Bioconjugation of Serratiopeptidase with Titanium Oxide Nanoparticles: Improving Stability and Antibacterial Properties.

IF 5 3区 医学 Q1 ENGINEERING, BIOMEDICAL
Jhon Jairo Melchor-Moncada, Santiago Vasquez-Giraldo, Augusto Zuluaga-Vélez, Lina Marcela Orozco, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias
{"title":"Bioconjugation of Serratiopeptidase with Titanium Oxide Nanoparticles: Improving Stability and Antibacterial Properties.","authors":"Jhon Jairo Melchor-Moncada, Santiago Vasquez-Giraldo, Augusto Zuluaga-Vélez, Lina Marcela Orozco, Luz Angela Veloza, Juan Carlos Sepúlveda-Arias","doi":"10.3390/jfb15100300","DOIUrl":null,"url":null,"abstract":"<p><p>Antimicrobial resistance (AMR) poses a significant global health threat, necessitating the development of novel antibacterial strategies. Serratiopeptidase (SP), a metalloprotease produced by bacteria such as <i>Serratia marcescens</i>, has gained attention not only for its anti-inflammatory properties but also for its potential antibacterial activity. However, its protein nature makes it susceptible to pH changes and self-proteolysis, limiting its effectiveness. This study aimed to increase both the enzymatic stability and antibacterial activity of serratiopeptidase through immobilization on titanium oxide nanoparticles (TiO<sub>2</sub>-NPs), leveraging the biocompatibility and stability of these nanomaterials. Commercial TiO<sub>2</sub>-NPs were characterized using TGA/DTG, FT-IR, UV-Vis, and XRD analyses, and their biocompatibility was assessed through cytotoxicity studies. Serratiopeptidase was produced via fermentation using the C8 isolate of <i>Serratia marcescens</i> obtained from the intestine of <i>Bombyx mori</i> L., purified chromatographically, and immobilized on carboxylated nanoparticles via EDC/NHS coupling at various pH conditions. The optimal enzymatic activity was achieved by using pH 5.1 for nanoparticle activation and pH 5.5 for enzyme coupling. The resulting bioconjugate demonstrated stable proteolytic activity at 25 °C for 48 h. Immobilization was confirmed by FT-IR spectroscopy, and the Michaelis-Menten kinetics were determined. Notably, the bioconjugate exhibited two-fold greater antibacterial activity against <i>E. coli</i> than the free enzyme or TiO<sub>2</sub>-NPs at 1000 µg/mL. This study successfully developed a serratiopeptidase-TiO<sub>2</sub> bioconjugate with enhanced enzymatic stability and antibacterial properties. The improved antibacterial activity of the immobilized enzyme presents a promising approach for developing new tools to combat antimicrobial resistance, with potential applications in healthcare, food safety, and environmental protection.</p>","PeriodicalId":15767,"journal":{"name":"Journal of Functional Biomaterials","volume":null,"pages":null},"PeriodicalIF":5.0000,"publicationDate":"2024-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11508812/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Functional Biomaterials","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.3390/jfb15100300","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ENGINEERING, BIOMEDICAL","Score":null,"Total":0}
引用次数: 0

Abstract

Antimicrobial resistance (AMR) poses a significant global health threat, necessitating the development of novel antibacterial strategies. Serratiopeptidase (SP), a metalloprotease produced by bacteria such as Serratia marcescens, has gained attention not only for its anti-inflammatory properties but also for its potential antibacterial activity. However, its protein nature makes it susceptible to pH changes and self-proteolysis, limiting its effectiveness. This study aimed to increase both the enzymatic stability and antibacterial activity of serratiopeptidase through immobilization on titanium oxide nanoparticles (TiO2-NPs), leveraging the biocompatibility and stability of these nanomaterials. Commercial TiO2-NPs were characterized using TGA/DTG, FT-IR, UV-Vis, and XRD analyses, and their biocompatibility was assessed through cytotoxicity studies. Serratiopeptidase was produced via fermentation using the C8 isolate of Serratia marcescens obtained from the intestine of Bombyx mori L., purified chromatographically, and immobilized on carboxylated nanoparticles via EDC/NHS coupling at various pH conditions. The optimal enzymatic activity was achieved by using pH 5.1 for nanoparticle activation and pH 5.5 for enzyme coupling. The resulting bioconjugate demonstrated stable proteolytic activity at 25 °C for 48 h. Immobilization was confirmed by FT-IR spectroscopy, and the Michaelis-Menten kinetics were determined. Notably, the bioconjugate exhibited two-fold greater antibacterial activity against E. coli than the free enzyme or TiO2-NPs at 1000 µg/mL. This study successfully developed a serratiopeptidase-TiO2 bioconjugate with enhanced enzymatic stability and antibacterial properties. The improved antibacterial activity of the immobilized enzyme presents a promising approach for developing new tools to combat antimicrobial resistance, with potential applications in healthcare, food safety, and environmental protection.

