School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India; Environment and Sustainability Group, J. Craig Venter Institute, La Jolla, CA, USA.
School of Environmental Sciences, Jawaharlal Nehru University, New Delhi, India.
Environ Res. 2021 Jan;192:110277. doi: 10.1016/j.envres.2020.110277. Epub 2020 Oct 16.
The present study discusses the genomic analysis of Bacillus sp. ISTL8 along with the production of EPS (Extracellular polymeric substances) using carbofuran, a toxic carbamate pesticide. Bacillus strain was isolated from landfill soil and evaluated for high growth rates and EPS production. One strain, renamed ISTL8 grew on a broad range of carbon sources, including toxic carbofuran, while producing copious EPS. Growth assays verified the strain to be thermophilic, low salt tolerant, and with a preference for neutral pH. SEM (Scanning Electron Microscopy) was used for morphological characterization of the EPS while the monomeric composition, bonding patterns and functional groups were deduced by GC-MS (Gas Chromatography-Mass Spectrometry), H and C NMR (Nuclear Magnetic Resonance) and FTIR (Fourier Transform Infrared Spectroscopy). The production of EPS using carbofuran (carbamate pesticide) as a carbon source was found to be 6.20 ± 0.29 g L containing 61.17% w/w carbohydrates, 29.72% w/w proteins and 6.11% w/w lipids (of dry EPS). The potential cytotoxicity of EPS was evaluated with 3- (4,5-dimethyl thiazol-2-Yl) -2,5-diphenyl tetrazolium bromide (MTT) assay and found non-toxic (2.25%). WGS (Whole genome sequencing) was performed for the strain Bacillus sp. ISTL8 producing EPS; an array of genes putatively involved in the EPS production were identified in several different genomic locations, guiding potential genetic manipulation studies in the future. The results highlight the potency of a bacterial isolate Bacillus sp. ISTL8 to produce non-cytotoxic EPS using carbofuran that can be further harnessed for environmental and commercial applications. Additionally, WGS revealed an array of EPS specific genes which can be effectively engineered for much enhanced production.
本研究讨论了芽孢杆菌 sp. ISTL8 的基因组分析以及利用毒carbamate 农药克百威生产 EPS（胞外聚合物）。芽孢杆菌菌株是从垃圾填埋场土壤中分离出来的，具有高生长率和 EPS 生产能力。一株命名为 ISTL8 的菌株能够在广泛的碳源上生长，包括有毒的克百威，同时产生大量的 EPS。生长实验证实该菌株为嗜热、耐低盐、偏好中性 pH。SEM（扫描电子显微镜）用于 EPS 的形态学表征，而单体组成、键合模式和官能团则通过 GC-MS（气相色谱-质谱联用）、H 和 C NMR（核磁共振）和 FTIR（傅里叶变换红外光谱）来推断。使用克百威（carbamate 农药）作为碳源生产 EPS 的产量为 6.20±0.29g/L，其中含有 61.17%w/w 的碳水化合物、29.72%w/w 的蛋白质和 6.11%w/w 的脂质（干 EPS）。通过 3-（4,5-二甲基噻唑-2-Yl）-2,5-二苯基四氮唑溴盐（MTT）测定法评估 EPS 的潜在细胞毒性，发现其无毒性（2.25%）。对产 EPS 的芽孢杆菌 sp. ISTL8 进行 WGS（全基因组测序）；在几个不同的基因组位置鉴定出了一系列与 EPS 生产相关的假定基因，为未来的潜在遗传操作研究提供了指导。研究结果突出了细菌分离株芽孢杆菌 sp. ISTL8 的潜力，它可以使用克百威生产非细胞毒性的 EPS，可进一步用于环境和商业应用。此外，WGS 揭示了一系列 EPS 特异性基因，可以有效地进行工程改造以实现更高的产量。
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