• Applied Biological Chemistry
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• v.14 no.3
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• pp.207-212
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• 1971

An enzyme preparation from a newly isolated Candida that showed a high lipase activity was subjected to examination of its reaction rate under various conditions. The original and a diluted enzyme solutions showed the zero order curve starting at the point of 50 minutes in time. When PVA was used as an emulsifyer more activity was observed than the case of gum arabic. The optimal temperature and pH were $37{\sim}40^{\circ}C$ and 7.0, respectively. Oleic acid as a fatty acid conferred on the enzyme an inhibitory action while calcium ion a positive one. Sodium cholate yielded an increase in reaction rate at the first stage.

• Food Science and Biotechnology
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• v.14 no.3
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• pp.368-370
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• 2005

Citronellyl valerate was synthesized by a lipase from a Rhizopus sp strain isolated and the lipase produced, at UNICAMP, Brazil. Direct esterification was performed in a solvent-free medium to produce the flavor ester. Response surface methodology was used to optimize the process with respect to the substrate molar ratio and lipase concentration. The results show that the synthesis of citronellyl valerate can be carried out in a solvent-free medium, the maximum ester conversion rate achieved being 91.5% after 48 hours of reaction time.

• Journal of Microbiology and Biotechnology
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• v.21 no.11
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• pp.1159-1165
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• 2011

Biodiesel is methyl and ethyl esters of long-chain fatty acids produced from vegetable oils or animal fats. Lipase enzymes have occasionally been used for the production of this biofuel. Recently, biodiesel production using immobilized lipase has received increased attention. Through enhanced stability and reusability, immobilized lipase can contribute to the reduction of the costs inherent to biodiesel production. In this study, methanol-tolerant lipase M37 from Photobacterium lipolyticum was immobilized using the cross-linked enzyme aggregate (CLEA) method. Lipase M37 has a high lysine content (9.7%) in its protein sequence. Most lysine residues are located evenly over the surface of the protein, except for the lid structure region, which makes the CLEA preparation yield quite high (~93%). CLEA M37 evidences an optimal temperature of $30^{\circ}C$, and an optimal pH of 9-10. It was stable up to $50^{\circ}C$ and in a pH range of 4.0-11.0. Both soluble M37 and CLEA M37 were stable in the presence of high concentrations of methanol, ethanol, 1-propanol, and n-butanol. That is, their activities were maintained at solvent concentrations above 10% (v/v). CLEA M37 could produce biodiesel from olive oil and alcohols such as methanol and ethanol. Additionally, CLEA M37 generated biodiesel via both 2-step methanol feeding procedures. Considering its physical stability and reusability, CLEA M37 may potentially be used as a catalyst in organic synthesis, including the biodiesel production reaction.

• KSBB Journal
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• v.18 no.1
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• pp.1-7
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• 2003

Screening of a strain was carried out to produce an enantioselective lipase toward the precursor of ltraconazole as azole-containg compounds, which are well known as antifungal drug agents. An Acinetobacter sp. SY-01 strain which can selectively hydrolyze the racemic substrates was isolated and the racemic substrate was resolved to the S-ester in 95.6% enantiomeric excess after 74.8% hydrolysis. The optimum temperature and pH for the conversion were $50^{\circ}C$, pH 7.0. However, the temperature and pH had no effect on the enantiomeric excess. Addition of solvents decreased the conversion and slightly increased the enantiomeric excess. However, the kind of solvents had no effect on enantiomeric excess. The substrate concentration decrease enantiomeric excess and this is confirmed by the products generated from hydrolysis, and also enantiomeric excess could be increased by the removal of reaction products.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1183-1188
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• 2005

Several lipases of commercial grade were screened to catalyze the methanolysis of castor oil, and an immobilized Candida antarctica (Novozym 435) had the highest activity among the lipases tested. To enhance the yield of methyl ricinoleate, several reaction parameters were optimized. The optimum temperature was $50^{\circ}C$, and the original water content of lipase was sufficient to maintain the activity of lipase, and additional water supplied inhibited the methanolysis of castor oil. Because the lipase was deactivated by methanol, the reaction was tested by three-step addition of 1 molar equivalent of methanol to the oil. However, the oil was not completely converted to its methyl esters. The final reaction mixture using 3 molar equivalents of methanol to the oil consisted of $70\%$ methyl ricinoleate, $18\%$ monoricinoleate, $11\%$ diricinoleate, and trace triricinoleate at the equilibrium state. The yield of methyl ricinoleate was $97\%$ at 6 molar ratio of methanol to the oil with 300g of castor oil and 6g of immobilized Candida antarctica at $50^{\circ}C$ within 24 h.

