• Journal of Microbiology and Biotechnology
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• 제15권1호
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• pp.155-160
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• 2005

Abstract A direct approach was used to retrieve active lipases from Paenibacillus polymyxa genome databases. Twelve putative lipase genes were tested using a typical lipase sequence rule built on the basis of a consensus sequence of a catalytic triad and oxyanion hole. Among them, six genes satisfied the sequence rule and had similarity (about 25%) with known bacterial lipases. To obtain the six lipase proteins, lipase genes were expressed in E. coli cells and lipolytic activities were measured by using tributyrin plate and pnitrophenyl caproate. One of them, contig 160-26, was expressed as a soluble and active form in E. coli cell. After purifying on Ni-NTA column, its detailed biochemical properties were characterized. It had a maximum hydrolytic activity at $30^{\circ}C$ and pH 7- 8, and was stable up to $40^{\circ}C$ and in the range of pH 5- 8. It most rapidly hydrolyzed pNPC$_6$ among various PNPesters. The other contigs were expressed more or less as soluble forms, although no lipolytic activities were detected. As they have many conserved regions with lipase 160-26 as well as other bacterial lipases throughout their equence, they are suggested as true lipase genes.

• Natural Product Sciences
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• 제18권2호
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• pp.116-120
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• 2012

Pancreatic lipase digests dietary fats by hydrolysis, which is a key enzyme for lipid absorption. Therefore, reduction of fat absorption by the inhibition of pancreatic lipase is suggested to be a therapeutic strategy for obesity. We previously reported coumarins and secoiridoids of Fraxinus rhynchophylla as inhibitory constituents on adipocyte differentiation. Further investigation on F. rhynchophylla led to the isolation of lignan derivatives such as lignans (1 - 10), sesquilignans (11 - 14) and coumarinolignans (15 - 17). Among them, coumarinolignans and sesquilignans were first reported from Fraxinus species. Among the constituents isolated, sesquilignans showed the significant inhibition on pancreatic lipase, whereas lignans and coumarinolignans exerted weak effects.

• Macromolecular Research
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• 제16권7호
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• pp.651-658
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• 2008

Poly(butylene succiate) (PBS), poly(butylene succinate-co-L-lactate) (PBSL), and poly(butylene succinate-co-6-hydroxycaproate) (PBSCL) polymers were degraded by lipase $PS^{(R)}$, and the enzymatic degradation mechanism of PBS was analyzed in detail. The enzymatic degradation of PBS gave 4-hydroxybutyl succinate (4HBS) as the main product. An exo-type hydrolysis mechanism was proposed based on this observation. The terminal chain of PBS had conformational similarity to ordinary tri- and diglycerides and could be incorporated as a substrate in the active site of this lipase. The surface adsorption of the lipase was much larger on PBS and its copolymer films than on the other polyester films because the lipase adhered quite strongly to the polymer terminal through a specific adsorption mechanism. Kinetic analysis showed that the total number of surface adsorption points per unit area of PBSL and PBSCL copolymers was larger than that of the PBS homopolymer.

• Natural Product Sciences
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• 제7권1호
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• pp.17-20
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• 2001

The activity of the lipase and phospholipase extracted from the seed meal of Nicotiana tabacum L. was studied with the help of Spectrophotometer at different pH, Temperatures and Solvents. Both lipase and phospholipase shown optimum activity at pH 6 and $45^{\circ}C$. Their activities also maximize when n-heptane was used as solvent media.

• 한국식품영양과학회지
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• 제44권12호
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• pp.1912-1917
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• 2015

천연식물에 광범위하게 존재하는 대표적인 페닐프로파노이드 화합물인 caffeic acid에 대해 배 유래의 polyphenol oxidase로 산화반응을 수행하여 상대적으로 높은 pancreatic lipase 저해 활성($IC_{50}$; $161.2{\pm}2.8{\mu}g/mL$)을 확인하였으며, 이는 caffeic acid와 비교하였을 경우 활성이 상승함을 알 수 있었다. Caffeic acid 산화반응물에 대해서 $C_{18}$ 겔을 활용한 column chromatography를 수행하여 4종의 리그난 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 phellinsin A(2), caffeicinic acid(3), isocaffeicinic acid(4), 7,8-erythro-caffeicin(5)으로 동정하였다. 이들 화합물중 phellinsin A(2)는 $IC_{50}$ 값이 $66.3{\pm}2.6{\mu}M$로 가장 강한 효능을 나타내었으며, 다음으로 caffeic acid 2분자의 산화 결합을 통해 생합성된 caffeicinic acid(3)의 $IC_{50}$ 값이 $109.6{\pm}3.7{\mu}M$의 저해능을 나타내었다. 배에 존재하는 polyphenol 산화효소에 의해 생합성된 caffeic acid 이량체가 pancreatic lipase 저해 활성 물질임을 확인하였으며, 이들 활성은 caffeic acid가 결합 양상에 따른 화합물의 구조에 따라 다름이 시사되었다. 향후 이들 활성물질의 활성 기작에 대한 연구가 필요하며 본 연구 결과는 보다 우수한 pancreatic lipase 저해능을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 이용될 수 있을 뿐만 아니라 항비만 물질의 상업화를 위한 기초자료로 이용될 수 있을 것으로 사료된다.

