• 한국수산과학회지
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• 제19권1호
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• pp.20-26
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• 1986

Very little information is available in the literature on storage of fish sauce. Therefore, microbiological and chemical chracteristics during storage and quality of fish sauce were investigated and discussed to present data about the optimum storage condition. The chopped sardine meat was mixed with equal amount of water and $9\%$(w/w) of $75\%$ vital wheat gluten and then hydrolyzed by addition of commercial proteolytic enzymes such as bromelain, papaya protease, ficin and a enzyme mixture (Pacific Chem. Co.) for 4 hours at $52.5^{\circ}C$. The reaction mixture was heated for 30 min at $100^{\circ}C$ for enzyme inactivation, pasteurization and color development and then centrifuged for 20 min at 4,000 rpm. Table salt and benzoic acid were added for bacteriostatic effect and stored for 80 days at $15{\pm}1^{\circ}C$ and $30{\pm}1^{\circ}C$. The results were summarized as follows: 1. The amount of amino-nitrogen and pH of fish sauce were almost unchanged during storage. 2. Mininum concentration of salt for bacteriostatic activity was $9\%$(w/w) regardless of addition of benzoic acid. 3. the yields of amino-nitrogen were $63.1\%$ for the hydrolysate prepared without enzyme, $79.7\%$ for that with bromelain, $69.9\%$ with ficin, $74.3\%$ with papaya pretense, and $78.1\%$ with enzyme mixture, respectively. 4. The contents of amino-nitrogen were $4510.0mg\%$ on the dry basis for the product prepared by autolysis, $5483.2mg\%$ for that prepared with bromelain, $5305.7mg\%$ with ficin, $4994.1mg\%$ with papaya protease and $5582.3mg\%$ with the enzyme mixture, respectively. 5. The contents of crude protein were $51.35\%$ on the dry basis for the product prepared by autolysis and 55 to $59\%$ for prepared with commercial enzymes. 6. The hydrolysate prepared with the enzyme mixture revealed a little stronger meaty taste than any other products. 7. The level of crude protein in residues was still high ($69.5{\sim}77.2\%$ on the dry basis) and might be originated from the added vital wheat gluten.

• 한국미생물·생명공학회지
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• 제14권3호
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• pp.225-232
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• 1986

보리고추장 양조시 고추씨 및 마늘을 첨가하여 제조한 고추장의 숙성과정중의 효소활성도와 이화학적 변화 및 관능검사를 조사하였다. 고추장숙성 중 효소의 활성은 전분액화·당화효소, 중성 protease, lipase모두 증가되었고 전분액화효소를 제외하고는 숙성후기에도 효소의 잔존활성이 높았으며, 고추씨의 첨가도 효소 활성유지에 효과적이었다. pH와 적정산도는 고추씨의 첨가로 변화가 적었고, 마늘의 첨가도 숙성후기에는 큰 차이가 없었다. 총질소, 아미노태질소, 총당, 환원당은 고추씨나 마늘의 첨가로 증가되었으며, 암모니아태질소는 이들의 첨가로 숙성중기에 증가하나 숙성후기에는 큰 차이가 없었다. 알코올의 생성은 고추씨나 마늘의 첨가로 적었으며, 조지방은 고추씨의 첨가로 증가하였다. 10주 숙성 고추장의 맛, 향기 및 색은 전반적으로 마늘첨가구가 무첨가구에 비하여 우수하였다. 따라서 통고추를 이용한 보리고추장은 원료에 2％의 마늘을 첨가하므로써 품질을 개선할 수 있다.

• 한국균학회지
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• 제16권3호
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• pp.175-184
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• 1988

