• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.666-670
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• 2003

Strains producing the protease, which is essential for growth of lactic acid bacteria and fermented kimchi, were screened and identified. Among five types of selected pulmuone kimchis (Jeonlado kimchi, ripened kimchi, yeolmu kimchi, kakdugi, and baechu kimchi), nine strains of bacteria were screened and identified by whole cell protein pattern and API test. The nine strains consisted of one of Lactobacillus sp., one of Leuconostoc sp., six of Bacillus sp., and one of Brevibacillus sp. The protease activities of these strains were compared with known strains (Bac. subtilis KCCM 12248 and Bac. licheniformis KCCM 11851) producing protease. Among tested strains, K-2 (Brevibacillus sp.) showed the highest value (0.11 unit/mg) in protease activity.

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.158-165
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• 2006

In this study, we investigated the biological activity of antioxidant and antibacterial activity of citrus by-product ferment. Among the six probiotic bacteria, Bacillus subtilis and Saccharomyces cerevisiae had the highest antioxidant activity. Hot water extracts from citrus by-product of ferment were screened for antibacterial activity fish pathogenic bacteria by paper disc method. Among the various hot water extracts, the Bacillus subtilis, Saccharomyces cerevisiae, Pediococcus pentosaceus, Lactobacillus rhamnosus, Lactobacillus plantarum, Enterococcus faecium showed relatively strong antibacterial activities in the order. The reducing activity on the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $O^{2-}$ and OH radical scavenging potential were sequentially screened, in search for antioxidant activities of citrus by-product ferment.

• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.347-356
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• 2019

Bacillus sp. BS2 showing strong fibrinolytic activity was isolated from sea squirt (munggae) jeotgal, a traditional Korean fermented seafood. BS2 was identified as B. velezensis by molecular biological methods. B. velezensis BS2 grows well at 15% NaCl and at $10^{\circ}C$. When B. velezensis BS2 was cultivated in TSB broth for 96 h at $37^{\circ}C$, the culture showed the highest fibrinolytic activity ($131.15mU/{\mu}l$) at 96 h. Three bands of 27, 35 and 60 kDa were observed from culture supernatant by SDS-PAGE, and fibrin zymography showed that the major fibrinolytic protein was the 27 kDa band. The gene (aprEBS2) encoding the major fibrinolytic protein was cloned, and overexpressed in heterologous hosts, B. subtilis WB600 and E. coli BL21 (DE3). B. subtilis transformant showed 1.5-fold higher fibrinolytic activity than B. velezensis BS2. Overproduced AprEBS2 in E. coli was purified by affinity chromatography. The optimum pH and temperature were pH 8.0 and $37^{\circ}C$, respectively. $K_m$ and $V_{max}$ were 0.15 mM and $39.68{\mu}M/l/min$, respectively, when N-succinyl-Ala-Ala-Pro-Phe-pNA was used as the substrate. AprEBS2 has strong ${\alpha}$-fibrinogenase and moderate ${\beta}$-fibrinogenase activity. Considering its high fibrinolytic activity, significant salt tolerance, and ability to grow at $10^{\circ}C$, B. velezensis BS2 can be used as a starter for jeotgal.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.880-888
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• 2016

Changes in quality characteristics of gochujang prepared with different ingredients (white rice, barley, sorghum and millet, bamboo salt) and meju starters were studied. The gochujang samples were fermented for 6 weeks at $40^{\circ}C$ and analyzed for changes in their physicochemical properties and sensory characteristics. Eight different gochujang samples were prepared: WR-SS-AB, WR-SS-ABL, WR-BS-AB, WR-BS-ABL, MG-SS-AB, MG-SS-ABL, MG-BS-AB, MG-BS-ABL (WR: white rice, MG: mixed grains, SS: solar salt, BS: bamboo salt, AB: Aspergillus oryzae/Bacillus subtilis starter meju, ABL: Aspergillus oryzae/Bacillus subtilis/Lactobacillus plantarum starter meju). There were significant differences between experimental groups in terms of moisture content, pH, acidity, and color value. All gochujang samples showed increased moisture contents and pH as well as increased acidity and amino-type nitrogen during fermentation. MG-BS-ABL showed the highest contents of amino-type nitrogen and free amino acids after 6 weeks of fermentation compared with other experiment groups. In addition, MG-BS-ABL showed the highest sensory analysis score, including appearance, flavor, taste, and overall acceptability after the fermentation of samples. Based on the results, MG-BS-ABL exhibited similar quality characteristics as general gochujang but showed the highest sensory scores and contents of amino-type nitrogen and free amino acids. These results might be due to the high protein contents of mixed grains and high mineral contents of bamboo salt and fermented products from mixed probiotic starters.

