• 한국미생물·생명공학회지
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• 제47권1호
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• pp.87-95
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• 2019

Esterase produced by Acinetobacter sp. B1 (strain B1) was optimized by means of one-variable-at-a-time and response surface methodologies. Results of the one-variable-at-a-time experiment showed that Tween 80 significantly increased esterase production of strain B1. The addition of Tween 80 to the culture medium increased the biomass and esterase activity of strain B1, stimulated content of total extracellular protein, and enhanced the oleic acid (C18:1) composition in the cell membrane of strain B1. The influence of eight culture variables on esterase production was evaluated by Plackett-Burman design. Results showed that Tween 80, pH, and $K_2HPO_4$ significantly affected the esterase production of strain B1. Tween 80, pH, and $K_2HPO_4$ were further optimized by central composite design. Under the optimized conditions (w/v, soluble starch 2.5%, tryptone 1.5%, Tween 80 0.8%, $K_2HPO_4$ 0.5%, NaCl 0.5%, pH 8.0, inoculum size 1%, and inoculum age 24 h), the maximum esterase activity of strain B1 was 152.13 U/ml, which was 10-fold higher than that of non-optimization after 36 h cultivation.

• KSBB Journal
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• 제27권6호
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• pp.352-360
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• 2012

Bacillus clausii I-52 which produced SDS- and $H_2O_2$-tolerant extracellular alkaline protease (BCAP) was isolated from heavily polluted tidal mud flat of West Sea in Incheon, Korea and stable strain (transformant C5) of B. clausii I-52 harboring another copy of BCAP gene in the chromosome was developed using the chromosome integration vector, pHPS9-fuBCAP. When investigated the production of BCAP using B. clausii transformant C5 through pilot-scale submerged fermentation (500 L) at $37^{\circ}C$ for 30 h with an aeration rate of 1 vvm and agitation rate of 250 rpm, protease yield of approximately 105,700 U/mL was achieved using an optimized medium (soybean meal 2%, wheat flour 1%, sodium citrate 0.5%, $K_2HPO_4$ 0.4%, $Na_2HPO_4$ 0.1%, NaCl 0.4%, $MgSO_4{\cdot}7H_2O$ 0.01%, $FeSO_4{\cdot}7H_2O$ 0.05%, liquid maltose 2.5%, $Na_2CO_3$ 0.6%). The enzyme stability of BCAP was increased by addition of polyols (10%, v/v) and also, the stabilities of BCAP towards not only the thermal-induced inactivation at $50^{\circ}C$ but also the SDS and $H_2O_2$-induced inactivation at $50^{\circ}C$ were enhanced. Among the polyols examined, the best result was obtained with propylene glycol (10%, v/v). The BCAP supplemented with propylene glycol exhibited extreme stability against not only the detergent components such as ${\alpha}$-orephin sulfonate (AOS) and zeolite but also the commercial detergent preparations. The granulized enzyme of BCAP was prepared with approximately 1,310,000 U/g of granule. Wash performance analysis using EMPA test fabrics revealed that BCAP granule exhibited high efficiency for removal of protein stains in the presence of anionic surfactants as well as bleaching agents. When compared to Savinase 6T$^{(R)}$ and Everlase 6T$^{(R)}$ manufactured by Novozymes, BCAP under this study probably showed similar or higher efficiency for the removal of protein stains. These results suggest that the alkaline protease produced from B. clausii transformant C5 showing high stability against detergents and high wash performance has significant potential and a promising candidate for use as a detergent additive.

• Preventive Nutrition and Food Science
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• 제9권1호
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• pp.14-20
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• 2004

Bacillus strains capable of producing fibrinolytic enzyme were isolated from traditional fermented Korean soybean paste and Japanese fermented soybean (Natto). Among the 16 strains, a selected Bacillus sp. was identified as bacillus firmus, with 80.7％ homology, by API kit analysis. Seed starter or B. firmus NA-1 was prepared with 5％ soymilk prepared from micronized soybean powder. To produce fibrinolytic enzyme by B. firmus NA-1 the liquid culture was performed with NB broth (pH 7.0) fortified with 1％ galactose, 0.1% tryptone, and 0.5% $K_2$HPO$_4$, by shaking with 180 rpm at 37$^{\circ}C$. Fibrinolytic enzyme activity reached the highest value at 7.8 unit/mL (plasmin unit) after fermentation for 72 hr. The crude fibrinolytic enzyme showed higher relative activity in the range of pH 7.0∼9.0. The activity of crude fibrinolytic enzyme was well maintained even after concentration by the vacuum evaporation at 5$0^{\circ}C$ for 1 hr.

