Borneo bioinformatics portal test is a critical element of SQA and represents a comprehensive review of specifications, design and coding. The test represents an abnormality in the development of the portal. A series of tests systematically reveals several different types of errors. This study aims to evaluate the performance and optimization of Borneo’s Bioinformatics portal with a series test activities using the Web Performance Optimization methodology. Testing query performance with measuring the response time and page loading timings from the object relationship mapping (ORM) model Laravel PHP framework in offline and online. For optimization, we set a pre-test and post-test scenario to evaluate the efficiency performance test results. The results study found that the query relation model, parsing script (javaScript and CSS), service scale and dimension images in the interaction process to the database are the dominant resources affecting the performance of the Bioinformatics portal. Performance optimization through determining the appropriate query relation model, minify and defer parsing script or combine images using CSS sprites to reduce scala image.