Pilasombut, Komkhae

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Preferred name
Pilasombut, Komkhae
Alternative Name
Pilasombut, K.
Main Affiliation
Email
komkhae.pi@kmitl.ac.th

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    Item type:Publication,
    Purification and amino acid sequence of a bacteriocins produced by Lactobacillus salivarius K7 isolated from chicken intestine
    (2006-01-01)
    Pilasombut, Komkhae
    ;
    Sakpuaram, Thavajchai
    ;
    Wajjwalku, Worawidh
    ;
    Nitisinprasert, Sunee
    ;
    Swetwiwathana, Adisorn
    A bacteriocin-producing strain, Lactobacillus K7, was isolated from a chicken intestine. The inhibitory activity was determined by spot-on-lawn technique. Identification of the strain was performed by morphological, biochemical (API 50 CH kit) and molecular genetic (16S rDNA) basis. Bacteriocin purification processes were carried out by amberlite adsorption, cation exchange and reverse-phase high perform ance liquid chromatography. N-terminal amino acid sequences were performed by Edman degradation. Molecular mass was determined by electrospray-ionization (ESI) mass spectrometry (MS). Lactobacillus K7 showed inhibitory activity against Lactobacillus sakei subsp. sakei JCM 1157<sup>T</sup>, Leuconostoc mesenteroides subsp. mesenteroides JCM 6124<sup>T</sup> and Bacillus coagulons JCM 2257<sup>T</sup>. This strain was identified as Lb. salivarius. The antimicrobial substance was destroyed by proteolytic enzymes, indicating its proteinaceous structure designated as a bacteriocin type. The purification of bacteriocin by amberlite adsorption, cation exchange, and reverse-phase chromatography resulted in only one single active peak, which was designated FK22. Molecular weight of this fraction was 4331.70 Da. By amino acid sequence, this peptide was homology to Abp 118 beta produced by Lb. salivarius UCC118. In addition, Lb. salivarius UCC118 produced 2-peptide bacteriocin, which was Abp 118 alpha and beta. Based on the partial amino acid sequences of Abp 118 beta, specific primers were designed from nucleotide sequences according to data from GenBank. The result showed that the deduced peptide was high homology to 2-peptide bacteriocin, Abp 118 alpha and beta.
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    Purification and characterization of a novel plantaricin, KL-1Y, from Lactobacillus plantarum KL-1
    (2015-06-13)
    Rumjuankiat, Kittaporn
    ;
    Perez, Rodney Horanda
    ;
    Pilasombut, Komkhae
    ;
    Keawsompong, Suttipun
    ;
    Zendo, Takeshi
    Three bacteriocins from Lactobacillus plantarum KL-1 were successfully purified using ammonium sulfate precipitation, cation-exchange chromatography and reverse-phase HPLC. The bacteriocin peptides KL-1X, -1Y and -1Z had molecular masses of 3053.82, 3498.16 and 3533.16 Da, respectively. All three peptides were stable at pH 2–12 and 25 °C and at high temperatures of 80 and 100 °C for 30 min and 121 °C for 15 min. However, they differed in their susceptibility to proteolytic enzymes and their inhibition spectra. KL-1Y showed broad inhibitory activities against Gram-positive and Gram-negative bacteria, including Salmonella enterica serovar Enteritidis DMST 17368, Pseudomonas aeruginosa ATCC 15442, P. aeruginosa ATCC 9027, Escherichia coli O157:H7 and E. coli ATCC 8739. KL-1X and -1Z inhibited only Gram-positive bacteria. KL-1X, KL-1Y and KL-1Z exhibited synergistic activity. The successful amino acid sequencing of KL-1Y had a hydrophobicity of approximately 30 % and no cysteine residues suggested its novelty, and it was designated “plantaricin KL-1Y”. Plantaricin KL-1Y exhibited bactericidal activity against Bacillus cereus JCM 2152<sup>T</sup>. Compared to nisin, KL-1Y displayed broad inhibitory activities of 200, 800, 1600, 800, 400 and 400 AU/mL against the growth of Bacillus coagulans JCM 2257<sup>T</sup>, B. cereus JCM 2152<sup>T</sup>, Listeria innocua ATCC 33090<sup>T</sup>, Staphylococcus aureus TISTR 118, E. coli O157:H7 and E. coli ATCC 8739, respectively, whereas nisin had similar activities against only B. coagulans JCM 2257<sup>T</sup> and B. cereus JCM 2152<sup>T</sup>. Therefore, the novel plantaricin KL-1Y is a promising antimicrobial substance for food safety uses in the future.