• KSBB Journal
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• v.14 no.4
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• pp.511-516
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• 1999

The effects of environmental and compositon factors, such as reaction time, metal ions, pH, agitation speed, the weight ratio of water to oil, and the weight of enzyme, on the hydrolysis of oils by Lipase-OF were investigated. In case of oils with low melting point, the optimum temperature of hydrolysis were the enzyme activity was maximum was 37$^{\circ}C$. However, when the melting temperature was higher than 4$0^{\circ}C$, the optimum temperature was around the fusion temperature. The activity of Lipase-OF decreased very rapidly with increase of temperature in the range of higher than 45$^{\circ}C$ and the activity perished above $65^{\circ}C$. The effect of agitation speed was investigated from 150 to 650 rpm. The hydrolysis of oils increased as the agitation speed increased up to 350 rpm, but it did not increase any more above 350 rpm. The weight ratio of water to oil was changed from 1 : 9 to 9 : 1 for the investigation of the effect on the hydrolusis. The weight ratio for maximum hydrolysis was 1 : 1. $Ca^{2+}\;and\;Mg^{2+}$ among various metal ions had some effect on the stimulation of hydrolysis. The optimum concentration of the ions was about 100ppm at which the hydrolysis increased, compared with that of distilled water, by 2 to 3%. The Optimum pH of Lipase-OF was 7. The hydrolysis decreased as the pH decreased as the pH decreased and also decreased as the pH increased. The content of enzyme affected the hydrolysis of oil. The hydrolysis increased with the content of Lipase-OF in the range of less than 0.013 wt% of substrate. However, the increase of hydrolysis with the content of Lipase-OF ceased above 0.013 wt%. The experiments investigating the effect of environmental and composition factors on the hydrolysis of oils showed that the optimum temperature was 37$^{\circ}C$, the pH 7, the concentration of $Ca^{2+}\;or\;Mg^{2+}$ 100 ppm, the agitation speed 350 rpm, the weight ratio of water to oil 1 : 1, and the content of Lipase-OF 0.013 wt% of substrate.

• Microbiology and Biotechnology Letters
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• v.24 no.2
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• pp.213-216
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• 1996

Optically active carboxylic acid, D-(-)-$\beta$-hydroxyisobutyric acid {(D)-(-)-HIBA} is a useful chiral starting material for the preparation of enantiomerically pure bioactive compounds which have a chiral methyl carbon center in the molecule such as captopril, $\alpha$-tocopherol, erythromycin A, muscone and so on. (S)-3-Acetoxy-2-methylpropanol can be used as the precursor of (D)-(-)-HIBA, that is, chemical oxidation of the hydroxyl group and subsequent hydrolysis of acyl group in (S)-3-acetoxy-2-methylpropanol affords D-(-)-$\beta$-hydroxyisobutyric acid. (S)-3-Acetoxy-2-methyl-propanol was prepared by lipase-catalyzed asymmetric hydrolysis. In the enzymatic hydrolysis system, lipase AY (Candida rugosa) provided the expected (S)-3-acetoxy-2-methylpropanol in 60% e.e. of the enantiomeric purity under the phosphate buffer and organic co-solvent system.

• 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%.

• KSBB Journal
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• v.23 no.6
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• pp.509-512
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• 2008

The hydrolysis reaction of soybean oil was conducted experimentally by various source enzymes. The analytical result of hydrolysate of soybean oil showed that the compositions were linoleic acid, olic acid, palmitic acid, and stearic acid in order. The enzymes CR-E and CC-E from Candida rufosa and Candida cylindracea had two hold or more hydrolysis conversions than those of Lipase 16, Novozyme 871, and Lipolase-100L under the same conditions. Therefore CR-E and CC-E were selected for further experiments. These two enzymes had similar ranges of optimun conditions as follows: pH 3-6, $35-45^{\circ}C$, and water to soybean oil ratio of 3.3 or above. They finally got conversions 95% above.

• 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.

• Applied Biological Chemistry
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• v.23 no.1
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• pp.52-57
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• 1980

This study was conducted to clarify the mode of action and the positional specificity of Trichosporon cutaneum lipase during the course of hydrolysis of triolein and monoolein mixture by thin-layer chromatography. 1. The hydrolytic activity of the lipase to oleyl glycerides was in the order of triolein>diolein>monoolein. 2. Both of triolein and diolein were hydrolyzed by the lipase at high and almost the same rate. 3. The hydrolysis of monoolein by the lipase was very slow compared to the other two oleyl glycerides. 4. This lipase appeared to have a very low specificity toward the outer chains of triolein.

