• Korean Chemical Engineering Research
• /
• v.50 no.2
• /
• pp.257-263
• /
• 2012

Biodiesel is a renewable energy which is nontoxic and acting as a replacement for conventional diesel which derived from fossil fuel. Classified biodiesel producing way such as acid, base, supercritical and enzyme methods, this study focused on eco-friendly production of biodiesel using supercritical and immobilized enzyme process. Assuming a plant with a production rate of 10,000 tons a year, a PRO II simulator program was used to simulate the product conversion rate and total energy consumption. The product conversion in supercritical process and immobilized enzyme was found to be 91.17% (including 0.9% glycerol) and 93.18% (including 1.0% glycerol) respectively. The result shows that the efficiency of immobilized enzyme process is higher compared to supercritical process but having lower end product purity. From the energy consumption point of view, supercritical process consume about 8.9 MW while immobilized enzyme process consume much lower energy which is 3.9 MW. Consequently, this study certifies that energy consumption of supercritical process is 2.3 times higher than immobilized enzyme process.

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

• 한국신재생에너지학회:학술대회논문집
• /
• 2005.11a
• /
• pp.688-688
• /
• 2005

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

• Biotechnology and Bioprocess Engineering:BBE
• /
• v.9 no.6
• /
• pp.465-470
• /
• 2004

Optimal conditions for the in situ immobilization of lipase in aldehyde-silica packed columns, via reductive amination, were investigated. A reactant mixture, containing lipase and sodium borohydride (NaCBH), was recirculated through an aldehyde-silica packed column, such that the covalent bonding of the lipase, via amination between the amine group of the enzyme and the aldehyde terminal of the silica, and the reduction of the resulting imine group by NaCBH, could occur inside the bed, in situ. Mobile phase conditions in the ranges of pH $7.0{\~}7.8$, temperatures between $22{\~}28^{circ}C$ and flow rates from $0.8{\~}1.5\;BV/min$ were found to be optimal for the in situ immobilization, which routinely resulted in an immobilization of more than 70 mg­lipase/g-silica. Also, the optimal ratio and concentration for feed reactants in the in situ immobilization: mass ratio [NaCBH]/[lipase] of 0.3, at NaCBH and lipase concentrations of 0.75 and 2.5 g/L, respectively, were found to display the best immobilization characteristics for concentrations of up to 80 mg-lipase/g-silica, which was more than a 2-fold increase in immobilization compared to that obtained by batch immobilization. For tributyrin hydrolysis, the in situ immobilized lipase displayed lower activity per unit mass of enzyme than the batch-immobilized or free lipase, while allowing more than a $45\%$ increase in lipase activity per unit mass of silica compared to batch immobilization, because the quantity of the immobilization on silica was aug­mented by the in situ immobilization methodology used in this study.

• 한국생물공학회:학술대회논문집
• /
• 2001.11a
• /
• pp.633-634
• /
• 2001

The in situ immobilization of lipase in an LC column with aldehyde-silica stationary phase was optimized. The effects of feed composition, temperature, mobile phase pH and now rate on immobilization efficiency were investigated. Lipase activity after in situ immobilization was compared with free enzyme and batch immobilized enzyme.

• Microbiology and Biotechnology Letters
• /
• v.48 no.4
• /
• pp.491-505
• /
• 2020

A newly isolated strain, Proteus vulgaris OR34, from olive mill waste was found to secrete an alkaline extracellular lipase at 11 U·ml-1 when cultivated on an optimized liquid medium. This lipase was purified 94.64-fold with a total yield of 9.11% and its maximal specific activity was shown to be 3232.58 and 1777.92 U·mg-1 when evaluated using the pH-stat technique at 55℃ and pH 9 and Tributyrin TC4 or olive oil as the substrate. The molecular mass of the pure OR34 lipase was estimated to be around 31 kDa, as revealed by SDS-PAGE and its substrate specificity was investigated using a variety of triglycerides. This assay revealed that OR34 lipase preferred short and medium chain fatty acids. In addition, this lipase was stable in the presence of high concentrations of bile salt (NaDC) and calcium ions appear not to be necessary for its activity. This lipase was inhibited by THL (Orlistat) which confirmed its identity as a serine enzyme. In addition, the immobilization of OR34 lipase by adsorption onto calcium carbonate increased its stability at higher temperatures and within a larger pH range. The immobilized lipase exhibited a high tolerance to organic solvents and retained 60% of its activity after 10 months of storage at 4℃. Finally, the OR34 lipase was applied in biodiesel synthesis via oleic acid mediated esterification of methanol when using hexane as solvent. The best conversion yield (67%) was obtained at 12 h and 40℃ using the immobilized enzyme and this enzyme could be reused for six cycles with the same efficiency.

• Applied Chemistry for Engineering
• /
• v.21 no.4
• /
• pp.452-456
• /
• 2010

It is known that 1,3-diglyceride (1,3-DG) hardly accumulates inside human body because the metabolism of 1,3-DG is entirely different from that of general fats such as triglycerides (TG). This research focuses on the selective synthesis of 1,3-DG by the esterification reaction using an immobilized lipase. For a reaction between glycerin and oleic acid (OA) with a mole ratio of 1 : 2 under vacuum, changes in the compositions of monoglyceride (MG), TG and DG and the contents of 1,3-isomers in DG were investigated, as a function of reaction temperature and the amount of lipase. The reactivities determined by the rate of the consumption of OA became higher with the increase in temperature and the amount of lipase. When the results were compared with those obtained in the earlier study where Novozym was applied as an immobilized lipase, the reactivity was higher for Novozym, on the other hand, selectivity to 1,3-DG was much higher for Lipozyme. Especially, it is remarkable that 1,3-DG content in total DG reached to 98% in the reaction carried out at $50^{\circ}C$ using Lipozyme.

• Journal of the Korean Chemical Society
• /
• v.41 no.9
• /
• pp.483-487
• /
• 1997

The whole cells of Chromobacterium chocolatum was immobilized in the matrix of polyacrylamide and then used for the hydrolysis of dimethyl-cis-1,3-dibenzyl-2-oxoimidazolidine-4,5-dicarboxylate. This hydrolysis yielded the optically active monoester ( > 96% ee) which is useful as an synthetic intermediate of (+)-biotin. We have studied the optimum condition of hydrolysis by using immobilized cells under variable concentration of substrate, reaction time and pH levels. The activity of lipase in immobilized cell was retained for longer than 4 weeks. The best conversion yield of product was obtained when 2 g of wet cell was immobilized and then reacted with 200 mg of substrate at pH 7.

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