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

• Microbiology and Biotechnology Letters
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• v.46 no.1
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• pp.51-58
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• 2018

The expression, purification, and characterization of cold-adapted lipase from the psychrophile, Janthinobacterium sp. were investigated. The gene encoding lipase from Janthinobacterium sp. PAMC 25641 was cloned into a pET28a(+) vector and heterologously expressed in Escherichia coli BL21 (DE3). The amino acid sequence deduced from the nucleotide sequence (930 bp) corresponded to a protein having 309 amino acid residues with a molecular weight of 32.7 kDa and a pI of 5.55. Recombinant E. coli harboring the Janthinobacterium lipase gene were induced by addition of isopropyl-${\beta}$-D-thiogalactopyranoside. $Ni^{2+}$-NTA affinity chromatography was used to purify the lipase, which had a specific activity of 107.9 U/mg protein. The effect of temperature and pH on the activity of lipase was measured using p-nitrophenyl octanoate as a substrate. The stability of the lipase at low temperatures indicated it is a cold-adapted enzyme. The lipase activity was increased by $Na^{2+}$, $Mg^{2+}$, and $Mn^{2+}$, and decreased by $Zn^{2+}$ and $Co^{2+}$. Analysis of the lipase activity using various p-nitrophenyl esters showed a strong preference toward short acyl chains of the esters, indicating the ability of the cold-adapted lipase to hydrolyze short-chain esters.

• Journal of Life Science
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• v.20 no.6
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• pp.902-906
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• 2010

A producer of thermophilic extracellular lipase, Geobacillus kaustophilus DSM 7263, was selected from various microorganisms of the Geobacillus genus. We investigated optimum conditions for mass production of G. kaustophilus lipase. Among the different natural oil media, olive oil was optimal for enzyme production. The maximum amount of enzyme production was obtained when G. kaustophilus was grown in a medium containing 0.5% olive oil as a carbon source. The pH and temperature for optimal growth were pH 8.0 and $55^{\circ}C$, respectively, while the optimum pH and temperature for lipase production were pH 6.0 and $50^{\circ}C$, respectively. In the presence of $Mg^{2+}$ and $Mn^{2+}$, lipase production was dramatically enhanced by 247% and 157%, respectively, whereas enzyme production was inhibited by $Zn^{2+}$, $Cu^{2+}$, and $Cd^{2+}$. The addition of 0.1% (v/v) triton X-100 increased lipase production and cell growth when compared to the negative control.

• Korean journal of applied entomology
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• v.46 no.1 s.145
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• pp.131-139
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• 2007

From the course of screening of useful enzyme producing microorganism from insect guts, we isolated 9 lipase producing strains and their lipase producing activities were tested. 16S rDNA sequence analysis showed that they were Gram negative bacteria grouped on Serratia sp., Pseudomonas sp., and Burkholderia sp.. Among them, an excellent lipase producing strain, Burkholderia sp. HY-10 identified by 16S rDNA analysis and biochemical methods, was further studied its lipase producing characteristics. It was isolated from a longcorm beetle, Prionus insularis and showed cell density dependent lipase producing activity in the culture media that contained olive oil as a carbon source. Maximum lipase production was achieved in the M9 media containing 0.5% yeast extract and 0.5% olive oil when cultured at $30^{\circ}C$ for 36-42 hrs.

• Korean Journal of Food Science and Technology
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• v.8 no.1
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• pp.33-41
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• 1976

These experiment were conducted to investigate the cultural condition of the lipase production by Rhizopus japonicus. The results obtained were as follows: 1. Soybean meal and ammonium sulfate were the most effective in the lipase production as organic and inorganic nitrogen sources, respectively. 2. The lipase production was strongly inhibited, when added as carbon sources xylose, glucose, fructose, galactose, maltose, soluble starch, and dextrin causing the lowering of pH of the medium during culture. Sucrose did not inhibit the lipase production, but not caused any effect when added. 3. $K_2HPO_4$ as phosphate salt and $MgSO_4{\cdot}7H_2O$ as magnesium salt were the most effective in the lipase production. 4. The addition of olive oil, soybean oil, and coconut oil respectively increased the enzyme production and especially 1% olive oil increased it by 50%. 5. The enzyme production increased slightly on the addition of yeast extract to $0.05{\sim}0.07%$. 6. The optimum composition of the medium for the lipase production by Rhizopus japonicus was in the composition of soybean meal 2%; $K_2HPO_4{\cdot}$ 0.5%; $(NH_4)_2SO_4$ 0.1%; $MgSO_4\;7H_2O$ 0.05%; yeast extract 0.05%; olive oil 1%. The maximum production of the lipase was attained by the incubation far 48hrs under the optimum incubation condition.