• 한국식품영양학회지
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• 제17권2호
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• pp.163-170
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• 2004

계란찜 제조시 물의 첨가량,가열 온도와 시간, pH, NaCl의 첨가량이 전란액의 점도와 색, 계란찜의 텍스쳐와 색에 미치는 영향을 조사하였다. 계란찜은 계란을 할난시켜 완전히 섞어 계란액을 만든 다음ultrasonicator로 탈기시키고 10$0^{\circ}C$의 증기로 7분간 가열한 다음 22$^{\circ}C$에서 90분간 냉각시켰다. 그 결과 물의 첨가량은 50% 이상부터 계란액의 점도와 계란찜의 견고성 및 항복응력에 현저한 감소가 있었다. 계란액의 색은 a와 b값의 감소가 현저하였고, 계란찜은 ( - )값 범위에서의 a값이 더 낮아져 녹색이 짙어짐을 보였다. 가열 온도의 증가는 응고시간의 단축과 높은 항복응력이 측정되었으나 변형도는 감소함을 보였다. 가열시간에 따른 계란찜의 L과 b값은 9$0^{\circ}C$와 10$0^{\circ}C$에서 감소함을 보였고, a값은 8$0^{\circ}C$에서 감소함을 보였다. 소금의 첨가를 1.3%까지 하였을 때 계란찜의 항복응력과 단단함이 크게 증가하였지만, 색은 약간의 b값 감소 외에는 큰 차이가 없었다. 계란액의 pH를 4.0에서 10.0으로 증가시켰을 때 pH 6.0에서 점도가 가장 낮았으나 계란찜은 항복응력과 단단함에서 가장 높은 값을 보여주었고 pH 8.0 이상에서는 L값과 b값의 감소가 현저하였다. 이상의 결과에서 물의 첨가량과 가열온도, 가열시간, 소금첨가량은 계란찜의 텍스쳐 및 색특성에 영향을 줌이 밝혀졌다.

• 한국식품저장유통학회지
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• 제22권5호
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• pp.727-734
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• 2015

본 연구에서는 새로운 프로바이오틱스를 선발하고자 protease를 생산하는 GRAS균주를 된장 및 청국장으로부터 분리하고 그 특성을 조사하였다. 분리한 균주를 분리 동정한 결과 각각 B. amyloliquefaciens CDD5, B. amyloliquefaciens CPD4 및 B. amyloliquefaciens CGD3로 명명하였다. 분리균주의 생육은 12시간 배양 시, 7.13~7.32 log CFU/mL로 최대 생육도를 보였으며 24시간까지 유지되다가 감소하는 경향을 나타내었고, 그 중 B. amyloliquefaciens CGD3의 protease 활성이 9.21 U/mL로서 가장 높게 나타났다. B. amyloliquefaciens이 생산하는 protease 활성은 pH 7.0~10.0, 온도는 $50^{\circ}C$에서 최적활성을 나타내었으며, casein에서 protease활성이 가장 우수하게 나타났다. pH, NaCl, glucose 농도를 달리한 배양조건에 따른 B. amyloliquefaciens의 생육특성으로는 pH 5.0~10.0의 넓은 범위에서 생육하였고, NaCl은 12% 농도까지 증식하였으며 glucose 5%까지는 생육하나 10% 농도부터는 현저하게 생육이 낮아짐을 확인하였다. 또한 혈당저하를 유도하는 ${\alpha}$-glucosidase의 저해활성을 측정한 결과, B. amyloliquefaciens CPD4 및 B. amyloliquefaciens CGD3에서는 각각 96.65% 및 97.85%의 저해율을 나타내었다. 이러한 결과는 특히 protease활성이 가장 우수하고 ${\alpha}$-glucosidase 또한 높은 저해활성을 가진 B. amyloliquefaciens CGD3 균주의 생육특성을 고려하여 단백질 분해 분야의 프로바이오틱 적용 및 당뇨예방용 기능성식품 소재로의 가능성이 기대된다.

• Archives of Pharmacal Research
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• 제21권4호
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• pp.429-435
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• 1998

Lectins and its A- and B-chains from Korean mistletoe (Viscum album var. coloratum) were isolated by affinity chromatography on the Sepharose 4B modified by lactose-BSA conjugate synthesized by reductive amination of ligand (lactose) to .epsilon.-amino groups of lysine residues of spacer (BSA) after reduction by $NaCNBH_3$. The lactose-BSA conjugate was coupled to Sepharose 4B activated by cyanogen bromide. The molecular weight determined by SDS-PAGE were a 31 kD of A-chain and a 35kD of B-chain. Amino acid analysis and N-terminal sequencing were performed. The effects of pH, temperature and guanidine chloride on the conformation of the lectin were investigated by measuring its intrinsic fluorescence and compared with its hemagglutinating activities. Blue shift was detected on the acidic pH and there was a close relationship between activities and conformation of the lectin. Under denaturing conditions, the tryptophan emission profile of lectin showed typical denaturaiional red shift which also correspond to the conformations and activity of lectin.

