• 한국미생물·생명공학회지
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• 제47권4호
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• pp.555-564
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• 2019

Biodiesel is produced through the transesterification process in the presence of alcohol and a catalyst that catalyzes the conversion of triglycerides to esters and glycerol compounds. A more optimal product conversion can be achieved using enzymes, such as lipase. Lipase is reported to be produced in osmophilic yeasts due to the low water content in their natural habitats. Wild forest honey is one of the osmophilic natural habitats in Indonesia. However, lipase-producing yeast has not been reported in the Indonesian honey. In this study, we screened the lipase-producing yeasts isolated from wild forest honey collected from Central Sulawesi. The production profile and activity of lipase were determined at different pH values and temperatures. One promising yeast was isolated from the honey, which was identified as Zygosaccharomyces mellis SG 1.2 based on ITS sequence. The maximum lipase production (24.56 ± 1.30 U/mg biomass) was achieved by culturing the strain in a medium containing 2% olive oil as a carbon source at pH 7 and 30℃ for 40 h. The optimum pH and temperature for lipase activity were 6 and 55℃, respectively. The enzyme maintained 80% of its activity upon incubation at 25℃ for 4 h. However, the enzyme activity decreased by more than 50% upon incubation at 35 and 40℃ for 2 h. This is the first study to report the lipase producing capability of Z. mellis. Further studies are needed to optimize the enzyme production.

• 한국미생물·생명공학회지
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• 제48권2호
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• pp.167-178
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• 2020

Lipase-producing fungi have been isolated from environments containing lipids. The non-dairy creamer industrial waste has a high amount of lipids so it is a potential source for the isolation of lipase-producing fungi. However, the study of fungi that secrete lipase from this industrial waste has not been reported. The purpose of this study was to obtain lipase-producing filamentous fungi from non-dairy creamer industrial waste. Mineral salt and potato dextrose agar were used as media for the isolation process. The qualitative screening was conducted using phenol red agar medium and the quantitative screening using broth medium containing glucose and olive oil. Isolates producing the highest amounts of lipase were identified with molecular methods. We found that 5 out of 19 isolated filamentous fungi are lipase producers. Further analysis showed that isolate Ms.11 produced the highest amount of lipase compared to others. Based on ITS sequence Ms.11 was identified as Aspergillus aculeatus. The lipase activity in medium containing 1% glucose + 1% olive oil at pH 7.0 and 30℃ after 96 and 120 h of incubation was 5.13 ± 0.30 U/ml and 5.22 ± 0.59 U/ml, respectively. The optimum lipase activity was found at pH 7.0, 30℃ and using methanol or ethanol in the reaction tube. Lipase was more stable at 20-30℃ and maintained 85% of its activity. It was concluded that isolate Ms.11 is a potential source of lipase that catalyzes transesterification reactions. Further studies are required to optimize lipase production to make the strain suitable for industry purposes.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.491-505
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• 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.

• Journal of Microbiology
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• 제44권6호
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• pp.583-590
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• 2006

In this study, an organic solvent tolerant bacterial strain was isolated. This strain was identified as Pseudomonas sp. strain S5, and was shown to degrade BTEX (Benzene, Toluene, Ethyl-Benzene, and Xylene). Strain S5 generates an organic solvent-tolerant lipase in the late logarithmic phase of growth. Maximum lipase production was exhibited when peptone was utilized as the sole nitrogen source. Addition of any of the selected carbon sources to the medium resulted in a significant reduction of enzyme production. Lower lipase generation was noted when an inorganic nitrogen source was used as the sole nitrogen source. This bacterium hydrolyzed all tested triglycerides and the highest levels of pro-duction were observed when olive oil was used as a natural triglyceride. Basal medium containing Tween 60 enhanced lipase production to the most significant degree. The absence of magnesium ions ($Mg^{2+}$) in the basal medium was also shown to stimulate lipase production. Meanwhile, an alkaline earth metal ion, $Na^+$, was found to stimulate the production of S5 lipase.

• KSBB Journal
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• 제14권6호
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• pp.696-701
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• 1999

본 연구에서는 lipase를 이용해서 ricinoleic acid를 대량생산하기 위해 피마자유의 완전가수분해 조건을 찾고자 하였다. 널리 알려진 lipase CR, lipase CC, lipase PP를 대상으로 피마자유의 가수분해의 가능성을 시험하고, 유기용매를 사용함으로써 가수분해도를 향상시킬 수 있음을 확인하였다. 일반적으로 lipase의 활성을 감소시키는 극성용매의 경우 피마자유의 가수분해에 있어서도 효소의 활성을 감소시켰고, 물과 섞이지 않는 hydrophobic solvent가 피마자유의 가수분해도를 크게 증가시키는 것을 확인하였다. 본 연구에서는 isopropyl ether의 효과가 가장 크며, 조건에 따라 가수분해도를 두 배 이상 증가시킨다는 것을 확인하였다. 그리고 유기용매를 사용함으로써 pH의 영향을 바꾸거나 감소시킬 수 있다는 사실도 확인하였다. 용매와 물의 부피비에 의해서 가수분해가 영향을 받는다는 사실과 특히 유기용매보다는 물의 양에 절대적으로 영향받는다는 사실을 발견하였다. 하지만, 물과 유기용매의 부피비와 함께 lipase와 피마자유의 무게비도 매우 중요하다는 것을 확인하였다. 30$^{\circ}C$에서 isopropyl ether를 사용할 경우 무게 비로 2 wt%일 때는 약 82%, 4 wt% 이상의 lipase CC나 lipase CR을 사용하면 피마자유가 완전히 가수분해되는 사실을 발견하였다.

