• 미생물학회지
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• 제38권3호
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• pp.230-233
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• 2002

활성산소는 많은 경우에 성인병, 염증, 암, 동맥경화 등과 관련되어 있다. 따라서 항산화물질의 섭취는 성인병 예방이나 치료에 도움이 될 수 있다. 보리, 쑥, 다시마, 대두는 자연식품으로 항산화물질을 포함하고 있다. 이들을 발효시켰을 때, 원재료에 비해 항산화도가 증가할 수 있는지 여부를 결정하였다. 항산화도를 결정하기 위해서는 1,1-diphenyl-2-picrylhydrazyl (DPPH)를 이용하였다. 보리, 쑥, 다시마, 대두를 Bacillus licheniformis Bl으로 발효 시켰을 때, ethanol 추출 발효산물의 항산화도는 원재료에 비해 각각, 2.6, 1.6, 2.7, 1.7배로 증가하였다. 또한,250∼300 nm에 걸쳐 발효대두의 ethanol 추출성분의 peak 영역이 대두의 것에 비해 뚜렷하게 높았으며, 이는 이런 구성성분의 차이가 양 항산화도의 차이와 관련되어 있을 가능성을 제시해 준다. Paraquat는 Escherichia coli내부에서 활성산소를 만들어 균의 성장을 억제하는 것으로 알려져 있다. Ethanol로 추출한 발효대두 성분은 paraquat 의한 균의 억제를 회복시킬 수 있었다. 이는 발효대두 성분이 체내에서도 이용될 수 있는 가능성을 보여준다.

• Asian-Australasian Journal of Animal Sciences
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• 제21권9호
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• pp.1312-1317
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• 2008

The objective of the present trial was to determine the effect of a thermostable probiotic containing Bacillus licheniformis and B. subtilis on health and production parameters of fattening rabbits from weaning until slaughter. In a rabbitry with average post-weaning mortality of 5-9%, 1,680 rabbits were supplied with: a) a basic feed, or b) the same basic feed supplemented with probiotic from the 4th day postweaning (41st day of age) up to 88th day of age. Clinical signs, microbiological status and growth performance were recorded for two distinct fattening periods, growing and finishing. A significant decrease in mortality of probiotic-treated rabbits when compared to the controls was observed during the growing and entire fattening periods. Within these periods, E. coli and C. perfringens - but not P. multocida - were isolated at a lower frequency from probiotic-treated rabbits (p<0.05). Compared to the control animals, probiotic-treated rabbits were 54 g and 123 g heavier at the end of the growing and finishing phases, respectively, and had significantly higher average daily gain and better feed conversion ratio (p<0.05).

• Journal of Microbiology and Biotechnology
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• 제34권5호
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• pp.1154-1163
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• 2024

Glucosylation is a well-known approach to improve the solubility, pharmacological, and biological properties of flavonoids, making flavonoid glucosides a target for large-scale biosynthesis. However, the low yield of products coupled with the requirement of expensive UDP-sugars limits the application of enzymatic systems for large-scale. C. glutamicum is a Gram-positive and generally regarded as safe (GRAS) bacteria frequently employed for the large-scale production of amino acids and biofuels. Due to the versatility of its cell factory system and its non-endotoxin producing properties, it has become an attractive system for the industrial-scale biosynthesis of alternate products. Here, we explored the cell factory of C. glutamicum for efficient glucosylation of flavonoids using apigenin as a model flavonoid, with the heterologous expression of a promiscuous glycosyltransferase, YdhE from Bacillus licheniformis and the endogenous overexpression of C. glutamicum genes galU1 encoding UDP-glucose pyrophosphorylase and pgm encoding phosphoglucomutase involved in the synthesis of UDP-glucose to create a C. glutamicum cell factory system capable of efficiently glucosylation apigenin with a high yield of glucosides production. Consequently, the production of various apigenin glucosides was controlled under different temperatures yielding almost 4.2 mM of APG1(apigenin-4'-O-β-glucoside) at 25℃, and 0.6 mM of APG2 (apigenin-7-O-β-glucoside), 1.7 mM of APG3 (apigenin-4',7-O-β-diglucoside) and 2.1 mM of APG4 (apigenin- 4',5-O-β-diglucoside) after 40 h of incubation with the supplementation of 5 mM of apigenin and 37℃. The cost-effective developed system could be used to modify a wide range of plant secondary metabolites with increased pharmacokinetic activities on a large scale without the use of expensive UDP-sugars.

