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( Karthiyaini Damodharan ),( Sasikumar Arunachalam Palaniyandi ),( Seung Hwan Yang ),( Joo Won Suh ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.10
We characterized the probiotic properties of Lactobacillus helveticus strains KII13 and KHI1 isolated from fermented cow milk by in vitro and in vivo studies. The strains exhibited tolerance to simulated orogastrointestinal condition, adherence to Caco-2 cells, and antimicrobial activity. Both L. helveticus strains produced bioactive tripeptides, isoleucylprolyl- proline and valyl-prolyl-proline, during fermentation of milk. KII13 showed higher in vitro cholesterol-lowering activity (47%) compared with KHI1 (28%) and L. helveticus ATCC 15009 (22%), and hence, it was selected for in vivo study of cholesterol-lowering activity in atherogenic diet-fed hypercholesterolemic mice. For the study, mice were divided into four groups (viz., normal diet control group, atherogenic diet control group (HCD), KII13- atherogenic diet group (HCD-KII13), and Lactobacillus acidophilus ATCC 43121-atherogenic diet group (HCD-L.ac) as positive control). The serum total cholesterol level was significantly decreased by 8.6% and 7.78% in the HCD-KII13 and HCD-L.ac groups (p < 0.05), respectively, compared with the HCD group. Low-density lipoprotein cholesterol levels in both HCD-KII13 and HCD-L.ac groups were decreased by 13% and 11%, respectively, compared with the HCD group (both, p < 0.05). Analysis of cholesterol metabolism-related gene expression in mice liver showed increased expression of LDLR and SREBF2 genes in mice fed with KII13. By comparing all the results, we conclude that L. helveticus KII13 could be used as a potential probiotic strain to produce antihypertensive peptides and reduce serum cholesterol.
Damodharan, Karthiyaini,Palaniyandi, Sasikumar Arunachalam,Yang, Seung Hwan,Suh, Joo-Won National Research Council 2015 Canadian journal of microbiology Vol.61 No.11
<P>In this study, we evaluated the probiotic properties of Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus fermentum strains isolated from fermented radish. All the strains survived the simulated oro-gastrointestinal transit condition and showed significantly higher adherence to Caco-2 cells compared with the probiotic strain Lactobacillus rhamnosus GG. The strains showed broad-spectrum antimicrobial activity, autoaggregation, and coaggregation capacity with pathogens. Furthermore, the Lactobacillus strains inhibited the adherence of Yersinia enterocolitica subsp. enterocolitica, Shigella boydii, and Salmonella choleraesuis to the Caco-2 cell line. The strains possessed bile salt hydrolase activity and their cholesterol-lowering activity in vitro was above 50% in the presence of bile. Strains of L. plantarum and L. pentosus possessed the plantaricin-encoding plnEF gene. In addition, the Lactobacillus strains maintained about 80% cell viability after freeze-drying in the presence of a combination of 5% skim milk and 5% maltodextrin as cryoprotectant, and 70% recovery of cell viability was observed in the absence of any cryoprotectant.</P>
Karthiyaini Damodharan,Sasikumar Arunachalam Palaniyandi,양승환,서주원 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.3
The aim of this study was to enhance the viability of probiotic strains Pediococcus pentosaceus KID7, Lactobacillus plantarum KII2, Lactobacillus fermentum KLAB6 and Lactobacillus helveticus KII13 in gastrointestinal transit, freeze-drying condition and during storage time by microencapsulation using a combination of alginate, fenugreek gum and locust bean gum. The microcapsules were prepared using various ratio of alginate to fenugreek gum to locust bean gum and tested for its dissolution in colonic fluid. The combination that efficiently dissolved in colonic fluid was selected for co-encapsulation of the probiotic strains and prebiotics to produce synbiotic microcapsules. Further, we observed that the bacteria encapsulated with alginate-fenugreek gum-locust bean gum (AFL) matrix tolerated gastrointestinal condition efficiently compared to non-encapsulated bacteria. The encapsulated bacterial cells retained higher viability than non-encapsulated cells during freeze-drying condition and subsequent storage for 3 months at 4°C. These results show the utility of AFL matrix in microencapsulation of probiotics for use in food industry.
