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Volume 12
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10.3390/microorganisms12071275
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Article

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

by
Meijuan Zhang
1,†,
Jie Zhen
2,3,4,†,
Jia Teng
2,5,
Xingya Zhao
2,3,
Xiaoping Fu
2,3,
Hui Song
2,3,4,
Yeni Zhang
5,
Hongchen Zheng
2,3,4,6,* and
Wenqin Bai
2,3,4,6,*
1
College of Life Science and Agriculture Forestry, Qiqihar University, Qiqihar 161006, China
2
Industrial Enzymes National Engineering Research Center, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
3
National Center of Technology Innovation for Synthetic Biology, Tianjin 300308, China
4
Tianjin Key Laboratory for Industrial Biological Systems and Bioprocessing Engineering, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
5
College of Food Science and Biotechnology, Tianjin Agricultural University, Tianjin 300392, China
6
Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work and share first authorship.
Microorganisms 2024, 12(7), 1275; https://doi.org/10.3390/microorganisms12071275
Submission received: 29 April 2024 / Revised: 19 June 2024 / Accepted: 20 June 2024 / Published: 23 June 2024
(This article belongs to the Section Microbial Biotechnology)
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Abstract

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 B. subtilis 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 Bacillus subtilis 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 B. subtilis, reaching 12,295 U/mL, which was 1.9-times higher than that expressed in the recombinant Escherichia coli 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.
Keywords: signal peptide; secretion; transcription; extracellular expression; N-terminal amino acid sequence; alkaline pectin lyase signal peptide; secretion; transcription; extracellular expression; N-terminal amino acid sequence; alkaline pectin lyase

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MDPI and ACS Style

Zhang, M.; Zhen, J.; Teng, J.; Zhao, X.; Fu, X.; Song, H.; Zhang, Y.; Zheng, H.; Bai, W. N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis. Microorganisms 2024, 12, 1275. https://doi.org/10.3390/microorganisms12071275

AMA Style

Zhang M, Zhen J, Teng J, Zhao X, Fu X, Song H, Zhang Y, Zheng H, Bai W. N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis. Microorganisms. 2024; 12(7):1275. https://doi.org/10.3390/microorganisms12071275

Chicago/Turabian Style

Zhang, Meijuan, Jie Zhen, Jia Teng, Xingya Zhao, Xiaoping Fu, Hui Song, Yeni Zhang, Hongchen Zheng, and Wenqin Bai. 2024. "N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis" Microorganisms 12, no. 7: 1275. https://doi.org/10.3390/microorganisms12071275

APA Style

Zhang, M., Zhen, J., Teng, J., Zhao, X., Fu, X., Song, H., Zhang, Y., Zheng, H., & Bai, W. (2024). N-Terminal Sequences of Signal Peptides Assuming Critical Roles in Expression of Heterologous Proteins in Bacillus subtilis. Microorganisms, 12(7), 1275. https://doi.org/10.3390/microorganisms12071275

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