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1.
Yomi Watanabe Toshihiro Nagao Yutaka Nishida Yoshiaki Takagi Yuji Shimada 《Journal of the American Oil Chemists' Society》2007,84(11):1015-1021
Acid oil, a by-product of vegetable oil refining, was enzymatically converted to fatty acid methyl esters (FAME). Acid oil
contained free fatty acids (FFA), acylglycerols, and lipophilic compounds. First, acylglycerols (11 wt%) were hydrolyzed at
30 °C by 20 units Candida rugosa lipase/g-mixture with 40 wt% water. The resulting oil layer containing 92 wt% FFA was used for the next reaction, methyl
esterification of FFA to FAME by immobilized Candida antarctica lipase. A mixture of 66 wt% oil layer and 34 wt% methanol (5 mol for FFA) were shaken at 30 °C with 1.0 wt% lipase. The degree
of esterification reached 96% after 24 h. The resulting reaction mixture was then dehydrated and subjected to the second esterification
that was conducted with 2.2 wt% methanol (5 mol for residual FFA) and 1.0 wt% immobilized lipase. The degree of esterification
of residual FFA reached 44%. The degree increased successfully to 72% (total degree of esterification 99%) by conducting the
reaction in the presence of 10 wt% glycerol, because water in the oil layer was attracted to the glycerol layer. Over 98%
of total esterification was maintained, even though the first and the second esterification reactions were repeated every
24 h for 40 days. The enzymatic process comprising hydrolysis and methyl esterification produced an oil containing 91 wt%
FAME, 1 wt% FFA, 1 wt% acylglycerols, and 7 wt% lipophilic compounds. 相似文献
2.
The purification of tocopherols and phytosterols (referred to as sterols) from soybean oil deodorizer distillate (SODD) was
attempted. Tocopherols and sterols in the SODD were first recovered by short-path distillation, which was named sODD tocopherol/sterol
concentrate (SODDTSC). The SODD-TSC contained MAG, DAG, FFA, and unidentified hydrocarbons in addition to the two substances
of interest. It was then treated with Candida rugosa lipase to convert sterols to FA steryl esters, acylglycerols to FFA, and FFA to FAME. Methanol (MeOH), however, inhibited
esterification of the sterols. Hence, a two-step in situ reaction was conducted: SODDTSC was stirred with 20 wt% water and 200 U/g mixture of C. rugosa lipase at 30°C, and 2 moles of MeOH per mole of FFA was added to the reaction mixture after 16h. The lipase treatment for
40 h in total achieved 80% conversion of the initial sterols to FA steryl esters, complete hydrolysis of the acylglycerols,
and a 78% decrease in the initial FFA content by methyl esterification. Tocopherols did not change throughout the process.
To enhance the degree of steryl and methyl esterification, the reaction products, FA steryl esters and FAME, were removed
by short-path distillation, and the resulting fraction containing tocopherols, sterols, and FFA was treated with the lipase
again. Distillation of the reaction mixture purified tocopherols to 76.4% (recovery, 89.6%) and sterols to 97.2% as FA steryl
esters (recovery, 86.3%). 相似文献
3.
Conversion of oleic acid phytosteryl esters (OASE) to free phytosterols (referred to as sterols) by an enzymatic process was
attempted. Enzymatic hydrolysis of OASE reached a steady state at 55–60% hydrolysis, but addition of methanol (MeOH) significantly
accelerated the conversion of OASE to sterols. Screening of commercially available enzymes indicated that Pseudomonas aeruginosa lipase was most effective for the conversion. Based on the study of several factors affecting the reaction, the optimal conditions
were determined as follows: ratio of OASE to MeOH, 1∶2 (mol/mol); water content, 10 wt%; lipase amount, 20 U/g by weight of
reaction mixture; temperature, 30°C. When the reaction was conducted for 48 h with stirring, the conversion reached 98%. FAME
accumulated in the reaction mixture, but FFA did not, indicating that the FAME was poorly recognized as a substrate in the
reverse conversion of sterols to OASE but the FFA was easily recognized as a substrate. The high conversion of OASE to sterols
was therefore due to elimination of FFA from the reaction system. After the enzymatic reaction, the oil layer was fractionated
at −20°C with 5 vol parts of n-hexane. Sterols were efficiently purified in the resulting precipitate (92% recovery, 99% purity). 相似文献
4.