• 한국식품영양과학회지
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• 제23권3호
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• pp.496-501
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• 1994

The synthesis of lauryl palmitate from palmitic acid and lauryl alcohol was investigated in organic solvents using lipase. Water-immiscible organic solvent such as hexane, toluenem cyclohexane, and isooctane were found to be suitable of ester synthesis . The effect of water content on the initial rate of conversion was examined . As the content increased, the reaction rate increased. But addition of water in organic solvent decreased therostability of enzyme . The best lauryl palmitate synthesis was achieved with water content of 0.2-0.4% reaction temperature of 4$0^{\circ}C$ and 45$^{\circ}C$ for Candida cylindracea lipase porcine, pancreatic lipase, respectively. when ester synthesis was carried out under the optimum conditions, the conversion yield of palmitate into lauryl palmitate after 70hrs reached 85% and 69 % for the Candida cylindracea lipase and porcine opancreatic lipase, respectivley.

• 한국의류학회지
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• 제35권6호
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• pp.670-677
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• 2011

This study is to optimize the enzymatic processing conditions of Polylactic Acid (PLA) fiber using lipase from Porcine pancreas as an environmental technology. Hydrolytic activity dependent on pH, temperature, enzyme concentration, and treatment time, and structural change of PLA fiber were evaluated. The PLA fiber hydrolysis by lipase was maximized at 50% (o.w.f) lipase concentration $50^{\circ}C$ for 120 minutes under pH 8.5. There was a change of the protein absorbance in the treatment solution before and after the lipase treatment. In addition, there was no substantial change in the molecular and crystalline structures of PLA by lipase treatment as confirmed by DSC, XRD, and FT-IR.

• 한국의류학회지
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• 제32권7호
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• pp.1075-1081
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• 2008

Eco-friendly processing using enzymes has been focused in textile industry in order to reduce environmental pollutions. This paper was suggested to hydrolyze non-cellulose, such as fats and waxes in cotton fabrics by lipase treatment. Enzymatic treatment conditions were controlled according to pH, temperature, enzyme concentration, and treatment time. The physical properties of the lipase-treated cotton fabrics were evaluated by measuring weight loss, moisture regain and dyeing properties. The surface morphology of lipase-treated cotton fabrics were observed by SEM. As a result, the optimum conditions for the lipase treatment were at pH 4.2, temperature 50$^{\circ}C$, concentration 50%, and treatment time 90 minutes. Calcium chloride and Triton X-100 were effective auxiliaries in lipase treatment.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.764-769
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• 2000

The lipase gene(lip) from Bacillus stearothermophilus was recombined in vitro by utilizing the DNA shuffling technique. After four rounds of shuffling, transformation, and screening based on the initial rate of clear zone formation on a tricaprylin plate, a clone (M10) was isolated, the cell extract of which showed about 2.8-fold increased lipase activity. The DNA sequence of the mutant lipase gene (m10) showed 3 base changes, resulting in two cryptic mutations and one amino acid substitution: S113($AGC{\rightarrow}AGT$), L252 ($TTG{\rightarrow}TTA$), and G275E ($GGA{\rightarrow}GAA$). SDS-PAGE analysis revealed that the increased enzyme activity observed in M10 was partly caused by high expression of the m10 lipase gene. The amount of the expressed G275E lipase was estimated to comprise as much as 41% of the total soluble proteins of the cell. The maximum velocity ($V_{max}$) of the purified mutant enzyme for the hydrolysis of olive oil was measured to be 3,200 U/mg, which was 10% higher than that of the parental (WT) lipase (2,900 U/mg). Its optimum temperature for the hydrolysis of olive oil was $68^{\circ}C$ and it showed a typical $Ca^{2+}$-dependent thermostability, properties fo which were the same as those of the WT lipase. However, the mutant enzyme exhibited a high enantiospecificity towards (S)-naproxen compared with the WT lipase. In addition, it showed increased hydrolytic activity towards triolein, tricaprin, tricaprylin, and tricaproin.

• 미생물학회지
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• 제33권4호
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• pp.237-241
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• 1997

Streptomyces coelicolor A3(2) 배양액의 lipase(EC 3.1.1.3)는 ${\alpha}$-naphthyl-butyrate에 대하여 활성을 나타내어 ${\alpha}$-naphthyl-acetate에 활성을 나타내는 esterase와 구분할 수 있었다. Streptomyces coelicolor A3(2) 세포외 lipase를 Sephadex G-100, DEAE-Cellulose 그리고 Phenyl-Sepharose CL4B 크로마토그래피의 과정을 통해 16% 수율로 15배 분리정제 하였다. SDS-폴리아크릴아마이드 겔 전기영동에서는 34.7 kDa정도의 분자량을 갖는 것으로 나타났다. Tributyrin를 기질로 사용하였을 때 이 lipase의 최적활성조건은 pH 8에서 9 그리고 $37^{\circ}C$였다. 기질로서 triacylglycerol의 지방산 길이가 증가할수록 활성은 감소하였다. A-factor에 의해 lipase활성이 억제되므로 배양초기의 낮은 lipase활성과 관련될 것으로 보인다.