[vc_empty_space][vc_empty_space]
Molecular cloning and expression of levansucrase gene from bacillus licheniformis BK1 isolated from bledug kuwu mud crater
Permatasari N.U.a,b, Ratnaningsih E.a, Hertadi R.a
a Biochemistry Research Division, Faculty of Mathematics and Natural Sciences, Institut Teknologi Bandung, Indonesia
b Chemistry Department, Faculty of Mathematics and Natural Sciences, Hasanuddin University, Indonesia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2019 Permatasari et al.Halophilic bacteria are halotolerant microorganisms commonly found in natural environments containing high NaCl concentration. Bacillus licheniformis BK1 is a moderate halophilic bacterium isolated from Bledug Kuwu mud crater, Central Java, Indonesia. This bacterium optimally grows in LB medium containing 15% (w/v) NaCl, and known to produce levansucrase. The levansucrase gene from this bacterium has been successfully isolated and sequence (accession number MF774877.1). The obtained pET-lsbl-bk1 recombinant clone was expressed in E. coli BL21 (DE3) and overexpressed by IPTG induction. The obtained Lsbl-bk1 levansucrase recombinant in the supernatant has a specific activity of 545.678 U/mg protein, in which the unit is defined as μmol of glucose released per minute in sucrose containing medium. This activity is 74% higher compared to those from B. licheniformis BK1 wild type. On the other hand, the cell lysate only showed an increase of 46%. The nucleotide sequence of lsbl-bk1 gene indicated that the open reading frame consists of 1,452 bases encoding 483 amino acid residues of Lsbl-bk1 protein with a cleaved signal peptide between Ala29 and Lys30. This protein is predicted as a member of glycoside hydrolase family, a typical for levansucrase. In addition, three conserved residues of the predicted catalytic triad were identified to be Asp93, Asp256, Glu352.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Bacillus licheniformis BK1,Glycoside hydrolase,Halophilic,Levansucrase[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text]This work is funded by the grant from doctoral dissertation research and the ministry of research and technology with the contract number #009/SP2H/LT/DRPM/IV/2017.[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]