In recent years, there has been tremendous progress in the development of nanobiosensors, which are a critical need of the hour and could be integrated with point-of-care testing for various applications, including healthcare, the environment, energy harvesting, electronics, and the food industry. In order to create biosensors with high selectivity, uniqueness, and sensitivity, nanomaterials play a crucial role in effectively sensing bioreceptors like cells, enzymes, and antibodies. Without nanomaterials, it is nearly impossible to carry out any application in science and technology. Nanomaterials’ unique electrical, thermal, chemical, optical, mechanical, and physical properties set them apart from tiny particles employed in a wide range of applications. There is a growing global demand for a secure workplace as people become more health conscious. This chapter briefly reviewed environmental pollution issues, such as the classification of pollutants and the specific nanosensors used to detect them. This will also encompass the recent breakthroughs in advanced nanotechnology and nanobiotechnology applications in environmental pollutant detection using nanobiosensors for air pollutants and sensing environmental pathogenic microbes that could help control environmental pollution and could reduce costs and save time. However, for large-scale applications, the nanotechnology industries should promote safety regulations, including developing biosources related to nanostructures, to make this technology commercially viable and to develop advanced technologies, such as environmentally friendly materials and green systems, that are compatible with environmental requirements. In order to address the demand for vast quantities of highly pure, structurally well-identified, and precisely functionalized nanomaterials, green nanofabrication also needs to be aggressively pursued.