• Bulletin of the Korean Chemical Society
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• v.33 no.2
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• pp.409-414
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• 2012

Xanthorrhizol is a bisabolane type of natural sesquiterpene, the major component of essential oils of Curcuma xanthorrhiza. 2-(3-Methoxy-4-methylphenyl)propan-1-ol and 2-(3-hydroxy-4-methyl phenyl)propan-1-ol could be essential building block for enantioselective synthesis of xanthorrhizol. Enantioselective (c = 53%, E = $80{\pm}3$) for R-(+)-2-(3-hydroxy-4-methylphenyl) propan-1-ol and (c = 58%, E = $27{\pm}1$) for R-(+)-2-(3-methoxy-4-methylphenyl) propan-1-ol resolution processes were developed via lipase-catalyzed reaction. We found lipase Aspergillus oryzae (AOL) and Porcine pancreas (PPL) are selective to transesterification and hydrolysis in organic and aqueous phase. Modified demethylated substrate is appropriate for enantioselective hydrolysis reaction without any additives. Enantiopure chiral alcohol was crystallized from ethyl acetate/n-hexane co-solvent system. Gram scale resolved chiral intermediate will facilitate the synthesis of the unnatural S-(+)-xanthorrhizol, the corresponding isomer of the natural one.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.367-370
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• 2002

Candida cylindracea lipase was immobilized by adsorption on acid washed glass beads. It was observed that protein loading of the support depends on the size of the particle, with smaller particle containing higher amount of protein per unit weight. Initial reaction rate linearly varied up to enzyme concentration of 17.25 U/mL. Amount of free fatty acids produced was linearly proportional up to the enzyme loading of 1650 $\mu$g/g of bead. Achievement of chemical equilibrium took longer time in the case of less protein loading. Degree of hydrolysis was found to decrease in second and third consecutive batch operations on repeated use of immobilized lipase.

• Journal of Microbiology and Biotechnology
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• v.11 no.6
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• pp.951-956
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• 2001

A 28-kDa extracellular lipase (pI 8.7) was purified to homogeneity from the culture supernatant of Trichosporon sp. Y- 11 by mmonium sulfate precipitation, DEAE-Sephadex A-50, Bio-Gel P-30, CM- Sephadex C-50, and Bio-Gel P- 10 chromatographies. The purified enzyme exhibited a specific activity of $2,741{\;}{\mu}mol/min/mg$ based on the hydrolysis of triolein, and the optimal hydrolysis activity was dentified at pH 8.0 and $40^{\circ}C$. The enzyme activity was inhibited by $Ag^+$ and enhanced by $Fe^{2+}$, $Fe^{3+}$, $Mg^{2+}$, $Mn^{2+}$, and $Li^{+}$. The enzyme activity exhibited for the hydrolysis of both tributyrin and trilinolein. The ester synthesis of unsaturated fatty acids with various alcohols catalyzed by the purified lipase in a nonaqueous medium or microaqueous system was also investigated. The esterification activity of the lipase increased with an increase of the carbon chain length in the alcohol. The synthesis rate of linoleic acid and oleyl alcohol was the highest with an optimal temperature and pH of $40^{\circ}C$ and 8.0, respectively. The water content and agitation also affected the esterification activity of the lipase.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.523.2-523
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• 1986

The hydrolysis of olive oil was attempted with immobilized C. rugosa lipase in the reverse phase solvent system. (i.e. immobilized wet particles is dispersed in continuous phase olive oil or organic solvents containing olive oil). Sephadex LH-20 and LH-60 were used as the supports that can be used in organic solvents. The water content of wet particles of sephadex LH-20 and LH-60 were about 72% (w/w) and 85% (w/w), respectively Both swollen gels with 0.05M buffers adsorbed about 18% of lipase dissolved. They were easily dispersed in liquid olive oil or in organic solvents. The effects of organic solvents on the stability and catalytic activity of the lipase have been examined. The results revealed that isooctane is superior to the other solvents examined for enzymatic fat spliting in reverse phase system. Kinetics of enzymatic hydrolys of olive oil by immobilized lipase has been investigated in a batch reactor. Effects of pH and temperature on the lipase were studied. The substrate concentration was influenced positively on the thermal stability.