메주로부터 분리한 항진균 및 항세균 활성의 Bacillus polyfermenticus CJ9

정지혜,장해춘,Jung, Ji-Hye,Chang, Hae-Choon 한국미생물·생명공학회 2009 한국미생물·생명공학회지 Vol.25 No.4

메주로부터 항진균 및 항세균 활성을 나타내는 균주를 분리하고 동정하여 B. polyfermenticus CJ9로 명명하였다. B. polyfermenticus CJ9의 생육에 따른 항균 활성을 측정한 결과 항세균 활성은 배양 12시간에 최대 활성을 나타내며 72시간까지 90% 이상 활성을 유지하다 120시간에 활성을 완전히 상실하였다. 항진균 활성은 배양 24시간 이후부터 최대 활성을 나타내었고, 사멸기 이후 활성이 다소 감소되었으나 배양 120시간까지 활성이 유지되었다. B. polyfermenticus CJ9은 식품과 인체에 유해한 곰팡이, 효모, 그람 양성 및 음성 세균에 대한 항진균 활성과 항세균 활성을 동시에 나타내었다. B. polyfermenticus CJ9의 항세균 활성은 $37^{\circ}C$ 에서 24시간 열처리 후에 활성을 상실하였으며, pH 5.0~9.0 구간에서는 안정한 활성을 나타내었으나 pH 3.0~4.0 구간에서 활성이 감소하였다. 항진균 물질은 $121^{\circ}C$에서 15분간 열처리시 활성이 감소되었으나 역가가 완전히 소실되지 않았으며, pH 3.0~9.0 구간에서 안정한 활성을 나타내었다. 항세균 물질과 항진균 물질은 proteinase K, protease, trypsin, $\alpha$-chymotrypsin 등의 단백분해효소 처리로 역가를 상실하거나 일부 감소되어 단백질성 물질임을 추정하였다. B. polyfermenticus CJ9의 항세균 물질과 항진균 물질을 $C_{18}$ Sep-Pak column에 흡착된 분획으로부터 역가를 확인하여 소수성 물질임을 알 수 있었으며, Tricine-SDS-PAGE 및 direct detection 실험을 통하여 분자량을 확인한 결과 항진균 물질은 약 1.4 kDa의 물질임을 확인하였다. 그러나 항세균 활성 물질은 열 불안정성 때문에 동 실험법상에서 그 분자량을 확인할 수 없었다. B. polyfermenticus CJ9이 생산하는 항균 물질은 항세균 및 항진균 활성을 동시에 가지는 단백질성 물질로서 천연 식품보존제 및 정장제재로 활용이 기대되며, 이를 위하여 항세균 물질과 항진균 물질의 정제 및 구조분석 등의 연구가 필요하다. A CJ9 bacterial strain, which showed antifungal and antibacterial activities, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, as well as its 16S rRNA sequence. B. polyfermenticus CJ9 showed the antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. The antibacterial activity was stable in the pH 5.0~9.0, but the activity was lost at $37^{\circ}C$ for 24 hr. The antifungal activity was stable in the pH range of 3.0~9.0 and reduced at $121^{\circ}C$ for 15 min, but antifungal activity was not completely destroyed. The antibacterial activity was completely inactivated by proteinase K, protease, trypsin, and $\alpha$-chymotrypsin. The antifungal activity was also completely inactivated by protease and $\alpha$-chymotrypsin, and reduced its activity by proteinase which indicated that the antifungal and antibacterial compounds have proteineous nature. The apparent molecular mass of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa. The molecular mass of the antibacterial compound could not be determined because of its heat-liable characteristic.

초등학교 남아의 인터넷 폭력게임 사용시간과 게임과몰입정도가 공격적 행동에 미치는 영향

정지혜(Ji Hae Jung),박주희(Ju Hee Park),임양미(Yang Mi Lim) 한국아동학회 2014 아동학회지 Vol.35 No.4

The purpose of this study was to examine the effects of the time spent on violent internet games and game overindulgence on aggressive behavior exhibited by elementary school-aged boys and to investigate the mediating effects of self-control on the relationship between children`s game overindulgence and their aggressive behavior. The subjects of this study were 118 fourth to sixth grade boys from five elementary schools. The instruments used were the aggression subscale of the Korean-Youth Self Report (Oh, Ha, Lee, & Hong, 2001), a questionnaire of the time spent playing violent internet games developed by the authors, the Internet Game Addiction Scale for Children (Kim, Lee, Kim, Kim, & Kim, 2006) and the Self-Control Scale (Nam, 1999). The data were analyzed by means of a Structural Equation Model (SEM). The results were as follows. First, children`s self-control mediated the relationship between game overindulgence and aggressive behavior. Second, the amount of time spent playing violent internet games had no effect on children`s aggressive behavior.

