Minimization and optimization of α-amylase terminator for heterologous protein production in Bacillus licheniformis

Abstract Terminators serve as the regulatory role in gene transcription termination; however, few researches about terminator optimization have been conducted, which leads to lack of available and universal for expression regulation Bacillus . To solve this problem expand synthetic biology toolbox licheniformis , T1 endogenous α-amylase ( amyL ) was characterized research, with a termination efficiency 87.81%. Then, we explored optimized strength from four aspects: distance between stop codon terminator, GC content at bottom stem structure, loop size, U-tract length, best T24 attained by combination strategy, increased 97.97%, better than commonly used T7 (T7 P Escherichia coli Finally, applied protein expression, which, respectively, led 33.00%, 25.93%, 11.78% increases green fluorescence intensity, red keratinase activity, indicating its universality expression. Taken together, research not only expands plug-and-play B. but also provides reference artificial design versatile intrinsic terminator. Graphical