Zhang Meijuan, Zhen Jie, Teng Jia, Zhao Xingya, Fu Xiaoping, Song Hui, Zhang Yeni, Zheng Hongchen, Bai Wenqin
College of Life Science and Agriculture Forestry, Qiqihar University, Qiqihar 161006, China.
Industrial Enzymes National Engineering Research Center, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China.
Microorganisms. 2024 Jun 23;12(7):1275. 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 (SPLipA) from lipase A play an important role in promoting the expression of APL. Furthermore, it was revealed that SPLipA 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.
蛋白质的N端序列及其相应的编码序列可能在异源表达中发挥关键作用。在本研究中，对碱性果胶裂解酶APL在[具体表达系统未提及]中的分泌表达进行了研究，以探讨不同信号肽的N端5 - 7个氨基酸序列对蛋白质表达和分泌的影响。首次发现来自脂肪酶A的信号肽（SPLipA）N端的前五个氨基酸序列在促进APL表达方面发挥重要作用。此外，本研究表明，与其他信号肽相比，SPLipA导致更高的分泌表达，这主要是由于其编码的N端氨基酸具有相对较高的转录水平以及其高效的分泌能力。在此基础上，本工作构建的重组菌株在[具体表达系统未提及]中实现了APL胞外产量的新纪录，达到12295 U/mL，比先前报道的重组[具体菌株未提及]菌株中表达的产量高出1.9倍。本研究中发现的新理论有望在增强细胞内外外源蛋白的表达方面发挥重要作用。
