SKRIPSI Jurusan Biologi - Fakultas MIPA UM, 2018

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CHARACTERIZATION OF Pseudomonas stutzeri Outer Membrane Protein and Response Profile of Antibiotics-Extracts of Piper betle Leaves





Cahya, Dini Aulia. 2018. Characterization of Pseudomonas stutzeri Outer Membrane Proteins and Response Profile of Antibiotics-Extracts of Piper betle Leaves. Bachelor Thesis, Departement of Biology,Faculty of Mathematics and Science, State University of Malang. Supervisior : (I) Abdullah DolaDalee, M.ScM., (II) Dr.Umie Lestari, M.Si., (III) Dr.EndangSuarsini, M.Ked.


Keywords :Outer Membrane Protein, Pseudomonas stutzeri, Antibiotics, Piper betle Extract, Response Profile.

Pseudomonas stutzeri is one of the cause nosocomial infection.Pseudomonas stutzeriis ubiquitous in hospital environments and it is an opportunistic, but rare, pathogen. It has been reported to cause bacteraemia, meningitis, pneumonia and osteomyelitis. This species was susceptible to a wide range of antibiotics, including ampicillin, tetracycline, streptomycin, nalidixic acid, neomycin, kanamycin, polymyxin, and gentamicin. The emergence of poly antimicrobial resistant strains of hospital pathogens has also presented a challenge in the provision of good quality inpatient care. At the global level, people have understood the bad effect of antibiotics and they are now shifting over to natural products.Therefore, it is of great interest to carry out a screening of these plants in order to validate their use in folk medicine and to reveal the active principle by isolation and characterisation of their constituents.

The aim of this research is to Characterized the outer membrane protein of Pseudomonas stutzeri and investigasting the antibacterial effect of antibiotics and the methanol,acetone and hexane extract of Piper betle leaves. SDS-PAGE electrophoresis was used in this research to isolate protein based on the molecular weight. Microdilution was used to determine the MIC and MBC of antibiotics and Piper betle leaves extracts. The extracts was used 3 solvents , methanol, aceton and hexane. Tetracycline, Gentamycin, Kanamycin and Penicillin was used.

The Quantitative assay of outer membrane protein of Pseudomonas stuzeri’s protein extractions showed concentration range 1.78 Mg/ml-2.06 Mg/ml. Meanwhile, the Quantitative assay  showed 13 bands protein from SDS-PAGE extraction. The bands protein are 127.8 kDa, 107.9 kDA, 91.2 kDa, 81.5 kDa, 68.8 kDa, 54.9 kDa, 49. 1 kDa, 43. 8 kDa, 39.2 kDa, 33.1 kDa, 24.9 kDa, 16.8 kDa, 14.2 kDa. MIC values were determined by two-fold serial dilution microplate bioassay and indicating growth with tetrazolium violet. MBC valus were determined by streaked the MIC from microplate to lactos agar and incubated in 24 hours.Piper betle L Acetone Extracts was the most active with average MIC value for Pseudomonas spp ( 31,25 µg/ml) followed by Piper betle L Methanol Extracts with values 62,5 µg/ml and Piper betle L Hexane Extracts with values 125 µg/ml. for the Antibiotics, Gentamycin was the most sensitive for Pseudomonas spp with values 0.048838 ug/ml followed by Kanamycin with values 12,5 µg/ml, Tetracycline and Penicilin showed that Pseudomonas spp is resistantance to those antibiotics.