고객지원 분야 최우수상 S-OiL 보너스 카드

장효일,민웅기 (사)한국인터넷전문가협회 2007 Web Korea Annual Vol.2007 No.-

Streptococcus salivarius subsp. thermophilus가 생산하는 Bacteriocin의 특성

이장혁,장효일 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.1

Antagonism assay를 하여 원유에서 억제환의 크기가 가장 크며 단백질 가수분해 효소에 의해 항균 능력이 상실되는 균을 분리하여 Streptococcus salivarius subsp.thermophilus로 동정하였다. 이 균주는 45℃에서 specific growth rate가 가장 높은 반면 bacteriocin을 생산하지 않았다. 이 bacteriocin은 고온에서도 상당한 열안정성을 보였으며 단백질 가수분해 효소에 의해 활성이 상실되었으나 α-amylase, DNaseI, RNase는 bacteriocin활성에 영향을 주지 못하였다. One bacterial strain, that had made the largest inhibition zone at the antagonism assay and also that lost the inhibition activity by the protease treatment, was isolated from raw milk. That strain was identified as Streptococcus salivarius subsp. thermophilus. The specific growth rate of this strain was maximum at 45℃. However, at this temperature the strain produced no bacteriocin. The bacteriocin activity was quite stable even at high temperature. Moreover, the activity of the bacteriocin was sensitive to proteases, but not to α-amylase, DNase I, or RNase.

농축산 식품으로 부터 Listeria monocytogens, Enterobacter sakazakii, Yersinia enterocolitica 를 검출하기 위한 DNA chip 개발

구만복,장효일,류지훈,방지현 한국육종학회 2009 한국육종학회 학술발표회 발표요지 Vol.41 No.1

핵산분해효소에 관한 연구 (제1보) Streptomyces 속 균주가 생산하는 Phosphomono , diesterase

이정치,장효일,김혁일,양한철 ( J . C . Lee,H . I . Chang,H . I . Kim,H . C . Yang ) 한국미생물생명공학회 1978 한국미생물·생명공학회지 Vol.6 No.3

Lactobacillus casei YIT 9018로부터 분리한 Prophage cured Strain의 특성

이정준,오태광,장효일,백영진 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.5

Prophage cured strain인 Lactobacillus casei HY 2782의 특성을 조사하여, L. casei HY 2782가 모균주인 L. casei YIT 9018의 특성을 유지하고 있는 안정된 균주임을 확인하였다. Temperate phage DNA를 probe로 한 southern hybridization을 통해 L. casei YIT 9018의 chromosomal DNA에 temperate phage가 prophage 상태로 incorporation되어 있음을 확인하였으며, L. casei HY 2782의 chromosomal DNA로부터 prophage genome이 완전히 curing 되었음을 알 수 있었다. L. casei HY 2782가 균증식 능력, 산생성 능력, 당이용성 그리고 효소활성에 있어서 L. casei YIT 9018과 차이가 없는 안정된 균주임을 알 수 있었다. Prophage cured strain인 L. casei HY 2782가 temperate phage의 숙주로 이용될 때, 숙주에 대한 temperate phage의 M.0.I. 값은 1.0∼5.0로 나타났다. L. casei HY 2782로부터 분리한 pLC90 plasmid의 제한효소절편 분석을 통해 pLC90 plasmid의 크기가 68.22 kb 임을 알 수 있었으며 L. casei HY 2782의 plasmid profiles, genomic DNA patterns 그리고 cellular fatty acids 구성성분이 모균주인 L. casei YIT 9018과 동일함을 확인하였다. 그리고 두 균주의 주요 fatty acids 구성성분은 C_(l4:O), C_(16:1), C_(16:0), C_(18:1), C_(19:cyclo)이었다. Arbitrarily primed polymerase chain reaction을 실행하여 두 균주간 차이를 확인하였다. b_5-l/l7-2 primer를 사용한 경우는 L. casei YIT 9018에만 나타나는 약 1.3 kb의 DNA band를 볼 수 있었으며, b_5-2/l7-2 primer를 사용하여 PCR을 실행하였을 때는 L. casei HY 2782에만 나타나는 약 1.0 kb의 DNA band를 볼 수 있었다. Lactobacillus casei HY 2782, prophage cured strain was characterized to be stable as much as L. casei YIT 9018, parent strain. By southern hybridization, it was confirmed that the temperate phage was incorporated in chromosomal DNA of L. casei YIT 9018 as a prophase. It was also proved that the prophage was cured from chromosomal DNA of L. casei HY 2782. The growth rate, lactic acid producing ability, carbohydrates fermentation, and ensymatic activity of L. casei HY 2782 were found to be similar to those of L. casei YIT 9018. When L. casei HY 2782 was used as a host, the multiplicity of infection (M.O.I.) of the temperate phage for L. casei HY 2782 was 1.0∼5.0. Restriction enzyme analysis of pLC90 plasmid from L. casei HY 2782 was shown that the size was an approximately 68.22 kb. The plasmid profiles, genomic DNA patterns, and cellular fatty acids composition of L. casei HY 2782 were similar to those of L. casei YIT 9018. And the major fatty acids composition of these strains were C_(l4:0), C_(l6:l), C_(16:0), C_(18:1) and C_(19:cyclo-) 10 sets of arbitrary primer in the PCR were screened to find differentiation against two strains of L. casei. Among them, b_5-l/l7-1 primer was produced an approximately 1.3 kb DNA band of only L. casei YIT 9018. And b_5-2/l7-2 primer was produced an approximately 1.0 kb DNA band of only L. casei HY 2782.

