Marey Moustafa A, Abozahra Rania, El-Nikhely Nefertiti A, Kamal Miranda F, Abdelhamid Sarah M, El-Kholy Mohammed A
Department of Microbiology and Biotechnology, Division of Clinical and Biological Sciences, College of Pharmacy, Arab Academy for Science, Technology and Maritime Transport (AASTMT), Abu Kir Campus, P.O. Box 1029, Alexandria, Egypt.
Microbiology and Immunology Department, Faculty of Pharmacy, Damanhour University, Damanhour, Egypt.
Microb Cell Fact. 2024 Jun 13;23(1):174. doi: 10.1186/s12934-024-02438-6.
The objectives of the current study were to extract pyocyanin from Pseudomonas aeruginosa clinical isolates, characterize its chemical nature, and assess its biological activity against different bacteria and cancer cells. Due to its diverse bioactive properties, pyocyanin, being one of the virulence factors of P. aeruginosa, holds a promising, safe, and available therapeutic potential.
30 clinical P. aeruginosa isolates were collected from different sources of infections and identified by routine methods, the VITEK 2 compact system, and 16 S rRNA. The phenazine-modifying genes (phzM, phzS) were identified using polymerase chain reaction (PCR). Pyocyanin chemical characterization included UV-Vis spectrophotometry, Fourier Transform Infra-Red spectroscopy (FTIR), Gas Chromatography-Mass Spectrometry (GC-MS), and Liquid Chromatography-Mass Spectrometry (LC-MS). The biological activity of pyocyanin was explored by determining the MIC values against different clinical bacterial strains and assessing its anticancer activity against A549, MDA-MB-231, and Caco-2 cancer cell lines using cytotoxicity, wound healing and colony forming assays.
All identified isolates harboured at least one of the phzM or phzS genes. The co-presence of both genes was demonstrated in 13 isolates. The UV-VIS absorbance peaks were maxima at 215, 265, 385, and 520 nm. FTIR could identify the characteristic pyocyanin functional groups, whereas both GC-MS and LC-MS elucidated the chemical formula CHNO, with a molecular weight 210. The quadri-technical analytical approaches confirmed the chemical nature of the extracted pyocyanin. The extract showed broad-spectrum antibacterial activity, with the greatest activity against Bacillus, Staphylococcus, and Streptococcus species (MICs 31.25-125 µg/mL), followed by E. coli isolates (MICs 250-1000 µg/mL). Regarding the anticancer activity, the pyocyanin extract showed IC values against A549, MDA-MB-231, and Caco-2 cancer cell lines of 130, 105, and 187.9 µg/mL, respectively. Furthermore, pyocyanin has markedly suppressed colony formation and migratory abilities in these cells.
The extracted pyocyanin has demonstrated to be a potentially effective candidate against various bacterial infections and cancers. Hence, the current findings could contribute to producing this natural compound easily through an affordable method. Nonetheless, future studies are required to investigate pyocyanin's effects in vivo and analyse the results of combining it with other traditional antibiotics or anticancer drugs.
本研究的目的是从铜绿假单胞菌临床分离株中提取绿脓菌素,表征其化学性质,并评估其对不同细菌和癌细胞的生物活性。绿脓菌素作为铜绿假单胞菌的毒力因子之一,因其多样的生物活性特性,具有安全且可用的治疗潜力。
从不同感染源收集30株临床铜绿假单胞菌分离株,通过常规方法、VITEK 2 compact系统和16S rRNA进行鉴定。使用聚合酶链反应(PCR)鉴定吩嗪修饰基因(phzM、phzS)。绿脓菌素的化学表征包括紫外可见分光光度法、傅里叶变换红外光谱法(FTIR)、气相色谱 - 质谱联用仪(GC - MS)和液相色谱 - 质谱联用仪(LC - MS)。通过测定对不同临床细菌菌株的最低抑菌浓度(MIC)值,并使用细胞毒性、伤口愈合和集落形成试验评估其对A549、MDA - MB - 231和Caco - 2癌细胞系的抗癌活性,来探索绿脓菌素的生物活性。
所有鉴定出的分离株均含有phzM或phzS基因中的至少一个。13株分离株中同时存在这两个基因。紫外可见吸收峰在215、265、385和520nm处为最大值。FTIR可鉴定绿脓菌素的特征官能团,而GC - MS和LC - MS均阐明其化学式为CHNO,分子量为210。这四种技术分析方法证实了所提取绿脓菌素的化学性质。提取物显示出广谱抗菌活性,对芽孢杆菌属、葡萄球菌属和链球菌属的活性最强(MIC为31.25 - 125μg/mL),其次是大肠杆菌分离株(MIC为250 - 1000μg/mL)。关于抗癌活性,绿脓菌素提取物对A549、MDA - MB - 231和Caco - 2癌细胞系的IC值分别为130、105和187.9μg/mL。此外,绿脓菌素显著抑制了这些细胞中的集落形成和迁移能力。
所提取的绿脓菌素已证明是对抗各种细菌感染和癌症的潜在有效候选物。因此,目前的研究结果有助于通过一种经济实惠的方法轻松生产这种天然化合物。尽管如此,未来还需要研究绿脓菌素在体内的作用,并分析将其与其他传统抗生素或抗癌药物联合使用的结果。