본(本) 연구(硏究)는 균사침투가 깊은 koji의 제조조건(製造條件)과 청주료중의 제효소(諸酵素)의 동태(動態)를 조사(調査)한 것으로 다음과 같은 결과(結果)를 얻었다. 1. 원료현미(原料玄米)를 30% 도감(搗減)한 결과(結果), 수분(水分)은 9%, 조단백(粗蛋白)은 26%, 조지방(粗脂肪)은 96% 감소(減少)되었으며 전분가(澱粉價)는 9% 상승(上昇)되었다. 2. 균사침투가 깊은 koji 제조(製造)의 최적조건(最適條件)은 제국시간(製麴時間) 4시간(時間), ${\alpha}$미수분(米水分) 40%, 제국(製麴) 전후반(前後半) 상대습도(相對濕度)가 각각(各各) 98% 및 80%, 접종국균(接種麴菌)의 포자수(胞子數)는 $1.0{\times}10^4\;spores/g{\cdot}{\alpha}$미(米)였다. 3. 청주발효중(淸酒醱酵中) ${\alpha}-Amylase$의 $23{\sim}27%$가 실활(失活)되었으며 $35{\sim}70%$가 흡착(吸着)되었다. 4. 청주발효중(淸酒醱酵中) Glucoamylase의 14%가 실활(失活)되었으며 $74{\sim}92%$가 흡착(吸着)되었다. 5. 청주발효중(淸酒醱酵中) Acid pretense (pH 3.0)의 $13{\sim}14%$가 실활(失活)되 었으며 $70{\sim}73%$가 흡착(吸着)되 었다. 6. 균사침투가 깊은 koji와 얕은 koji 효소(酵素)의 청주발효중(淸酒醱酵中) 뚜렷한 효소적(酵素的) 차이점은 발견되지 않았다.

• 한국식품영양과학회지
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• 제38권1호
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• pp.76-82
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• 2009

본 연구에서는 발효과정 중 누룩의 일반성분 및 효소 활성변화를 측정하고, 단백질의 분해정도와 밀 단백질에 민감한 환자의 혈청과 반응성을 관찰하였다. 누룩의 수분함량은 발효 1일 24.20%였으며 발효가 진행됨에 따라 미생물의 이용과 증발로 인해 감소되어 발효 15일에는 13.60%를 나타내었다. 누룩 발효 중 총당과 환원당의 함량은 발효시간이 경과함에 따라 증가하는 경향을 나타내었다. 누룩의 발효 초기 조단백질 함량은 1.58%였으나 발효가 진행됨에 따라 다소증가하여 발효 15일에는 2.05%로 나타났다. 또한 누룩의 조지방 함량은 발효 초기 1.32%였으며 발효 15일에는 2.24%로 다소 증가함을 나타내었다. ${\alpha}$-Amylase 활성은 발효 초기 1,416.67 U/g에서 발효 15일에는 2,833.00 U/g으로 급격히 증가하였으며, glucoamylase 활성은 발효 초기 497.90 U/g에서 발효 10일 경과 후 705.40 U/g으로 최대 활성을 보였으나 그 후 완만한 감소를 나타내었다. 누룩의 산성 protease 활성은 발효 초기에는 검출되지 않았으나 발효 10일에 75.00 U/g으로 급격히 증가하기 시작하여 발효 15일에는 327.00 U/g에 도달하였으며 중성 protease 활성도 발효 10일에 90.00 U/g으로 급격히 증가하기 시작하여 15일에는 354.00 U/g로 검출되었으며 이후에도 계속 증가하였다. 반면에 알칼리성 protease 활성은 검출되지 않았다. 발효가 진행됨에 따라 누룩 중 고분자 단백질이 각종 미생물에 의해 분해되어 저분자의 펩타이드 밴드가 생성되었으며 밀 민감성 환자의 혈청과 반응하는 밴드도 점차 감소하여 발효 15일에는 거의 검출되지 않았으므로 누룩 중 밀 단백질의 항원성이 감소하였음을 확인할 수 있었다. 따라서 본 연구를 통해 누룩은 영양학적 측면뿐만 아니라 효소학적 측면의 장점까지 겸비한 효율적인 한국 고유의 발효 식품용 스타터(starter)로써 전통주 제조에 필수적이며, 누룩의 고분자 단백질이 발효에 의해 저분자 펩타이드로 분해됨을 확인함으로써 밀 민감성 환자들을 위한 다양한 가공식품용 소재로 활용이 기대된다.

