靛蓝
靛玉红
生物催化
化学
单加氧酶
酶
饱和突变
漆酶
有机化学
环境友好型
生化工程
组合化学
生物化学
生物
细胞色素P450
基因
催化作用
反应机理
艺术
视觉艺术
突变体
工程类
生态学
作者
Nicolás Núñez-Navarro,Javier Salazar,Francisco Castillo,César A. Ramírez‐Sarmiento,Ignacio Poblete‐Castro,Flavia C. Zacconi,Loreto P. Parra
标识
DOI:10.3390/ijms232012544
摘要
Indigoids are natural pigments obtained from plants by ancient cultures. Romans used them mainly as dyes, whereas Asian cultures applied these compounds as treatment agents for several diseases. In the modern era, the chemical industry has made it possible to identify and develop synthetic routes to obtain them from petroleum derivatives. However, these processes require high temperatures and pressures and large amounts of solvents, acids, and alkali agents. Thus, enzyme engineering and the development of bacteria as whole-cell biocatalysts emerges as a promising green alternative to avoid the use of these hazardous materials and consequently prevent toxic waste generation. In this research, we obtained two novel variants of phenylacetone monooxygenase (PAMO) by iterative saturation mutagenesis. Heterologous expression of these two enzymes, called PAMOHPCD and PAMOHPED, in E. coli was serendipitously found to produce indigoids. These interesting results encourage us to characterize the thermal stability and enzyme kinetics of these new variants and to evaluate indigo and indirubin production in a whole-cell system by HPLC. The highest yields were obtained with PAMOHPCD supplemented with L-tryptophan, producing ~3000 mg/L indigo and ~130.0 mg/L indirubin. Additionally, both enzymes could oxidize and produce several indigo derivatives from substituted indoles, with PAMOHPCD being able to produce the well-known Tyrian purple. Our results indicate that the PAMO variants described herein have potential application in the textile, pharmaceutics, and semiconductors industries, prompting the use of environmentally friendly strategies to obtain a diverse variety of indigoids.
科研通智能强力驱动
Strongly Powered by AbleSci AI