• Microbiology and Biotechnology Letters
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• v.28 no.4
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• pp.223-227
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• 2000

A novel two-step acetone treatment method was developed to enhance the enantioselectivity of Candida rugosa lipase (CRL) toward the hydrolysis of racemic naproxen methyl ester. The acetone-teated CRL was considerably more enantioselective than the crude CRL, yielding an enantiomeric excess of 98~100%. The crude and acetone-treated CRLs were subjected to anion exchange chromatography, and their chromatography profiles were compared. In consequence, both chromatography profiles were found to be almost identical, resulting in two separate lipase peaks (lipase A and B). The lipase B, which is known to be less enantioselective, was treated with acetone using a two-step treatment method. The enantioselectivity of acetone-treated lipase B was dramatically increased, yielding an enantiomeric excess of 99%.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.6
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• pp.522-525
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• 2006

Biodiesel conversion from soybean oil reached a maximum of 70% at 18 h using immobilized 1,3-specific Rhizopus oryzae lipase alone. Biodiesel conversion failed to reach 20% after 30 h when immobilized nonspecific Candida rugosa lipase alone was used. To increase the biodiesel production yield, a mixture of immobilized 1,3-specific R. oryzae lipase and nonspecific C. rugosa lipase was used. Using this mixture a conversion of greater than 99% at 21 h was attained. When the stability of the immobilized lipases mixture was tested, biodiesel conversion was maintained at over 80% of its original conversion after 10 cycles.

• KSBB Journal
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• v.14 no.2
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• pp.225-229
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• 1999

Enzymatic resolution reactions were investigated using Candida rugosa lipase for the production of potically pure S-(+)-Ketoprofen. When the enzymatic hydroysis (and esterification) of recemic ketoprofen esters (and recemic ketoprofen with alcohol) was investigated comparatively, aqueous media was more specific for S-enantiomer than organic media. In the enzymatic hydrolysis of racemic ketoprofen ethyl ester in aqueous media, optimal temperature and pH for enantioselectivity were $37^{\circ}C$ and 4, respectively. The stereoselectivity of the enzyme was enhanced by adding dialcohols such as ethylene glycol and propylene glycol. The enantiomeric ratio obtained in the 40 %(v/v) ethylene glycol was 2-fold higher than that without the additive. By adding $CH_2Cl_2$, $CHCl_3$ and $CCl_4$ (5%,v/v), the enantioselectivity was reversed. A dramatic increase in the stereoselectivity was achieved using lipase purified by anion exchange chromatography. The type A lipase(the first eluted lipase fraction) showed an enantiomeric ratio of >100, whereas the type B lipase(the second eluted lipase fraction) exhibited enantimomer ratio of 9.0 in the hydrolysis of racemic ketoprofen ethyl ester.

• Analytical Science and Technology
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• v.29 no.3
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• pp.105-113
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• 2016

The present study investigated the immobilization of lipases on silica nanoparticles and silica-coated magnetite nanoparticles as supports with a functional group to enhance the stability of lipase. The influence of functional groups, such as the epoxy group and the amine group, on the activity and stability of immobilized lipase was also studied. The epoxy group and the amino group were introduced onto the surface of nanoparticles by glycidyl methacrylate and aminopropyl triethoxysilane, respectively. Immobilized Candida rugosa lipase on silica nanoparticles and silica-coated magnetite nanoparticles with a functional group showed slightly lower initial enzyme activities than free enzyme; however, the immobilized Candida rugosa lipase retained over 92 % of the initial activity, even after 3 times reuse. Lipase was also immobilized on the silica-coated magnetite nanoparticles by cross-linked enzyme aggregate (CLEA) using glutaraldehyde and covalent binding, respectively, were also studied. Immobilized Candida rugosa lipase on silica nanoparticles and silica-coated magnetite nanoparticles by CLEA and covalent binding showed higher enzyme activities than free enzyme, while immobilized Candida rugosa lipase retained over 73 % of the initial activity after 5 times reuse.