Lcatobacillus casei YIT 9018로 부터 분리한 Prophage Cured Strain의 특성

이정준,오태광,장효일,백영진,Lee, Jeong-Jun,Oh, Tae-Kwang,Chang, Hyo-Ihl,Baek, Young-Jin 한국미생물 · 생명공학회 1994 한국미생물·생명공학회지 Vol.22 No.5

Lactobacillus casei HY 2782, prophage cured strain was characterized to be stable as much as L casei YIT 9018, parent strain. By southern hybridization, it was confirmed that the temperate phage was incorporated in chromosomal DNA of L. casei YIT 9018 as a prophage. It was also proved that the prophage was cured from chromosomal DNA of L casei HY 2782. The growth rate, lactic acid producing ability, carbohydrates fermentation, and enzymatic activity of L. casei HY 2782 were found to be similar to those of L. casei YIT 9018. When L casei HY 2782 was used as a host, the multiplicity of infection (M.O.I.) of the temperate phage for L. casei HY 2782 was 1.0~5.0. Restriction enzyme analysis of pLC90 plasmid from L. casei HY 2782 was shown that the size was an approximately 68.22 kb. The plasmid profiles, genomic DNA patterns, and cellular fatty acids composition of L. casei HY 2782 were similar to those of L casei YIT 9018. And the major fatty acids composition of these strains were C$_{14;0}$,C$_{16;1}$, C$_{16;0}$, C$_{18;1}$ and C$_{19;cyclo-}$ 10 sets of arbitrary primer in the PCR were screened to find differentiation against two strains of L. casei. Among them, b$_{5}-1/17-1 primer was produced an approximately 1.3 kb DNA band of only L casei YIT 9018. And b$_{5}-2/17-2 primer was produced an approximately 1.0 kb DNA band of only L casei HY 2782.

Screening of Immunostimulatory Probiotic Lactic Acid Bacteria from Chicken Feces as Animal Probiotics

이은경,이나경,이시경,장효일,백현동 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.4

The principal objective of this study was to screen and select acid-tolerant Lactobacillus strains from chicken feces, feeds,and other sources. Fourty six strains evidencing acid tolerance (pH 3.5) were isolated in this study. Among them, nine strains exhibited marked immunostimulatory effects. Therefore, nine candidate strains were characterized for probiotic use. In order to evaluate macrophage activation, NO production was measured using RAW 264.7 cells. In particular, three strains (FC812, FC222, and FC113) evidenced the highest levels of NO production measured at 38.39±20.01, 35.06±27.73, and 33.88±15.99 μM, respectively, at a concentration of 108 CFU/mL. The majority of strains, with the exception of strain FC322, evidenced marked resistance to artificial gastric juice (pH 2.5 with 1%(w/v) pepsin). Additionally, strains FC222,FC421, FC511, and FC721 were highly resistant to artificial bile acid (0.1%(w/v) oxgall), whereas strains FC113, FC322,FC422, FC621, and FC812 were the least resistant to bile. All nine strains exerted antimicrobial effects against chickenrelated pathogens. Additionally, all nine strains were found to be resistant to several antibiotics. The isolated strains, except for strain FC322, were tentatively identified as Lactobacillus salivarius, using an API 50 CHL kit. These results demonstrate that some probiotic organisms may potentially probiotic properties, and thus may serve as an effective alternative to antibiotics in animal applications.

