• Title/Summary/Keyword: lipase B26

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Biodiversity of Bacterial lipase genes

  • Kim, Hyung-Kwoun
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2001.06a
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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.

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High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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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.

Polymorphisms of the Lipoprotein Lipase Gene of Red Seabream, Pagrus major (참돔의 lipoprotein lipase 유전자 다형성)

  • Jang, Yo-Soon;Hong, Kyung-Pyo;Noh, Choong-Hwan
    • Ocean and Polar Research
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    • v.26 no.4
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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.

Effects of Coffee and Aflatoxin B1 on the Pancreatic Exocrine Function and Structure (Coffee와 Aflatoxin B1이 췌장의 외분비 기능 및 조직에 미치는 영향)

  • 안혜선
    • Journal of Nutrition and Health
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    • v.26 no.3
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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.

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Antibacterial Effect of Fructose Laurate Synthesized by Candida antarctica B Lipase-Mediated Transesterification

  • Lee, Ki Ppeum;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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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 (Mr 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.

Postprandial Plasma Lipid Levels and Digestive Enzyme Activities After High Fat Meal in Rats Adapted to Dietary Fiber (식이섬유 식이에 적응된 흰쥐에서 고지방식이가 식후 혈장지질농도와 소화효소활성에 미치는 영향)

  • 양정례;서명자;송영선
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.116-122
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    • 1997
  • Rats were adapted to diets containing 10% cellulose,10% sodium alginate and fiber-free diet for 5 weeks. Following a 14 hour fasting, rats were fed 5g of a test meal that provided 50% energy from fat, then killed at 4 hour postprandially. Plasma and lipoprotein fraction-cholesterol levels were lower in sodium alginate-fed animals than in rats fed other diets. Plasma TG did not differ among diet treatments. Increase in TG content of HDL fraction occurred in dietary fiber groups. Intestinal apolipoprotein B level and lipase activity were lower in sodium alginate-fed group than in other dietary groups. These results suggest that chronic consumption of sodium alginate affects plasma cholesterol level as in the case of fiber supplemetation, but is less likely to modify the acute Plasma TG response to high fat meal than if a fiber supplement is incorporated into the meal.

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The Optimal Enrichment Condition of Rotifer Brachionus rotundiformis (소화효소 활성으로 본 rotifer Brachionus rotundiformis의 적정 영양강화 조건)

  • Kwon, O-Nam;Park, Heum-Gi
    • Journal of Aquaculture
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    • v.21 no.1
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    • pp.41-46
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    • 2008
  • The purpose of the study was to suggest the optimal lipid enrichment conditions used digestive enzyme activity of rotifer changing due to water temperature and salinity. The high population growth appeared at the experiment temperature more than 28 degrees highly on the culture temperature(maximum 32 degrees, 1,453 individual/mL). The fecundity was low at high temperature, and the egg ratio was high at low temperature. Population growth of 10 and 15 ppt appeared in most highly, but the fecundity and the egg ratio were high most significantly appeared in natural seawater(32 psu). The digestive enzyme activity by the culture environment mainly showed high activity in natural seawater(amylase exclusion, 15 psu). However, the TAP activity by the water temperature showed highly at the more high temperature, but the amylase and the lipase appeared at low temperature. We carried out the lipid enrichment at 20 degrees and 26 degrees in a condition of the natural seawater. Total protein, the total essential amino acids differed not significantly. The methionine content that was essential amino acids, a total lipid content, unsaturated index of fatty acids, DHA and the DHA/EPA ratio were high significantly each in $20^{\circ}C$ enrichment trial. Therefore, we could suggest the $20^{\circ}C$ and natural seawater for the optimal lipid enrichment condition in aquaculture, because methionine contents, several indexes by the lipid, TG-lipase activity, fecundity and egg ratio are high.

Production of Structured Lipid Containing Capric and Conjugated Linoleic Acids and Studies of Physicochemical Characteristics (Capric acid와 conjugated linoleic acid를 함유한 재구성 지질의 합성 및 특성연구)

  • Shin, Jung-Ah;Lee, Ki-Teak
    • Korean Journal of Food Science and Technology
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    • v.36 no.4
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    • pp.537-541
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    • 2004
  • Based on reaction time and substrate molar ratio, structured lipid (SL-corn) was produced at 1:2:2(corn oil/capric acid/CLA) and 4% immobilized lipase from Rhizomucor miehei (RM IM). Reaction was carried out for 24 hr at $55^{\circ}C$ in 1-L stirred-batch reactor. After reaction, 13.3 mol% capric acid and 8.9%, CLA were incorporated into corn oil. Iodine and saponification values of SL-corn were 68 and 202, respectively. Tocopherol content decreased after reaction (about 39%). SL-corn showed more yellowish color than corn oil (p<0.05). Reversed-phase HPLC indicated triacylglycerol species containing capric acid in SL-corn resulted in faster crystallization than that of corn oil.

Expression and Biochemical Characterization of Cold-Adapted Lipases from Antarctic Bacillus pumilus Strains

  • Litantra, Ribka;Lobionda, Stefani;Yim, Joung Han;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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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.

The Development of Gastrointestinal Tract and Pancreatic Enzymes in White Roman Geese

  • Shih, B.L.;Yu, B.;Hsu, J.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.6
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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.