Usage of Mobile banking services increases rapid as possible all over the world and also it intends to enhance the quality of banking transaction. But people around Sri Lanka still reluctant to use mobile banking services instead of traditional banking modes. This study attempts to investigate impact of E-service quality and interaction effect of gender on customer satisfaction from mobile banking services. Although there were lots of researches had been conducted related to the Moderating role of gender in the relationship between E-Service quality and customer satisfaction in foreign countries, the researches relating to Sri Lanka particularly to Trincomalee district are limited. So, it is needed to find out the moderating role of gender in the relationship of E-service quality and customer satisfaction in Trincomalee. Therefore, Convenient sampling and self-administered questionnaires were used to collect responses from one hundred eighty-nine sample from Trincomalee district, Sri Lanka were collected and investigated thoroughly. Perceived Ease of use and Perceived usefulness has been regarded as the dimensions which determines E-Service quality (Davis, 1989). To identify the level, relationship, impact and the moderation effect Mean scores, Pearson correlation co-efficient, Regression Analysis and Process v3. 4 by Andrew F. Hayes were used respectively. This study concluded that there is a strong positive relationship and the impact on customer satisfaction due to the e-service quality. But the moderating role of gender tend to become insignificant. Thereby it can be concluded that banking strategies need not to be focused separately on each gender instead whole respondents can be viewed as a whole without any gender-based groups. But improving E-Service quality will have significant positive impact on customer satisfaction on mobile banking services.