• Applied Biological Chemistry
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• v.40 no.3
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• pp.173-177
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• 1997

Bacillus licheniformis 9945a releases a natural ${\gamma}-poly(glutamic\;acid)({\gamma}-PGA)$ into fermentation broth and shows a mucoid phenotype on the solid agar medium. Transformation of mucoid cells of Bacillus species has not been simple and straightforward. The transpositional activity of Tn10 in B. licheniformis also has not been own either. Thus, a spontaneous non-mucoid derivative of the B. licheniformis was obtained first. Shuttle vector pHV1248 containing mini-Tn10 was introduced into the non-mucoid derivative by the method of protoplast transformation. The resulting transformant was reverted to the wild mucoid phenotype, and then mutated randomly with the mini-transposon by heat induction. Auxotrophs requiring arginine, lysine, or tryptophan were isolated by replica plating method. Southern blotting and DNA-DNA hybridzation analysis showed that these auxotrophs were generated by mini-Tn10 insertion into the chromosomal DNA. This method of transformation and mutation using pHV1248 would be useful for the generation of diverse mutants of B. licheniformis 9945a.(Received January 24,1997; accepted March 10, 1997)

• Journal of Life Science
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• v.16 no.2 s.75
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• pp.175-179
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• 2006

A bacterium that produced a large amount of poly-$\gamma$-glutamate (PGA) was isolated from the compost and designated as Bacillus subtilis CH-10. The optimum temperature and pH for PGA production were at $37^{\circ}C$ and 7.5, respectively. The maximum amount of PGA production (18.84 mg/ml) was obtained when it was grown in a medium containing 3% L-glutamate and 5% sucrose at $37^{\circ}C$ with shaking. The result that the L-glutamate significantly induced PGA production indicates that it produces a PGA by the glutamate dependent manner. Some properties of the PGA obtained at different times of cultivation were investigated by SDS-PAGE and ninhydrin analysis. The PGA production was elongated along with cultivation time and maximum amount was achieved at 96 h. Average molecular weight of PGA was estimated to be 1100 kDa by FDNB method.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.436-436
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• 2005

• Journal of Veterinary Clinics
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• v.29 no.4
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• pp.283-296
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• 2012

Allergic contact dermatitis (ACD) is an inflammatory skin disease and regarded as a prototype of T-cell mediated delayed-type hypersensitivity reactions. Poly-${\gamma}$-glutamic acid (PGA) is a biodegradable polymer that is produced by Bacillus subtilis. This study was performed to assess the effects of PGA in a canine model of ACD. ACD was induced on the back of dogs induced by sensitization and repeated application by 2,4-dinitro-1-chlorobenzene (DNCB). Topical treatment of PGA was applied once a day for 12 days and skin biophysical parameters including transepidermal water loss (TEWL), skin hydration, skin pH, skin thickness and erythema index, were measured every two days during experimental periods. Histopathology and immunohistochemistry were performed to evaluate the antiinflammatory effect. In skin biophysical parameters, TEWL, skin hydration, skin thickness and erythema index were significantly increased, with a maximum increase appeared on day 2 (p < 0.05). On the other hand, skin pH was significantly decreased, with a maximum decrease appeared on day 2 (p < 0.01). After the completion of PGA treatment, skin biophysical parameters were significantly reached those of baseline in a time-dependent manner (p < 0.05). In histopathology, marked increases of epidermal thicknesses were induced after DNCB challenge with numerous inflammatory cell infiltrations and edematous changes, decreases of connective tissue occupied regions in dermis. In addition, marked increases of cytokine - tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$)-immunoreactivities in the dermis and of apoptotic markers - caspase-3 and PARP-immunoreactivities in the epidermis were observed in DNCB-PBS control as compared with intact control, respectively (p < 0.01). It means, the ACD and related apoptotic changes were induced by DNCB in the present study. However, these ACD induced by DNCB and related apoptosis in epidermis were significantly inhibited by treatment of PGA treated skin, the decreases of infiltrated inflammatory cells and related decreases of pro-inflammatory cytokine immunoreactivities were also observed (p < 0.01). Based on these findings, PGA may have anti-inflammatory and alleviatory effects in the allergic contact dermatitis.

