• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.195-203
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• 2015

Two bacterial strains, Bacillus licheniformis YB-1413 and YB-1414, producing extracellular α-galactosidase, were obtained from homemade Doenjang. On the basis of their biochemical properties, 16S rRNA sequences and random amplified polymorphic DNA patterns by polymerase chain reaction, they were found to be somewhat different from one another. α-Galactosidase productivities of the two isolates were increased by wheat bran, but drastically decreased by melibiose, raffinose and sucrose which were used as carbon sources. The enzyme productivities were increased by yeast extract as a nitrogen source with maximum levels of 1.87 U/ml for YB-1413 and 1.69 U/ml for YB-1414, respectively. The enzymes of both isolates exhibited maximum activity for hydrolysis of para-nitrophenyl-α-D-galactopyranoside (pNP-αGal) under reaction conditions of pH 6.0 and 45℃. Their hydrolyzing activities for pNP-αGal were drastically decreased by the addition of low concentrations of ribose and galactose. They were capable of hydrolyzing completely α-1,6 linked galactosyl residue in melibiose, raffinose and stachyose, which are known to be anti-nutritional factors in products of soybean and legume. In relation to the latter, the isolates YB-1413 and YB-1414 have potential applicability in improving soybean-fermented foods and the nutritional value of soybean feed.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.2
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• pp.231-237
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• 2006

The extracellular enzyme alginase produced by Bacillus licheniformis AL-577 was purified by ion chromatography on CM-Cellulose column, DEAE-Sepharose column, and followed by gel filtration on Sephadx G-100 column. The optimum pH and temperature for the activity of the purified enzyme were 6.0 and $35^{\circ}C$, respectively. The enzyme was stable at the pH range of $6.0\~9.0$ and at $20^{\circ}C$. The molecular weight of the enzyme was estimated to be about 25,500 daltons by SDS-polyacrylamide gel electrophoresis. NaCl was required for high activity of the enzyme. The enzyme was inhibited by $Ba^{2+},\;Co^{2+},\;Cu^{2+},\;Fe^{2+},\;Mg^{2+},\;Zn^{2+},\;NH_4^+$, EDTA, L-cysteine, and 2-mercaptoethanol, while stimulated by DTT, O-phenanthroline, $K^+$ and $Li^+$. This enzyme was proposed to be an alginase specifically degrading alginic acid.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.328-333
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• 2004

The maximum productivity of ${\alpha}-galactosidase,$ capable of hydrolyzing completely ${\alpha}-D-l,6-galactopyranosyl$ linkages within oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, was reached to 718 mU/ml in the culture filtrate of Bacillus licheniformis at death phase. The ${\alpha}-galactosidase$ was identified to show different efficiencies for hydrolyzing the ${\alpha}-galactooligosaccharides$ according to analysis of reaction products by both TLC and quantification of the liberated reducing sugars. The enzyme was active on ${\alpha}-galactooligosaccharides$ in the order of melibiose, raffinose, and stachyose. Though the hydrolyzing activity of enzyme was faintly inhibited by reaction products such as galactose, glucose and sucrose with amounts of five folds more than the added substrates (20 mM), the largest inhibition of enzyme activity was caused by galactose among the end products. Unknown compound, which migrated slower than melibiose on TLC, was detected during hydrolysis reaction of melibiose, suggesting that the ${\alpha}-galactosidase$ has a glycosyl transferase activity. In addition, the enzyme was able to hydrolyze efficiently raffinose and stachyose existed in the soluble extract of soybean meal.

• Korean Journal of Food Science and Technology
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• v.31 no.6
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• pp.1570-1576
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• 1999

Quality characteristics of 3 kinds of kochujang were investigated during 6 months of fermentation in order to improve the quality of industrial kochujang to that of traditional one. Three different kinds of kochujang were prepared using Aspergillus oryzae, Aspergillus oryzae plus Bacillus licheniformis and Aspergillus oryzae, Bacillus licheniformis plus Saccharomyces rouxii. Kochujang prepared with all three microorganisms contained lesser amounts of free sugar than the other preparations and glucose was the most abundant free sugar found. Ethanol was the most abundant amount among alcohols in kochujang. Kochujang prepared with A. oryzae, B. licheniformis plus S. rouxii has more amino nitrogen content than that prepared with A. oryzae only. Capsaicin contents of kochujang decreased slightly in all three kochujang, but there was no significant differences. The results of sensory evaluation showed statistically significant difference among three samples and panel members preferred the taste and flavor of kochujang prepared with all three microorganisms to those of the other samples.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.406-411
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• 2008

This study was conducted to examine the quality enhancement of fermented soybean pastes (cheonggukjang) using microorganisms with high enzyme activities and proper experimental design. The microorganisms for soybean paste fermentation were selected from a specific area of Gyeonggi and were idenlified by 16S rDNA sequence analysis. To prepare the cheonggukjang, an optimum mixing ratio of selected microorganisms was determined using contour plots and numerical optimization methods. A total of 39 microorganisms were isolated from the soybean paste, consisting primarily of Bacillus subtilis and Bacillus licheniformis, and no mold was found. Three microorganisms showing high enzyme activities were selected and used to formulate an optimum mixing ratio for cheonggukjang preparation. Based on levels of amino-nitrogen, ammonium-nitrogen, antioxidant activity values, and sensory preference results, the optimum mixing ratio of 50% of Bacillus sp. SC-1 and 50% SC-3 was suggested for the manufacture of high quality of cheonggukjang.

