Penetration testing and vulnerability assessment have become pivotal in strengthening corporate IT environments against an ever-expanding threat landscape. This systematic review critically examines the application of two widely utilized tools, Nessus and Wireshark, for enhancing cybersecurity resilience within corporate infrastructures. Nessus, a leading vulnerability scanner, enables organizations to proactively identify, prioritize, and remediate weaknesses in their systems. Wireshark, a powerful network protocol analyzer, assists in scrutinizing traffic for anomalies that could signal ongoing attacks or security misconfigurations. Through a structured analysis of peer-reviewed articles, technical reports, and case studies, this review categorizes findings across multiple corporate settings, highlighting best practices, challenges, and gaps in implementation. The study employs PRISMA methodology to ensure rigorous selection, assessment, and synthesis of 52 relevant publications from 2015 to 2021. Results indicate that while Nessus excels in vulnerability identification across operating systems and web applications, its effectiveness depends significantly on regular updates and proper configuration. Meanwhile, Wireshark’s packet-level inspection provides unparalleled visibility into network behavior but demands expert knowledge for accurate interpretation and actionable insights. The integration of Nessus and Wireshark in a comprehensive security strategy leads to measurable improvements in incident response times, compliance adherence, and risk mitigation outcomes. However, reliance on manual configurations and the potential for overwhelming data volumes remain persistent challenges. Emerging trends reveal that coupling Nessus with machine learning techniques for automated prioritization, and combining Wireshark with AI-driven anomaly detection, is gaining traction among advanced corporate security teams. Furthermore, organizational culture, training, and continuous monitoring were identified as critical enablers for maximizing the effectiveness of these tools. This review concludes with recommendations for developing adaptive frameworks that synergize Nessus and Wireshark capabilities with evolving cybersecurity needs. Future research should explore the scalability of hybrid automation models and the integration of these tools within zero-trust architectures to sustain proactive defense mechanisms in dynamic corporate environments.