Cloning of polyhydroxyalkanoate synthesis genes and expression in recombinants

• Main Authors: Li-Ying Kuo, 郭俐櫻
• Other Authors: Yu-Tzu Huang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/41674371373733377662

碩士 === 中原大學 === 土木工程研究所 === 97 === It is economically efficiency to treat wastewater by activated sludge. However, disposal of waste sludge which provide by activated sludge treatment is still a major issue for environment. Recently, many research focus on the study of waste reduction and generation . In this study, PHA synthesis genes of specific strains Ralstonia eutropha in activated sludge system were cloned to Escherichia coli. Next, different temperature and carbon source were optimized to maximize the PHA content of recombinant bacteria. The results showed that the highest PHB contents of wild-type fed with glucose, fructose and, acetic acid as carbon source were 25.9％, 30.2％, and 17.8％, respectively. Moreover, this study constructed recombinant E.coli harboring plasmid pET-phaCABRe and cultured at the optimize temperature 30℃ with glucose, fructose and, acetic acid. The PHB content were 37.7％, 37％,and 14.6％, respectively. Furthermore, inducer was added and PHB content of recombinant E.coli was the highest, 73.4%, while glucose was used as carbon source. The recombinant E.coli constructed in this study have ability to synthesize PHA, we will apply this recombinant by feeding supernatant of sludge treatment as the carbon source to synthesize PHA in the future.