메주로부터 분리한 Bacillus polyfermenticus CJ6의 항진균 활성

정지혜(Ji Hye Jung),장해춘(Hae Choon Chang) 한국식품영양과학회 2009 한국식품영양과학회지 Vol.38 No.4

메주로부터 곰팡이 및 세균 등에 생육 저해활성을 나타내는 균주 B. polyfermenticus CJ6을 분리ㆍ동정하였다. 분리균주 B. polyfermenticus CJ6는 2단 대수기를 나타내는 생육곡선상 특이점을 나타내었으며 배양 30시간 이후부터 최대활성을 나타내었고 사멸기 이후 활성이 다소 감소되었으나 120시간까지 활성을 유지하였다. B. polyfermenticus CJ6의 항진균 활성 물질은 70℃ 이상에서 활성이 감소되었으나 121℃에서 15분간 열처리 시 역가가 완전히 소실되지 않았다. pH 안정성 실험에서는 pH 3.0~9.0 구간에서 안정한 활성을 나타내었으며, 각종 효소에 대한 영향에서 항진균 활성물질은 proteinase K, protease, α-chymotrypsin 등의 단백분해효소 처리로 역가를 상실하거나 일부 감소되어 단백질성 물질임을 추정하였다. 균주의 항진균 활성 물질을 C18 Sep-Pak column으로 부분 정제한 후 Tricine-SDS-PAGE 및 direct detection 실험을 통하여 분자량이 약 1.4 kDa의 물질임을 확인하였다. B. polyfermenticus CJ6가 생산하는 항진균 활성 물질은 기존에 거의 보고되지 않은 B. polyfermenticus 유래의 단백질성 항진균 활성 물질로서 천연보존제 및 천연항균제재로 사용이 기대되며, 이를 위하여 항진균 활성 물질들의 정제 및 구조분석 등의 연구가 필요하다. The CJ6 bacterial strain, which possesses strong antifungal activity, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, and 16S rRNA gene sequencing. B. polyfermenticus CJ6 showed antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. Antifungal activity from B. polyfermenticus CJ6 was reduced after 24 hr at 70℃ but antifungal activity was not completely destroyed. The antifungal activity was stable in the pH range of 3.0~9.0, and inactivated by proteinase K, protease, and α-chymotrypsin, which indicate its proteinaceous nature. The apparent molecular masses of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa.