• 한국식품과학회지
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• 제34권5호
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• pp.867-872
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• 2002

정미성이 높은 효모 추출물을 얻고자 각종 효소의 사용에 대한 최적 조합 및 공정법을 알아보기 위하여 맥주 폐효모박을 각종 효소로 처리하여 효모추출물 중의 정미성분(IMP, GMP 및 유리아미노산)을 측정하여 비교, 분석하였다. Glucanase(0.5%) 처리에 의한 효모추출물중의 조단백질 함량은 33.6% 이었다. Tunicase(1%) 28.0% 와 무처리구 21.1%에 비해 최고 1.6배의 증가를 보였다. 단백질 분해효소처리에 의한 조단백질의 함량은 bromelin(1%), protamex(1%) 처리에서 각각 30.8%, 29.8%로 무처리구에 비해 최고 1.4배의 증가를 보였다. 효소 복합처리에 의한 상승효과는 glucanase(0.5%)+protamex(1%) 처리구에서 조단백질의 함량이 34.4%로 나타나 glucanase 단독처리구의 33.6%보다 높은 함량을 나타냈다. IMP+GMP 총함량은 glucanase + phophodiesterase + adenyldeaminase (G+P+A) 혼합 처리구에서 1,066 mg/100 g, glucanase + ptotamex + phophodiesterase + adenyldeaminase (G+Pro+P+A) 혼합 처리구에서는 1,047 mg/100 g으로 비슷하였다. 유리아미노산의 함량은 protamex가 첨가된 G+Pro+P+A 혼합처리구에서 2,302 mg/100 g으로 가장 높게 나타났다. 따라서 IMP, GMP 및 유리아미노산의 함량을 모두 고려해 볼 때 세포벽 분해효소 (gulcanase 0.5%, 12시간), 단백질 분해효소 (protamex 1%, 3시간), 핵산 분해효소 (phosphodiesterase 0.1%, 3시간) 및 핵산 전이효소 (adenyldeaminase 1%, 1.5시간)를 순차적으로 적용시켜 가수분해시키는 것이 효모 추출물의 정미성을 높이는 최적의 효소 복합 사용공정으로 판단되었다.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1281-1287
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• 2019

Nattokinase (NK, E.C. 3.4.21.62) is a serine protease produced by Bacillus subtilis natto that shows promise for the treatment of thrombotic disease. In this study, we assessed the effects of NK on the development of hepatocellular carcinoma (HCC), a principal malignancy of the liver that causes morbidity and mortality worldwide. Crude extracts of NK (NCE) were isolated from fermentation medium by centrifugation and separated into three fractions (<10 K, 100~30 K and >30K). Orthotopic HCC mouse models were established and NCE was administered by oral gavage. H&E staining was performed to examine the pathology of HCC livers. Immunohistochemistry and immunofluorescence were used to evaluate FOXM1, CD31, CD44 and vimentin expression in the liver. Compared to PBS groups, NCE increased the survival rates of HCC-bearing mice to 31% and decreased ascites. Low-intensity ultrasound imaging showed that the hypoechoic mass area was lower in NCE-treated mice and that tumor growth significantly decreased. IHC staining showed that the expression of FOXM1 was inhibited by NCE treatment. Immunofluorescence results revealed lower levels of CD31, CD44 and vimentin in the NCE groups. Taken together, these data demonstrate that NCE from Bacillus subtilis natto improves survival and inhibits tumor growth in HCC mice.

• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.364-370
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• 2008

Chemical analysis and in vitro studies were conducted to investigate the nutritive value for ruminants of cell mass from lysine production (CMLP) which is a by-product of the lysine manufacturing process. Proximate analysis, protein fractionation, and in vitro protein degradation using protease from Streptomyces griseus and strained ruminal fluid were carried out to estimate ruminal protein degradability of CMLP with two reference feedstuffs-soybean meal (SBM) and fish meal (FM). Amino acid composition and pepsin-HCl degradability were also determined to evaluate postruminal availability. CMLP contained 67.8% crude protein with a major portion being soluble form (45.4% CP) which was composed of mainly ammonium nitrogen (81.8% soluble CP). The amount of nucleic acids was low (1.15% DM). The total amount of amino acids contained in CMLP was 40.60% DM, which was lower than SBM (47.69% DM) or FM (54.08% DM). CMLP was composed of mainly fraction A and fraction B2, while the protein fraction in SBM was mostly B2 and FM contained high proportions of B2 and B3 fractions. The proportion of B3 fraction, slowly degradable protein, in CP was the highest in fish meal (23.34%), followed by CMLP (7.68%) and SBM (1.46%). CMLP was degraded up to 51.40% at 18 h of incubation with Streptomyces protease, which was low compared to FM (55.23%) and SBM (83.01%). This may be due to the insoluble portion of CMLP protein being hardly degradable by the protease. The in vitro fermentation by strained ruminal fluid showed that the amount of soluble fraction was larger in CMLP (40.6%) than in SBM (17.8%). However, because the degradation rate constant of the potentially degradable fraction of CMLP (2.0%/h) was lower than that of SBM (5.8%/h), the effective ruminal protein degradability of CMLP (46.95%) was slightly lower than SBM (53.77%). Unavailable fraction in the rumen was higher in CMLP (34.0%) compared to SBM (8.8%). In vitro CP degradability of CMLP by pepsin was 80.37%, which was lower than SBM (94.42%) and FM (89.04%). The evaluation of protein degradability using different approaches indicated that soluble protein in CMLP may supply a large amount of ammonia in the rumen while insoluble protein can be by-passed from microbial attacks due to its low degradability. The results from this study suggest that CMLP can be used as a protein supplement to ruminants for supplying both non-protein nitrogen to rumen microbes and rumen undegradable protein to the host animal.