Yomi Watanabe Yuji Shimada Akio Sugihara Hideo Noda Hideki Fukuda Yoshio Tominaga 《Journal of the American Oil Chemists' Society》2000,77(4):355-360
Candida antarctica lipase is inactivated in a mixture of vegetable oil and more than 1∶2 molar equivalent of methanol against the total fatty
acids. We have revealed that the inactivation was eliminated by three successive additions of 1∶3 molar equivalent of methanol
and have developed a three-step methanolysis by which over 95% of the oil triacylglycerols (TAG) were converted to their corresponding
methyl esters (ME). In this study, the lipase was not inactivated even though 2∶3 molar equivalent of methanol was present
in a mixture of acylglycerols (AG) and 33% ME (AG/ME33). This finding led to a two-step methanolysis of the oil TAG: The first-step
was conducted at 30°C for 12 h with shaking in a mixture of the oil, 1∶3 molar equivalent of methanol, and 4% immobilized
lipase; the second-step reaction was done for 24 h after adding 2∶3 molar equivalent of methanol (36 h in total). The two-step
methanolysis achieved more than 95% of conversion. When two-step reaction was repeated by transferring the immobilized lipase
to a fresh substrate mixture, the enzyme could be used 70 cycles (105 d) without any decrease in the conversion. From the
viewpoint of the industrial production of biodiesel fuel production, the two-step reaction was conducted using a reactor with
impeller. However, the enzyme carrier was easily destroyed, and the lipase could be used only several times. Thus, we attempted
flow reaction using a column packed with immobilized Candida lipase. Because the lipase packed in the column was drastically inactivated by feeding a mixture of AG/ME33 and 2∶3 molar
equivalent of methanol, three-step flow reaction was performed using three columns packed with 3.0 g immobilized lipase. A
mixture of vegetable oil and 1∶3 molar equivalent of methanol was fed into the first column at a constant flow rate of 6.0
mL/h. The eluate and 1∶3 molar equivalent of methanol were mixed and then fed into the second column at the same flow rate.
The final step reaction was done by feeding a mixture of eluate from the second column and 1∶3 molar equivalent of methanol
at the same flow rate. The ME content in the final-step eluate reached 93%, and the lipase could be used for 100 d without
any decrease in the conversion. 相似文献
5.
Enzymatic conversion of waste edible oil to biodiesel fuel in a fixed-bed bioreactor 总被引:9,自引:0,他引:9
Yomi Watanabe Yuji Shimada Akio Sugihara Yoshio Tominaga 《Journal of the American Oil Chemists' Society》2001,78(7):703-707
The conversion of waste edible oil to biodiesel fuel in a fixed-bed bioreactor was investigated. Three-step methanolysis of
waste oil was conducted using three columns packed with 3 g of immobilized Candida antarctica lipase. A mixture of waste oil and 1/3 molar equivalent of methanol against total fatty acids in the oil was used as substrate
for the first-step reaction, and mixtures of the first- and second-step eluates and 1/3 molar equivalent of methanol were
used for the second- and third-step reactions, respectively. Ninety percent of waste oil was converted to the corresponding
methyl esters (ME) by feeding substrate mixtures into the first, second, and third reactors at flow rates of 6, 6 and 4 mL/h,
respectively. We also attempted one-step methanolysis of waste oil. When a mixture of waste oil and 90% ME-containing eluate
(1∶3, wt/wt) and an equimolar amount of methanol against total fatty acids in the waste oil was fed into a reactor packed
with 3 g of immobilized C. antarctica lipase at a flow rate of 4 mL/h, the ME content in the eluate reached 90%. The immobilized biocatalyst could be used for
100 d in the two reaction systems without significant decrease in its activity. Waste oil contained 1980 ppm water and 2.5%
free fatty acids, but these contaminants had little influence on enzymatic production of biodiesel fuel. 相似文献
6.