• Korean Journal of Food Science and Technology
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• v.17 no.2
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• pp.75-80
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• 1985

To utilize lipase obtained from Candida cylindracea for lipid hydrolysis, methods to immobilize lipase by adsorption and reaction characteristics of the immobilized lipase by adsorption were investigated. Among the tested adsorbents, silica gel was selected as a suitable adsorbent. The optimum condition for adsorption of lipase was when 47.5 units of lipase were adsorbed to 1.6g of silica gel at pH7.0 and $5^{\circ}C$ for 100 min. Optimum pH and temperature for activity of the immobilized lipase were at $37^{\circ}C$ and pH7.0, which were same as the soluble lipase. Optimum enzyme concentration of the immobilized lipase were 30g for milk fat and 80g for olive oil, whereas those of the soluble lipase were 800 units for milk fat and 1200 units for olive oil. The optimum substrate concentrations of the immobilized and soluble lipases were 20% lipid, regardless of lipid types. Rapid hydrolysis of milk fat was observed with the soluble lipase for the initial 4 hours and with the immobilized lipase for the initial 8 hours. The immobilized lipase produced same amount of capric acid as the soluble lipase, but more myristic acid and less butyric acid than the soluble lipase.

• Korean Chemical Engineering Research
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• v.47 no.5
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• pp.546-552
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• 2009

Enzymatic ethanolysis of wheat germ oil with immobilized lipase was investigated for enhancing the function of wheat germ oil. Ethanolysis reactions were carried out in two different systems; non-pressurized and pressurized system. In non-pressurized system, the enzymatic ethanolysis was carried out in an erlenmeyer flask(25 ml) containing a mixture of wheat germ oil and 99.90% ethanol using 1~5 wt% immobilized lipase as Lipozyme TL-IM and Lipozyme RM-IM and the reaction mixtures were incubated at $40{\sim}70^{\circ}C$ with 120 rpm shaking. In pressurized system, the enzymatic ethanolysis was carried out at various condition; immobilized lipase concentration(2 wt%), reaction time(24 h), reaction temperature($40{\sim}60^{\circ}C$) and reaction pressure(75, 100, 150, 200 bars). The samples obtained from each fraction were analyzed by HPLC for analysing contents of monoglyceride, diglyceride, and triglyceride. The conversion of wheat germ oil relied on the reaction temperature and the concentration of immobilized lipase. The optimum condition of enzymatic ethanolysis in non-pressurized and pressurized systems was at $50^{\circ}C$ and 100 bar.

• Journal of the Korean Applied Science and Technology
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• v.22 no.2
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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.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.1
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• pp.99-102
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• 2005

Organic solvents are widely used in biotransformation systems. There are many efforts to reduce the consumption of organic solvents because of their toxicity to the environment and human health. In recent years, several groups have started to explore novel organic solvents called room temperature ionic liquids in order to substitute conventional organic solvents. In this work, lipase-catalyzed transesterification in several uni- and bi-phasic systems was studied. Two representative hydrophobic ionic liquids based on 1-butyl-3-methylimidazolum coupled with hexafluorophosphate ([BMIM][$PF_6$]) and bis[{trifluoromethylsulfonyl} imide] ([BMIM] [$Tf_{2}N$]) were employed as reaction media for the transesterification of n-butanol. The commercial lipase, Novozym 435, was used for the transesterification reaction with vinyl acetate as an acyl donor. The conversion yield was increased around $10\%$ in a water/[BMIM][$Tf_{2}N$] bi-phasic system compared with that in a water/hexane system. A higher distribution of substrates into the water phase is believed to enhance the conversion yield in a water/[BMIM][$Tf_{2}N$] system. Partition coefficients of the substrates in the water/[BMIM][$Tf_{2}N$] bi-phasic system were higher than three times that found in the water/hexane system, while n-butyl acetate showed a similar distribution in both systems. Thus, RTILs appear to be a promising substitute of organic solvents in some biotransformation systems.

• Bulletin of the Korean Chemical Society
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• v.31 no.3
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• pp.650-652
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• 2010

A lipase from Pseudomonas cepacia was immobilized onto single walled carbon nanotubes (SWNTs) in two different ways in each of two solvent systems (buffer and ionic liquid). The most efficient immobilization was achieved in ionic liquid (1-butyl-3-methylimidazolium tetrafluoroborate, BMIM-$BF_4$). In this procedure, carbon nanotubes were first functionalized noncovalently with 1-pyrenebutyric acid N-hydroxysuccinimide ester and then subject to the coupling reaction with the lipase in ionic liquid. The resulting immobilized enzyme displayed the highest activity in the transesterification of 1-phenylethyl alcohol in the presence of vinyl acetate in toluene.