• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1827-1834
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• 2015

The SMG1 lipase from Malassezia globosa is a newly found mono- and diacylglycerol (DAG) lipase that has a unique lid in the loop conformation that differs from the common alpha-helix lid. In the present study, we characterized the contribution of three residues, L103 and F104 in the lid and F278 in the rim of the binding site groove, on the function of SMG1 lipase. Site-directed mutagenesis was conducted at these sites, and each of the mutants was expressed in the yeast Pichia pastoris, purified, and characterized for their activity toward DAG and p-nitrophenol (pNP) ester. Compared with wild-type SMG1, F278A retained approximately 78% of its activity toward DAG, but only 11% activity toward pNP octanoate (pNP-C8). L103G increased its activity on pNP-C8 by approximately 2-fold, whereas F104G showed an approximate 40% decrease in pNP-C8 activity, and they both showed decreased activity on the DAG emulsion. The deletion of 103-104 retained approximately 30% of its activity toward the DAG emulsion, with an almost complete loss of pNP-C8 activity. The deletion of 103-104 showed a weaker penetration ability to a soybean phosphocholine monolayer than wild-type SMG1. Based on the modulation of the specificity and activity observed, a pNP-C8 binding model for the ester (pNP-C8, N102, and F278 form a flexible bridge) and a specific lipid-anchoring mechanism for DAG (L103 and F104 serve as "anchors" to the lipid interface) were proposed.

• KSBB Journal
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• v.19 no.2
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• pp.148-153
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• 2004

Lipase catalyzes the hydrolysis of glycerides into fatty acids and glycerol. The study of microbial lipases has been stimulated in resent years. It is due to the potential uses of lipases in esterification of oils to glycerol, alcohols and carbohydrates. Development of lipase producing yeast has been focused concerning to the utilization of yeast culture for animal feed. In this study, yeast like cells was isolated from a waste oil and sludge. A strain having higher lipase activity was selected by random mutagenesis using UV-radiation. The optimal cultivation conditions in submerged culture were examined in terms of lipase production. 2.0％ of high fructose syrup, 1,0％ of CSL, and 1.0％ of olive oil were selected as the nutritional media for the production of lipase. The maximum lipase activity of 1.12 U/ml and viable cell number of 8.8${\times}$10$\^$7/ cells/mL were obtained at 27$^{\circ}C$ with an initial pH of 5.0.

• KSBB Journal
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• v.15 no.1
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• pp.72-75
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• 2000

The hydrolysis characteristics of each fatty acid composing the fish oil by means of the lipase from Candida cylindracea was investigated. The saturated fatty acids, C14:0, C16:0 and C18:0, and the unsaturated fatty acids with one double bond, C16:1, C18:1(n-7), C18:(n-9), C20:1 and C22:1 were more easily hydrolyzed than the $\omega$-3 polyunsaturated fatty acids. when the number of carbon of the $\omega$-3 fatty acids was same but that of double bond was different, the hydrolysis of the $\omega$-3 fatty acids having lower number of double bond was more rapidly carried out. When the degree of polyunsaturation was same but the number of carbon was different, the lipase acted more rapidly upon the $\omega$-3 fatty acids with lower number of carbon. Docosahexaenoic acid(DHA) was most highly concentrated in the glyceride mixture after 120hr hydrolysis among the various $\omega$-3 polyunsaturated fatty acids. The weight percentage of DHA to the $\omega$-3 fatty acids in the fish oil was 31.87% but that in the glyceride mixture after 120hr hydrolysis was 51.89%.

