Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater
AMA 10th edition
In-text citation: (1), (2), (3), etc.
Reference: Permatasari NU, Ratnaningsih E, Hertadi R. Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater. Eurasia J Biosci. 2019;13(1), 223-230.

APA 6th edition
In-text citation: (Permatasari et al., 2019)
Reference: Permatasari, N. U., Ratnaningsih, E., & Hertadi, R. (2019). Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater. Eurasian Journal of Biosciences, 13(1), 223-230.

Chicago
In-text citation: (Permatasari et al., 2019)
Reference: Permatasari, Nur Umriani, Enny Ratnaningsih, and Rukman Hertadi. "Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater". Eurasian Journal of Biosciences 2019 13 no. 1 (2019): 223-230.

Harvard
In-text citation: (Permatasari et al., 2019)
Reference: Permatasari, N. U., Ratnaningsih, E., and Hertadi, R. (2019). Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater. Eurasian Journal of Biosciences, 13(1), pp. 223-230.

MLA
In-text citation: (Permatasari et al., 2019)
Reference: Permatasari, Nur Umriani et al. "Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater". Eurasian Journal of Biosciences, vol. 13, no. 1, 2019, pp. 223-230.

Vancouver
In-text citation: (1), (2), (3), etc.
Reference: Permatasari NU, Ratnaningsih E, Hertadi R. Molecular cloning and expression of levansucrase gene from Bacillus licheniformis BK1 isolated from Bledug Kuwu Mud Crater. Eurasia J Biosci. 2019;13(1):223-0.

Abstract

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.

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