Manoharan Chitra, Thomas Dyna Susan, Yashwant Rasalkar Sandhya, Roy Gourab, Kunjupillai Vijayan, Mishra Rakesh Kumar, Nongthomba Upendra, Gopalapillai Ravikumar
Seri-Biotech Research Laboratory, Central Silk Board, Kodathi, Bengaluru, 560 035 India.
Department of Biotechnology, St Joseph's University, Lalbagh Road, Bengaluru, 560 027 India.
3 Biotech. 2025 May;15(5):132. doi: 10.1007/s13205-025-04293-7. Epub 2025 Apr 17.
silk fibroin and the antimicrobial peptide cecropin B are promising biomolecules for biomedical applications due to their unique and complementary properties. In this study, we successfully expressed recombinant silk fibroin (R. fib) alone and in fusion with cecropin B (R. fib-cec) in , with yields of 6 mg/L for R. fib and 1.3 mg/L for the R. fib-cec fusion protein. Characterization of the proteins through SDS-PAGE, Western blotting, and MALDI-TOF confirmed the successful expression and purity of the recombinant proteins. Notably, the fusion protein exhibited potent broad-spectrum antibacterial activity against Gram-negative and Gram-positive , highlighting its potential as an antimicrobial agent. Furthermore, the R. fib and the R. fib-cec demonstrated significant protective effects against HO and UVB-induced oxidative damage in human adult dermal fibroblast cells. Pretreatment with R. fib and R. fib-cec significantly improved cell viability and morphology. R. fib and R. fib-cec increased viable cell numbers in H₂O₂-treated cells (61.8% and 83.5%, respectively) compared to the control (33%), and in UVB-irradiated cells (67.7% and 87.3%, respectively) compared to the control (38%). Both proteins also significantly reduced LDH release, a marker of cell damage. These results demonstrate that R. fib, especially R. fib-cec, protects against oxidative stress-induced cellular damage, promoting cell proliferation and reducing cytotoxicity. These findings highlight the multifunctionality of the silk-cecropin B fusion protein, making it a promising candidate for diverse biomedical applications, including antimicrobial therapies, skin protection, and wound healing.
The online version contains supplementary material available at 10.1007/s13205-025-04293-7.
丝素蛋白和抗菌肽天蚕素B因其独特且互补的特性，是生物医学应用中很有前景的生物分子。在本研究中，我们成功地在[具体表达系统未给出]中单独表达了重组丝素蛋白（R. fib）以及与天蚕素B融合表达（R. fib-cec），R. fib的产量为6 mg/L，R. fib-cec融合蛋白的产量为1.3 mg/L。通过SDS-PAGE、蛋白质印迹和基质辅助激光解吸电离飞行时间质谱对蛋白质进行表征，证实了重组蛋白的成功表达和纯度。值得注意的是，融合蛋白对革兰氏阴性菌和革兰氏阳性菌均表现出强大的广谱抗菌活性，突出了其作为抗菌剂的潜力。此外，R. fib和R. fib-cec对人成人皮肤成纤维细胞中过氧化氢（HO）和紫外线B（UVB）诱导的氧化损伤显示出显著的保护作用。用R. fib和R. fib-cec预处理可显著改善细胞活力和形态。与对照组（33%）相比，R. fib和R. fib-cec使过氧化氢处理的细胞中活细胞数量增加（分别为61.8%和83.5%），与对照组（38%）相比，在紫外线B照射的细胞中活细胞数量也增加（分别为67.7%和87.3%）。两种蛋白还显著降低了乳酸脱氢酶（LDH）释放，这是细胞损伤的一个指标。这些结果表明，R. fib，尤其是R. fib-cec，可保护细胞免受氧化应激诱导的损伤，促进细胞增殖并降低细胞毒性。这些发现突出了丝素蛋白 - 天蚕素B融合蛋白的多功能性，使其成为包括抗菌治疗、皮肤保护和伤口愈合在内的多种生物医学应用的有前途的候选物。
在线版本包含可在10.1007/s13205-025-04293-7获取的补充材料。
