| abstract |
The invention relates to a method for expression optimization of multi-subunit enzyme and biosynthesis of a high value-added compound by utilizing a lignin monomer, wherein an RBS library of pE 1K-GcobA-M3 is constructed by a Golden gate cloning method, the RBS library of the library G/C is screened by researching the growth rate, the conversion rate, the expression ratio of the enzyme, the enzyme activity of crude enzyme liquid and the like of the library, the growth rate of a strain is high, and the enzyme activity is also high. Thereafter, the RBS library was used for enzymatic reaction pathway construction, catalyzing monolignol compounds guaiacol (guaiacol) and pyrogallol (pyrogallol) to produce high value-added compounds such as catechol, catechol-O-sulfate, and erythrophenol. The invention utilizes the constructed biocatalyst to realize the high-efficiency utilization of aromatic compounds (guaiacol and 3-methoxy catechol) from lignin through biotransformation, and constructs the biosynthesis route of catechol-O-sulfate and rhodophenol for the first time. |