Engineering microbes for phosphate removal

• Main Author: Chua, Chistopher Kok Leng.
• Other Authors: Evan Abelard Laksmana
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Engineering
• Online Access: http://hdl.handle.net/10356/51820
• Institution: Nanyang Technological University

In this project, a solution was proposed to remove and recycle phosphate from wastewater. Experiments were conducted to find a viable solution. Genetic modifications were carried out by transforming plasmids containing phosphate specific transport (pst) and polyphosphate kinase (ppk), into Escherichia coli. Ascorbic acid method was used to measure the content of phosphate in the medium and the plasmid pBC29 carrying the ppk gene was found to have the best results. The growth of M.g was experimented and the protocol that offers M.g with the best growth rate and magnetic property was adopted. In order to insert ppk gene into M.g, it has to be inserted into the pBBR1MCS-2 vector first, before into M.g, as it contains the necessary tra genes required for conjugation. Digestive enzymes, BamHI and XohI were used to insert the ppk gene into the pBBR1MCS-2. The plasmid pBBR1MCS-2 is then inserted into E.coli. An established cloning protocol was followed to transfer ppk gene into M.g, and resulted in strain of M.g that is resistant to kanamycin and rifampicin.