Bacillus polyfermenticus CJ6가 생산하는 항세균 물질의 특성 및 병원성 식중독 미생물의 성장 억제 효과

정지혜(Ji Hye Jung),장해춘(Hae Choon Chang) 한국식품영양과학회 2011 한국식품영양과학회지 Vol.40 No.6

B. polyfermenticusCJ6가 생산하는 항세균 활성 물질의 특성을 규명하고 정제하였다. 생육에 따른 항세균 활성은 배양 4시간부터 나타내어 정지기인 배양 8시간에 최대 활성(102,400 AU/㎖)을 나타내고 이후 급격히 활성이 감소되었다. B. polyfermenticusCJ6의 항세균 활성 물질은 37℃에서 24시간 처리 시 활성의 50% 저하되고 70℃에서 24시간 처리시 활성이 완전히 소실되어 열에 불안정하였으며, pH 3.0∼9.0 구간에서 안정한 활성을 나타내었다. B. polyfermenticusCJ6의 항세균 활성 물질은 protease, α-chymotrypsin을 처리 시 역가가 크게 감소하였으며 proteinase K의 단백분해 효소에 역가를 완전히 상실하여 단백질성 물질임을 추정하였다. B. polyfermenticusCJ6는 E. coliO157:H7, S. aureus, S.Typhi, L. monocytogenes의 식중독 세균과의 각각 공동 배양에서 배양 3∼6시간 이내에 식중독 세균의 생육을 완전히 저해하는 것으로 나타났다. B. polyfermenticusCJ6가 생산하는 항세균 활성 물질은 SPE, recycling preparative HPLC, analytical HPLC을 이용한 4단계의 정제과정을 통하여 최종 항세균 활성을 나타내는 3개의 분획을 분리하였다. 정제된 물질의 분자량을 결정하기 위하여 UPLC를 이용한 ESI-MS/MS를 시행한 결과 3개의 분획에서 분자량이 다른 5개의 peptide(757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, 1617.8170 Da)를 확인하였으며 이는 기존에 보고된 bacteriocin 분자와는 다른 분자량을 지니는 것으로 나타났다. B. polyfermenticusCJ6와 식중독 세균과의 공동배양 실험에서 B. polyfermenticusCJ6는 식중독 세균에 대하여 매우 강력한 항세균 활성을 나타내며 그 원인물질이 되는 새로운 항 미생물 peptides 물질은 천연식품 보존제 및 사료 보존제뿐만 아니라 정장용 제재, 식중독 예방 및 치료용 생균제재 등으로도 활용이 기대되며, 이를 위하여 분리된 5개의 peptide의 정확한 구조 및 특성 규명 등의 연구가 앞으로 필요할 것이다. In this study, Bacillus polyfermenticus CJ6 harboring antibacterial activity was isolated from meju. The antibacterial activity of Bacillus polyfermenticus CJ6 was stable in the pH range of 3.0∼9.0, but it disappeared after culture at 70℃ for 24 hr. Antibacterial activity was inactivated by proteinase K, protease, and α-chymotrypsin, indicating its proteinaceous nature. The growth inhibitory effects of B. polyfermenticus CJ6 culture on food-borne pathogens such as Staphylococcus aureus, Salmonella Typhi, Listeria monocytogenes, and Escherichia coli O157:H7 were examined in this study. Approximately 6∼6.2 log CFU/㎖ of each pathogen was co-cultured with B. polyfermenticus CJ6 in a 50 ㎖ culture volume for 24 hr. Growth of S. aureus and L. monocytogenes was completely inhibited after 3 hr of incubation. Growth of S. Typhi and E. coli O157:H7 was also completely inhibited after 6 hr of incubation. The antibacterial compounds from B. polyfermenticus CJ6 were purified by solid phase extraction (C18 Sep-pak cartridge), recycling preparative HPLC, and analytical HPLC. Ultra-high performance liquid chromatography and electrospray ionization tandem mass spectrometry analysis were used to identify the purified antibacterial compounds, which were confirmed to be five peptides (757.4153 Da, 750.3444 Da, 1024.5282 Da, 1123.6083 Da, and 1617.8170 Da).

유전자조작, 균주분리 메주로부터 분리한 항진균 및 항세균 활성의 Bacillus polyfermenticus CJ9

정지혜 ( Ji Hye Jung ),장해춘 ( Hae Choon Chang ) 한국미생물생명공학회 2009 한국미생물·생명공학회지 Vol.37 No.4

농구 지도자의 지도행동 유형 분석

나정선(Jung Sun Na),정지혜(Ji Hae Jung),유정애(Jung Ae Yoo),이기봉(Gee Bong Lee) 한국여성체육학회 2002 한국여성체육학회지 Vol.16 No.2

The purpose of this study was to examine leader behavior types of basketball team coach`s for high school and university. A questionnaire measuring Leadership Sports(LSS) was used in this study. Three hundred and seventy nine athletes of high school and university were selected as subjects in the area of Seoul, Incheon and Kyongee province. In this study I used one~way ANOVA, independent t-test for statistical analysis. The findings from the analysis results were followings. 1) Korean basketball team coaches are prefer to use Training and Instruction behavior. 2) Athletes of university basketball team prefer to coach`s Autocratic behavior compare to athletes of high school basketball team coaches prefer to Training and Instruction behavior, and coaches of university basketball team coaches prefer to Autocratic behavior. 3) Male basketball athletes prefer to coach`s Social Support behavior, but female basketball athletes prefer to Positive Feedback behavior. Otherwise basketball athletes mainly used Training and Instruction behavior. 4) In this case, the athletes with more career prefer to Autocratic behavior from their coaches. And actually the coaches who train athletes with a lot of career mainly use Autocratic behavior.

