In Vitro Cytotoxicity and Hemolysis Effect of Poly-Gamma-Glutamic Acid Nano-Polymer Biosynthesized Using Some Isolates of Bacillus spp.

IF 2.1 Q3 MICROBIOLOGY

Microbiology Research Pub Date : 2023-10-29 DOI:10.3390/microbiolres14040118

Eman M. Elsayed, Ahmed A. Farghali, Mohamed I. Zanaty, Medhat Abdel-Fattah, Dalal Hussien M. Alkhalifah, Wael N. Hozzein, Ahmed M. Mahmoud

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引用次数: 0

Abstract

Natural polymers have good biological, biocompatible properties and a low degradation rate, so they can be used in medical applications. This study demonstrates an optimized biosynthesis for poly-gamma-glutamic acid nano-polymer (ɣ-PGA NP) using three bacterial isolates encoded as B4, B5, and B6. The 16S rRNA gene sequence of the isolates showed 98.5, 99, and 99.8% similarity with Bacillus sp., with accession numbers MZ976778, MZ976779, and MZ956153, respectively. The optimal conditions of the biosynthesis were 35 °C, followed by incubation for 30 h, 7 pH, 0.7 O.D600 of bacterial broth, carbon sources of glutamic acid and glucose, and ammonium sulfate as the nitrogen source. Biosynthesized ɣ-PGA NPs were characterized using X-ray diffraction (XRD), Zeta sizer, Fourier transform infrared (FT-IR), transmission electron microscopy (TEM), and nuclear magnetic resonance (1HNMR). The TEM study showed the formation of oval-shaped ɣ-PGA NPs with sizes 31.75–32.69 nm, 18.63–21.66 nm, and 37.57–39.22 nm for B4, B5, and B6, respectively. XRD, FT-IR, and 1HNMR confirmed the purity of the polymer. The hemolysis rates of the biosynthesized ɣ-PGA NPs were below the acceptable value. Moreover, the cytotoxicity test conducted using an MTT assay on peripheral blood mononuclear cells (PBMCs) revealed no significant effect on the cell viability within a dose range of up to 512 μg/mL.

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利用芽孢杆菌分离物合成的聚γ -谷氨酸纳米聚合物的体外细胞毒性和溶血作用。

天然聚合物具有良好的生物相容性和低降解率，可用于医疗领域。本研究利用编码为B4、B5和B6的三种细菌分离物优化了聚γ -谷氨酸纳米聚合物(γ -PGA NP)的生物合成。分离菌株的16S rRNA基因序列与芽孢杆菌相似度分别为98.5%、99%和99.8%，登录号分别为MZ976778、MZ976779和MZ956153。生物合成的最佳条件为35℃，7 pH, 0.7 O.D600菌液，谷氨酸和葡萄糖为碳源，硫酸铵为氮源，孵育30 h。利用x射线衍射(XRD)、Zeta粒度仪、傅里叶变换红外(FT-IR)、透射电子显微镜(TEM)和核磁共振(1HNMR)对生物合成的α -PGA NPs进行了表征。透射电镜研究表明，B4、B5和B6分别形成了尺寸为31.75 ~ 32.69 nm、18.63 ~ 21.66 nm和37.57 ~ 39.22 nm的椭圆形的α -PGA NPs。XRD, FT-IR和1HNMR证实了该聚合物的纯度。生物合成的α -PGA NPs溶血率低于可接受值。此外，采用MTT法对外周血单个核细胞(PBMCs)进行的细胞毒性试验显示，在高达512 μg/mL的剂量范围内，对细胞活力无显著影响。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microbiology Research MICROBIOLOGY-

CiteScore

1.90

自引率

6.70%

发文量

审稿时长

10 weeks

期刊介绍： Microbiology Research is an international, online-only, open access peer-reviewed journal which publishes original research, review articles, editorials, perspectives, case reports and brief reports to benefit researchers, microbiologists, physicians, veterinarians. Microbiology Research publishes ‘Clinic’ and ‘Research’ papers divided into two different skill and proficiency levels: ‘Junior’ and ‘Professional’. The aim of this four quadrant grid is to encourage younger researchers, physicians and veterinarians to submit their results even if their studies encompass just a limited set of observations or rely on basic statistical approach, yet upholding the customary sound approach of every scientific article.