• Microbiology and Biotechnology Letters
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• v.40 no.2
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• pp.98-103
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• 2012

For screening of useful enzymes producing microorganisms from Meju, we isolated high lipase producing strains and their lipolytic enzyme activities were then tested. The lipolytic enzyme activities of isolated microorganisms were therefore tested on the Y124 strain. The gene sequence analysis of ITS from Y124 strain revealed Yarrowia lipolytica. Lipase production by the Y124 strain was studied in media containing various carbon sources. The Y124 strain drastically increased lipolytic enzyme activity in YPO media containing olive oil, as well as in YPDO media containing both olive oil and glucose. Maximal lipase production was achieved in YPD (yeast extract-peptone-D-glucose) media containing 0.7% olive oil when cultured at $30^{\circ}C$ for 8 hrs. The lipase produced from the Y124 strain showed the highest activity in p-NPO (p-nitrophenyl octanoate ($C_8$)), amongst the various p-nitrophenyl esters.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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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.

• BMB Reports
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• v.34 no.5
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• pp.457-462
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• 2001

Using the monomolecular film technique, we compared the interfacial properties of Staphylococcus simulans lipase (SSL) and Staphylococcus aureus lipase (SAL). These two enzymes act specifically on glycerides without any detectable phospholipase activity when using various phospholipids. Our results show that the maximum rate of racemic dicaprin (rac-dicaprin) hydrolysis was displayed at pH 8.5, or 6.5 with Staphylococcus simulans lipase or Staphylococcus aureus lipase, respectively The two enzymes interact strongly with egg-phosphatidyl choline (egg-PC) monomolecular films, evidenced by a critical surface pressure value of around $23\;mN{\cdot}m^{-1}$. In contrast to pancreatic lipases, $\beta$-lactoglobulin, a tensioactive protein, failed to inhibit Staphylococcus simulans lipase and Staphylococcus aureus lipase. A kinetic study on the surface pressure dependency, stereoselectivity, and regioselectivity of Staphylococcus simulans lipase and Staphylococcus aureus lipase was performed using optically pure stereoisomers of diglycerides (1,2-sn-dicaprin and 2,3-sn-dicaprin) and a prochiral isomer (1,3-sn-dicaprin) that were spread as monomolecular films at the air-water interface. Both staphylococcal lipases acted preferentially on distal carboxylic ester groups of the diglyceride isomer (1,3-sn-dicaprin). Furthermore, Staphylococcus simulans lipase was found to be markedly stereoselective for the sn-3 position of the 2,3-sn-dicaprin isomer.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.101-104
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• 1977

Lipase from Geotrichum candidum was heat inactivated in 0.1M phosphate buffer solution. The thermal inactivation followed first order kinetics for the range of temperatures $50^{\circ}-80^{\circ}C$ except at $50^{\circ}C$. The changes in enthalpy, entropy and Gibbs free energy at $60^{\circ}C$ were 120.4 kJ/mol, 73.0 J/mol K and 96.9 kJ/mol respectively a value of $19^{\circ}C$(Geotrichum candidum lipase) is greater than that of lipases from milk and pancreas. The effect of detergents, lecithin and linoleic acid or the thermal inactivation of lipase was found to be negligible.

• Microbiology and Biotechnology Letters
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• v.50 no.1
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• pp.71-80
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• 2022

Lipases (triacylglycerol acylhydrolases [EC 3.1.1.3]) are water-soluble enzymes. They catalyze the hydrolysis of fats and oils. A cold-active lipase from marine Bacillus cereus HSS, isolated from the Mediterranean Sea, Alexandria, Egypt, was purified and characterized. The total purification depending on lipase activity was 438.9 fold purification recording 632 U/mg protein. The molecular weight of the purified lipase was estimated to be 65 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The optimum substrate concentration, enzyme concentration, pH, and temperature were 1.5 mM, 100 µl, pH 6 and 10℃, respectively. The lipase was tolerant to NaCl concentrations ranging from 1.5 to 4.5%. The lipase was affected by the tested metal ions, and its activity was inhibited by 16% in the presence of 0.05 M SDS. The application of the cold-active lipase for the removal of an oil stain from a white cotton cloth showed that it is a promising biological agent for the treatment of oily wastes and other related applications. To the best of our knowledge, this is the first report of the purification and characterization of a lipase from marine B. cereus HSS isolated from the Mediterranean Sea.