氧化钛纳米颗粒与塞拉提肽酶的生物共轭:提高稳定性和抗菌性
抗菌药耐药性(AMR)对全球健康构成严重威胁,因此有必要开发新型抗菌策略。沙雷氏肽酶(SP)是一种由沙雷氏菌(Serratia marcescens)等细菌产生的金属蛋白酶,它不仅具有抗炎特性,而且还具有潜在的抗菌活性,因此备受关注。然而,其蛋白质性质使其易受 pH 值变化和自身蛋白水解的影响,从而限制了其有效性。本研究旨在利用纳米材料的生物相容性和稳定性,通过将其固定在氧化钛纳米颗粒(TiO2-NPs)上,提高血清肽酶的酶稳定性和抗菌活性。利用 TGA/DTG、FT-IR、UV-Vis 和 XRD 分析对商用 TiO2-NPs 进行了表征,并通过细胞毒性研究对其生物相容性进行了评估。利用从蚕宝宝肠道中分离出的 C8 沙雷氏菌发酵生产沙雷氏肽酶,经色谱纯化,并在不同 pH 值条件下通过 EDC/NHS 偶联固定在羧基化纳米粒子上。纳米颗粒活化的 pH 值为 5.1,酶偶联的 pH 值为 5.5,从而获得了最佳的酶活性。通过傅立叶变换红外光谱确认了固定化,并测定了迈克尔斯-门顿动力学(Michaelis-Menten kinetics)。值得注意的是,在 1000 µg/mL 的浓度下,生物共轭物对大肠杆菌的抗菌活性是游离酶或 TiO2-NPs 的两倍。本研究成功开发了一种血清肽酶-二氧化钛生物共轭物,它具有更高的酶稳定性和抗菌特性。固定化酶抗菌活性的提高为开发抗菌剂抗药性的新工具提供了一种前景广阔的方法,有望应用于医疗保健、食品安全和环境保护领域。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Journal of Functional Biomaterials
Journal of Functional Biomaterials Engineering-Biomedical Engineering
CiteScore
4.60
自引率
4.20%
发文量
226
审稿时长
11 weeks
期刊介绍: Journal of Functional Biomaterials (JFB, ISSN 2079-4983) is an international and interdisciplinary scientific journal that publishes regular research papers (articles), reviews and short communications about applications of materials for biomedical use. JFB covers subjects from chemistry, pharmacy, biology, physics over to engineering. The journal focuses on the preparation, performance and use of functional biomaterials in biomedical devices and their behaviour in physiological environments. Our aim is to encourage scientists to publish their results in as much detail as possible. Therefore, there is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. Several topical special issues will be published. Scope: adhesion, adsorption, biocompatibility, biohybrid materials, bio-inert materials, biomaterials, biomedical devices, biomimetic materials, bone repair, cardiovascular devices, ceramics, composite materials, dental implants, dental materials, drug delivery systems, functional biopolymers, glasses, hyper branched polymers, molecularly imprinted polymers (MIPs), nanomedicine, nanoparticles, nanotechnology, natural materials, self-assembly smart materials, stimuli responsive materials, surface modification, tissue devices, tissue engineering, tissue-derived materials, urological devices.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信