Isolasi, Karakterisasi, dan Kloning Gen Penyandi α α α α α-AmilaseudBakteri Halofil Moderat asal Bledug Kuwu

摘要

A moderately halophilic bacteria, BK-05, was isolated from Bledug Kuwu, a saline terrestrial area at Central Java. Basedudon partial sequence of 16S-rRNA gene, the isolate was closely related to Halobacillus litoralis. This isolate showed amylolyticudactivity when grown on saline media [15% (w/v) NaCl] suplemented with starch. A pair of primer was designed based on theudsequence of amyH gene from Halomonas meridiana and Pseudoalteromonas haloplanktis. PCR amplification using these primersudshowed three DNA bands with each size approximately 1.50, 1.00, and 0.75 kb. Partial DNA sequencing analysis based on itsuddeduced protein sequence revealed that the 1.50 kb band was closely related to the sequence of metalloprotease from Bacillusudsubtilis (approximately 54.3% identity in 184 amino acid overlap). Southern hybridization analysis showed that the 1.50 kbudfragment was located within a 4.0 kb fragment of BamHI, 4.8 kb of EcoRI, 4.3 kb of HindIII, and 4.0 kb of XhoI digestion ofudBK-05 genomic DNA, respectively.