• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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• pp.163-164
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• 2001

A number of bacterial species produce extracellular lipases. Among them, many lipase genes have been cloned and sequenced. A comparison of primary sequences revealed only very limited sequence homology among them. Based on the sequence homologies and molecular sizes (Mr), bacterial lipases were classified into four discrete groups. From soil samples taken around Taejon, five different lipase-producing bacteria were isolated; Proteus vulgaris K80, Bacillus stearothermophilus Ll, B. pumilus B26, Staphylococcus haemolyticus L62, S. aureus B56. Nucleotide sequence analysis showed that Staphylococcus lipase genes (L62 and B56) composed of pre-pro-mature parts, Bacillus lipase genes (Ll and B26) pre-mature parts, and Proteus lipase gene (K80) mature part only. In addition, the molecular sizes of their mature parts were quite different from 19,000 to 45,000. Finally, they had very little homology (less than 20％) in their amino acid sequences. Judging from the above results, lipase K80 belonged to bacterial lipase Group I, lipase L1 and lipase B26 Group III, and lipase L62 and lipase B56 Group IV. This diversity in their primary structures was also reflected in their enzymatic properties; temperature effects, pH effects, substrate specificity, detergent effects, and so on.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.892-896
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• 2003

High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

• Ocean and Polar Research
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• 제26권4호
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• pp.551-557
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• 2004

Polymorphism of the lipoprotein lipase (LPL) gene which plays an important role in regulation of lipid deposition was analysed in two red seabream (pagrus major) populations (KF4, cultured KORDI line, n=100 : JPN, imported from Japan, n=100). We amplified a DNA fragment (1,091 bp) including the exon 2 region of the LPL gene, and conducted PCR-RFLP analysis using MspI and AluI. The PCR products were also sequenced. Two alleles (A and B) were found in MspI digestion and Sve alleles (A, B, C, D and E) in AluI digestion. The sequenced data revealed four nucleotide substitutions including one transversion at the MspI recognition site (nt 2,235, $C{\rightarrow}10$) and three transitions at the AluI recognition sites (nt 1,721, $A{\rightarrow}G;$ nt 2,319, $C{\rightarrow}T;$ nt 2,319, $T{\rightarrow}C$). Among them, substitutions at the nt 2,235 and 2,319 sites which are located in the exon 2 were proved to be silent point mutations. MspI polymorphism resulted in 3 genotypes, and the allele frequency was significantly different between the two fish populations, KF4 and JPN. In the case of AluI polymorphism, the 5 alleles (A, B, C, D, E) comprised 12 genotypes of the 5 alleles. KF4 population, alleles D and I were specific to the LPL gene Polymorphisms would be useful DNA markers for red seabream population.

• Journal of Nutrition and Health
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• 제26권3호
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• pp.268-276
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• 1993

Coffee is known to increase pancreatic secretion of digestive enzymes. The mutagen, aflatoxin B1(AFB1) is contained in fermented foods and known to increase the specific activities of pancreatic chymotrypsin, trypsi, amylase, and lipase. Nowadays, coffee intake is increased among Koreans who have consumed relatively high amount of traditional fermented foods. Therefore, this study was performed to examine the effect of coffee and AFB1 on pancreatic exocrine function and structure. Rats were divided into 10 experimental groups. The first five groups were W(control group), LD(0.2g decaffeinated coffee/Kg B.W), HD(3g decaffeinated coffee/Kg B.W), LC(0.2g coffee/Kg B.W), and HC(3g coffee/Kg B.W). The second five groups were WA, LDA, HDA, LCA, HCA, same as first five groups in caffieine level but treated with AFB1. The result of this experiment showed that the caffeine intake did not influence significantly on the growth and feed efficiency. But water intake was increased by caffeine intake and AFB1 treatment. The weights of pancreas and liver were increased as the caffeine intake was increased. Trypsin activities were tend to increase in concentrated coffee groups(HD, HC). AFB1 treated groups showed the higher trypsin level than the AFB1 untreated groups. Amylase activities were tend to increase in concentrated coffee groups(HD, HC) of AFB1 untreated animals. AFB1 treated did not show the additional effect on the stimulated amylase secretion by coffee. Lipase activities were tend to decrease in concentrated coffee groups(HD, HC) of AFB1 untreated animals. Lipase activities were increased in the order named WA group, coffee groups, decaffeinated coffee groups in AFB1 treated animals. AFB1 treated groups showed the higher lipase level than AFB1 untreated groups. In the histologic observation of pancreas HCA group showed more dense compound tubuloalveolar glands and proliferation of nuclei than normal. The result suggested a development of a atypia which is ongoing phase to a cancer.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1579-1585
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• 2016

