• Asian-Australasian Journal of Animal Sciences
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• v.33 no.8
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• pp.1352-1359
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• 2020

Objective: Lactobacilli in rabbit intestine is rare and its function in rabbit gut health is not fully understood. The present study aimed to evaluate in vivo the probiotic potential of Lactobacillus casei for suckling rabbits. Methods: Two healthy 5-day-old suckling rabbits with similar weights from each of 12 New Zealand White litters were selected and disturbed to control group and treatment group. All rabbits were artificially fed. The treatment group had been supplemented with live Lactobacillus casei in the milk from the beginning of the trial to 13 days of age. At 15 days of age, healthy paired rabbits were slaughtered to collect intestinal samples. Results: i) Oral administration of Lactobacillus casei significantly increased the proportion of Lactobacilli in the total intestinal bacteria (p<0.01) and obviously reduced that of Escherichia-Shigella (p<0.01); ii) treatment increased the length of vermiform appendix (p<0.05); iii) a higher percentage of degranulated paneth cells was observed in the duodenum and jejunum when rabbits administered with Lactobacillus casei (p<0.01); and iv) the expression of toll-like receptor 9, lysozyme (LYZ), and defensin-7-like (DEFEN) in the duodenum and jejunum was stimulated by supplemented Lactobacillus casei (p<0.05). Conclusion: Orally administered Lactobacillus casei could increase the abundance of intestinal Lactobacilli, decrease the relative abundance of intestinal Escherichia-Shigella, promote the growth of appendix vermiform, stimulate the degranulation of paneth cells and induce the expression of DEFEN and LYS. The results of the present study implied that Lactobacillus casei exhibited probiotic potential for suckling rabbits.

• Journal of Microbiology
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• v.45 no.5
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• pp.409-417
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• 2007

Burkholderia sp. HY-10 isolated from the digestive tracts of the longicorn beetle, Prionus insularis, produced an extracellular lipase with a molecular weight of 33.5 kDa estimated by SDS-PAGE. The lipase was purified from the culture supernatant to near electrophoretic homogenity by a one-step adsorption-desorption procedure using a polypropylene matrix followed by a concentration step. The purified lipase exhibited highest activities at pH 8.5 and $60^{\circ}C$. A broad range of lipase substrates, from $C_4\;to\;C_{18}$ p-nitrophenyl esters, were hydrolyzed efficiently by the lipase. The most efficient substrate was p-nitrophenyl caproate ($C_6$). A 2485 bp DNA fragment was isolated by PCR amplification and chromosomal walking which encoded two polypeptides of 364 and 346 amino acids, identified as a lipase and a lipase foldase, respectively. The N-terminal amino acid sequence of the purified lipase and nucleotide sequence analysis predicted that the precursor lipase was proteolytically modified through the secretion step and produced a catalytically active 33.5 kDa protein. The deduced amino acid sequence for the lipase shared extensive similarity with those of the lipase family 1.2 of lipases from other bacteria. The deduced amino acid sequence contained two Cystein residues forming a disulfide bond in the molecule and three, well-conserved amino acid residues, $Ser^{131},\;His^{330},\;and\;Asp^{308}$, which composed the catalytic triad of the enzyme.

• The Korean Journal of Food And Nutrition
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• v.29 no.2
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• pp.162-170
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• 2016

The effects of dietary ${\beta}$-glucan, obtained from bacterial fermentation, on the intestinal mass, short chain fatty acids, lactate production and pH in Sprague-Dawley (SD) rats were evaluated. SD rats fed with 0% (control group), 1% or 5% ${\beta}$-glucan supplemented diets (w/w) for 3 weeks. The presence of ${\beta}$-glucan in the diets resulted in a significant increase in colonic contents in a dose dependent manner. The amount of short chain fatty acids increased in rats fed ${\beta}$-glucan diets. Rats fed the 5% ${\beta}$-glucan diets had higher levels of acetate, propionate and butyrate by 1.8, 1.7 and 3.0 fold of the control group in the cecum, and 2.2, 2.9 and 3.1 fold of the control group in the colon, respectively. The ${\beta}$-glucan diets also significantly increased the levels of cecal and colonic lactate by 1.4~3.4 fold, when compared to the control diet, indicating that dietary ${\beta}$-glucan stimulated the growth of lactic acid bacteria within the intestine. These results suggest that dietary ${\beta}$-glucan, by providing short chain fatty acids and reducing the cecal and colonic pH, may be beneficial in improving gut health, and provide evidence for the use of ${\beta}$-glucan as a dietary supplement for human consumption.