Conversion of vegetable oil to biodiesel using immobilized Candida antarctica lipase 总被引:11,自引:8,他引:3
Yuji Shimada Yomi Watanabe Taichi Samukawa Akio Sugihara Hideo Noda Hideki Fukuda Yoshio Tominaga 《Journal of the American Oil Chemists' Society》1999,76(7):789-793
Biodiesel derived from vegetable oils has drawn considerable attention with increasing environmental consciousness. We attempted
continuous methanolysis of vegetable oil by an enzymatic process. Immobilized Candida antarctica lipase was found to be the most effective for the methanolysis among lipases tested. The enzyme was inactivated by shaking
in a mixture containing more than 1.5 molar equivalents of methanol against the oil. To fully convert the oil to its corresponding
methyl esters, at least 3 molar equivalents of methanol are needed. Thus, the reaction was conducted by adding methanol stepwise
to avoid lipase inactivation. The first step of the reaction was conducted at 30°C for 10 h in a mixture of oil/methanol (1:1,
mol/mol) and 4% immobilized lipase with shaking at 130 oscillations/min. After more than 95% methanol was consumed in ester
formation, a second molar equivalent of methanol was added and the reaction continued for 14 h. The third molar equivalent
of methanol was finally added and the reaction continued for 24 h (total reaction time, 48 h). This three-step process converted
98.4% of the oil to its corresponding methyl esters. To investigate the stability of the lipase, the three-step methanolysis
process was repeated by transferring the immobilized lipase to a fresh substrate mixture. As a result, more than 95% of the
ester conversion was maintained even after 50 cycles of the reaction (100 d). 相似文献
7.
Lipase-catalyzed interesterification between fish oil and medium-chain TAG has been investigated in a packedbed reactor with
a commercially immobilized enzyme. The enzyme, a Thermomyces lanuginosa lipase immobilized on silica by granulation (lipozyme TL IM; Novozymes A/S, Bagsvaerd, Denmark), has recently been developed
for fat modification. This study focuses on the new characteristics of the lipase in a packed-bed reactor when applied to
interesterification of TAG. The degree of reaction was strongly related to the flow rate (residence time) and temperature,
whereas formation of hydrolysis by-products (DAG and FFA) were only slightly affected by reaction conditions. The degree of
reaction reached equilibrium at 30–40 min residence time, and the most suitable temperature was 60°C or higher with respect
to the maximal degree of reaction. The lipase was stable in a 2-wk continuous operation without adjustment of water content
or activity of the column and the substrate mixture. 相似文献
8.
Purification of docosahexaenoic acid from tuna oil by a two-step enzymatic method: Hydrolysis and selective esterification 总被引:3,自引:0,他引:3
Yuji Shimada Kazuaki Maruyama Akio Sugihara Shigeru Moriyama Yoshio Tominaga 《Journal of the American Oil Chemists' Society》1997,74(11):1441-1446
Purification of docosahexaenoic acid (DHA) was attempted by a two-step enzymatic method that consisted of hydrolysis of tuna
oil and selective esterification of the resulting free fatty acids (FFA). When more than 60% of tuna oil was hydrolyzed with
Pseudomonas sp. lipase (Lipase-AK), the DHA content in the FFA fraction coincided with its content in the original tuna oil. This lipase
showed stronger activity on the DHA ester than on the eicosapentaenoic acid ester and was suitable for preparation of FFA
rich in DHA. When a mixture of 2.5 g tuna oil, 2.5 g water, and 500 units (U) of Lipase-AK per 1 g of the reaction mixture
was stirred at 40°C for 48 h, 83% of DHA in tuna oil was recovered in the FFA fraction at 79% hydrolysis. These fatty acids
were named tuna-FFA-Ps. Selective esterification was then conducted at 30°C for 20 h by stirring a mixture of 4.0 g of tuna-FFA-Ps/lauryl alcohol (1:2, mol/mol), 1.0 g water, and 1,000 U of Rhizopus delemar lipase. As a result, the DHA content in the unesterified FFA fraction could be raised from 24 to 72 wt% in an 83% yield.