Enterococcus faecolis KBL703 Plasmid p703/5의 Replication Origin의 Cloning

전영욱,정세영,김영우,장효일 한국산업미생물학회 1994 한국미생물·생명공학회지 Vol.22 No.1

Enterococcus faecalis KBL 703은 서로 다른 크기를 갖는 3개의 plasmid들을 가지고 있다(p703/9, p703/5, p703/4). 이 plasmid 들중 하나인 p703/5로부터 plasmid 복제에 필요한 부위(regplicaiton origin)를 분리하였다. 이를 위한 En. faecalis 형질전환실험의 숙주로 사용하기 위하여 두개의 plasmid-cured 변이주를 얻었으며, 각 p703/5의 DNA 절편과 CAT(chloramphenicol acetyl transferase) 유전자를 가진 4개의 DNA 구조물들로 이들을 형질전환시킨 결과 제한효소 SalI으로 절단된 p703/5의 3.6 kb DNA 절편을 가진 DNA 구조물만이 이 균주내에서 plasmid로서 안정하게 유지됨을 알 수 있었다. En. faecalis ATCC 29212를 형질전환하였을 때에도 이 새로운 재조합 plasmid가 세포내에서 안정하게 유지됨을 알 수 있었다. Enterococcus faecalis KBL703 has three plasmids(p703/9, p703/5 and p703/4). Within p703/5, the specific DNA region that would confer replication function(replication origin) was searched by transformation experiments. In order to use as the recipient of transformation, two plasmid-cured strains were made from this strain. Four recombinant DNA constructs, each containing fragment of p703/5 and CAT(chloramphenicol acetyl transferase) gene were also made. And they were used to transform the plasmid-cured strains. Only one DNA construct containing 3.6 kb Sall fragment was stably maintained as plasmid in these strains. Additional experiment using another Enterococcus faecalis strain(ATCC 29212) as a recipient was successfully done and it was confirmed that this newly constructed recombinant plasmid contained the replication origin from p703/5 plasmid.

Medium Optimization for the Production of Probiotic Lactobacillus acidophilus A12 Using Response Surface Methodology

이나경,박여랑,최가진,장효일,백현동 한국축산식품학회 2010 한국축산식품학회지 Vol.30 No.3

Lactobacillus acidophilus A12 was isolated from chicken feces for use as an immunostimulating livestock probiotic. The purpose of this study was to optimize the production of L. acidophilus A12 using response surface methodology (RSM). Initially, the influence of growth medium was studied in terms of carbon sources (glucose, fructose, lactose, glycerol,sucrose, ethanol, and mannitol), nitrogen sources (beef extract, yeast extract, malt extract, and tryptone), and inorganic salts (CaCl2, MgSO4, KH2PO4, (NH4)2SO4, FeSO4, and NaCl). Through one factor-at-a time experiment, lactose, yeast extract,and CaCl2 were determined to be the best sources of carbon, nitrogen, and inorganic salt, respectively. The optimum composition was found to be 17.7 g/L lactose, 18.6 g/L yeast extract, and 0.9 g/L CaCl2. Under these conditions, a maximum cell density of 9.33 Log CFU/mL was produced, similar to the predicted value.

Shikonin Induces Apoptotic Cell Death via Regulation of p53 and Nrf2 in AGS Human Stomach Carcinoma Cells

고현석,김선중,심소희,장효일,하창훈 한국응용약물학회 2016 Biomolecules & Therapeutics(구 응용약물학회지) Vol.24 No.5

Shikonin, which derives from Lithospermum erythrorhizon, has been traditionally used against a variety of diseases, including cancer, in Eastern Asia. Here we determined that shikonin inhibits proliferation of gastric cancer cells by inducing apoptosis. Shikonin’s biological activity was validated by observing cell viability, caspase 3 activity, reactive oxygen species (ROS) generation, and apoptotic marker expressions in AGS stomach cancer cells. The concentration range of shikonin was 35-250 nM with the incubation time of 6 h. Protein levels of Nrf2 and p53 were evaluated by western blotting and confirmed by real-time PCR. Our results revealed that shikonin induced the generation of ROS as well as caspase 3-dependent apoptosis. c-Jun-N-terminal kinases (JNK) activity was significantly elevated in shikonin-treated cells, thereby linking JNK to apoptosis. Furthermore, our results revealed that shikonin induced p53 expression but repressed Nrf2 expression. Moreover, our results suggested that there may be a co-regulation between p53 and Nrf2, in which transfection with siNrf2 induced the p53 expression. We demonstrated for the first time that shikonin activated cell apoptosis in AGS cells via caspase 3- and JNK-dependent pathways, as well as through the p53-Nrf2 mediated signal pathway. Our study validates in partly the contribution of shikonin as a new therapeutic approaches/ agent for cancer chemotherapy.