- 期刊
乳酸菌細胞表層展示系統之研究
Study on the Cell Surface Display System of Lactic Acid Bacteria
摘要
表層蛋白質有維持細胞形態、做為酵素的骨架及細胞附著和感染等功能。本研究以聚合酶連鎖反應法，夾取嗜酸乳桿菌ATCC 4356表層蛋白質靜默基因C端123個胺基酸(SBC)之DNA序列(sbc)，SBC可由誘導表現後的大腸桿菌利用親和性層析純化之，用以製作抗體。融合乳酸菌胞外蛋白酶USP45 訊息胜肽、乘載蛋白及表層蛋白質靜默基因C端123個胺基酸，表現於副乾酪乳桿菌或乳酸鏈球菌。在乳酸鏈球菌中雖伴隨降解現象，但有較佳錨定效果。而副乾酪乳桿菌中，錨定效果較差，且被剪切掉落於胞外。本試驗結果顯示，表層蛋白質靜默基因C端123個胺基酸具有作為乳酸鏈球菌細胞表層展示系統錨定用攜帶蛋白之潛力。
並列摘要
Surface layer (S-layer) protein is characterized with biological functions such as maintenance of cell shape, cell adhesion, molecular sieve, and scaffolding for enzyme. In this study, the DNA sequence sbc of silent S-protein C-terminal 123 amino acids (SBC) was cloned from Lactobacillus acidophilus ATCC4356 chromosome by polymerase chain reaction. The SBC was purified from induced expression Escherichia coli culture by affinity chromatography and subjected to anti-SBC antibody preparation. The fusion DNA fragment of the lactococcal extracellular protease USP45 signal peptide, passenger protein and SBC were expressed in Lactococcus lactis and Lactobacillus paracasei. Results showed that the fusion protein anchored onto the cell wall of L. lactis with degraded form. In L. paracasei, the fusion protein was not well anchored onto the cell wall and SBC was detected in the extracellular fraction. In conclusion, the SBC is considered as a potent tool for L. lactis cell surface anchoring.