To elevate the DHA content further, the FFA were extracted from the reaction mixture with n-hexane and esterified again under the same conditions. The DHA content was raised to 91 wt% in 88% yield by the repeated
esterification. Because selective esterification of fatty acids with lauryl alcohol proceeded most efficiently in a mixture
that contained 20% water, simultaneous reactions during the esterification were analyzed qualitatively. The fatty acid lauryl
esters (L-FA) generated by the esterification were not hydrolyzed. In addition, L-FA were acidolyzed with linoleic acid, but
not with DHA. These results suggest that lauryl DHA was generated only by esterification. 相似文献
9.
Per Langholz Peter Andersen Torben Forskov Walther Schmidtsdorff 《Journal of the American Oil Chemists' Society》1989,66(8):1120-1123
Lipases catalyze the reesterification or hydrolysis of fats and oils and may show specificity towards certain fatty acids.
An immobilized lipase from the fungusMucor miehei shows specificity towards docosahexaenoic acid (DHA) (DHA being a poor substrate), whether the DHA is used as a free fatty
acid (FFA) as substrate in esterification with methanol, or fatty acid methyl ester (FAME) is used as a substrate for hydrolysis.
The specificity of the lipase fromM. miehei may be applied to concentrate DHA originating from a marine oil in either the FAME or the FFA fraction, which can be separated. 相似文献
10.
TAG (MLM) with medium-chain FA (MCFA) at the 1,3-positions and long-chain FA (LCFA) at the 2-position, and TAG (LMM) with
LCFA at the 1(3)-position and MCFA at 2,3(1)-positions are a pair of TAG regioisomers. Large-scale preparation of the two
TAG regioisomers was attempted. A commercially available FFA mixture (FFA-CLA) containing 9-cis, 11-trans (9c, 11t)- and 10t,12c-CLA was selected as LCFA, and caprylic acid (C8FA) was selected as MCFA. The MLM isomer was synthesized by acidolysis of acyglycerols (AG) containing two CLA isomers with
C8FA: A mixture of AG-CLA/C8 FA (1∶10, mol/mol) and 4 wt% immobilized Rhizomucor miehei lipase was agitated at 30°C for 72 h. The ratio of MLM to total AG was 51.1 wt%. Meanwhile, LMM isomer was synthesized by
acidolysis of tricaprylin with FFA-CLA: A mixture of tricaprylin/FFA-CLA (1∶2, mol/mol) and 4 wt% immobilized R. miehei lipase was agitated at 30°C for 24 h. The ratio of LMM to total AG was 51.8 wt%. MLM and LMM were purified from 1,968 and
813 g reaction mixtures by stepwise short-path distillation, respectively. Consequently, MLM was purified to 92.3% with 49.1%
recovery, and LMM was purified to 93.2% with 52.3% recovery. Regiospecific analyses of MLM and LMM indicated that the 2-positions
of MLM and LMM were 95.1 mol% LCFA and 98.3 mol% C8 FA, respectively. The results showed that a process comprising lipase reaction and short-path distillation is effective for
large-scale preparation of high-purity regiospecific TAG isomers. 相似文献
11.