Karthiyaini Damodharan,Sasikumar Arunachalam Palaniyandi,Bao Le,서주원,양승환 한국미생물학회 2018 The journal of microbiology Vol.56 No.10
A novel actinobacterium, strain SK68, was isolated from the rhizosphere of peanut plant and its salinity stress alleviation ability was studied using tomato (Solanum lycopersicum cv. Micro-Tom) plants. Based on 16S rDNA based phylogenetic analysis, strain SK68 has been identified as a Streptomyces sp. Strain SK68 had branched substrate mycelium bearing smooth surfaced spores and the spore colour is brownish grey on ISP4 medium. It exhibited enzyme activities such as xylanase, cellulase, amylase, and pectinase and degraded hypoxanthine, casein, and L-tyrosine. The strain SK68 differed in its banding pattern in BOX-PCR and RAPD fingerprinting compared to the closely matching type strains Streptomyces erythrochromogenes NBRC 3304T (AB184746), S. flavotricini NBRC 12770T (AB184132), S. racemochromogenes NBRC 12906T (AB184235), and S. polychromogenes NBRC 13072T (NR041109). Strain SK68 was evaluated for its salinity stress-alleviating activity in tomato plants with 180 mmol/L NaCl under gnotobiotic condition. A significant increase in plant biomass was observed in strain SK68-inoculated tomato plants under salt stress compared to control and salt-stressed non-inoculated plants.
팔라니얀디사시쿠마,Karthiyaini Damodharan,이기원,양승환,서주원 한국생물공학회 2015 Biotechnology and Bioprocess Engineering Vol.20 No.3
Ginsenoside Rd, a minor ginseng saponin, has several pharmacological activities. Traditionally, saponins are extracted using organic solvents or hot water extraction. However, both of these methods have disadvantages such as formation of artefacts and compound decomposition. Additionally, the use of organic solvents for extraction is hazardous to the environment. Therefore, we aimed to produce ginsenoside Rd without using organic solvents or hot extraction. We developed a simultaneous extraction and transformation process for higher yields of ginsenoside Rd using a combination of high hydrostatic pressure (HHP) and enzymes. Several commercial glycosidases in various combinations were studied for the enrichment of ginsenoside Rd from major ginsenosides by enzymatic transformation and HHP. We found that treatment with a combination of cellulase (2 U/mL), cellobiase (4 U/mL) and HHP of 100 Mpa at pH 4.8 and 45°C for 24 h resulted in a ginsenoside Rd content of 3.47 ± 0.35 mg/g of fresh ginseng. This yield is 2.1-fold higher than that of the corresponding enzyme treatment at atmospheric pressure (AP, 0.1 Mpa) at pH 4.8, 45°C and for 24 h. This simultaneous extraction and transformation process can be used for the preparation of Rd enriched ginseng beverage without using hazardous organic solvents.
( Sasikumar Arunachalam Palaniyandi ),( Karthiyaini Damodharan ),( Joo-won Suh ),( Seung Hwan Yang ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.5
The present study focused on the production, characterization, and in vitro prebiotic evaluation of an exopolysaccharides (EPS) from Bacillus sonorensis MJM60135 isolated from ganjang (fermented soy sauce). Strain MJM60135 showed the highest production (8.4 ± 0.8 g/l) of EPSs compared with other isolates that were screened for EPS production based on ropy culture morphology. Furthermore, MJM60135 was cultured in 5 L of medium and the EPS was extracted by ethanol precipitation. The emulsification activity of the EPS was higher in toluene than in o-xylene. Fourier transform infrared spectroscopy analysis showed the presence of hydroxyl and carboxyl groups and glycosidic linkages. The isolated EPS contained mannose and glucose, as observed by thin-layer chromatography analysis of the EPS hydrolysate. Lactic acid bacteria (LAB) and pathogenic E. coli K99 and Salmonella enterica serovar Typhimurium were tested for their growth utilizing the EPS from B. sonorensis MJM60135 as the sole carbon source for its possible use as a prebiotic. All the tested LAB exhibited growth in the EPSsupplied medium compared with glucose as carbon source, whereas the pathogenic strains did not grow in the EPS-supplied medium. These findings indicate that the EPS from B. sonorensis MJM60135 has potential application in the bioremediation of hydrocarbons and could also be used as a prebiotic.
