聚羟基丁酸酯
电穿孔
质粒
生物
基因组编辑
Cas9
基因组
清脆的
羟基烷酸
盐单胞菌属
突变体
基因
转化(遗传学)
计算生物学
细菌
遗传学
16S核糖体RNA
作者
Tong Xu,Junyu Chen,Ruchira Mitra,Lin Lin,Zhengwei Xie,Guo‐Qiang Chen,Hua Xiang,Jing Han
标识
DOI:10.1038/s42003-022-03570-y
摘要
Halomonas bluephagenesis, a haloalkaliphilic bacterium and native polyhydroxybutyrate (PHB) producer, is a non-traditional bioproduction chassis for the next generation industrial biotechnology (NGIB). A single-sgRNA CRISPR/Cas9 genome editing tool is optimized using dual-sgRNA strategy to delete large DNA genomic fragments (>50 kb) with efficiency of 12.5% for H. bluephagenesis. The non-essential or redundant gene clusters of H. bluephagenesis, including those encoding flagella, exopolysaccharides (EPSs) and O-antigen, are sequentially deleted using this improved genome editing strategy. Totally, ~3% of the genome is reduced with its rapid growth and high PHB-production ability unaffected. The deletion of EPSs and O-antigen gene clusters shows two excellent properties from industrial perspective. Firstly, the EPSs and O-antigen deleted mutant rapidly self-flocculates and precipitates within 20 min without centrifugation. Secondly, DNA transformation into the mutant using electroporation becomes feasible compared to the wild-type H. bluephagenesis. The genome-reduced H. bluephagenesis mutant reduces energy and carbon source requirement to synthesize PHB comparable to its wild type. The H. bluephagenesis chassis with a reduced genome serves as an improved version of a NGIB chassis for productions of polyhydroxyalkanoates (PHA) or other chemicals.
科研通智能强力驱动
Strongly Powered by AbleSci AI