• Journal of Ginseng Research
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• v.42 no.1
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• pp.57-67
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• 2018

Background: Ginseng contains many small metabolites such as amino acids, fatty acids, carbohydrates, and ginsenosides. However, little is known about the relationships between microorganisms and metabolites during the entire ginseng fermentation process. We investigated metabolic changes during ginseng fermentation according to the inoculation of food-compatible microorganisms. Methods: Gas chromatography mass spectrometry (GC-MS) datasets coupled with the multivariate statistical method for the purpose of latent-information extraction and sample classification were used for the evaluation of ginseng fermentation. Four different starter cultures (Saccharomyces bayanus, Bacillus subtilis, Lactobacillus plantarum, and Leuconostoc mesenteroide) were used for the ginseng extract fermentation. Results: The principal component analysis score plot and heat map showed a clear separation between ginseng extracts fermented with S. bayanus and other strains. The highest levels of fructose, maltose, and galactose in the ginseng extracts were found in ginseng extracts fermented with B. subtilis. The levels of succinic acid and malic acid in the ginseng extract fermented with S. bayanus as well as the levels of lactic acid, malonic acid, and hydroxypruvic acid in the ginseng extract fermented with lactic acid bacteria (L. plantarum and L. mesenteroide) were the highest. In the results of taste features analysis using an electronic tongue, the ginseng extracts fermented with lactic acid bacteria were significantly distinguished from other groups by a high index of sour taste probably due to high lactic acid contents. Conclusion: These results suggest that a metabolomics approach based on GC-MS can be a useful tool to understand ginseng fermentation and evaluate the fermentative characteristics of starter cultures.

• Microbiology and Biotechnology Letters
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• v.51 no.2
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• pp.147-156
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• 2023

Tetragenococcus halophilus CY54, an isolate from jeotgal, grows best in media with 5% NaCl and can grow at 18% and higher salt concentration. Three different doenjang samples were prepared with multiple starters including T. halophilus CY54. TBZA doenjang was prepared with T. halophilus, Bacillus subtilis, Zygosaccharomyces rouxii and Aspergillus oryzae. BZA doenjang was prepared with the same 3 starters except T. halophilus. KACC doenjang was prepared with a single starter, B. subtilis KACC16750. During 16 weeks of fermentation at 25℃, the viable counts were maintained in the range of 7-8 log CFU/g in all 3 samples. As fermentation progressed, pH decreased and titratable acidity (TA) gradually increased. Crude protein contents decreased slightly. TBZA doenjang showed higher amino-type nitrogen (ANN) and volatile basic nitrogen (VBN) contents, and KACC doenjang showed higher ammonia-type nitrogen (AMN) content. TBZA doenjang showed higher fibrinolytic and protease activity than other doenjang samples. Metabolites analyses by GC/MS showed that doenjang samples were separated from each other by partial least squares-discriminant analysis (PLS-DA) analysis. Seventeen major metabolites involved in the differences between samples were identified and they included organic acids, amino acids, sugars, fatty acids and alcohols. TBZA doenjang showed higher contents for most metabolites responsible for flavor and taste of fermented foods including doenjang. These results showed that T. halophilus could be useful as a starter for doenjang and can improve the product quality by accelerating the fermentation processes.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.5
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• pp.643-649
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• 2011

The objective of this study was to determine the effect of feeding direct-fed microbials (DFM) on the growth performance and prophylaxis of calf diarrhea during the pre-weaning period as an alternative to antibiotics. A multi-species DFM was formulated including three lactic acid bacteria (Lactobacillus salivarius Ls29, Pediococcus acidilactia Pa175, and L. plantarum Lp177), three Bacillus strains (B. subtilis T4, B. polymyxa T1 and SM2), one yeast, Saccharomyces boulardii, and a nonpathogenic E. coli Nissle 1917. Lactic acid bacteria and Bacillus strains were selected based on the antibacterial activity against various animal pathogens, especially pathogenic E. coli using agar diffusion methods in vitro. Test and control groups were fed milk replacer and calf starter supplemented with DFM ($10^9$ cfu each of eight species/d/head, n = 29) or with antibiotics (0.1% neomycin sulfate in milk replacer and Colistin 0.08% and Oxyneo 110/110 0.1% in calf starter, n = 15), respectively. Overall fecal score and the incidence rate of diarrhea were reduced in the DFM group compared to the antibiotics one. About 40% of calves in antibiotic group suffered from diarrhea while in DFM group only 14% showed diarrhea. There was no difference in the average daily gain and feed efficiency of two groups. The hematological levels of calves were all within the normal range with no significant difference. In conclusion, the feeding of multispecies DFM during the pre-weaning period could reduce calf diarrhea and there was no difference in the growth performance between the groups, thus showing the potential as an alternative to antibiotics.

