Ribosome-Engineered Lacticaseibacillus rhamnosus Strain GG Exhibits Cell Surface Glyceraldehyde-3-Phosphate Dehydrogenase Accumulation and Enhanced Adhesion to Human Colonic Mucin.
Ribosome-Engineered Lacticaseibacillus rhamnosus Strain GG Exhibits Cell Surface Glyceraldehyde-3-Phosphate Dehydrogenase Accumulation and Enhanced Adhesion to Human Colonic Mucin.
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DOI：
10.1128/aem.01448-20
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发表时间：
2020-10-01
影响因子：
4.4
通讯作者：
Shimosato T
中科院分区：
文献类型：
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作者：
Ishida M;Namai F;Shigemori S;Kajikawa S;Tsukagoshi M;Sato T;Ogita T;Shimosato T
我们试图将核糖体工程(RE)应用于益生菌乳酸菌，并验证RE的影响。在这里，我们发现RE Lacticaseibacillusrhamnosus GG(LGG-MTK56N)的一个突变体在细胞表面带有GAPDH；GAPDH是通过ABC转运蛋白输出的。与野生型亲本相比，LGG-MTK56N对人结肠粘蛋白的粘附性更强，细胞大小和形状明显不同。这些发现表明，LGG-MTK56N中的RE在蛋白质合成、蛋白质转运和细胞形态方面发生了戏剧性的变化，并影响了对人结肠粘蛋白的黏附。关于益生菌的健康益处，个体宿主反应的差异已经成为一个问题。在这里，我们将在放线菌研究中开发的核糖体工程(RE)技术应用于鼠李糖乳杆菌(LGG)。RE可以通过使用抗生素诱导核糖体和/或RNA聚合酶的自发突变，有效地提高微生物的潜力。在本研究中，我们在编码核糖体蛋白S12的lgG rpsL基因中发现了八种链霉素耐药突变。值得注意的是，携带K56N突变体的LGG(LGG-MTK56N)与野生型(LGG-WT)相比，在细胞表面表达高水平的甘油醛-3-磷酸脱氢酶(GAPDH)。GAPDH在结肠粘蛋白黏附中起关键作用。事实上，在使用Biacore系统的实验中，与LGG-WT相比，LGG-MTK56N显著增加了A型人结肠粘蛋白的粘附性。黏附在结肠上的能力是益生菌的一个重要特性；因此，这些结果表明，RE是一种有效的益生菌乳酸菌育种策略。重要性我们试图将核糖体工程(RE)应用于益生菌乳酸菌，并验证RE的影响。在这里，我们发现RE Lacticaseibacillusrhamnosus GG(LGG-MTK56N)的一个突变体在细胞表面带有GAPDH；GAPDH是通过ABC转运蛋白输出的。与野生型亲本相比，LGG-MTK56N对人结肠粘蛋白的粘附性更强，细胞大小和形状明显不同。这些发现表明，LGG-MTK56N中的RE在蛋白质合成、蛋白质转运和细胞形态方面发生了戏剧性的变化，并影响了对人结肠粘蛋白的黏附。
We sought to apply ribosome engineering (RE) to probiotic lactic acid bacteria and to verify RE’s impact. Here, we showed that one mutant of RE Lacticaseibacillus rhamnosus GG (LGG-MTK56N) bore a GAPDH on the cell surface; the GAPDH was exported via an ABC transporter. Compared to the wild-type parent, LGG-MTK56N adhered more strongly to human colonic mucin and exhibited a distinct cell size and shape. These findings demonstrate that RE in LGG-MTK56N yielded dramatic changes in protein synthesis, protein transport, and cell morphology and affected adherence to human colonic mucin. Differences in individual host responses have emerged as an issue regarding the health benefits of probiotics. Here, we applied ribosome engineering (RE) technology, developed in an actinomycete study, to Lacticaseibacillus rhamnosus GG (LGG). RE can effectively enhance microbial potential by using antibiotics to induce spontaneous mutations in the ribosome and/or RNA polymerase. In this study, we identified eight types of streptomycin resistance mutations in the LGG rpsL gene, which encodes ribosomal protein S12. Notably, LGG harboring the K56N mutant (LGG-MTK56N) expressed high levels of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) on the cell surface compared with the LGG wild type (LGG-WT). GAPDH plays a key role in colonic mucin adhesion. Indeed, LGG-MTK56N significantly increased type A human colonic mucin adhesion compared to LGG-WT in experiments using the Biacore system. The ability to adhere to the colon is an important property of probiotics; thus, these results suggest that RE is an effective breeding strategy for probiotic lactic acid bacteria. IMPORTANCE We sought to apply ribosome engineering (RE) to probiotic lactic acid bacteria and to verify RE’s impact. Here, we showed that one mutant of RE Lacticaseibacillus rhamnosus GG (LGG-MTK56N) bore a GAPDH on the cell surface; the GAPDH was exported via an ABC transporter. Compared to the wild-type parent, LGG-MTK56N adhered more strongly to human colonic mucin and exhibited a distinct cell size and shape. These findings demonstrate that RE in LGG-MTK56N yielded dramatic changes in protein synthesis, protein transport, and cell morphology and affected adherence to human colonic mucin.