Sasikumar Arunachalam Palaniyandi,손병모,Karthiyaini Damodharan,서주원,양승환 한국생물공학회 2016 Biotechnology and Bioprocess Engineering Vol.21 No.5
Lactic acid bacteria (LAB) were screened for ginsenoside transforming activity using crude ginseng extract. Thin-layer chromatography analysis of fermented ginseng extract showed that LAB strain MJM60396 possessed higher ginsenoside transformation ability than other strains. It converted major ginsenosides into minor ginsenosides such as Rg3 and Rh2. MJM60396 also showed high β-glucosidase activity. Strain MJM60396 was identified as Lactobacillus paracasei subsp. tolerans based on 16S rRNA gene sequence. To delineate the pathway involved in the production of the minor ginsenosides Rg3 and Rh2, strain MJM60396 was incubated with pure ginsenoside Rb1. HPLC analysis revealed the appearance of Rg3 and Rh2 peak from the incubation mixture containing Rb1 and strain MJM60396. Furthermore, β-glucosidase enzyme was prepared from strain MJM60396. To achieve its maximum activity, we optimized the pH and temperature conditions. Cell-free β-glucosidase enzyme hydrolyzed ginsenoside Rb1 through the following pathway: ginsenoside Rb1 → Rd → Rg3 → Rh2. This is the first report on the transformation of ginsenosides Rb1 to Rg3 and Rh2 by a Lac. paracasei subsp. tolerans strain. Our results indicate that Lac. paracasei subsp. tolerans MJM60396 has the potential to be used for preparing ginsenosides Rg3 and Rh2 as nutraceuticals.
( Vellingiri Manon Mani ),( Arockiamjeyasundar Parimala Gnana Soundari ),( Damodharan Karthiyaini ),( Kathirvel Preethi ) 한국균학회 2015 Mycobiology Vol.43 No.3
The increasing emergence of lead drugs for the resistance produced by the pathogenic strains and arrival of new diseases have initiated the need for searching novel metabolites with best anticancer and antimicrobial properties than the existing one. With this view, the investigation was conducted for the isolation, identification, and biological evaluation of potential endophytic fungi of Aegle marmelos, a medicinal tree used for more than three decades, for curing various disorders. A total of 169 endophytic fungal strains obtained from sampling and among those 67 were pigmented strains. Upon antagonistic screening, five endophytic fungal strains exhibited antagonistic potentiality by inhibiting the pathogens. These five potent strains were characterized at molecular level by sequencing the amplified internal transcribed spacer (ITS) 1 and ITS 4 regions of rDNA and they were grouped under order Pleosporales, Eurotiales, and Capnodiales. The metabolites from the respective strains were produced in fungal culturing media and extracted using polar solvents. Further, the extracts of five endophytes manifested antimicrobial activity against tested clinical pathogens and Alternaria alternata (FC39BY), Al. citrimacularis (FC8ABr), and Curvularia australiensis (FC2AP) exhibited significant antimicrobial profile against 9 of 12 tested pathogens, showing broad spectrum activity. The antioxidant levels of all the five endophytes revealed the highest activity at least concentrations, and major activity was unveiled by the members of order Pleosporales FC2AP and FC8ABr. This research explains the value of endophytic fungal extracts and its significance of antimicrobial and antioxidant properties.