• Korean Journal of Food Science and Technology
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• v.48 no.2
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• pp.153-158
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• 2016

The principal objective of this study was to produce a chitosan nanoparticle (NP) system for improving blood circulation. Chitosan NPs were prepared using fucoidan and $poly-{\gamma}-glutamic$ acid (PGA), denoted as CS/Fu and CS/Fu/PGA NPs, respectively. As the chitosan concentration was increased, the activated partial thromboplastin time (APTT) of the NPs significantly increased (p<0.05). When the concentration of fucoidan and ${\gamma}-PGA$ was 5-20 and $1-10{\mu}g/mL$, respectively, the size of the CS/Fu and CS/Fu/PGA NPs was approximately 200 and 100 nm, respectively. With an increase in the fucoidan and PGA concentration, the APTT of CS/Fu and CS/Fu/PGA NPs significantly increased (p<0.05). These results suggest that CS/Fu and CS/Fu/PGA NPs could be used as a potent NP system for improving blood circulation.

• Nutrition Research and Practice
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• v.15 no.3
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• pp.309-318
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• 2021

BACKGROUD/OBJECTIVES: Data regarding the effects of poly-γ-glutamic acid (γ-PGA) on sleep status are limited. This study aimed to test whether γ-PGA and vitamin B₆ (VitB6) supplements improve sleep duration and quality. SUBJECTS/METHODS: A factorial randomized, double-blinded, placebo-controlled crossover study included 47 adults (25 men and 22 women) who were free of chronic disease. Stratified randomized allocation considered age and gender for three interventions, group A (supplementation with γ-PGA 600 mg; n = 16), group B (supplementation with VitB6 100 mg; n = 14), and group C (dual supplementation of both γ-PGA 600 mg and VitB6 100 mg; n = 17). Participants underwent a 1-mon intervention period, followed by a 1-mon washout period, and then a second 1-mon intervention period. Differences (mean ± SD) in nighttime sleep status before and after supplementation were compared between the placebo and intervention groups using nonparametric tests. RESULTS: Significant changes in sleep duration (0.27 ± 0.98 h, P < 0.05) and the Pittsburgh Sleep Quality Index global score (-0.52 ± 1.58, P < 0.05) indicating improved sleep status were observed in the intervention compared with the placebo of group C while no significant changes were observed in groups A and B. No statistical significance was detected between the intervention and the placebo; however, there was a greater increase in the group C intervention (4.59 ± 38.5 ng/mL) in serum serotonin concentrations than the groups A and B interventions. No side effects were observed. CONCLUSIONS: On the basis of these findings, the dual supplementation of γ-PGA and VitB6 may be effective as functional food components to improve nighttime sleep status.

• Korean Journal of Food Science and Technology
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• v.50 no.6
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• pp.572-580
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• 2018

Biologically active substances including gamma-aminobutryric acid (GABA) were added into whey by co fermentation using Bacillus subtilis HA and Lactobacillus plantarum EJ2014. The first fermentation using B. subtilis HA with 5% monosodium glutamate (MSG) and 2% glucose enhanced the production of poly-${\gamma}$-glutamic acid (PGA), resulting in higher consistency of $4.09Pas^n$ as well as whey protein peptides. After the second fermentation using L. plantarum EJ2014, the remaining MSG (3.40%) as a precursor was completely converted to 2.21% GABA. Furthermore, the lactose content in whey decreased from 6.73 to 3.68% after co-fermentation, and the tyrosine content increased from 20.47 to 38.24%. Peptides derived of whey proteins were confirmed by SDS-PAGE. Viable cell counts of B. subtilis and L. plantarum were 5.83 log CFU/mL and 9.20 log CFU/mL, respectively. Thus, co-fermentation of whey could produce the novel food ingredient fortified with biologically active compounds including GABA, ${\gamma}$-PGA, peptides, and probiotics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.11
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• pp.1544-1551
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• 2016

