Department of Biotechnology & Biochemistry, Sardar Bhagwan Singh University, Balawala, Dehradun, Uttarakhand, 248161, India.
Department of Biotechnology, Shri Guru Ram Rai University, Patel Nagar, Dehradun, Uttarakhand, 248001, India.
World J Microbiol Biotechnol. 2023 Aug 10;39(10):274. doi: 10.1007/s11274-023-03711-3.
The lead is poisonous metal and because of its chemical nature it acts as an environmental contaminant through the water or soil and it becomes toxic to humans. The toxicity of Pb occurs as a change in the conformation of nucleic acid and protein, inhibition of enzyme activity, disruption of membrane function and oxidative phosphorylation. For protoplast preparation, the removal of the cell wall and protoplast formation obtained by specific lytic enzyme. In cytoplasmic membrane, the envelope of bacteria consists of overlying cell wall. From hypertonic environment, the complete cell wall removal occurs due to which it maintains the osmotic integrity of the cell and produces the protoplast. In current work, protoplasts were produced by specific lytic enzyme (lysozyme and macerozyme), chemo fused (with the help of Polyethylene Glycol) and regenerated from strains Staphylococcus sp. and Bacillus sp. The fused protoplast was spherical in shape observed under microscopy. Colonies were screened on specific medium supplemented with Pb (Concentration at the rate of 1.5mM). One resistant colony (MICBT-1) was selected and further examined and applied for the transformation of Pb in the broth medium. The strain removed 98% of Pb at 1mM concentration. Next, sucrose containing medium was best which gives maximum protoplast regeneration. From various organisms, fusion technique has been used to combine the genes to create the strains having desired properties. This is a significant technique for engineering of bacterial strains for advantageous applied properties. Further MICBT-1 applied in artificially contaminated soil and removed maximally in exchangeable fraction (remains up to 0.05 mM). An efficient bioremediating agent for lead transformation from soil and water is expected to ease the ever-increasing problem. Further, it is needful to obtain new strain with the help of protoplast technology which can reduce the pollutant. This lead tolerant strain can be applied for bioremediation purposes in the Pb contaminated soil and water environment.
铅是一种有毒的金属，由于其化学性质，它会通过水或土壤成为环境污染物，对人类有毒。Pb 的毒性表现为核酸和蛋白质构象的改变、酶活性的抑制、膜功能的破坏和氧化磷酸化的中断。对于原生质体制备，通过特定的裂解酶去除细胞壁并形成原生质体。在细胞质膜中，细菌的包膜由细胞壁覆盖。由于高渗环境，细胞壁完全去除，从而保持细胞的渗透完整性并产生原生质体。在当前的工作中，通过特定的裂解酶（溶菌酶和纤维素酶）、化学融合（使用聚乙二醇）并从葡萄球菌属和芽孢杆菌属菌株中再生原生质体。融合的原生质体在显微镜下观察呈球形。在补充有 Pb（浓度为 1.5mM）的特定培养基上筛选出菌落。选择一个抗性菌落（MICBT-1）并进一步检查和应用于在肉汤培养基中转化 Pb。该菌株在 1mM 浓度下去除了 98%的 Pb。接下来，含有蔗糖的培养基是最好的，它可以最大程度地再生原生质体。从各种生物体中，融合技术已被用于组合基因，以创建具有所需特性的菌株。这是对具有有利应用特性的细菌菌株进行工程改造的重要技术。进一步将 MICBT-1 应用于人工污染的土壤中，并在可交换部分（最多保留 0.05mM）中最大程度地去除。一种有效的生物修复剂，可以从土壤和水中转化 Pb，有望缓解日益严重的问题。此外，有必要借助原生质体技术获得新的菌株，以减少污染物。这种耐铅菌株可应用于 Pb 污染土壤和水环境的生物修复。
