Image of Cloning, Expression, and Partial Purification of Plantaricin W Locus Produced by Lactobacillus plantarum S34

Publikasi Staf Biotek

Cloning, Expression, and Partial Purification of Plantaricin W Locus Produced by Lactobacillus plantarum S34



Lactobacillus plantarum S34 was reported to have antibacterial activities which associated to bacteriocins production. Part of the gene encoding one of the bacteriocin loci that produced by L. plantarum S34, termed plantaricin W (plnW), was amplified from plasmid and cloned into Escherichia coli DH5α using pGEM® -T Easy vector system. PlnW nucleotide sequence (± 405 bp) was identified as a putative integral membrane protein. Moreover, plnW was heterologously expressed as a fusion protein with His(6)-tagged thioredoxin using pET-32a(+) expression vector into E. coli BL21 (DE3) pLysS. Recombinant plnW fusion protein was accumulated in the cell cytoplasm, however, along with soluble fractions, insoluble aggregates identified as inclusion bodies were also exhibited. Partial purifications were conducted for the soluble and insoluble fractions using ligand Co2+ affinity chromatography and polyacrylamide gel electroelution method, respectively. Molecular mass of approximately 33 kDa was detected based on SDS-PAGE separation and confirmed by Western blot as a recombinant plnW fusion protein. The purified protein will be useful for deciphering the bacteriocin structural and functional relationships.

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Informasi Detil

Judul Seri
Berita Biologi
No. Panggil
-
Penerbit Pusat Penelitian Biologi LIPI. : Cibinong, Bogor.,
Deskripsi Fisik
Vol. 16 (1): 59-67.
Bahasa
English
ISBN/ISSN
0126-1754
Klasifikasi
NONE
Tipe Isi
text
Tipe Media
other
Tipe Pembawa
online resource
Edisi
-
Subyek
Info Detil Spesifik
-
Pernyataan Tanggungjawab

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