N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis

• Published in: Microorganisms (Basel) Vol. 12; no. 7; p. 1275
• Main Authors: Zhang, Meijuan, Zhen, Jie, Teng, Jia, Zhao, Xingya, Fu, Xiaoping, Song, Hui, Zhang, Yeni, Zheng, Hongchen, Bai, Wenqin
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 23.06.2024
• Subjects: alkaline pectin lyase; Amino acids; Bacillus subtilis; Coding; E coli; Enzymes; extracellular expression; LipA protein; N-terminal amino acid sequence; Optimization; Pectin; Pectin lyase; Peptides; Plasmids; Protein expression; Protein synthesis; Proteins; Secretion; signal peptide; Signal peptides; transcription; Work capacity; United States > US; China; signal peptide; extracellular expression; secretion; N-terminal amino acid sequence; transcription; alkaline pectin lyase
• ISSN: 2076-2607; 2076-2607
• DOI: 10.3390/microorganisms12071275

The N-terminal sequences of proteins and their corresponding encoding sequences may play crucial roles in the heterologous expression. In this study, the secretory expression of alkaline pectin lyase APL in was investigated to explore the effects of the N-terminal 5-7 amino acid sequences of different signal peptides on the protein expression and secretion. It was identified for the first time that the first five amino acid sequences of the N-terminal of the signal peptide (SP LipA) from lipase A play an important role in promoting the expression of APL. Furthermore, it was revealed that SP LipA resulted in higher secretory expression compared to other signal peptides in this study primarily due to its encoding of N-terminal amino acids with relatively higher transcription levels and its efficient secretion capacity. Based on this foundation, the recombinant strain constructed in this work achieved a new record for the highest extracellular yields of APL in , reaching 12,295 U/mL, which was 1.9-times higher than that expressed in the recombinant strain previously reported. The novel theories uncovered in this study are expected to play significant roles in enhancing the expression of foreign proteins both inside and outside of cells.