Sugar esters are valuable compounds composed of various sugars and fatty acids that can be used as antibacterial agents and emulsifiers in toothpaste and canned foods. For example, fructose fatty acid esters suppress growth of Streptococcus mutans, a typical pathogenic bacterium causing dental caries. In this study, fructose laurate ester was chosen as a target material and was synthesized by a transesterification reaction using Candida antarctica lipase B. We performed a solvent screening experiment and found that a t-butanol/dimethyl sulfoxide mixture was the best solvent to dissolve fructose and methyl laurate. Fructose laurate was synthesized by transesterification of fructose (100 mM) with methyl laurate (30 mM) in t-butanol containing 20% dimethyl sulfoxide. The conversion yield was about 90%, which was calculated based on the quantity of methyl laurate using high-performance liquid chromatography. Fructose monolaurate (M_r 361) was detected in the reaction mixture by high-resolution mass spectrometry. The inhibitory effect of fructose laurate on growth of oral or food spoilage microorganisms, including S. mutans, Bacillus coagulans, and Geobacillus stearothermophilus, was evaluated.

• 한국식품영양과학회지
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• 제26권1호
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• pp.116-122
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• 1997

식이섬유에 5주간 적응된 횐쥐 (S.D., male)를 14시간 절식시킨 뒤 총 에너지의 50%가 되는 고지방식이를 급여하고 4시간 뒤의 혈장 지질 농도와 소화효소활성을 조사하여 다음과 같은 결과를 얻었다. 혈장과 지단백의 콜레스테롤 함량은 sodium alginate군에서 가장 낮았다(p<0.05). 혈장 중성지방 함량은 식이군간에 차이가 없었으나 LDL-중성지방은 sodium alginate군에서 가장 낮았다(p<0.05). HDL-중성지방 함량은 식이섬유 군에서 높았다 (p<0.05). 소장 apolipoprotein B는 무섬 유식이군과 cellulose군에 비해 sodium alginate군에서 유의적으로 높았다(p<0.05). 소장 내용물에서의 amylase활성은 각 식이군간에 유의적인 차이는 없었으나 식이 섬유 첨가군에서 다소 높았으며, lipase 활성은 무섬유 식이군과 cellulose군에 비해 sodium alginate군에서 유의적으로 높았다(p<0.05). 소장내용물에서 pretense 활성과 담즙산 함량은 식이군간에 유의적인 차이가 없었으나, 단백질 함량은 무섬유 식이군에 비해 식이섬유 첨가군에서 유의적으로 높았다(p<0.05). 이상의 결과를 종합해 볼 때 장기간의 식이 섬유 섭취는 식이섬유가 배제된 고지방식 후에도 혈장 콜레스테롤 농도를 낮추는 효과가 있으나, 혈장 중성지방 농도는 식이 섬유가 함유된 식이에 비해 영향을 받지 않는 것으로 보인다.

• 한국양식학회지
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• 제21권1호
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• pp.41-46
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• 2008

본 연구의 목적은 rotifer Brachionus rotundiformis의 수온과 염분농도의 변화에 따른 이들의 소화효소 활성을 파악하고, 소화효소 활성을 기초로 적정 영양강화 조건을 찾는 것이다. 다른 온도에서 24시간 배양된 rotifer 밀도는 $32^{\circ}C$에서 1,453 개체/mL로 가장 높았다(P<0.05). 그리고 배양온도 $28^{\circ}C$에서 염분농도에 따른 rotifer 밀도는 15 에서 2,147 개체/mL로 가장 높게 나타났다(P<0.05). 배양환경에 따른 rotifer의 소화효소 활성에서 TAP 개체당 활성은 $24-32^{\circ}C$에서 유의적으로 높게 나타났으며(P<0.05), 단백질 비활성은 $32-36^{\circ}C$에서 높게 조사되었다(P<0.05). 그리고 TG-lipase 활성에서는 개체당 활성과 단백질 비활성 모두 $20^{\circ}C$에서 가장 높은 활성을 보였다(P<0.05). 또한 염분별 실험에서 TAP와 TG-lipase 활성은 32 psu에서 가장 높은 활성을 보였다(P<0.05). 32 psu에서 온도별 영양강화한 rotifer의 methionine 함량(% in protein), 지질 함량과 지방산 불포화도이 $20^{\circ}C$에서 유의적으로 높게 나타났다. 특히, DHA와 DHA/EPA 비가 $20^{\circ}C$ 실험구에서 12.6%와 12.5의 비로 각각 높았다(P<0.05). 따라서 rotifer B. rotundiformis의 지질 영양강화는 특히 TGlipase 활성을 높여 줄 수 있는 32 psu, $20^{\circ}C$에서 실시하는 것이 지방산 불포화도, DHA 및 DHA/EPA 비를 높여 줄 수 있는 효과적인 방법이라고 판단된다.