• Nutrition Research and Practice
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• 제9권6호
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• pp.569-578
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• 2015

BACKGROUND/OBJECTIVES: Fermentation can increase functional compounds in fermented soybean products, thereby improving antioxidant and/or anti-inflammatory activities. We investigated the changes in the contents of phenolics and isoflavones, antioxidant activity and anti-inflammatory activity of Doenjang during fermentation and aging. MATERIALS/METHODS: Doenjang was made by inoculating Aspergillus oryzae and Bacillus licheniformis in soybeans, fermenting and aging for 1, 3, 6, 8, and 12 months (D1, D3, D6, D8, and D12). Doenjang was extracted using ethanol, and sequentially fractioned by hexane, dichloromethane (DM), ethylacetate (EA), n-butanol, and water. The contents of total phenolics, flavonoids and isoflavones, 2,2-diphenyl-1 picryl hydrazyl (DPPH) radical scavenging activity, and ferric reducing antioxidant power (FRAP) were measured. Anti-inflammatory effects in terms of nitric oxide (NO), prostaglandin (PG) E2 and pro-inflammatory cytokine production and inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 expressions were also measured using LPS-treated RAW 264.7 macrophages. RESULTS: Total phenolic and flavonoid contents showed a gradual increase during fermentation and 6 months of aging and were sustained thereafter. DPPH radical scavenging activity and FRAP were increased by fermentation. FRAP was further increased by aging, but DPPH radical scavenging activity was not. Total isoflavone and glycoside contents decreased during fermentation and the aging process, while aglycone content and its proportion increased up to 3 or 6 months of aging and then showed a slow decrease. DM and EA fractions of Doenjang showed much higher total phenolic and flavonoid contents, and DPPH radical scavenging activity than the others. At $100{\mu}g/mL$, DM and EA fractions of D12 showed strongly suppressed NO production to 55.6% and 52.5% of control, respectively, and PGE2 production to 25.0% and 28.3% of control with inhibition of iNOS or COX-2 protein expression in macrophages. CONCLUSIONS: Twelve month-aged Doenjang has potent antioxidant and anti-inflammatory activities with high levels of phenolics and isoflavone aglycones, and can be used as a beneficial food for human health.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1154-1162
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• 2021

The transcriptional capacities of target genes are strongly influenced by promoters, whereas few studies have focused on the development of robust, high-performance and cross-species promoters for wide application in different bacteria. In this work, four novel promoters (P_k.rtufB, P_k.r1, P_k.r2, and P_k.r3) were predicted from Ketogulonicigenium robustum and their inconsistency in the -10 and -35 region nucleotide sequences indicated they were different promoters. Their activities were evaluated by using green fluorescent protein (gfp) as a reporter in different species of bacteria, including K. vulgare SPU B805, Pseudomonas putida KT2440, Paracoccus denitrificans PD1222, Bacillus licheniformis and Raoultella ornithinolytica, due to their importance in metabolic engineering. Our results showed that the four promoters had different activities, with P_k.r1 showing the strongest activity in almost all of the experimental bacteria. By comparison with the commonly used promoters of E. coli (tufB, lac, lacUV5), K. vulgare (Psdh, Psndh) and P. putida KT2440 (JE111411), the four promoters showed significant differences due to only 12.62% nucleotide similarities, and relatively higher ability in regulating target gene expression. Further validation experiments confirmed their ability in initiating the target minCD cassette because of the shape changes under the promoter regulation. The overexpression of sorbose dehydrogenase and cytochrome c551 by P_k.r1 and P_k.r2 resulted in a 22.75% enhancement of 2-KGA yield, indicating their potential for practical application in metabolic engineering. This study demonstrates an example of applying bioinformatics to find new biological components for gene operation and provides four novel promoters with broad suitability, which enriches the usable range of promoters to realize accurate regulation in different genetic backgrounds.

• 한국축산식품학회지
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• 제38권4호
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• pp.749-758
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• 2018

The objective of this study was to provide preliminary data for food industry by investigating the distribution of microorganisms in raw materials and sausage examining the effect of heating temperature on sausage quality. Total microbes in sausage ranged 2.21-3.11 Log CFU/g. Bacillus pumilus, B. licheniformis, Staphylococcus saprophyticus, and Enterococcus faecalis were detected on sausage. Total microbes in raw materials was 1.59-7.16 Log CFU/g. Different types of microorganisms were found depending on raw materials, with B. pumilus and B. subtilis were being detected in both raw materials and sausage. Total microbes in sausage after heating was in the range of 1.10-2.22 Log CFU/g, showing the trend of decrease in total microbe with increasing heating temperature, although the decrease was not significant. With increasing heating temperature, pH and hardness were also increased. The yield of sausage manufactured at $85^{\circ}C$ was 95.42% while that manufactured at $65^{\circ}C$ was 96.67%. Therefore, decreasing heating temperature during sausage production might increase yield and save energy without microbiological effect.