• Journal of Veterinary Science
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• 제22권4호
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• pp.43.1-43.10
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• 2021

Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016-2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions: These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

• 한국소음진동공학회:학술대회논문집
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• 한국소음진동공학회 2010년도 춘계학술대회 논문집
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• pp.698-699
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• 2010

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1323-1329
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• 2005

A mathematical model was developed for predicting the growth kinetics of Listeria monocytogenes in tryptic soy broth (TSB) as a function of combined effects of temperature, pH, and NaCl. The TSB containing four different concentrations of NaCl (2, 4, 5, and $10\%$) was initially adjusted to six different pH levels (pH 5, 6, 7, 8, 9, and 10) and incubated at 4, 10, 25, or 37$^{circ}C$. In all experimental variables, the primary growth curves were well fitted ($r^{2}$=0.982 to 0.998) to a Gompertz equation to obtain the lag time (LT) and specific growth rate (SGR). Surface response models were identified as appropriate secondary models for LT and SGR on the basis of coefficient determination ($r^{2}$=0.907 for LT, 0.964 for SGR), mean square error (MSE=3.389 for LT, 0.018 for SGR), bias factor ($B_{1}$B,=0.706 for LT, 0.836 for SGR), and accuracy factor ($A_{f}$=1.567 for LT, 1.213 for SGR). Therefore, the developed secondary model proved reliable predictions of the combined effect of temperature, NaCl, and pH on both LT and SGR for L. monocytogenes in TSB.

• 한국식품영양과학회지
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• 제45권4호
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• pp.551-556
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• 2016

본 연구는 돈육 유화물을 저염 육제품 개발의 기초자료로 활용하고자 돈육 유화물 제조 시 NaCl 첨가량에 따른 이화학적 특성을 조사하였고, 처리구별로 돈육 유화물 제조 시 NaCl 첨가량을 0%, 0.3%, 0.6%, 0.9%, 1.2%, 1.5% 첨가하여 6개의 처리구로 분류하여 제조하였다. 수분 함량은 NaCl 1.5% 첨가구가 대조구와 다른 처리구들에 비해 유의적으로 높은 값을 가졌다(P<0.05). 회분 함량은 NaCl 첨가율이 증가할수록 상승하는 추세를 보였으며, 단백질 함량은 NaCl 첨가에 따라 감소하는 경향을 나타내었다. 가열 전 pH는 NaCl 0.9%, 1.2%, 1.5% 첨가구가 유의적으로 낮았으며 (P<0.05), 가열 후에는 모든 NaCl 첨가구가 대조구에 비해 유의적으로 낮은 pH를 나타내었다(P<0.05). 색도 측정 결과 가열 전 명도는 모든 NaCl 처리구가 대조구에 비해 유의적으로 높은 수치를 나타내었으며(P<0.05), 적색도와 황색도는 대조구에 비해 유의적으로 낮은 수치를 나타내었다(P<0.05). 가열 후 명도와 황색도는 NaCl 첨가량이 증가할수록 감소하는 경향을 나타내었으며, 적색도는 증가 추세를 보였다. 보수력과 가열수율은 NaCl 첨가량이 증가함에 따라 증가하는 경향을 보였으며, 0.9% 처리구부터 유의적으로 높은 값을 나타내었다(P<0.05). 점도는 NaCl 1.2%, 1.5% 처리구가 69.60~71.41 Pa s로, 대조구와 NaCl 0.3% 처리구의 51.13~51.61 Pa s보다 높은 점도를 가진 것으로 나타내었다. 물성 측정 결과 탄력성과 응집성은 대조구와 처리구간에 유의적인 차이가 없었으며(P>0.05), 경도와 검성, 씹음성은 NaCl 1.5% 처리구가 대조구와 다른 처리구보다 유의적으로 높은 수치를 나타내었다(P<0.05). 이상의 연구 결과를 종합하면 돈육 유화물에 NaCl이 저염 수준인 0.9~1.2% 첨가되었을 때 NaCl 1.5% 첨가구와 이화학적으로 유사한 특성이 있는 것으로 나타나 돈육 유화물을 이용한 저염육제품 제조 시 염 농도를 0.9%, 1.2% 첨가하는 것이 바람직할 것으로 생각한다.