• 한국응용과학기술학회지
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• 제22권2호
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• pp.106-115
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• 2005

Bio-diesel as fatty acid methyl ester was derived from such oils as soybean, peanut and canola oil by lipase catalyzed continuous trans-esterification. So the activation of lipase(Novozym - 435) was kept to be up to 4:1, the limiting molar ratio of methanol to oil under one-step addition of methanol due to the miscibility of oil and methanol through the static mixer for 4hrs and the elimination of glycerol on the surface of lipase by 7wt% silica gel. Therefore the overall yield of fatty acid methyl ester from soybean oil appeared to be 98% at 50$^{\cdot}C$ of reaction temperature under two-steps addition of methanol with 2${\times}$2:1 of methanol to oil molar ratio at an interval of 5.5hrs, 7wt% of lipase, 24 number of mixer elements, 0.2ml/min of flow rate and 7wt% of silica gel.

• 한국미생물·생명공학회지
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• 제19권4호
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• pp.377-382
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• 1991

우지를 lipase에 의해 지방산과 글리세린으로 분해하는 반응을 액상 및 고상에서 수행하였다. 올리브유를 기질고 lipase OF 360(일본 메이토사 제품)의 특성을 조사한 결과 최적 pH는 6, 최적 온도는 $37^{\circ}C$이었다. 우지를 기질로 액상 효소반응을 수행한 결과는 물사용량 80 wt/wt, 온도 $37^{\circ}C$, 효소사용량 200unit/g tallow 조건에서 93의 가수분해율을 얻을 수 있었다.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.480-490
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• 2020

A novel lipase gene, Lip-1420, was isolated from a metagenomic library constructed from reed marsh from Mt. Jumbong in Korea, comprising 112,500 members of recombinant plasmids. The DNA sequence of Lip-1420-subclone (5,513 bp) was found to contain at least 11 ORFs according to the GenBank database. The ORF-3 gene was inserted into the pET21a plasmid containing the C-terminal 6-His tag and transformed into E. coli BL21(DE3) to express the recombinant lipase protein. Lip-1420 was purified using a fast protein liquid chromatography system. The gene was registered in GenBank (MH628529). The values of Km and Vmax were determined as 0.268 mM and 1.821 units, respectively, at 40℃ and pH 8.0, using p-nitrophenyl palmitate as the substrate. This lipase belongs to family IV taxonomically because it has conserved HGGG and GDSAG motifs in the constitutive amino acid sequence. According to the predicted structural model, the binding sites are represented by residues H78, G81, D150, S151, A152, V181, and D236. Finally, Lip-1420 showed triolein selectivity for methanolysis between triolein (18:1) and tristearin (18:0) substrates. Further study of the selective mechanism and structure-function relationship of this new lipase could be useful for more practical applications.

• Journal of Microbiology and Biotechnology
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• 제17권1호
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• pp.74-80
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• 2007

An enantioselective lipase gene (esf) for the kinetic resolution of optically active (S)-flurbiprofen was cloned from the new strain Serratia marcescens ES-2. The esf gene was composed of a 1,845-bp open reading frame encoding 614 amino acid residues with a calculated molecular mass of 64,978 Da. The lipase expressed in E. coli was purified by a three-step procedure, and it showed preferential substrate specificity toward the medium-chain-length fatty acids. The esf gene encoding the enantioselective lipase was reintroduced into the parent strain S. marcescens ES-2 for secretory overexpression. The transformant S. marcescens BESF secreted up to 217kU/ml of the enantioselective lipase, about 54-fold more than the parent strain, after supplementing 3.0% Triton X-207. The kinetic resolution of (S)-flurbiprofen was carried out even at an extremely high (R,S)-flurbiprofen ethyl ester [(R,S)-FEE] concentration of 500 mM, 130 kU of the S. marcescens ES-2 lipase per mmol of (R,S)-FEE, and 1,000 mM of succinyl ${\beta}-cyclodextrin$ as the dispenser at $37^{\circ}C$ for 12h, achieving the high enantiomeric excess and conversion yield of 98% and 48%, respectively.

• Journal of Applied Biological Chemistry
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• 제52권3호
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• pp.151-156
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• 2009

As part of an investigation to identify microorganisms that are biotechnologically interesting for industrial application, we isolated a bacterial strain from Chungkookjang that produces extracellular neutral lipase. In addition, the crude enzyme was characterized. This isolated strain, designated as JKA-3 was identified as Bacillus subtilis JKA-3 based on morphological, physiological and biochemical characteristics, as well as phylogenetic analysis using 16S rRNA gene sequence. The cells were rod-shaped and $0.6-0.8{\times}2.0-2.3\;{\mu}m$ in size. Optimal growth conditions were $35-40^{\circ}C$ and pH 6.0-8.0. The isolate was able to grow in up to 0-10.0% (w/v) NaCl. Optimal activity conditions of the crude lipase fraction of B. subtilis JKA-3 were pH of 7.0 at $35^{\circ}C$. This enzyme was stable in the pH ranging 6.0-8.0.