Document Details
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Document Type |
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Thesis |
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Document Title |
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ISOLATION AND CHARACTERIZATION OF CHITIN AND CHITINASE WITH THE MOLECULAR STRUCTURE SCREENING OF CHITINASE GENE FROM LOCAL STRAINS, SAUDI ARABIA عزل ووصف خصائص الكيتين و الكايتنيز و فحص التركيب الجزيئي لجين الكايتنيز من سلالات محلية, المملكة العربية السعودية |
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Subject |
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faculty of science |
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Document Language |
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Arabic |
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Abstract |
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Chitin is a natural biopolymer, and considered as the most abundant material in earth after cellulose. This biopolymer could be degraded into low molecular weight products through chitinolytic enzymes. In this research, the chitin have been extracted from shrimp shell wastes and cockroaches obtained from local area of Saudi Arabia. The chitin percent were found to be 18.9 and 1.4 % for shrimp shell wastes, and cockroach, respectively. The extracted chitin was characterized using infrared spectroscopy, X-ray diffractometry, and scanning electron microscope. The results were compared with previously isolated chitin in the literatures, and they were quite similar. In other hand, the production of the chitinolytic enzymes implies change in the activity of different species. In this research, a new isolates belonging to Streptomyces laurentii strain ATCC 31255 and Cellulosimicrobium funkei strain W6122 have been characterize for the first time for their chitinase activity. They were identified using 16S rRNA gene analysis, and in the liquid medium, the two isolates have an enzyme activity of 0.533 and 0.537 (U/ml), respectively. The maximum chitinase production were obtained when those bacterial strains were grown in Luria-Bertani (LB) broth amended with 1% colloidal chitin, for 1 day, and at temperature of 30 ⁰ C. The optimum pH value were found to be 4 for S.laurentii and 7 for C.funkei. The enzyme have been purified using Sephadex G-100 and DEAE-Cellulose chromatography column, and found to have a similar molecular size of ~50 kDa. Also, the chitinase encoding gene were screened for S.laurentii using bioinformatics based tools, since there complete genome is available. The identified gene were found to have an ORF of 687 bp, encoding 221 amino acid, and a percentage of 68.2 % for G+C. However, those bacterial species could be used to test the different applications of chitinase enzyme and in recycling of disposable chitin wastes. |
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Supervisor |
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Dr. Maaly Hassan Ali |
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Thesis Type |
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Master Thesis |
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Publishing Year |
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1441 AH
2019 AD |
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Co-Supervisor |
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Dr. Jehan Khan |
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Added Date |
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Sunday, December 8, 2019 |
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Researchers
| سجى الحسين الجدعاني | Aljadaani, Saja Alhussain | Researcher | Master | |
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