Toshihiro?Nagao Akiko?Kawashima Motoo?Sumida Yomi?Watanabe Kengo?Akimoto Harukazu?Fukami Akio?Sugihara Yuji?Shimada
Two oils containing a large amount of 2-arachidonoyl-TAG were selected to produce structured TAG rich in 1,3-capryloyl-2-arachidonoyl
glycerol (CAC). An oil (TGA58F oil) was prepared by fermentation of Mortierella alpina, in which the 2-arachidonyoyl-TAG content was 67 mol%. Another oil (TGA55E oil) was prepared by selective hydrolysis of a
commercially available oil (TGA40 oil) with Candida rugosa lipase. The 2-arachidonoyl-TAG content in the latter was 68 mol%. Acidolysis of the two oils with caprylic acid (CA) using
immobilized Rhizopus oryzae lipase showed that TGA55E oil was more suitable than TGA58F oil for the production of structured TAG containing a higher
concentration of CAC. Hence, a continuous-flow acidolysis of TGA55E oil was performed using a column (18×125 mm) packed with
10 g immobilized R. oryzae lipase. When a mixture of TGA55E oil/CA (1∶2, w/w) was fed at 35°C into the fixed-bed reactor at a flow rate of 4.0 mL (3.6
g)/h, the degree of acidolysis initially reached 53%, and still achieved 48% even after continuous operation for 90 d. The
reaction mixture that flowed from the reactor contained small amounts of partial acylglycerols and tricaprylin in addition
to FFA. Molecular distillation was used for purification of the structured TAG, and removed not only FFA but also part of
the partial acylglycerols and tricaprylin, resulting in an increase in the CAC content in acylglycerols from 44.0 to 45.8
mol%. These results showed that a process composed of selective hydrolysis, acidolysis, and molecular distillation is effective
for the production of CAC-rich structured TAG. 相似文献
12.
Akiko Kawashima Yuji Shimada Toshihiro Nagao Akihiro Ohara Tsugio Matsuhisa Akio Sugihara Yoshio Tominaga 《Journal of the American Oil Chemists' Society》2002,79(9):871-877
γ-Linolenic acid (GLA) has the physiological functions of modulating immune and inflammatory responses. We produced structured
TAG rich in 1,3-dicapryloyl-2-γ-linolenoyl glycerol (CGC) from GLA-rich oil (GLA45 oil; GLA content, 45.4 wt%), which was
prepared by hydrolysis of borage oil with Candida rugosa lipase having weak activity on GLA. A mixture of GLA45 oil/caprylic acid (CA) (1∶2, w/w) was continuously fed into a fixed-bed
bioreactor (18×180 mm) packed with 15 g immobilized Rhizopus oryzae lipase at 30°C, and a flow rate of 4 g/h. The acidolysis proceeded efficiently, and a significant decrease of lipase activity
was not observed in full-time operation for 1 mon. GLA45 oil contained 10.2 mol% MAG and 27.2 mol% DAG. However, the reaction
converted the partial acylglycerols to structured TAG and tricaprylin and produced 44.5 mol% CGC based on the content of total
acylglycerols. Not only FFA in the reaction mixture but also part of the tricaprylin and partial acylglycerols were removed
by molecular distillation. The distillation resulted in an increase of the CGC content in the purified product to 52.6 mol%.
The results showed that CGC-rich structured TAG can efficiently be produced by a two-step process comprising selective hydrolysis
of borage oil using C. rugosa lipase (first step) and acidolysis of the resulting GLA-rich oil with CA using immobilized R. oryzae lipase (second step). 相似文献
13.
Purification of arachidonic acid (AA) from Mortierella alpina single-cell oil was attempted. The process comprised three steps: (i) preparation of FFA by nonselective hydrolysis of the
oil with Alcaligenes sp. lipase; (ii) elimination of long-chain saturated FA from the resulting FFA by urea adduct fractionation; and (iii) enrichment
of AA through lipase-catalyzed selective esterification with lauryl alcohol (LauOH). In the third step, screening of industrially
available lipases indicated that Burkholderia cepacia lipase (Lipase-PS, Amano Enzyme Inc., Aichi, Japan) acted on AA more weakly than on other FA and was the most effective for
enrichment of AA in the FFA fraction. When the FFA obtained by urea adduct fractionation were esterified with 2 molar equivalents
of LauOH at 30°C for 16 h in a mixture with 20% water and 20 units (U)/g-mixture of Lipase-PS, the esterification reached
39% and the content of AA in the FFA fraction was raised from 61 to 86 wt%. To further increase the content of AA, unesterified
FFA were allowed to react again under the same conditions as those in the first selective esterification except for the use
of 50 U/g Lipase-PS. A series of procedures raised the content of AA to 97 wt% with a 49% recovery based on the initial content
in the single-cell oil. These results indicated that the three-step process for selective esterification with Lipase-PS was
effective for purifying AA from the single-cell oil. 相似文献
14.