• 생명과학회지
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• 제20권2호
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• pp.183-189
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• 2010

혈전용해효소를 생산하는 Bacillus sp. J-19가 보편적인 한국의 조미제인 멸치젓갈에서 분리되었다. 그 혈전용해 효소는 에탄올 침전, Sephadex G-50을 이용한 젤 여과법 등을 포함하는 일련의 크로마토그래피 방법으로 순수분리 되었다. 조효소 추출액과 비교해서, 그 효소의 비활성은 1021배 증가하였고, 수율은 23%이었다. 순수분리한 효소의 분자량은 SDS-PAGE 상 약 45 kDa이었다. 특히, 합성기질인 serine protease (H-D-Ile-Pro-Arg-pNA,S-2288)에 대한 아미드활성은 약 17 U/mg이었다. 또한, 그 45 kDa 혈전용해효소의 60% 이상의 활성이 30%(w/v) sodium chloride 의 존재 하에서도 유지되었다. 이러한 발견들은 특이한 혈전용해효소를 제공해서, 실용적인 혈전용해제 개발을 유도할 수 있다.

• Preventive Nutrition and Food Science
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• 제21권1호
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• pp.52-56
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• 2016

This study was performed to investigate the antifungal substances and the antifungal activity against fungi of lactic acid bacteria (LAB) isolated from kimchi. LAB from kimchi in Imsil showed antifungal activity against Penicillium brevicompactum strain FI02. LAB LI031 was identified as Lactobacillus sakei subsp. Antifungal substances contained in L. sakei subsp. ALI033 culture media were unstable at high pH levels. Both, the control and proteinase K and protease treated samples showed clear zones, suggesting that the antifungal substances produced by ALI033 were non-protein substances unaffected by protesases. Both, the control and catalase showed clear zones, suggesting that the antifungal metabolite was not $H_2O_2$. The molecular weights of the antifungal substances were ${\leq}3,000Da$. The organic acid content of crude antifungal substances produced by L. sakei subsp. ALI033 showed high concentrations of lactic acid (502.47 mg/100 g). Therefore, these results suggest that antifungal substance produced by L. sakei subsp. ALI033 is most likely due to its ability in producing organic acid.

• 한국축산식품학회지
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• 제18권1호
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• pp.19-26
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• 1998

In order to study the tenderizing effect of the proteolytic enzyme, ficin, from fig fruit (Ficus carica L), the enzyme was purified from fig latex by precipitation and chromatography. The ficin separated from Bongraesi showed single band on SDS-PAGE. However, the ficin from Masui showed tow bands. The specific activity of ficin purified from Bongraesi species was 2.8 unit/mg protein and that from Masui species was 6.5 unit / mg protein. The amounts of ficin purified from 50 mL of crude latex of Bongraesi and Masui were 1,760 mg and 657 mg, respectively. the water holding capacity of beef decreased to the large extent, when sugar Bongraesi latex and Masui latex were added. The hardness of beef showed decreasing tendency with the time, however, after 60 min, it decreased and thereafter increased a little after 120 min. the hardness of beef decreased sharply with addition of the latex of Bongraesi and Masui. The Masui has more tenderizing effect than the Bongraesi. When meat was mixed with tenderizing agent(ficin) and not heated, the change of color showed significant difference (p＜0.01). when meat was mixed with tenderizing agent(ficin) and heated, the toughness showed significant difference (p＜0.01) and the softness showed significant difference (p＜0.001).