建立一个人工途径,用于生物合成的八胺和synephrine
Jiao Feng1, Runyuan Jin1, Shasha Cheng1
1State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, People's Republic of China.
这项研究在大肠杆菌中设计了一条合成途径,用于从铁氨酸中产生胺和synephrine. 研究人员优化了胺β-氧化酶 (TBH) 和乙醇胺N-甲基转移酶 (PNMT) 的酶,实现了这两种化合物的显著标位.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 合成生物学 合成生物学
背景情况:
- 八胺和synephrine是具有各种生理作用的生物氨基.
- 这些化合物的生物合成通常涉及来自真核生物源的酶.
- 有效的微生物生产需要异质表达和途径优化.
研究的目的:
- 在大肠杆菌中*设计和建造一种人工生物合成途径,用于*Escherichia coli*中的八胺和synephrine生产.
- 识别和优化关键酶,胺β-氧酶 (TBH) 和乙醇胺N-甲基转移酶 (PNMT),用于异构的表达和活性.
- 建立一个单级联反应,以有效合成这些生物性氨基.
主要方法:
- 在大肠杆菌中对TBH和PNMT酶的查和异质表达.
- 优化酶表达策略和特定活动.
- 开发和优化单个反应步骤和一个一级联系统.
- 合成的八胺和synephrine的定位和量化.
主要成果:
- 在大肠杆菌中成功表达可溶性*Drosophila melanogaster*TBH,优化活性至229.50U·g-1 .
- 在第一步中达到0.10g/L的八胺和第二步的2.02g/L的synephrine的产量.
- 构建了一个单级联反应,从胺中产生30.05毫克/升的synephrine.
- 进一步优化的标位达到0.45g/L的八胺和0.20g/L的synephrine.
结论:
- 这项研究展示了使用工程微生物通路的第一个成功的八胺和synephrine生物合成.
- 开发的系统证明了通过合成生物学方法生产有价值的生物原胺的可行性.
- 进一步优化有潜力用于可扩展的胺和synephrine的工业生产.
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