Simple alkyl FA esters have numerous uses, including serving as biodiesel, a fuel for compression ignition (diesel) engines.
The use of acid-catalyzed esterification for the synthesis of FAME from acid oil, a by-product of edible vegetable oil refining
that is produced from soapstock, was investigated. Soybean acid oil contained 59.3 wt% FFA, 28.0 wt% TAG, 4.4 wt% DAG, and
less than 1% MAG. Maximum esterification occurred at 65°C and 26 h reaction at a molar ratio of total FA/methanol/sulfuric
acid of 1∶15∶1.5. Residual unreacted species under these conditions, as a fraction of their content in unesterified acid oil,
were FFA, 6.6%; TAG, 5.8%; and DAG, 2.6%. This corresponds to estimated concentrations of FFA, 3.2%; TAG, 1.3%; and DAG, 0.2%,
on a mass basis, in the ester product. In an alternative approach, the acylglycerol species in soapstock were saponified prior
to acidulation. High-acid (HA) acid oil made from this saponified soapstock had an FFA content of 96.2 wt% and no detectable
TAG, DAG, or MAG. Optimal esterification conditions for HA acid oil at 65°C were a mole ratio of FFA/methanol/acid of 1∶1.8∶0.17,
and 14 h incubation. FAME recovery under these conditions was 89% of theoretical, and the residual unesterified FFA content
was approximately 20 mg/g. This was reduced to 3.5 mg/g, below the maximum FFA level allowed for biodiesel, by washing with
NaCl, NaHCO3, and Ca(OH)2 solutions. Alternatively, by subjecting the unwashed ester layer to a second esterification, the FFA level was reduced to
less than 2 mg/g. The acid value of this material exceeded the maximum allowed for biodiesel, but was reduced to an acceptable
value by a brief wash with 0.5 N NaOH. 相似文献
15.
Yuji Shimada Akio Sugihara Masahiro Shibahiraki Hiroyuki Fujita Hirofumi Nakano Toshihiro Nagao Tadamasa Terai Yoshio Tominaga 《Journal of the American Oil Chemists' Society》1997,74(11):1465-1470
γ-Linolenic acid (GLA) was purified from borage oil by a two-step enzymatic method. The first step involved hydrolysis of
borage oil (GLA content, 22.2 wt%) with lipase, Pseudomonas sp. enzyme (LIPOSAM). A mixture of 3 g borage oil, 2 g water, and 5000 units (U) LIPOSAM was incubated at 35°C with stirring
at 500 rpm. The reaction was 91.5% complete after 24 h. The resulting free fatty acids (FFA) were extracted from the reaction
mixture with n-hexane (GLA content, 22.5 wt%; recovery of GLA, 92.7%). The second step involved selective esterification of borage-FFA with
lauryl alcohol by using Rhizopus delemar lipase. A mixture containing 4 g borage-FFA/lauryl alcohol (1:2, mol/mol), 1 g water, and 1000 U lipase was incubated at
30°C for 20 h with stirring at 500 rpm. Under these conditions, 74.4% of borage-FFA was esterified, and the GLA content in
the FFA fraction was enriched from 22.5 to 70.2 wt% with a recovery of 75.1% of the initial content. To further elevate the
GLA content, unesterified fatty acids were extracted, and esterified again in the same manner. By this repeated esterification,
GLA was purified to 93.7 wt% with a recovery of 67.5% of its initial content. 相似文献
16.