• Journal of Veterinary Science
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• v.25 no.3
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• pp.48.1-48.12
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• 2024

Importance: Early diagnosis of canine pancreatitis is challenging due to non-specific clinical signs. Currently, abdominal ultrasonography and measurement of canine pancreatic lipase (cPL) have been employed for the diagnosis of pancreatitis. Objective: Many qualitative and quantitative commercial cPL tests have been developed and used in veterinary clinics. This study aimed to compare three different methodologies SNAP cPL, Spec cPL, and Vcheck cPL tests to assess the concordance of these assays. Methods: Fifty serum samples were collected from 36 dogs with or without pancreatitis and subjected to SNAP cPL, Spec cPL, and Vcheck cPL tests. Agreement and correlation coefficients were calculated between the test results, and correlations were determined during the management of the patients. Results: The results of the three cPL assays were strongly correlated in 47/50 serum samples (94%). Cohen's kappa analysis between the Spec cPL and Vcheck cPL showed near perfect agreement (κ = 0.960, p < 0.001), SNAP cPL and Vcheck cPL (κ = 0.920, p < 0.001), and Spec cPL and SNAP cPL (κ = 0.880, p < 0.001). The correlation coefficients (r) between data from Spec cPL and Vcheck cPL tests was calculated by Spearman's correlation test (r = 0.958, p < 0.001). Furthermore, the patterns of change in serum cPL concentrations determined using Spec cPL and Vcheck cPL were significantly consistent during the monitoring period in 11 patients. Conclusions and Relevance: Our data illustrated that Spec cPL and Vcheck cPL tests are compatible for clinical use in the diagnosis and monitoring of canine pancreatitis.

• Journal of Mushroom
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• v.13 no.1
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• pp.56-62
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• 2015

A lipase producing bacterium was isolated from button mushroom bed, which showing high clear zone on agar media containing Tributyrin as the substrate. The strain was identified as Burkholderia cepacia by analysis of 16S rDNA gene sequence. Crude lipase (CL) was partially purified from 70% ammonium sulfate precipitation using the culture filtrate of B. cepacia. Immobilized lipases were prepared by cross-linking method with CL from B. cepacia and Novozyme lipase (NL) onto silanized Silica-gel as support. Residual activitiy of the immobilized CL (ICL) and immobilized NL (INL) was maintained upto 61% and 72%, respectively. Biodiesel (Fatty acid methyl ester, FAME) was recovered by transesterification and methanolysis of Canola oil using NaOH, CL and ICL as the catalysts to compare the composition of fatty acids and the yield of FAME. Total FAME content was NaOH $781mg\;L^{-1}$, CL $681mg\;L^{-1}$ and ICL $596mg\;L^{-1}$, in which the highest levels of FAME was observed to 50% oleic acid (C18:1) and 22% stearic acid (C18:0). In addition, the unsaturated FAME (C18:1, C18:2) decreased, while saturated FAME (C16:0, C18:0) increased according to increasing the reaction times with both CL and ICL, supporting CL possess both transesterification and interesterification activity. When reusability of ICL and INL was estimated by using the continuous reaction of 4 cycles, the activity of ICL and INL was respectively maintained 66% and 79% until the fourth reaction.

• Fisheries and Aquatic Sciences
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• v.12 no.1
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• pp.16-23
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• 2009

Lipids were extracted from marine rotifer, Brachionus rotundiformis in order to examine the functionality of lipid enzymatic modification. The fatty acids, palmitic, linoleic, oleic and stearic acids were the dominant forms accounting for approximately 35.8%, 21.5%, 15.9% and 7.7% of the total lipid content, respectively. Lipid fractions were categorized as neutral lipids (38.5%), glycolipids (45.9%) and phospholipids (17.6%), and after extraction from the rotifer were isolated by thin-layer chromatography (TLC) as free fatty acids (FFA), monoacylglycerol (MAG), diacylglycerol (DAG) and triacylglycerol (TAG). The production of polyunsaturated fatty acid (PUFA) concentrate from rotifer lipids was studied using lipase-catalyzed hydrolysis. In addition, rotifer lipids were modified by hydrolysis using lipases such as porcine pancreas, Candida rugosa and Rhizomucor miehei. The lipase from Rhizomucor miehei was effective in extracting linoleic acid (C 18:2), while the lipase from Candida rugosa was effective in palmitic acid (C16:0) extraction.