• KSBB Journal
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• v.17 no.6
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• pp.543-548
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• 2002

The enantioselective esterification of racemic ibuprofen catalyzed by a Candida rugosa lipase was studied according to reaction conditions such as a lipase concentration, reaction temperature, alcohol chain length and alcohol concentration. The S-(+)-ibuprofen alkyl esters prepared were converted to S-(+)-ibuprofen by hydrolysis with sulfuric acid as a catalyst. High conversions in the esterifications were obtained at 60$^{\circ}C$ and an equimolar ratio of octanol to ibuprofen. The initial reaction rate of the esterification decreased with increasing octanol concentration. Conversion and initial reaction rate increased with increasing alcohol chain length. Values of enantiomeric excess(ee) according to esterification reaction conditions did not change below 60$^{\circ}C$. On the other hand, values of conversion and ee for the chemical hydrolysis of S-(+)-ibuprofen alkyl esters were independent of alcohol alkyl chain length. Optical resolution of racemic ibuprofen was achieved by lipase catalyzed esterification and chemical hydrolysis. The separation method provided a high yield and enantioselectivity for the production of S-(+)-ibuprofen from racemic ibuprofen.

• 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.

• Journal of Microbiology and Biotechnology
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• v.18 no.12
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• pp.1927-1931
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• 2008

In this study, the enzymatic process for biodiesel production was optimized using a mixture of immobilized Rhizopus oryzae and Candida rugosa lipases. The optimal temperature and agitation speed for biodiesel production were $45^{\circ}C$ and 300 rpm, respectively. The optimal ratio of R. oryzae and C. rugosa lipases in the mixture was 3:1 (w:w). When 3 mmol of methanol was the initial reaction medium and 3 mmol of methanol was added every 1.5 h during biodiesel production, biodiesel conversion was over 98% at 4 h. In addition, when the immobilized lipase mixture was reused, biodiesel conversion exceeded 80% after 5 reuses.

• 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 Microbiology and Biotechnology
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• v.18 no.4
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• pp.735-741
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• 2008

In our previous study, a surfactant-coated Candida rugosa lipase immobilized in microemulsion-based organogels was exploited for the synthesis of ethyl isovalerate. In the present study, we are focusing on the effective reuse of lipase immobilized in microemulsion-based organogels (MBGs) in terms of retainment of the catalytic activity. As water is one of the co-products in esterification reactions, the removal of water becomes a priority to allow the reaction to work in the forward direction and to prevent back hydrolysis. Taking this fact into consideration, the lipase-containing microemulsion-based organogels were given pretreatment and/or several intermittent treatments with dry reverse micellar solution of AOT in organic solvent during repeated cycles of ester synthesis. The pretreated MBGs with dry reverse micellar solution exhibited lower water content and higher initial rates of esterification in comparison with untreated freshly prepared MBGs. The esterification efficiency of untreated MBGs started decreasing after 5 cycles of reuse and was almost completely lost by the end of the $8^{th}$ cycle. In contrast, pretreated MBGs exhibited a gradual decrease in esterification efficiency after 5 cycles and retained about 80% of the initial activity at the end of the $8^{th}$ cycle. The intermittent treatment of MBGs after every 3 cycles resulted in enhanced reusability of immobilized lipase for up to 9 cycles without significant loss in esterification activity, after which it resulted in a slow decrease in activity with about 27% lower activity at the end of the $12^{th}$ cycle. Furthermore, the treatment conditions such as concentration of AOT in liquid dessicant and time of treatment were optimized with respect to our system. The granulated MBGs proved to be better in terms of initial esterification rates (1.2-fold) as compared with the pelleted MBGs.

• Korean Journal of Chemical Engineering
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• v.35 no.11
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• pp.2220-2231
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• 2018

Lipase from Candida rugosa was immobilized on $MgO{\cdot}SiO_2$ hybrid grafted with amine, thiol, cyano, phenyl, epoxy and carbonyl groups. The products were analyzed using Fourier transform infrared spectroscopy, nuclear magnetic resonance, low-temperature $N_2$ sorption and elemental analysis. Additionally, the degree of coverage of the oxide material surface with different functional groups and the number of surface functional groups were estimated. The Bradford method was used to determine the quantity of immobilized enzyme. The largest quantity of enzyme (25-28 mg/g) was immobilized on the hybrid functionalized with amine and carbonyl groups. On the basis of hydrolysis reaction of p-nitrophenyl palmitate to p-nitrophenol, it was determined how the catalytic activity of the obtained biocatalysts is affected by pH, temperature, storage time, and repeated reaction cycles. The best results for catalytic activity were obtained for the lipase immobilized on $MgO{\cdot}SiO_2$ hybrids with amine and carbonyl groups. The biocatalytic system demonstrated activity above 40% in the pH range 4-10 and in the temperature range $30-70^{\circ}C$. Lipase immobilized on the $MgO{\cdot}SiO_2$ systems with amine and epoxy groups retains, respectively, around 80% and 60% of its initial activity after 30 days of storage, and approximately 60-70% after 10 reaction cycles.