토종효모를 이용한 블루베리 발효주의 발효 특성 및 항산화 활성

윤해훈(Hae Hoon Yoon),채규서(Kyu Seo Chae),손락호(Rak Ho Son),정지혜(Ji Hye Jung) 한국식품영양과학회 2015 한국식품영양과학회지 Vol.44 No.6

본 연구에서는 국내 복분자로부터 분리된 토종효모 Saccharomyces cerevisiae M-5와 수입 시판건조효모 Fermivin을 이용한 블루베리의 발효 품질 특성을 확인하였고, 항산화 활성을 확인하기 위해 총 폴리페놀 및 총 플라보노이드, DPPH 라디칼 소거 활성, ABTS 라디칼 소거 활성, 환원력을 측정하였다. S. cerevisiae M-5와 Fermivin을 블루베리에 1×10<SUP>9</SUP> CFU/kg이 되도록 접종하여 25°C에서 39일간 발효를 진행하였다. S. cerevisiae M-5를 이용한 블루베리 발효주(BBM)의 최종 품질은 알코올 13.10%, 당도 8.42°Bx, 산도 1.183%였으며, Fermivin을 이용한 블루베리 발효주(BBF)는 알코올 14.20%, 당도 8.2°Bx, 산도 1.153%로 발효가 완료되었다. 발효 특성은 BBM이 BBF와 비교 시 초기발효 속도가 빠르며 최종 알코올 생성량은 약 1% 낮은 것으로 확인되었다. 총 폴리페놀 함량은 BBJ가 0.74±0.07 TAE mg/mL, BBM이 1.34±0.20 TAE mg/mL, BBF가 1.09±0.01 TAE mg/mL로 발효 시 증가되었으며, BBM의 총 폴리페놀 함량이 BBF보다 약 1.2배 높게 확인되었다. 총 플라보노이드 함량은 BBM이 0.61±0.20 QUE mg/mL, BBF가 0.55±0.08 QUE mg/mL로 발효하였을 때 증가하며 BBM의 총 플라보노이드 함량이 높음을 확인하였다. 항산화 활성확인을 위한 DPPH 라디칼 소거 활성, ABTS 라디칼 소거활성 그리고 환원력 또한 총 폴리페놀 및 총 플라보노이드 함량이 높았던 BBM이 더 높은 항산화 활성을 나타내었다. 효모를 달리하여 제조된 블루베리 발효주의 관능검사를 실시한 결과 색상의 경우 BBF가 높았으나 향미, 맛, 전체적인 기호도는 BBM이 우수한 기호도를 보였다. 결과적으로 토종효모 S. cerevisiae M-5를 이용한 발효주는 수입 시판건조효모 Fermivin을 이용한 발효주보다 항산화 활성 및 관능적 기호도에서 우수하게 평가되어 토종효모 S. cerevisiae M-5는 와인 제조 등과 같은 산업적 활용이 가능하고 이용가치가 높을 것으로 판단된다. We investigated blueberry wine made using traditional yeast (Saccharomyces cerevisiae M-5) and Saccharomyces cerevisiae Fermivin which is widely used in wine manufacturing, and measured its fermentative characteristics and antioxidant activity. S. cerevisiae M-5 is a traditional yeast isolated from domestically grown Black raspberry (Rubus occidentalis). Both S. cerevisiae M-5 and Fermivin were inoculated into blueberry juice (BBJ) up to 1×10<SUP>9</SUP> CFU/kg, followed by incubation at 25°C for 39 days. Final fermentation products of blueberry fermented with S. cerevisiae M-5 (BBM) presented 13.10% alcohol, 8.42°Bx of sugar, and 1.183% acidity, and final fermentation products of blueberry fermented with Fermivin (BBF) presented 14.20% alcohol, 8.2°Bx of sugar, and 1.153% acidity. The contents of total polyphenol and flavonoid compounds of BBM were higher than those of BBF and BBJ. DPPH and ABTS radical scavenging activities and reducing power of BBM were higher than those of BBF. The sensory evaluation revealed that BBM showed excellent flavor, taste, and overall acceptability compared with BBF. Based on these results, the possibility of industrial utilization of S. cerevisiae M-5 as traditional yeast was confirmed by sensory evaluation and antioxidant activity. Fermentation rate of S. cerevisiae M-5 was similar to Fermivin, which is used in the food industry.

