| Summary: | 碩士 === 國立高雄海洋科技大學 === 海洋環境工程研究所 === 96 === The sea transportation is important at present, the ports is the place of trade and ship berthing. The ship fuel relies mainly on diesel which is the main resource of maritime oil pollution.
This study used Acinetobacter sp., collected from the Chien-chen fishing harbor in Kaohsiung, confirmed to be the Gram-negative bacteria and be capable to degrade diesel in the sea water although the degrading mechanism is still not clear. Therefore, this study focused on separating the functional protein of degrading diesel from A. sp..
In his study, experiment culture medium was designed to add diesel into sea water culture medium while contrast culture medium was designed to add LB into sea water culture medium. Using three stages in succession extract method, ECP、CMP and OMP were obtained respectively. By using SDS-PAGE to separate proteins and using Quantity One software to match the proteins band of experiment group and contrast group, it was obtained that OMP showed an obvious difference in 62 KDa、46 KDa and 28 KDa protein molecular weights. After cutting gels and extracting the three proteins, reverse phase high performance liquid chromatography (RP-HPLC) was used to separate and sublime the function proteins. Results showed that 46 KDa molecular weight had a signal at retention time of 63.21 min (the signal intensity was 4711 mmV), as well as 28 KDa molecular weight had a signal at retention time of 62.85 min (the signal intensity was 4010 mmV). It was demonstrated that 46 KDa and 28 KDa molecular weights were the diesel functional proteins of A. sp..
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