Yuji Shimada Norihito Sakai Akio Sugihara Hiroyuki Fujita Yo Honda Yoshio Tominaga 《Journal of the American Oil Chemists' Society》1998,75(11):1539-1544
γ-Linolenic acid (GLA) is a physiologically valuable fatty acid, and is desired as a medicine, but a useful method available
for industrial purification has not been established. Thus, large-scale purification was attempted by a combination of enzymatic
reactions and distillation. An oil containing 45% GLA (GLA45 oil) produced by selective hydrolysis of borage oil was used
as a starting material. GLA45 oil was hydrolyzed at 35°C in a mixture containing 33% water and 250 U/g-reaction mixture of
Pseudomonas sp. lipase; 91.5% hydrolysis was attained after 24 h. Film distillation of the dehydrated reaction mixture separated free
fatty acids (FFA; acid value 199) with a recovery of 94.5%. The FFA were selectively esterified at 30°C for 16 h with two
molar equivalents of lauryl alcohol and 50 U/g of Rhizopus delemar lipase in a mixture containing 20% water. The esterification extent was 52%, and the GLA content in the FFA fraction was
raised to 89.5%. FFA and lauryl esters were not separated by film distillation, but the FFA-rich fraction contaminated with
18% lauryl esters was recovered by simple distillation. To further increase the GLA content, the FFA-rich fraction was selectively
esterified again under similar conditions. As a result, the GLA content in the FFA fraction was raised to 97.3% at 15.2% esterification.
After simple distillation of the reaction mixture, lauryl esters contaminating the FFA-rich fraction were completely eliminated
by urea adduct fractionation. When 10 kg of GLA45 oil was used as a starting material, 2.07 kg of FFA with 98.6% GLA was obtained
with a recovery of 49.4% of the initial content. 相似文献
17.
Roxana Irimescu Yugo Iwasaki Ching T. Hou 《Journal of the American Oil Chemists' Society》2002,79(9):879-883
Regiospecific ethanolysis of homogenous TAG with immobilized Candida antarctica lipase (Novozym 435) was studied using trioleoylglycerol (TO) as a model substrate. Optimization of the reactant weight ratio
revealed that the 2-MAG reaction yield increased when a larger amount of ethanol was used. These results suggested that Novozym
435 showed strict regiospecificity in an excess amount of ethanol. The process optimization (reaction temperature and reactant
molar ratio) and a study of lipase specificity for various substrates were performed. Under the optimized conditions (ethanol/TO
molar ratio=77∶1 and 25°C), 2-monooleoylglycerol (2-MO) was obtained in more than 98% content among glycerides of the reaction
mixture and approximately 88% reaction yield in 4 h. The above reaction conditions were applied for ethanolysis of tridocosahexaenoylglycerol,
trieicosapentaenoylglycerol, triarachidonoylglycerol, tri-α-linolenoylglycerol, and trilinoleoylglycerol. Reaction yields
ranging from 71.9 to 93.7% were obtained in short reaction times (2.5 to 8 h). Purified (>98%) 2-MO and 2-monodocosahexaenoylglycerol
(2-MD) were reesterified with caprylic acid by immobilized Rhizomucor miehei lipase (Lipozyme IM) to afford symmetrical structured TAG. At a stoichiometric ratio of 2-MAG/caprylic acid, 25°C and 2–5
mm Hg vacuum, the glyceride composition of the esterification mixture was approximately 95% 1,3-dicapryloyl-2-oleoylglycerol
(COC) at 4 h, and 96% 1,3-dicapryloyl-2-docosahexaenoylglycerol (CDC) at 4 h, and 96% 1,3-dicapryloyl-2-docosahexaenoylglycerol
(CDC) at 8 h. The regioisomeric purity of both COC and CDC was 100%. 相似文献
18.