• Korean Journal of Microbiology
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• v.49 no.3
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• pp.286-291
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• 2013

Soybean pastes with 8% (w/w) salinity were prepared instead of soybean paste with 14% (w/w) salinity to meet the growing demands of Korean's low sodium diet. After aging, white films had appeared on the surface of all low-salted soybean pastes [8% (w/w) salinity] unlike high-salted soybean pastes [14% (w/w) salinity]. All of eight microbes isolated from the surface film were identified as Pichia kudriavzevii. Eleven Bacillus strains with good characteristics of fermentation were isolated from traditionally fermented soybean pastes in order to preserve their unique flavors and aromas after aging, and as a result of analyzing the biochemical characteristics and 16S rRNA sequences, those were identified as B. subtilis, B. licheniformis, and B. methylotrophicus. All of the Bacillus isolates had antagonistic activities against 8 isolates of the film-forming yeasts and harbored the genes for synthesis of antimicrobial surfactants including lichenysin and/or surfactin.

• Microbiology and Biotechnology Letters
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• v.27 no.2
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• pp.113-117
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• 1999

As a basic study for quality improvement of Korean soybean paste and soybean sauce, we investigated on microflora of soybean paste and soybean sauce fermentation. Major Microorganisms were isolated from the sample pastes and sauces, and identified systematically. Selected Microorganisms were identified by MIS whole cell fatty acid analysis by gas chromatography. Identification results showed that Bacillus licheniformis, bacillus pumilis and Bacillus subtilis were dominant in soybean paste and Staphylococcus vitulus, Bacillus subtilis, Bacillus pumilus, and Lactobacillus fermentum were dominant in soybean sauce. It seemed that these Microorganisms played an important role in soybean paste and soybean sauce fermentation and could be used for the further studies such as protease and amylase activities.

• Food Quality and Culture
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• v.3 no.1
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• pp.40-44
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• 2009

To evaluate the hypolipidemic effects of Cheonggukjang (CGJ), which is frequently used in Korea similar to Natto in Japan and Douchi in China like a dairy product, boiled soybeans were fermented with two Bacillus strains, B. subtilis and B. licheniforms, isolated from rice straw and their antihyperlipidemic effects of their products were investigated. Treatment with the CGJs significantly reduced blood triglyceride (TG) and total cholesterol (TC) levels and increased HDL cholesterol levels in Triton WR-1339-induced hyperlipidemic mice. The treatment of non-fermented soybeans alone also reduced blood TG and TC levels, but not significantly. Feeding the CGJs significantly lowered high blood TG and TC levels as well as body and epididymal mass weights in hyperlipidemic mice induced by the long-term feeding of a high-fat diet that increased blood HDL cholesterol levels. The B. subtilis-fermented CGJ products more potently reduced TG and TC levels, although the differences between the starters were not significant. These finding suggest that CGJ products may be effective as hypolipidemic foods by the synergistic interaction of soy and Bacillus strains.

• Food Science and Biotechnology
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• v.18 no.3
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• pp.594-603
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• 2009

The genus Bacillus includes a variety of diverse bacterial species, which are widespread throughout the environment due to their ubiquitous nature. A well-known member of the genus, Bacillus cereus, is a food poisoning bacterium causing both emetic and diarrhoeal disease. Other Bacillus species, particularly B. subtilis, B. licheniformis, B. pumilus, and B. thuringiensis, have also recently been recognized as causative agents of food poisoning. However, reviews and research pertaining to bacilli have focused on B. cereus. Here, we review the literature regarding the potentially toxigenic Bacillus species and the toxins produced that are associated with food poisoning.

• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1469-1476
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• 2007

Jeot-gal is a traditional Korean fermented seafood and has long been used for seasoning. We isolated 188 strains from shrimp, anchovy, and yellow corvina Jeot-gal, and screened sixteen strains that showed strong fibrinolytic activities on a fibrin plate. Among those strains, the strain that had the largest halo zone was chosen and identified as Bacillus licheniformis by using 16S rDNA sequencing and an API CHB kit. The fibrinolytic activity of Bacillus licheniformis was characterized and designated as bpKJ-31. The active component of bpKJ-31 was identified as a 37 kDa protein, designated bacillopeptidase F, by internal peptide mapping and N-terminal sequencing. The optimum activity of bpKJ-31 was shown at pH 9 and $40^{\circ}C$, with a chromogenic substrate for plasmin. It had high degrading activity for the $B{\beta}$-chain and $A{\alpha}$-chain of fibrin(ogen), and also acted on thrombin, but not skim milk and casein. The amidolytic activity of bpKJ-31 was inhibited by 1 mM phenylmethanesulfonyl fluoride, but 1 mM EDTA did not affect the enzyme activity, indicating that bpKJ-31 is an alkaline serine protease, like a plasmin. The bpKJ-31 showed approximately 14.3% higher fibrinolytic activity than the plasmin. These features of bpKJ-31 make it attractive as a health-promoting biomaterial.