• Journal of Microbiology
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• v.43 no.6
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• pp.493-498
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• 2005

The safety assessment of Bifidobacterium longum SPM1205 isolated from healthy Koreans and this strain's inhibitory effects on fecal harmful enzymes of intestinal microflora were investigated. The overall safety of this strain was investigated during a feeding trial. Groups of SD rats were orally administered a test strain or commercial reference strain B. longum $1\times10^9\;CFU/kg$ body weight/day for four weeks. Throughout this time, their feed intake, water intake and live body weight were monitored. Fecal samples were periodically collected to test harmful enzyme activities of intestinal microflora. At the end of the four-week observation period, samples of blood, liver, spleen, kidney, and gut tissues were collected to determine for hematological parameters and histological differences. The results obtained in this experiment demonstrated that four weeks of consumption of this Bifidobacterium strain had no adverse effects on rat's general health status, blood biochemical parameters or histology. Therefore, it is likely to be safe for human use. Fecal harmful enzymes such as $\beta-glucosidase,\;\beta-glucuronidase$, tryptophanase and urease, were effectively inhibited during the administration of the B. longum SPM1205. These results suggested that this B. longum SPM 1205 could be used for humans as a probiotic strain.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1266-1271
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• 2017

Lactobacillus fermentum strain JDFM216, isolated from a Korean infant feces sample, possesses the ability to enhance the longevity and immune response of a Caenorhabditis elegans host. To explore the characteristics of strain JDFM216 at the genetic level, we performed whole-genome sequencing using the PacBio system. The circular draft genome has a total length of 2,076,427 bp and a total of 2,682 encoding sequences were identified. Five phylogenetically featured genes possibly related to the longevity and immune response of the host were identified in L. fermentum strain JDFM216. These genes encode UDP-N-acetylglucosamine 1-carboxyvinyltransferase (E.C. 2.5.1.7), ErfK/YbiS/YcfS/YnhG family protein, site-specific recombinase XerD, homocysteine S-methyltransferase (E.C. 2.1.1.10), and aspartate-ammonia ligase (E.C. 6.3.1.1), which are involved in peptidoglycan synthesis and amino acid metabolism in the gut environment. Our findings on the genetic background of L. fermentum strain JDFM216 and its potential candidate genes for host longevity and immune response provide new insight for the application of this strain in the food industry as newly isolated functional probiotic.

• Preventive Nutrition and Food Science
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• v.18 no.4
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• pp.273-279
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• 2013

Protease widely exists in the digestive tract of animals and humans, playing a very important role in protein digestion and absorption. In this study, a high protease-producing strain Planomicrobium sp. L-2 was isolated and identified from the digestive tract of Octopus variabilis. The strain was identified by physiological and biochemical experiments and 16S rDNA sequences analysis. A protease was obtained from the strain Planomicrobium sp. L-2 through ammonium sulfate precipitation, dialysis and enrichment, DEAE-Sephadex A50 anion-exchange chromatography, and Sephadex G-100 gel chromatography. The molecular weight and properties of the protease were characterized, including optimum temperature and pH, thermal stability, protease inhibitions and metal ions. According to our results, the protease from Planomicrobium sp. L-2 strain designated as F1-1 was obtained by three-step separation and purification from crude enzyme. The molecular weight of the protease was 61.4 kDa and its optimum temperature was $40^{\circ}C$. The protease F1-1 showed a broad pH profile for casein hydrolysis between 5.0~11.0. No residual activity was observed after incubation for 40 min at $60^{\circ}C$ and 60 min at $50^{\circ}C$. F1-1 protease was inhibited by $Mn^{2+}$, $Hg^{2+}$, $Pb^{2+}$, $Zn^{2+}$, and $Cu^{2+}$ ions, as well as PMSF, indicating that the protease F1-1 was a serine protease. Additionally, research basis provided by this study could be considered for industrial application of octopus intestinal proteases.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.638-644
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• 2022

Probiotics modulate the gut microbiota, which in turn regulate immune responses to maintain balanced immune homeostasis in the host. However, it is unclear how probiotic bacteria regulate immune responses. In this study we investigated the immunomodulatory effects of heat-killed probiotics, including Lactiplantibacillus plantarum KC3 (LP3), Lactiplantibacillus plantarum CKDB008 (LP8), and Limosilactobacillus fermentum SRK414 (LF4), via phagocytosis, nitric oxide (NO), and pro-inflammatory cytokine production in macrophages. We thus found that heat-killed LP8 could promote the clearance of foreign pathogens by enhancing the phagocytosis of macrophages. Treatment with heat-killed LP8 induced the production of NO and pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β. In addition, heat-killed LP8 suppressed the production of NO and cytokines in LPS-induced RAW264.7 cells, suggesting that heat-killed LP8 exerts immunomodulatory effects depending on the host condition. In sum, these results indicate that heat-killed LP8 possesses the potential for immune modulation while providing a molecular basis for the development of functional probiotics prepared from inactivated bacterial cells.

