Construction and Characterization of a Gradient Strength Promoter Library for Fine-Tuned Gene Expression in Bacillus licheniformis

• DOI: 10.1021/acssynbio.1c00242
• Publication Date: 2021-08-27T17:30:35Z
• Authors: Yi Rao; Peifen Li; Xinxin Xie; Jiemin Li; Yongqing Liao; Xin Ma; Dongbo Cai; Shouwen Chen
• Keywords: Bacillus licheniformis; Corynebacterium glutamicum

Bacillus licheniformis DW2 is an important industrial strain for bacitracin production, and it also used biochemical however, the lack of effective toolkit precise regulation gene expression hindered its application seriously. Here, a gradient strength promoter library was constructed based on synthetase cluster PbacA. First, different PbacA variants were via coupling with various 5′-UTRs, ranges 32.6–741.8% attained among these promoters. Then, three promoters, PUbay (strong), (middle), PUndh (weakest), applied red fluorescent protein (RFP) keratinase assays, promoters proven to have good universality proteins. Second, replaced by bacitraicn titer enhanced 14.62% when applied, which decreased 98.05% under mediation compared that original DW2. Third, PUbay, PUyvgO, selected regulate levels critical genes are responsible pucheriminic acid synthesis, yield increased 194.1% manipulating synthetic competitive pathways. Finally, PbacA, green (GFP) RFP in Escherichia coli, consistent effects our results. Taken together, this research, provided fine-tuning reprogramming metabolite metabolic flux B. licheniformis.