Polyethylene (PE) is the second most abundant plastic worldwide due to its broad application in manufacturing disposable materials such as bags and bottles. Since it is highly resistant to natural biodegradation, it can accumulate in landfills, causing various ecological and toxicological consequences. Even though microbial degradation of PE has been extensively investigated, complete biodegradation of PE has yet to be achieved, and comparisons of PE biodegradation experiment findings are not practical. This may be due to the wide variety of PE substances used, and a wide variety of culture conditions, and many of the published findings still need key chemistry of PE materials, as well as basic biochemistry and microbiology. This review highlights the need for standardization in PE biodegradation studies and defining key biochemical terms. It aims to identify deficiencies and challenges in PE degradation experiments, enhancing future research for scientific and technological advancement. This review provides a biochemically based definition of PE biodegradation and summarizes microorganisms responsible for PE biodegradation. We also caution readers about the chemical makeup and structure of the PE, as well as the approaches utilized in microbes’ isolation. This review defines the fundamental elements required to conduct an effective PE biodegradation investigation.