Cloning and expression of plantaricin E and F genes of Lactobacillus plantarum S34 isolated from Indonesia traditional-fermented meat (Bekasam)

Umami, R.N.; Kusdianawati; Budiarto, R.B.; Mustopa, A.Z.; Fatimah; Danuri, H.

Cloning and expression of plantaricin E and F genes of Lactobacillus plantarum S34 isolated from Indonesia traditional-fermented meat (Bekasam)

2016

Heterologous protein expression has been used in attemp to increase bacteriocins yields by less laborius process. In this study, the plnEF genes (530 bp) encoding plantaricin S34 have been identified and cloned to pGEMTeasy s vector. Plantaricin S34 gene had 99% similarity with all those plnEF locus of Lactobacillus references strain aligned. Furthermore, plnE peptide of plantaricin S34 had unique one amino acid substitution at position 33 (lysine > cysteine), while two amino acids were substituted at position 14 (alanine > serine) and 42 (valine > isoleucine) appeared on plnF peptide. PCR amplification of mature fragment of plnE and plnF gene produced the bands with length of approximatly 102 bp and 105 bp respectively. Moreover, Both of fused recombinant plnE and plnF peptide have been expressed heterologously in E. coli as protein fusion with thioredoxin-(His)6tag with given molecular mass of approximatly 21 kDa for each peptides. Overall, the partial operon of plnEFI loci of L. plantarum S34 has been characterizied and the plnEF genes that composed this operon was successfully cloned and produced as hetelogous recombinant peptides.

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