title: Efficient Secretion of Recombinant Proteins from Rice Suspension-Cultured Cells Modulated by the Choice of Signal Peptide.
abstract: Plant-based expression systems have emerged as a competitive platform in the large-scale production of recombinant proteins. By adding a signal peptide, αAmy3sp, the desired recombinant proteins can be secreted outside transgenic rice cells, making them easy to harvest. In this work, to improve the secretion efficiency of recombinant proteins in rice expression systems, various signal peptides including αAmy3sp, CIN1sp, and 33KDsp have been fused to the N-terminus of green fluorescent protein (GFP) and introduced into rice cells to explore the efficiency of secretion of foreign proteins. 33KDsp had better efficiency than αAmy3sp and CIN1sp for the secretion of GFP from calli and suspension-cultured cells. 33KDsp was further applied for the secretion of mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) from transgenic rice suspension-cultured cells; approximately 76%-92% of total rice-derived mGM-CSF (rmGM-CSF) was detected in the culture medium. The rmGM-CSF was bioactive and could stimulate the proliferation of a murine myeloblastic leukemia cell line, NSF-60. The extracellular yield of rmGM-CSF reached 31.7 mg/L. Our study indicates that 33KDsp is better at promoting the secretion of recombinant proteins in rice suspension-cultured cell systems than the commonly used αAmy3sp.
authors.affiliation: Graduate School of Biotechnology and Bioengineering, Yuan Ze University, 135 Yuan-Tung Road, Taoyuan, Taiwan, ROC.
authors.forename: Li-Fen
authors.initials: LF
authors.lastname: Huang
authors.affiliation: Graduate School of Biotechnology and Bioengineering, Yuan Ze University, 135 Yuan-Tung Road, Taoyuan, Taiwan, ROC.
authors.forename: Chia-Chun
authors.initials: CC
authors.lastname: Tan
authors.affiliation: Department of Life Science, National Central University, Taoyuan, Taiwan, ROC.
authors.forename: Ju-Fang
authors.initials: JF
authors.lastname: Yeh
authors.affiliation: Department of Life Science, National Central University, Taoyuan, Taiwan, ROC.
authors.forename: Hsin-Yi
authors.initials: HY
authors.lastname: Liu
authors.affiliation: Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan, Taiwan, ROC.
authors.forename: Yu-Kuo
authors.initials: YK
authors.lastname: Liu
authors.affiliation: Department of Agronomy, National Chi-Yi University, Chiayi, Taiwan, ROC.
authors.forename: Shin-Lon
authors.initials: SL
authors.lastname: Ho
authors.affiliation: Department of Life Science, National Central University, Taoyuan, Taiwan, ROC.
authors.forename: Chung-An
authors.initials: CA
authors.lastname: Lu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D012275:Oryza / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D059828:Plant Cells / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000502:physiology*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor