微空气环境下,大肠杆菌BL21细胞在发酵器规模生产复合β-曼酶
Anjali Purohit1, Lata Pawar1, Sudesh Kumar Yadav2
1Center of Innovative and Applied Bioprocessing (CIAB), Sector-81, Knowledge City, Mohali, 140306, India.
Carbohydrate research
|May 24, 2024
概括
研究人员优化了beta-mannanase的生产,在E. coli中使用了具有成本效益的最小介质. 这种扩大规模的工艺实现了纯化β-曼酶的高产量,适用于水解曼诺酸糖.
科学领域:
- 生物技术是生物技术.
- 酵素工程是什么? 酶工程是什么
- 微生物发酵 微生物发酵
背景情况:
- β-曼酶是一种酶,对降解曼至关重要.
- 对于工业应用而言,有效且具有成本效益的β-曼酶生产是必不可少的.
- 异质表达为高水平酶生产提供了可行的途径.
研究的目的:
- 优化和扩大使用成本效益较低的最小介质生产β-曼酶的生产.
- 通过对大肠杆菌的异质表达,实现复合β-曼酶的高产量.
- 描述该酶在化mannooligosaccharides中的活性.
主要方法:
- 从微细菌camelliasinensis CIAB417通过异质表达在大肠杆菌BL21 (DE3) 中优化β-曼酶的产生.
- 从摇瓶扩大生产规模到5L发酵器,使用无维生素的成本效益较低的最小介质 (M9+e).
- 评估了氧气可用性和pH稳定性对细胞生长和酶生产的影响.
主要成果:
- 一种具有成本效益的最小介质 (M9+e) 被确定为培养大肠杆菌和生产β-mannanase的最佳培养基.
- 在整个发酵过程中,在摇瓶和发酵器尺度上保持稳定的pH值.
- 生产产出1.8g的净化重组β-曼酶每升的介质从2.0314g干细胞重量.
- 纯化的酶有效地将虫豆水解成mannooligosaccharides.
结论:
- 优化的最小介质和发酵过程使得可再生β-mannanase的经济高效,大规模生产成为可能.
- 生产的β-曼酶表现出显著的酶活性,在各种行业中具有潜在的应用.
- 这项研究为工业生产有价值的酶提供了一个强大的平台.
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