Lactobacillus plantarum GBL17 균주를 이용한 복분자 유산발효 특성

류은혜,윤해훈,정지혜,Ryu, Eun-Hye,Yoon, Hae-Hoon,Jung, Ji-Hye 한국식품조리과학회 2015 한국식품조리과학회지 Vol.31 No.6

Lactic acid fermentation of black raspberry (Rubus occidentalis) juice was carried out by using the Lactobacillus plantarum GBL17 strain. The sterilized black raspberry juice was fermented using the L. plantarum GBL17 strain at $30^{\circ}C$ for 72 hours after which the total acidity increased and the pH value decreased. In addition, the highest total acidity content (2.38%) was reached, the lowest pH value (3.22) was observed, and the sugar content decreased by $9.8^{\circ}Brix$ after the 72 hour fermentation. The number of viable cells rapidly increased up until 24 hours, after which it gradually decreased. HPLC analysis of the organic acids showed 14.51 mg/g of lactic acid content in the fermented black raspberry juice, which was not detected in the non-fermented black raspberry juice (control). The content of fructose and glucose slightly decreased after fermentation. The total polyphenol and flavonoid contents of the fermented black raspberry juice increased significantly after fermentation. The DPPH radical scavenging activity of fermented black raspberry juice (70.92%) was higher than that of the control (62.96%). After lactic acid fermentation, there was no significant increase in ABTS radical scavenging activity. These results confirm that lactic acid bacteria, such as L. plantarum GBL17, showed generally higher activities with a potential as a functional beverage.

토종효모를 이용한 복분자 발효주의 바이오제닉아민 함량 및 향기성분 분석

윤해훈,손락호,류은혜,정지혜,Yoon, Hae-Hoon,Son, Rak-Ho,Ryu, Eun-Hye,Jung, Ji-Hye 한국식품조리과학회 2015 한국식품조리과학회지 Vol.31 No.3

We investigate black raspberry (Rubus occidentalis) wine made using traditional yeast (Saccharomyces cerevisiae A8, B6, GBY2, GBY3) and S. cerevisiae Fermivin (FM), which is widely used in wine manufacturing, and analyze the biogenic amine content and the volatile flavor compounds. Black raspberries were separately inoculated with yeast up to $1{\times}10^9CFU/kg$, followed by incubation at $25^{\circ}C$ for 7 days. FM produced the highest alcohol content, however the final fermentation characteristics of the wine made using four different yeasts were similar. S. cerevisiae A8 had a large biogenic amine (BA) content, specifically tryptamine, thus we excluded this yeast from fermentation. S. cerevisiae GBY3 was selected for black raspberry wine fermentation as a result of sensory evaluation. The volatile flavor compounds of two wines (S. cerevisiae GBY3 and FM) were analyzed by gas chromatography and mass spectrometry. 37 compounds in the samples were separated, and several ester compounds were identified in greater amounts in the wine made with S. cerevisiae GBY3 than in the wine made with FM. A greater amount of the major compound, ethyl benzoate, giving the sweet and fruity flavor, was identified in wine made with S. cerevisiae GBY3 than in the wine made with FM. In conclusion, S. cerevisiae GBY3 was confirmed to produce no major BAs and a better flavored wine. These results give new leads in the production of high quality wine.

토종발효미생물을 이용한 오디 발효주의 항산화 활성 및 향기성분 분석

채규서(Kyu Seo Chae),정지혜(Ji Hye Jung),윤해훈(Hae Hoon Yoon),손락호(Rak Ho Son) 한국식품영양과학회 2014 한국식품영양과학회지 Vol.43 No.7

This study was carried out to develop mulberry wines fermented with traditional microorganisms (Saccharomyces cerevisiae B-8). S. cerevisiae B-8 is a traditional fermentation microorganism isolated from domestically grown Rubus occidentalis. Each S. cerevisiae B-8 and Fermivin was inoculated into mulberry up to 1×10<SUP>9</SUP> CFU/kg, followed by incubation at 25°C for 10 days. Mulberry fermented with S. cerevisiae B-8 (MBB) had a high alcohol content (16.47%), and the fermentation rate of MBB was faster than that of mulberry fermented with Fermivin (MBF). The total polyphenol and flavonoid contents of MBB were higher than those of MBF. DPPH radical scavenging activity of MBB was as high as that of MBF. ABTS radical scavenging activity of MBF was higher than those of MBB and mulberry juice (MBJ). In addition, reducing power of MBB was much higher than other samples. Flavor constituents of the two fermented wines were analyzed by gas chromatography and mass spectrometry. Twenty-three compounds from the sample were separated and identified as fifteen esters, six alcohols, an aldehyde, and an acetate. Particularly, tetradecanoic acid, ethyl ester of orris and violet flavor were ten times more abundant in MBB than in MBF. Several ester components were two times more abundant in MBB than in MBF. In conclusion, current findings indicate that MBB might have better antioxidant activities with flavor, which contributes to improved wine production with high quality and function.