Optimization of intracellular biosynthesis process involving regulation of multiple gene expressions is dependent on the efficient and accurate expression of each expression unit independently. However, challenges of analyzing intermediate products seriously hinder the application of high throughput assays. This study aimed to develop an engineering approach for unsterile production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) or (P3HB4HB) by recombinant Halomonas bluephagenesis (H. bluephagenesis) constructed via coupling the design of GFP-mediated transcriptional mapping and high-resolution control of gene expressions (HRCGE), which consists of two inducible systems with high- and low-dynamic ranges employed to search the exquisite transcription level of each expression module in the presence of γ-butyrolactone, the intermediate for 4-hydroxybutyrate (4HB) synthesis. It has been successful to generate a recombinant H. bluephagenesis, namely TD68-194, able to produce over 36 g/L P3HB4HB consisting of 16 mol% 4HB during a 7-L lab-scale fed-batch growth process, of which cell dry weight and PHA content reached up to 48.22 g/L and 74.67%, respectively, in 36 h cultivation. HRCGE has been found useful for metabolic pathway construction.

涉及调节多个基因表达的细胞内生物合成过程的优化取决于每个表达单元的有效和准确表达。然而，分析中间产物的挑战严重阻碍了高通量分析的应用。本研究旨在开发用于生产未消毒的聚（3-羟基丁酸酯的工程方法共-4-羟基丁酸酯）或（P3HB4HB）通过重组盐单胞菌bluephagenesis（H. bluephagenesis）是通过结合GFP介导的转录图谱设计和基因表达的高分辨率控制（HRCGE）设计而构建的，HRCGE由两个可诱导系统组成，该系统具有高和低动态范围，用于搜索每个表达模块的精确转录水平。 γ-丁内酯是4-羟基丁酸酯（4HB）合成的中间体。它已经成功地产生了重组蓝细菌，即TD68-194，能够在7升实验室规模的分批补料生长过程中产生超过36 g / L的P3HB4HB，其中16 mol％的4HB组成，其中的细胞干燥在36 h的栽培中，重和PHA含量分别达到48.22 g / L和74.67％。已经发现HRCGE对于代谢途径的构建是有用的。