Regiospecific analysis by ethanolysis of oil with immobilized <Emphasis Type="Italic">Candida antarctica</Emphasis> lipase 总被引:1,自引:1,他引:1
Shimada Y Ogawa J Watanabe Y Nagao T Kawashima A Kobayashi T Shimizu S 《Lipids》2003,38(12):1281-1286
A mixture of oil/ethanol (1∶3, w/w) was shaken at 30°C with 4% immobilized Candida antarctica lipase by weight of the reaction mixture. The reaction regiospecifically converted FA at the 1- and 3-positions to FA ethyl
esters, and the lipase acted on C14−C24 FA to a similar degree. The content of 2-MAG reached a maximum after 4 h; the content was 28–29 mol% based on the total amount
of FA in the reaction mixture at 59–69% ethanolysis. Only 2-MAG were present in the reaction mixture during the first 4 h,
and 1(3)-MAG were detected after 7 h. After removal of ethanol from the 4-h reaction mixture by evaporation, 2-MAG were fractionated
by silica gel column chromatography. The contents of FA in the 2-MAG obtained by ethanolysis of several oils coincided well
with FA compositions at the 2-position, which was analyzed by Grignard degradation. It was shown that ethanolysis of oil with
C. antarctica lipase can be applied to analysis of FA composition at the 2-position in TAG. 相似文献
19.
Seung In Hong Yangha Kim Sung Won Yoon Seung Yong Cho In‐Hwan Kim 《European Journal of Lipid Science and Technology》2012,114(9):1044-1051
Conjugated linoleic acid (CLA)‐enriched triacylglycerol (TAG) of 90 wt% was successfully synthesized in 10 h by direct esterification of glycerol and CLA using an immobilized lipase from Candida antarctica under vacuum. The best operating conditions for the synthesis of TAG were investigated according to the three parameters of temperature, enzyme loading, and vacuum. The synthesis of TAG increased with increasing temperature but it did not significantly change above 60°C (p>0.05). The increase of enzyme loading lead to an enhanced conversion of TAG, but enzyme loading of more than 10% (based on the total weight of the substrates) was not effective. Moreover, when vacuum increased, the conversion of TAG increased, but the conversion rate decreased when the vacuum level was too high. The best combination of temperature, enzyme loading, and vacuum level were 60°C, 10% of the total weight of the substrates, and 0.4 kPa, respectively. During the initial 6 h of reaction, Candida antarctica lipase had more selectivity for 10t,12c‐CLA than 9c,11t‐CLA onto the glycerol backbone, and a preference for the incorporation at the sn‐1,3 positions of glycerol rather than at the sn‐2 position. 相似文献
20.
Yomi?Watanabe Yoshie?Yamauchi-Sato Toshihiro?Nagao Takaya?Yamamoto Kentaro?Tsutsumi Akio?Sugihara Yuji?Shimada
We attempted to produce MAG of CLA through lipase-catalyzed esterification of a FFA mixture containing CLA (referred to as
FFA-CLA) with glycerol. Screening of lipases showed that MAG-CLA was produced efficiently at 5°C with Penicillium camembertii, Rhizopus oryzae, and Candida rugosa lipases. Among them, C. rugosa lipase was selected because the lipase is widely used as a catalyst for oils and fats processing. The reaction was conducted
with agitation of a 300-g mixture of FFA-CLA/glycerol (1∶5, mol/mol), a 200-U/g mixture of C. rugosa lipase, and 2% water. When the reaction was conducted at 30°C, the esterification scarcely proceeded, owing to inhibition
of the reaction by glycerol. But the reaction at 5°C eliminated the inhibition and produced MAG efficiently: The degree of
esterification reached 93.8% after 58 h, and MAG content in the reaction mixture was 88.4 wt%. To reduce the reaction time,
the reactor was connected with a vacuum pump after 24 h, and the reaction was continued with dehydration at 5 mm Hg. The degree
of esterification reached 94.7% after 24 h of dehydration (48 h in total), and MAG content increased to 93.0 wt%. Candida rugosa lipase acted a little more strongly on cis-9, trans-11 CLA than on trans-10,cis-12 CLA, but the contents of the two isomers in MAG obtained from a 48-h reaction were the same as the contents in FFA-CLA. 相似文献