• Molecules and Cells
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• 제37권2호
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• pp.172-177
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• 2014

A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-${\alpha}$-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the $5^{th}$ hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-${\beta}$-D-glucoside, genistein 7-O-${\beta}$-D-glucoside (genistin), genistein 4',7-O-${\beta}$-D-diglucoside, biochanin A-7-O-${\beta}$-D-glucoside (sissotrin), daidzein 4'-O-${\beta}$-D-glucoside, daidzein 7-O-${\beta}$-D-glucoside (daidzin), daidzein 4', 7-O-${\beta}$-D-diglucoside, and formononetin 7-O-${\beta}$-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOF-ESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli $BL21(DE3)/{\Delta}pgi{\Delta}zwf{\Delta}ushA$ overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of $200{\mu}M$ of supplemented isoflavonoids, without any additional UDP-${\alpha}$-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.

• 한국식품과학회지
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• 제37권6호
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• pp.1012-1017
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• 2005

게맛살로부터 분리한 주요 부패세균은 내열성 포자를 형성하는 Bacillus subtilis와 Bacillus licheniformis로 동정되었다. 게맛살의 제조 공정상 가열 처리 과정에서 B. subtilis와 B. Licheniformis 등 내열성 포자를 형성하는 균을 완전히 사멸시키기는 어려우며, 살아남은 포자는 유통과정 중, 적정 온도와 시간이 경과함에 따라, 영향 세포로 발아하여 게맛살의 부패에 영향을 미친다. 이러한 부패세균의 증식에 있어서 초기균수와 온도의 영향을 조사한 결과, 초기균수에 따른 최대증식속도상수(k)와 유도기(LT), 세대시간(GT)은 유의적인 차이가 없었으며, 온도의 영향이 지배적인 것으로 나타났다. 또한 본 실험에서 유도기(LT)와 온도의 관계는 $L(hr)=2.5219e^{-0.2467{\cdot}T}$의 관계가 성립하며, square root model과 polynomial model을 이용, 온도와 초기균수에 대한 최대증식속도상수(k)를 정량화한 정량평가모델을 개발하였으며, 그 식은 다음과 같다. $$Square\;root\;model:\;{\sqrt{k}}=0.0267\;(T-3.5089)$$ $$Polynomial model:\;k=-0.2160+0.0241T-0.01999A_0$$ 온도와 초기균수에 대한 최대증식속도상수(k)의 정량평가모델로부터 특정온도와 초기 균수에서 최대증식속도상수(k)를 계산할 수 있으며, 계산된 최대증식속도상수(k)를 균의 기본 증식 모델인 Gomperz model에 적용하여 균의 성장을 예측할 수 있었다.

• Journal of Animal Science and Technology
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• 제66권2호
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• pp.326-339
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• 2024

This study was conducted to investigate the effects of protease supplementation and different nutrient density of diets in growing-finishing pigs. A total of one hundred-eight crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial body weight (BW; 18.74 ± 3.46 kg) were used for 15 weeks. Pigs were randomly assigned to six dietary treatments with 6 replicates of 3 pigs per pen in a 3 × 2 factorial through the following arrangement: Three groups of protease (1, Basal diets; 2, Protease A: 125 mg/kg protease derived from Streptomyces sps; 3, Protease B: 100 mg/kg protease derived from Bacillus licheniformis) at two different nutrient density diets (1, Basal requirement; 2, 0.94%-0.98% higher than requirement in dietary protein and 50 kcal/kg in energy). High nutrient (HN) diets showed higher average daily gain (ADG) (p < 0.05) and apparent total tract digestibility (ATTD) of crude protein (CP) (p < .0001) compared to basal nutrient (BN) diets during growing periods. Supplementation of protease showed higher BW (p < 0.05) and ADG (p < 0.05) compared to non-supplementation of protease during growing periods. Also, supplementation of protease showed higher ATTD of CP (p < 0.01), ATTD of gross energy (p < 0.05) and decreased blood urea nitrogen (BUN) level (p = 0.001) compared to non-supplementation of protease during finishing periods. Pigs which fed the protease showed decreased ammonia (NH₃) emissions (p < 0.05) during experiment periods and decreased hydrogen sulfide (H₂S) emissions (p < 0.01) during finishing periods. Interactions between nutrient density and protease were observed, which decreased the feed conversion ratio (p < 0.05) in HN diets without protease compared to BN diets without protease during weeks 4 to 6. Also, interaction between nutrient density and protease was observed, which resulted in improved ATTD of CP (p < 0.01) in response to PTA supplementation with HN diets during the finishing period. In conclusion, supplementation of protease reduces NH₃ in feces and BUN in whole blood by increasing the digestibility of CP and improves growth performance. Also, diets with high nutrient density improved growth performance and nutrient digestibility in growing periods.