Information security has become an important issue in the world. Various information and communication technologies, such as the Internet of Things, big data, cloud, and artificial intelligence, are developing, and the need for information security is increasing. Although the necessity of information security is expanding according to the development of information and communication technology, interest in information security investment is insufficient. In general, measuring the effect of information security investment is difficult, so appropriate investment is not being practice, and organizations are decreasing their information security investment. In addition, since the types and specification of information security measures are diverse, it is difficult to compare and evaluate the information security countermeasures objectively, and there is a lack of decision-making methods about information security investment. To develop the organization, policies and decisions related to information security are essential, and measuring the effect of information security investment is necessary. Therefore, this study proposes a method of constructing an investment portfolio for information security measures using game theory and derives an optimal defence probability. Using the two-person game model, the information security manager and the attacker are assumed to be the game players, and the information security countermeasures and information security threats are assumed as the strategy of the players, respectively. A zero-sum game that the sum of the players' payoffs is zero is assumed, and we derive a solution of a mixed strategy game in which a strategy is selected according to probability distribution among strategies. In the real world, there are various types of information security threats exist, so multiple information security measures should be considered to maintain the appropriate information security level of information systems. We assume that the defence ratio of the information security countermeasures is known, and we derive the optimal solution of the mixed strategy game using linear programming. The contributions of this study are as follows. First, we conduct analysis using real performance data of information security measures. Information security managers of organizations can use the methodology suggested in this study to make practical decisions when establishing investment portfolio for information security countermeasures. Second, the investment weight of information security countermeasures is derived. Since we derive the weight of each information security measure, not just whether or not information security measures have been invested, it is easy to construct an information security investment portfolio in a situation where investment decisions need to be made in consideration of a number of information security countermeasures. Finally, it is possible to find the optimal defence probability after constructing an investment portfolio of information security countermeasures. The information security managers of organizations can measure the specific investment effect by drawing out information security countermeasures that fit the organization's information security investment budget. Also, numerical examples are presented and computational results are analyzed. Based on the performance of various information security countermeasures: Firewall, IPS, and Antivirus, data related to information security measures are collected to construct a portfolio of information security countermeasures. The defence ratio of the information security countermeasures is created using a uniform distribution, and a coverage of performance is derived based on the report of each information security countermeasure. According to numerical examples that considered Firewall, IPS, and Antivirus as information security countermeasures, the investment weights of Firewall, IPS, and Antivirus are optimized to 60.74%, 39.26%, and 0%, respectively. The result shows that the defence probability of the organization is maximized to 83.87%. When the methodology and examples of this study are used in practice, information security managers can consider various types of information security measures, and the appropriate investment level of each measure can be reflected in the organization's budget.