Phosphorylation exerts a crucial role in multiple biological cellular processes which is catalyzed by protein kinases and closely related to many diseases. Identification of kinase-substrate relationships is important for understanding phosphorylation and provides a fundamental basis for further disease-related research and drug design. In this study, we develop a novel computational method to identify kinase-substrate relationships based on multiple kernel learning. The comparative analysis is based on a 10-fold cross-validation process and the dataset collected from the Phospho.ELM database. The results show that ksrMKL is greatly improved in various measures when compared with the single kernel support vector machine. Furthermore, with an independent test dataset extracted from the PhosphoSitePlus database, we compare ksrMKL with two existing kinase-substrate relationship prediction tools, namely iGPS and PKIS. The experimental results show that ksrMKL has better prediction performance than these existing tools.