• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.283-289
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• 2002

This study was performed to search for potential microorganism that has rapid fermenting and physiological function from anchovy sauce. We isolated three bacterial strains, JM-1, JM-2, and JM-3 with proteolytic and fibrinolytic activity from anchovy sauce. Among the 3 bacterial strains, JM-3 showed the strongest proteolytic and fibrinolytic activity. Bacterial strain JM-3 was gram-positive rod, motile and formed endospore. The 16S rRNA of bacterial strain JM-3 was amplified by PCR and then its sequence was determined by ABI 310 genetic analyzer. The 16S rRNA sequence of bacterial strain JM-3 was compared to BLAST DNA database and identified to Bacillus subtilis with 99% of homology. The optimum temperature, pH and NaCl concentration for growth of B. subtilis JM-3 were $40^{\circ}C$, 5.0 and 0%, respectively. The optimum temperature, pH and NaCl concentration for proteolytic and fibrinolytic enzyme production of B. subtilis JM-3 were same as optimum conditions for growth. At 20% of NaCl concentration which is common NaCl concentration of fish sauce, B. subtilis JM-3 showed about 60% of proteolytic and fibrinolytic activity of 0% NaCl concentration. From above results, we found that B. subtilis JM-3 will be able to used for starter of functional fish sauce.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.684-689
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• 2003

In previous paper, we isolated the bacteria, Bacillus subtilis JM-3, with proteolytic and fibrinolytic activity for candidate microorganisms that have rapid fermenting and physiological functions from anchovy sauce. This study was carried out to search physiological functions of Bacillus subtilis JM-3, such as antimicrobial, antioxidative, antimutagenic, angiotensin-converting enzyme inhibition, and anticarcinogenic activity in vitro. The cell free culture of Bacillus subtilis JM-3 showed strong antibacterial activity against Listeria monocytogenes, antioxidative activity with 87% of inhibition rate against linoleic acid, 50% of antimutagenic activity against N-nitrosodimethylamine and N-nitrosomorpholine, and 88.9% of growth inhibition rate against SNU-1 cell line (stomach cancer cell of human). However, Bacillus subtilis JM-3 did not show angiotensin-converting enzyme inhibition activity.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.827-832
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• 2005

Bacillus subtilis JM3 protease from naturally fermented anchovy sauce was partially purified in 40-60% ammonium sulfate fraction. To accelerate fermentation of anchovy sauce, 2 and 4% crude B. subtilis JM3 proteases were added to 6 month-ripened anchovy sauce, and their hydrolysis degrees and amino-nitrogen contents were investigated at different storage times. Low molecular weight (LMW) peptide was purified by ultrafiltration ana gel permeation chromatography from anchovy sauce manufactured with B, subtilis JM3 protease. Anchovy sauces with 2 and 4% proteases increased hydrolysis rate by 27 and 32%, respectively. Amino-nitrogen contents of anchovy sauces fermented with 2 and 4% proteases were twofold higher than that of control. Control showed five peptide peaks on Bio-Rad P2 gel permeation chromatography spectrum, whereas anchovy sauces with 2 and 4% B. subtilis JM3 proteases showed six and seven peaks, respectively. ACE inhibitory activity was highest in peak 6 (43.75%) of anchovy sauce with 2% protease, followed by peak 5 (34.82%) of control. DPPH radical-scavenging effect was higher than 50% in all samples. Cytotoxicity was highest in peak 3 (44.12%) of control, fellowed by peak 5 (42.04%) of anchovy sauce with 4% protease.

• Journal of Mushroom
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• v.21 no.3
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• pp.140-144
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• 2023

The objective of this study was to achieve biological control of green mold disease in Pyogo mushrooms using antagonistic microorganisms. Bacillus subtilis BSM320 cells inhibited mycelial growth by 48-60% against three Trichodermaisolates including T. hazianumisolated from the substrates of Lentinula edodes, showing their antifungal activity.The bacteria were cultured to a high density of 4.2 × 10⁹±113.7 cfu/mlin aqueous extract of composted spent mushroom substrates of L. edodes containing 1% glucose and showed a higher growth rate than that observed when using the commercial medium, Luria-Bertani broth. The bacterial culture showed a 75% protective effect without damaging the mushroom fruiting bodies. These results suggest that B. subtilis BSM320culture is suitable for biological control of green mold disease during mushroom cultivation.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.508-514
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• 2001

A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.297-303
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• 1989

Attempts to isolate the naturally occurring ultra-violet resistant bacteria from environmental sources were made. The isolates, designated No.29, 100, and 107, among numbers of bacterial isolates revealed a remarkable resistance to UV ray, whose degree of resistance in dose/response kinetics was comparable to that of an endospore-former, Bacillus subtilis. In a range of 100-300 $Jm^{-2}$/min of UV irradiation, the isolates exhibited 500-1000 fold resistance compated with E. coli. The isolated appeared to possiss cell-bound pigment of organge or crimson-red. The isolate 29 is spherical in pairs or tetrads, whereas the isolates 100 and 107 are rod. All are Gram-gositive bacteria and seemed to be non-endospore-bearer. A number of biochemical studies pursued on the isolates suggested that they are quite different to each other. Electron microscopic examination and the physiological characters of the isolate 29 suggested that this UV resistant spherical bacterium might be one species of Deinococcus, probably Deinococus radiophilus. Since there is no documents on UV resistant, Gram-positive, non-sporeformer bacillus so far, the isolates 100 and 107 might be turned out as new kinds of UV resistant bacteria occurring in nature by further investigation.

• Journal of Mushroom
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• v.12 no.4
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• pp.350-356
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• 2014

This study was conducted to evaluate the feeding value of the spent mushroom (Hypsizygus marmoreus) substrates (SMS) in laying hens (Hy-Line Brown). The fresh spent mushroom (Hypsizygus marmoreus) substrates collected from the Namkyung farm were fermented with Bacillus subtilis EJ3 for 2 weeks. A total of twenty-four laying hens were fed corn-soy based experimental diets containing 0% (T0), 5% (T1), 10% (T2) and 15%(T3) fermented SMS for 12 weeks. There were no significant differences among the treatments in egg production, egg weight, egg mass, feed conversion and viability during the experimental period. Feed intake was significantly lowered in T0 (118.3 g) than T1 (121.9 g), T2 (120.3 g) and T3 (122.4 g). There were no significant differences among the treatments eggshell breaking strength, thickness and haugh unit, whereas the yolk color of T1, T2 and T3 were significantly heavy than T0. The palatability of boiled meat was significantly better in the T3 laying hens than in the T0 laying hens. In conclusion, fermented SMS can be used as resource of feed in laying hen feed at 5.0-15% level without effect on performance and egg qualify.