#TITLE_ALTERNATIVE#

• Main Author: DEWI ROSAHDI, TINA
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/9485
• Institution: Institut Teknologi Bandung

&#945;-Amylases (1,4-&#945;-D-glucan glucanohydrolase, E.C. 3.2.1.1) are endo-acting enzymes that catalyze the hydrolysis of &#945;-1,4-glycosidic linkages of amylose and amylopectin to produce &#945;-limit dextrins, linier olygosaccharides, and a small amount of maltose and glucose. Industrial applications of &#945;-amylases are dominated by &#945;-amylases from bacterial sources. Marine bacteria are one of the potential sources of &#945;-amylases yet to be further explored. The purpose of this research is to gain information about the presence of the local marine Vibrio sp. SFNB3 &#945;-amylase gene. <br /> <br /> <br /> EcoRI digested Vibrio sp. SFNB3 chromosomal DNA fragments with a size of 4.5 to 6 kb were ligated to a pUC19 vector which had been digested by a restriction enzyme EcoRI. Screening of the transformant carrying the &#945;-amylase gene was done based on the presence of a halo area in agar media containing starch after iodine addition. Restriction analysis of the recombinant plasmid from E. coli transformant showing halo area (Vsp.7) showed the present of chromosomal DNA fragment with the size of 4.7 kb. Nucleotide sequence analysis of the Vsp.7 insert DNA showed a similarity with a hypothetical protein from Vibrio harveyi ATCC BAA-1116 (accession number CP000789.1), a flavodoxin 2, and a single-stranded-DNA-specific exonuclease RecJ from Vibrio parahaemolyticus RIMD (accession number BA000031.2). <br /> <br /> <br /> The &#945;-amylase gene was not present on the Vsp.7 insert DNA fragments. However, valuable information was obtained in which V. parahaemolyticus RIMD has a putative gene encoding &#945;-amylase (accession number BA000031) with a size of 2.085 kb and consist of 695 amino acids residues. The size of this V. parahaemolyticus RIMD &#945;-amylase gene is not common for a gene encoding &#945;-amylase, thus indicates that the &#945;-amylase may possess a unique property. For further study, a primer can be designed from the nucleotide sequence of the V. parahaemolyticus RIMD &#945;-amylase gene and the local marine Vibrio sp. SFNB3 &#945;-amylase gene can be amplified by Polymerase Chain Reaction.