In a traditional classroom environment, instructor enthusiasm has been shown to enhance student’s emotion, affective perceptions, intrinsic motivation, and cognitive outcome. Additionally, emotional response theory argues that enthusiastic verbal and nonverbal cues of an instructor will induce positive emotional states in learners, which in turn, enact learners’ approach behaviours in the learning process. Therefore, should a pedagogical agent convey enthusiastic behaviours in a multimedia learning environment? Literature and theoretical reviews offer two competing views. The first view, based on emotional response theory, predicts that enthusiastic verbal and nonverbal cues of a pedagogical agent can induce higher positive emotions in learners, which in turn, enhance affective perceptions, intrinsic motivation, and cognitive outcome. However, the second view, based on cognitive load theory, suggests that pedagogical agent enthusiasm may increase extraneous cognitive load (additional processing in the mind), which negatively impact emotion, affective perceptions, intrinsic motivation, and cognitive outcome. To investigate the effects of agent enthusiasm, seventy-two university freshmen interacted with either an enthusiastic agent or a neutral agent (operationalized through vocal tones, facial expression, gestures, and remarks) that simulates the instructional role of a virtual tutor that delivers narrative demonstrations on how to predict the outputs of C-Programming algorithms. The results of our study showed that pedagogical agent enthusiasm significantly enhanced emotion, intrinsic motivation, affective perceptions, and cognitive outcome. Moreover, mediation analyses revealed that the facilitating effects of agent enthusiasm on intrinsic motivation, affective perceptions of the learning environment, and affective perceptions of the pedagogical agent were fully mediated by a learner’s positive emotion, thus demonstrating that the framework of emotional response theory can be applied to learner-agent interaction in a multimedia learning environment. Implications and suggestions for future research related to pedagogical agent enthusiasm are discussed in this paper.