Pilasombut, Komkhae

Preferred name

Pilasombut, Komkhae

Alternative Name

Pilasombut, K.

Main Affiliation

School of Agricultural Technology

komkhae.pi@kmitl.ac.th

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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 1157T, Leuconostoc mesenteroides subsp. mesenteroides JCM 6124T and Bacillus coagulons JCM 2257T. 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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In vitro screening of potential probiotic lactic acid bacteria isolated from intestinal contents and gills of Nile tilapia
(2020-07-01)
Prachom, N.
;
Rumjuankiat, K.
;
Sanguankiat, A.
;
Boonyoung
;
Pilasombut, K.
The potential probiotic lactic acid bacteria (LAB) were isolated from intestinal contents and gills of Nile tilapia (Oreochromis niloticus). Those properties included antagonistic activity against pathogenic bacteria, tolerance to pH, NaCl, fresh bile from Nile tilapia. Moreover, drug resistance profiles were examined. A total of forty isolates were collected. LAB isolated from the intestinal contents showed inhibitory activity against Bacillus coagulans TISTR1447, Pseudomonas fluorescens TISTR358 and Salmonella Typhimurium TISTR292, whereas those from the gills revealed only antagonistic activity against Bacillus coagulans TISTR1447, S. Typhimurium TISTR292 and Escherichia coli TISTR780. Isolates LI10 and LG5 were selected for further evaluation of probiotic properties. Both isolates could grow in pH 4-10, NaCl concentration up to 10% and fresh bile concentration up to 10%. These isolates were resistant to cefovecin, marbofloxacin, clindamycin and trimethoprim/sulfamethoxazole, meanwhile they were susceptible to benzylpenicillin, amoxicillin/clavulanic acid, vancomycin, tetracycline, nitrofurantoin and chloramphenicol. In addition, both isolates were identified by Matrix-assisted lazer desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) as Enterococcus feacalis. Therefore, these two isolates could potentially be used as probiotics.
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In vitro characterization of bacteriocin produced by lactic acid bacteria isolated from Nem Chua, a traditional Vietnamese fermented pork
(2015-08-01)
Pilasombut, Komkhae
;
Rumjuankiat, Kittaporn
;
Ngamyeesoon, Nualphan
;
Duy, Le Nguyen Doan
The aim of this study was to screen and In vitro characterize the properties of bacteriocin produced by lactic acid bacteria isolated from Vietnamese fermented pork (Nem chua). One hundred and fifty LAB were isolated from ten samples of Nem chua and screened for bacteriocin-producing lactic acid bacteria. Antimicrobial activity of bacteriocin was carried out by spot on lawn method against both gram positive and gram negative bacteria. One isolate, assigned as KL-1, produced bacteriocin and showed inhibitory activity against Lactobacillus sakei, Leuconostoc mesenteroides and Enterococcus faecalis. To characterize the bacteriocin-producing strain, optimum temperature, incubation period for maximum bacteriocin production and identification of bacteriocin-producing strain were determined. It was found that the optimum cultivation temperature of the strain to produce the maximum bacteriocin activity (12,800 AU/mL) was obtained at 30°C. Meanwhile, bacteriocin production at 6,400 AU/mL was found when culturing the strain at 37°C and 42°C. The isolate KL-1 was identified as L. plantarum. Antimicrobial activity of cell-free supernatant was completely inhibited by proteolytic enzyme of trypsin, alpha-chymotrypsin and proteinase K. Bacteriocin activity was stable at high temperature up to 100°C for 10 min and at 4°C storage for 2 d. However, the longer heating at 100°C and 4°C storage, its activity was reduced.
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Encapsulation of Pediococcus pentosaceus RSU-Nh1 into pectin-sodium alginate and chitosan coating
(2020-01-01)
Yodsenee, K.
;
Showpanish, K.
;
Sonhom, N.
;
Pilasombut, K.
;
Prachom, N.
Lactic acid bacteria (LAB) are beneficial to the human intestines, they inhibit the growth of pathogenic microorganisms and are used for lactic acid fermentation. LAB are partially digested when they pass through the digestive system, resulting in reduced survival rate. Encapsulation can be used to enhance the survival of probiotic bacteria as protection against harsh conditions in the gastrointestinal tract. Results indicated that the initial cell numbers of Pediococcus pentosaceus RSU-Nh1, free cells survived with 7.68 log CFU/ml, whereas encapsulation using sodium alginate combined with various pectins extracted from pomelo and passion fruit peel exhibited free cell survival with 7.35 and 8.12 log CFU/ml, respectively. Encapsulation by combining chitosan with shrimp shells and fish scales showed cell survival rates of 8.59 and 8.08 log CFU/ml, respectively, while encapsulation with pectin from passion fruit coated with chitosan from shrimp shells showed the highest number of Ped. pentosaceus RSU-Nh1 with 9.24 log CFU/ml in gastrointestinal simulation. It revealed that encapsulation improved the survival of Ped. pentosaceus RSU-Nh1 in simulated gastrointestinal conditions.
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Effect of extracted pectin from fruit wastes on growth of Pediococcus pentosaceus RSU-Nh1 and Lactobacillus plantarum RSU-SO2
(2020-01-01)
Showpanish, K.
;
Sonhom, N.
;
Pilasombut, K.
;
Woraprayote, W.
;
Prachom, N.
Two strains of lactic acid bacteria identified as Pediococcus pentosaceus RSU-Nh1 and Lactobacillus plantarum RSU-SO2 were isolated from Thai fermented pork sausage (Nham) and fermented spring onion (Ton-Hom-Dong) respectively. The culture supernatant (CFS) of both strains exhibited antimicrobial activity against both Gram-positive and Gramnegative pathogenic bacteria. After being treated with alpha-chymotrypsin and trypsin, the antimicrobial activity of their CFS was eliminated, which suggests that the antimicrobial compound produced was antimicrobial peptide or bacteriocin. Pectin extracted from different fruit peels was supplemented in the bacterial growth medium to investigate the effect of pectin on the growth and antimicrobial production of Ped. pentosaceus RSU-Nh1 and Lb. plantarum RSU-SO2. Pectin was extracted from pomelo, durian, passion fruit and salacca fruit peels with a pectin yield of 5.02 to 26.44%, degree of esterification of 80.84 to 86.68%, and galacturonic acid content of 52.36 to 64.23%. Supplementation with pomelo and passion fruit pectin exhibited the highest improvement of bacterial growth and antimicrobial production of Ped. pentosaceus RSU-Nh1 and Lb. plantarum RSU-SO2. These findings suggested the potential application of these pectin extracts as prebiotics to promote the growth and antimicrobial production of lactic acid bacteria.

Pilasombut, Komkhae

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