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Cloning and Expression of Plantaricin W Produced by Lactobacillus plantarum U10 Isolate from “Tempoyak” Indonesian Fermented Food as Immunity Protein in Lactococcus lactis

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Abstract

Plantaricins, one of bacteriocin produced by Lactobacillus plantarum, are already known to have activities against several pathogenic bacterium. L. plantarum U10 isolated from “tempoyak,” an Indonesian fermented food, produced one kind of plantaricin designated as plantaricin W (plnW). The plnW is suggested as a putative membrane location of protein and has similar conserved motif which is important as immunity to bacteriocin itself. Thus, due to study about this plantaricin, several constructs have been cloned and protein was analyzed in Lactococcus lactis. In this study, plnW gene was successfully cloned into vector NICE system pNZ8148 and created the transformant named L. lactis NZ3900 pNZ8148-WU10. PlnW protein was 25.3 kDa in size. The concentration of expressed protein was significantly increased by 10 ng/mL nisin induction. Furthermore, PlnW exhibited protease activity with value of 2.22 ± 0.05 U/mL and specific activity about 1.65 ± 0.03 U/mg protein with 50 ng/mL nisin induction. Immunity study showed that the PlnW had immunity activity especially against plantaricin and rendered L. lactis recombinant an immunity broadly to other bacteriocins such as pediocin, fermentcin, and acidocin.

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Acknowledgments

This study was supported by Competitive Program 2014 from Indonesian Institute of Science (LIPI). The authors would like to thank Dr. Andi Utama (Stem Cell and Cancer Institute, Kalbe Farma) for kindly supplying E.coli MC1061 and L.lactis NZ3900. They also thank Prof. Dae-Kyung Kang (Dankook University, Korea) for kindly supplying Lactobacillus acidophilus C99.

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Authors and Affiliations

  1. Research Center for Biotechnology, Indonesian Institute of Science (LIPI), Cibinong, Bogor, 16911, West Java, Indonesia

    Aksar Chair Lages, Apon Zaenal Mustopa & Linda Sukmarini

  2. School of Biotechnology, Bogor Agricultural University, Darmaga, Bogor, 16680, West Java, Indonesia

    Aksar Chair Lages &  Suharsono

  3. Research Center of Bioresources and Biotechnology, Bogor Agricultural University, Darmaga, Bogor, 16680, West Java, Indonesia

    Suharsono

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  1. Aksar Chair Lages
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  2. Apon Zaenal Mustopa
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Correspondence to Apon Zaenal Mustopa.

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Lages, A.C., Mustopa, A.Z., Sukmarini, L. et al. Cloning and Expression of Plantaricin W Produced by Lactobacillus plantarum U10 Isolate from “Tempoyak” Indonesian Fermented Food as Immunity Protein in Lactococcus lactis . Appl Biochem Biotechnol 177, 909–922 (2015). https://doi.org/10.1007/s12010-015-1786-9

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