• 한국식품과학회지
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• 제36권4호
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• pp.537-541
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• 2004

1-L stirred-batch type reactor에서 기질들의 몰 비율을 1:2:2(corn oil:capric acid:CLA)하고 RM IM lipase을 사용하여 재구성 지질을 합성하였다. 합성된 재구성 지질은 총 capric acid 와 CLA을 각각 13.3 mol%와 8.9 mol% 함유하였다. 기능성 지방산은 주로 sn-1,3 position에 분포하였고, sn-2 position에는 linoleic acid가 대부분이었다. 재구성 지질의 요오드가와 비누화가는 68과 202이었고, 이는 재구성 지질에 함유된 capric acid 때문이었다. 반응 후 재구성 지질의 총 tocopherol 함량은 약 39% 감소하였다. 색도분석결과, 재구성 지질과 옥배유 서로간에 백색도$(L^*)$와 적색도$(a^*)$에서는 유의적 차이가 없었으나, 황색도$(b^*)$에서는 재구성 지질이 옥배유보다 yellowness를 보이며 유의적 차이를 보였다(p<0.05). RP-HPLC 분석결과 재구성 지질에서 capric acid를 함유하고 있는 TAG형태의 PN은 26-44로 전체의 약 34.2 area% 차지함을 알 수 있었다. 또한 옥배유가 $-56.77^{\circ}C$에서 주된 crystallization peak 정점을 보인 반면에, 재구성 지질은 $-46.2^{\circ}C$에서 주된 peak 정점을 나타내었다.

• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1221-1228
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• 2013

Two lipase genes (bpl1 and bpl3) from Antarctic Bacillus pumilus strains were expressed in Bacillus subtilis. Both recombinant lipases BPL1 and BPL2 were secreted to the culture medium and their activities reached 3.5 U/ml and 5.0 U/ml, respectively. Their molecular masses apparent using SDS-PAGE were 23 kDa for BPL1 and 19 kDa for BPL3. Both lipases were purified to homogeneity using ammonium sulfate precipitation and HiTrap SP FF column and Superose 12 column chromatographies. The final specific activities were estimated to be 328 U/mg for BPL1 and 310 U/mg for BPL3. Both lipases displayed an optimum temperature of $35^{\circ}C$, similar to other mesophilic enzymes. However, they maintained as much as 70% and 80% of the maximum activities at $10^{\circ}C$. Accordingly, their calculated activation energy at a temperature range of $10-35^{\circ}C$ was 5.32 kcal/mol for BPL1 and 4.26 kcal/mol for BPL3, typical of cold-adapted enzymes. The optimum pH of BPL1 and BPL3 was 8.5 and 8.0, respectively, and they were quite stable at pH 7.0-11.0, showing their strong alkaline tolerance. Both lipases had a preference toward medium chain length ($C_6-C_{10}$) fatty acid substrates. These results indicate the potential for the two Antarctic B. pumilus lipases as catalysts in bioorganic synthesis, food, and detergent industries.

• Asian-Australasian Journal of Animal Sciences
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• 제18권6호
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• pp.841-847
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• 2005

The objective of this experiment was to investigate the development of gastrointestinal tract and activities of pancreatic enzymes in White Roman geese. Thirty developing embryos at the 22th, 24th and 26th day of incubation and at hatching, and sixteen or eight goslings, half males and half females, at the 1, 3, 7 or 11, 14, 21 and 28 days of age were sampled, respectively. The weights of the yolk, gastrointestinal tract and intestinal length, and the activities of pancreatic enzymes were measured. Residual yolk weight decreased rapidly during late incubation and was nearly depleted at 3 days of age. The protein and energy contents in the residual yolk of goslings at 3 days of age were significantly (p<0.05) less than those at the late incubation. From 6 days before hatching to 28 days of age, the absolute weights of gizzard, proventriculus, liver, pancreas, small intestine and large intestine in goslings increased by 48, 457, 94, 2334, 89 and 76 times, respectively. The relative weights of proventriculus, gizzard, liver, pancreas, small intestine and large intestine reached peaks at 3, 3, 14, 14, 11 and 11 days of age, respectively, and then decreased gradually. However, the relative lengths of small intestine and large intestine reached peaks at 3 days of age and at hatching, respectively. The activities of pancreatic trypsin and chymotrypsin increased sharply from hatching to 14 day of age, and then decreased gradually until 21 days of age. The activity and specific activity of pancreatic amylase were increased following by age and peaked at 7 to 11 and 21 days of age, respectively. The activity and specific activity of pancreatic lipase reached a plateau from 11 to 28 days of age. These results indicate that the gastrointestinal tract and activities of pancreatic enzymes developed more rapidly than body weight through the early growing period of goslings.