Various types of fermented soybean foods are consumed in various Asian countries, including China, Indonesia, Japan, Korea, and Vietnam. Cheonggukjang is a Korean whole soybean paste fermented by Bacillus subtilis and regarded as a healthy food. The objective of this study was to investigate the Ca-bioavailability and anti-osteoporotic effects of cheonggukjang (CGJ) containing high poly-gamma glutamate (PGA) contents in an animal model. Blood samples were collected from the jugular vein at 0, 0.5, 1, 2, 3, 4, and 5 h after a single oral dose in male rats. Ca-bioavailability in CGJ containing high PGA contents was approximately 3~5 times higher than that of natural CGJ. Female rats were either sham-operated (sham; n=5) or surgically ovariectomized (OVX; five animals per group) and then administered to OVX control, raloxifene hydrochloride (RLX) 1 mg/kg/d, or CGJ containing high contents of PGA (20 and 200 mg/kg/d) for 12 weeks. Serum osteocalcin concentration was significantly lower in the CGJ 200 mg/kg/d group compared with the OVX control group. Serum progesterone concentration was significantly higher in the CGJ 200 mg/kg/d group compared with the OVX control group. Reduction grade of the trabecular bone decreased in the RLX 1 and CGJ 200 mg/kg/d group compared with that of the OVX control group. In conclusion, CGJ 200 mg/kg/d may have inhibitory effects on osteoporosis in OVX rats, and Ca-bioavailability was improved in CGJ containing high PGA contents.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.664-670
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• 2016

Cheonggukjang (CKJ) is a Korean traditional food made of fermented soybeans. In comparison to normal intake of soybeans, Cheonggukjang has high digestibility with bioactive, antioxidant substances, and thrombolytic enzymes. Recent studies have reported anti-oxidant, anti-cancer, anti-inflammatory, anti-obesity activities as well as inhibitory activities against osteoporosis for CKJ. In this study, we identified the effects of CKJ on osteoblast differentiation by increasing the polyglutamic acid (PGA) content of CKJ. Alkaline phosphatase (ALP) activity and mineralization significantly increased in response to treatment with both natural CKJ (CKJ A) and PGA-increased CKJ (CKJ B). However, CKJ B exhibited higher ALP activity and mineralization than CKJ A. Real-time reverse transcription PCR demonstrated that mRNA expression of osteoblastic-associated genes such as type I collagen, alkaline phosphatase, osteocalcin, and osteopontin in C2C12 cells was significantly up-regulated by CKJ A or B treatment. These results indicate that treatment with CKJ has an anabolic effect on bone by increasing osteoblastic differentiation and ALP activity. Increasing PGA content in CKJ had a greater effect than CKJ A on up-regulation of osteoblastic gene expression in osteoblast cells.

• Food Science and Preservation
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• v.31 no.1
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• pp.138-148
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• 2024

The production of poly-γ-glutamic acid (γ-PGA) and γ-aminobutyric acid (GABA) was optimized by serial fermentation of Dendropanax morbiferus extract (DME) using Bacillus subtilis HA and Lactobacillus plantarum KS2020. The 1st alkaline fermentation was performed on 60% DME including 2% glucose and 10% monosodium ʟ-glutamate (MSG) as a precursor. The 1^st fermented DME had 57 mg% tyrosine. Consequently, the 2^nd lactic acid fermentation for 5 days increased the tyrosine content of 106 mg%. The mucilage containing γ-PGA showed a high content of 3.50% on the first day of alkaline fermentation and then increased to 4.10% after 2 days. The precursor (MSG) remaining in the 1^st fermented DME was efficiently converted to GABA by the 2^nd lactic acid fermentation in the presence of 5% skim milk, 1.5% glucose and 0.5% yeast extract, resulting in the production of 18.29 mg/mL GABA. The viable cells of lactic acid bacteria increased and indicated 9.49 log CFU/mL on the fermentation for 5 days, and the acidity of co-fermented DME indicated the highest value of 1.55%. Conclusively, the serial fermented DME has multi-functional ingredients containing γ-PGA, GABA, peptides and probiotics.