• Journal of Microbiology and Biotechnology
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• v.32 no.7
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• pp.877-884
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• 2022

Probiotics are microorganisms that can benefit host health when ingested in a live state, and lactic acid bacteria are the most common type. Among fungi, Saccharomyces boulardii (SB) is the only strain known to have a probiotic function with beneficial effects on colitis; however, information on other probiotic yeast strains is limited. Therefore, this study aimed to discover yeast strains expressing intestinal anti-inflammatory activities by exhibiting probiotic properties in dextran sodium sulfate (DSS)-induced colitis mice model. Nuruk (Korean traditional fermentation starter) containing various microbial strains was used as a source for yeast strains, and S. cerevisiae 28-7 (SC28-7) strain was selected with in vitro and in vivo characteristics to enable survival in the intestines. After 14 days of pretreatment with the yeast strains, DSS was co-administered for six days to induce colitis in mice. The results revealed that the disease activity index score was lowered by SC28-7 treatment compared to the DSS group, and the colon length and weight/length ratio were recovered in a pattern similar to that of the normal group. SC28-7 administration significantly reduced the secretion of pro-inflammatory cytokines in the serum and modified the mRNA expression of inflammatory cytokines (interleukin-1β, transforming growth factor-β, and interferon-γ) and proteins involved in gut barrier functions (mucin 2, mucin 3, zonula occludens-1, and occludin) in colon tissues. These results indicate that SC28-7 attenuates DSS-induced colon damage and inflammation, supporting its future use as a probiotic yeast for treating and preventing intestinal inflammatory diseases such as inflammatory bowel disease.

• Journal of Microbiology and Biotechnology
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• v.32 no.7
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• pp.892-901
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• 2022

The rising demand for carotenoids can be met by microbial biosynthesis as a promising alternative to chemical synthesis and plant extraction. Several species of lactic acid bacteria (LAB) specifically produce C₃₀ carotenoids and offer the added probiotic benefit of improved gut health and protection against chronic conditions. In this study, the recently characterized Lactiplantibacillus plantarum subsp. plantarum KCCP11226^T produced the rare C₃₀ carotenoid, 4,4'-diaponeurosporene, and its yield was optimized for industrial production. The one-factor-at-a-time (OFAT) method was used to screen carbon and nitrogen sources, while the abiotic stresses of temperature, pH, and salinity, were evaluated for their effects on 4,4'-diaponeurosporene production. Lactose and beef extract were ideal for optimal carotenoid production at 25℃ incubation in pH 7.0 medium with no salt. The main factors influencing 4,4'-diaponeurosporene yields, namely lactose level, beef extract concentration and initial pH, were enhanced using the Box-Behnken design under response surface methodology (RSM). Compared to commercial MRS medium, there was a 3.3-fold increase in carotenoid production in the optimized conditions of 15% lactose, 8.3% beef extract and initial pH of 6.9, producing a 4,4'-diaponeurosporene concentration of 0.033 A₄₇₀/ml. To substantiate upscaling for industrial application, the optimal aeration rate in a 5 L fermentor was 0.3 vvm. This resulted in a further 3.8-fold increase in 4,4'-diaponeurosporene production, with a concentration of 0.042 A₄₇₀/ml, compared to the flask-scale cultivation in commercial MRS medium. The present work confirms the optimization and scale-up feasibility of enhanced 4,4'-diaponeurosporene production by L. plantarum subsp. plantarum KCCP11226^T.

• Journal of the Korean Applied Science and Technology
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• v.39 no.5
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• pp.632-643
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• 2022

Although probiotics have been shown to improve health when consumed, recent studies have reported that they can cause unwanted side effects due to bacterial-human interactions. Therefore, the importance of prebiotics that can form beneficial microbiome in the gut has been emphasized. This study isolated and identified bacteria capable of producing biopoymer as a candidate prebiotic from traditional fermented foods. The isolated and identified strain was named WCYSK01 (Wissella sp. strain YSK01). The composition of the medium for culturing this strain was prepared by dissolving 3 g K₂HPO₄, 0.2 g MgSO₄, 0.05 g CaCl₂, 0.1 g NaCl in 1 L of distilled water. The LMBP(low molecular weight biopoymers) produced when fermentation was performed with sucrose and maltose as substrates were mainly consisted of DP3 (degree of polymer; isomaltotriose), DP4 (isomaltotetraose), DP5 (isomaltopentaose), and DP6 (isomaltoheptaose). The optimization of LMBP (low molecular weight of biopolymer) production was performed using the response surface methodology. The fermentation process temperature range of 18 to 32℃, the fermentation medium pH in the range of 5.1 to 7.9. The yield of LMBP production by the strain was found to be significantly affected by q fermentation temperature and pH. The optimal fermentation conditions were found at the normal point, and the production yield was more than 75% at pH 7.5 and temperature of 23℃.