• Journal of the Korean Society of Clothing and Textiles
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• v.36 no.6
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• pp.653-662
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• 2012

This study measured the effect of general pre-treatment on PLA fabrics to confirm the benefits of enzymatic processing on PLA fabrics in the textile industry as well as evaluated the hydrolytic activities of three lipases. The effects of lipase hydrolysis were analyzed through moisture regain, dyeing ability, tensile strength, and surface morphology. As a result, PLA fibers were easily damaged by a low concentration of sodium hydroxide and a low treatment temperature. The optimal treatment conditions of Lipase from Candida cylindracea were pH 8.0, $40^{\circ}C$, and 1,000 U. The optimal treatment conditions for Lipase from Candida rugosa were pH 7.2, $37^{\circ}C$, and 1,000 U. The optimal treatment conditions for Lipase from Porcine pancreas were pH 8.0, $37^{\circ}C$, and 2,000 U. The moisture regain and dyeing ability of PLA fabrics increased and the tensile strength of PLA fabrics decreased. The results of surface morphology revealed that there were some cracks due to hydrolysis on the surface of the fiber.

• The Korean Journal of Food And Nutrition
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• v.24 no.3
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• pp.291-294
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• 2011

A lipase inhibitor from Desmodium oxyphyllum DC. was purified by methanol extraction, systematic solvent extraction, silica gel column chromatography, $C_{18}$ solid phase extraction chromatography and RP-HPLC. We obtained the purified lipase inhibitor with 182 ng($IC_{50}$) of lipase inhibitory activity for a 0.06% yield. Its molecular weight was estimated to be 655.37 Da from an instrumental analysis of MALDI-TOF-MS and it was identified copper-3,5-dibromo-2-hydroxybenzoic acid ($C_{14}H_8Br_4CuO_6$) by $^1H$, $^{13}C$ NMR analysis.

• Microbiology and Biotechnology Letters
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• v.15 no.2
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• pp.122-128
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• 1987

Lipases from Candida rogosa and Rhizopus arrhizus were immobilized by entrapment with photo-crosslinkable resin prepolymer for the study of fat splitting and interesterification in isooctane-two phase system. Dioctylsulfosuccinate was selected as the most suitable surfactant during the immobilization. Lipase entrapped with hydrophobic photo-crosslinkable resin prepolymer(ENTP-3000) exhibited the highest activity, whereas lipase entrapped with hydrophilic gel(ENT-4000) was more stable in organic solvent. As the degree of hydrophobicity of the immobilization matrix was increased, Vm(app) of the lipase entrapped was increased, but Km(app) was approximately constant. While the optimum pH of the lipases entrapped on hydrophilic gel (ENT-4000) were around pH 7.0 for Candida lipase and Rhizopus lipase, the reaction rate of the lipases entrapped on hydrophobic gel were less dependent on pH variations for short reaction time. However, for longer reaction time, the lipnses from C. rugosa and R. arrhizus entrapped on hydrophobic gel yielded maximum rate at pH 6.0 and 6.5, respectively, Entrapment method endowed the lipase with thermal stability.

• Food Science and Preservation
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• v.17 no.5
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• pp.727-732
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• 2010

Activity-guided fractionation of an ethyl acetate (EtOAc)-soluble portion of an ethanolic extract from the leaves of cultivated mountain ginseng, using pancreatic lipase inhibition assay, led to the isolation and identification of three flavonoids of a previously described structure, kaempferol-3-O-sophoroside (I), kaempferol-3-O-${\beta}$-Dglucopyranoside (astragalin, II) and kaempferol (III). All compounds (I.III) showed pancreatic lipase inhibitory activities, with $IC_{50}$ values ranging from $20.3{\pm}2.2$ to $9.1{\pm}1.5$ ${\mu}M$, kaempferol (III) showed the most potent inhibitory activity with an $IC_{50}$ of $9.1{\pm}1.5$ ${\mu}M$. The level of activity may depend on the number of C-3 glucosyl group(s) linked to the kaempferol backbone, and the isolated compounds may have promise as pancreatic lipase inhibitors.