• Journal of Life Science
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• v.33 no.6
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• pp.481-489
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• 2023

The taste and quality of soy sauce, a fermented liquid condiment, is greatly influenced by microbial metabolism during fermentation. To investigate the microbiological characteristics of ganjang during the initial fermentation process, we prepared meju (fermented soybean) blocks fermented with starter cultures and solar salts and analyzed the microbial community quantitively using 16S rRNA gene profiling from ganjang that had been fermented over a five-week period. The ganjang samples were collected and analyzed after soaking for week one (1W), three (3W), and five (5W) weeks. We found that Halomonadaceae was significantly higher in the 1W group (89.83%) than the 3W and 5W groups (14.46%, and 13.78%, respectively). At a species level, Chromohalobacter beijerinckii and Chromohalobacter canadensis were the dominant species in the 1W group but several taxa such as Bacillus subtilis, Pediococcus acidilactici, and Enterococcus faecalis were more abundant in the 3W and 5W groups. Pearson correlation analysis of the relative abundance of the bacteria showed a negative correlation between Chromohalobacter and two bacterial genera Bacillus and Enterococcus. Beta-diversity showed a statistical distinction between the 1W and the 3W and 5W groups, while no significance was evident between the 3W and 5W groups. Linear discriminant effect size analysis was used to identify biomarkers and significant differences in the relative abundance of several halophilic bacteria, Bacillus sp. and lactic acid bacteria at 1W, 3W, and 5W, recpectively, which indicates the important role of the bacterial community at these time points.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.4
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• pp.540-548
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• 2015

This study examined the cultural characteristics and biological activities of fermented rice and amaranth with Bacillus subtilis KMKW4. These samples were made with various amounts of rice and amaranth [100:0 (R100), 90:10 (R90), 80:20 (R80), and 70:30 (R70)]. B. subtilis KMKW4 was used as starter for the fermentation, and its cultures at the lated logarithmic growth were inoculated for final concentration of 2% (v/v). Number of viable cells of fermented R80 (7.67 log CFU/mL) was greater than those of R100, R90, and R70 (7.48 log CFU/mL, 7.38 log CFU/mL, and 7.09 CFU/mL, respectively) during the fermentation period (120 h). Amylase activities of fermented R80 and R100 were 57.77 U/mL and 19.91 U/mL, respectively. Furthermore, amylase activities of fermented freeze-dried powders of R100 and R80 were 24.31 U/g and 9.12 U/g, respectively. Free sugar contents of R100 and R80 increased after fermentation, and that of R80 (5,454.15 mg/100 g) significantly increased compared to that of R100 (4,274.85 mg/100 g). The free amino acid content of R80 was higher than that of R100. DPPH and superoxide radical scavenging activities of 5 mg/mL of fermented freeze-dried powder (R80) were 44.21% and 89.76%, respectively. ACE inhibition rates and ${\alpha}$-glucosidase inhibitory activities were significantly higher in R80 than R100. This study suggested that fermentation of R80 might be a new potential source of antioxidant, anti-diabetic, and anti-hypertensive agents applicable to grain enzyme-containing foods.

• Journal of Microbiology and Biotechnology
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• v.29 no.4
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• pp.527-537
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• 2019

Three types of meongge (Halocynthia roretzi) jeotgal (MJ) were prepared with 3 different types of salts (12%, w/v): purified salt (PS), solar salt aged for 3 years (SS), and bamboo salt that had been recrystalized 3 times (BS). One set of MJ was fermented with starters, Bacillus subtilis JS2 and Tetragenococcus halophilus BS1-37 (each 6 log CFU/g), and another set without starters for 42 days at $10^{\circ}C$. The LAB count of the SSMJ (non-starter) was highest at day 28 (2.30 log CFU/g). The pH of the PSMJ and SSMJ was 5.72-5.77 at day 0, and 5.40-5.50 at day 42. BSMJ showed higher pH and lower titratable acidities than other samples. Amino-type nitrogen (ANN) increased continuously, and SSMJ showed higher values than other samples from day 14. Bacterial species of non-starter MJ were examined by culture independent method. Clone libraries of 16S rRNA genes were constructed in Escherichia coli from total DNA from non-starter MJ samples at day 0, 14, and 28. Thirty clones per each sample were randomly selected and DNA sequences were analyzed. Variovorax sp., uncultured bacterium, and Acidovorax sp. were the most dominant group at day 0, 14, and 28, respectively. Lactobacillus sakei and Streptococcus sp. were the next dominant group in SSMJ at day 28. A Streptococcus sp. was detected from PSMJ at day 28. Sensory evaluation for MJ samples at day 28 showed that SSMJ got higher overall acceptability scores. These results showed that solar salt can cause desirable changes in the microbial community of fermented foods, thereby positively affecting their overall quality.