• Microbiology and Biotechnology Letters
• /
• v.37 no.2
• /
• pp.118-124
• /
• 2009

Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

• Journal of the Korean Chemical Society
• /
• v.34 no.6
• /
• pp.629-636
• /
• 1990

Dephosphorylation of p-nitrophenyldiphenylphosphate(p-NPDPP) mediated by anions of sodium 2-alkylbenzimidazole-5-sulfonate(R-BI-SO$_3$Na) in CTABr micellar solutions are obviously slower than that by anion of sodium benzimidazole-5-sulfonate(BI-SO$_3$Na), and the reation rates were decreased with increase of lengths of alkyl groups. This presents a striking contrast to the reactions in aqueous solutions without added CTABr, of which the reaction rates are on approximately same levels. It seems due to steric effect of alkyl groups of R-BI$^-$SO$_3$Na in the Stern layer of micelle, and it is supported by measured activation parameters(△H$^\neq$/TEX>, △G$^\neq$/TEX> and △S$^\neq$/TEX>) of the reactions in aqueous and micellar solutions. In addition to nucleophilic ability of benzimidazole moiety of R-BI$^-$SO$_3$Na on the reactions, these compounds with long alkyl groups(nonyl to pentadecyl) are micellized for themseleves, and increase the reaction rates due to their micellar catalyses in aqueous solutions, not including CTABr.

• Journal of Life Science
• /
• v.25 no.9
• /
• pp.1021-1026
• /
• 2015

Invertase (β-D-fructosfuranosidase, EC 3.2.1.26) catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Three biochemical subgroups of invertases have been investigated in plants: vacuolar (soluble acid), cytoplasmic (soluble alkaline), and cell wall-bound (insoluble acid) invertases. An isoform of neutral invertase was purified from pea seedlings (Pisum sativum L.) and treated with gibberellic acid (GA) by sequential procedures consisting of ammonium sulfate precipitation, ion-exchange chromatography, absorption chromatography, and reactive green-19 affinity chromatography. The results of the overall insoluble invertase purification were a 430-fold increase. The purified neutral invertase was not glycosylated and had an optimum pH between neutral and alkaline (pH 6.8-7.5). It was inhibited by Tris, as well as by heavy metals, such as Hg²⁺ and Cu²⁺. Typical Michaelis–Menten kinetics were observed when the activity of the purified invertase was measured, with sucrose concentrations up to 100 mM. The K_m and V_max values were 12.95 mM and 2.98 U/min, respectively. The molecular mass was around 20 kDa. The sucrose-cleaving enzyme activity of this enzyme is similar to that of sucrose synthase and fructosyltransferase, but its biochemical characteristics are different from those of sucrose synthase and fructosyltransferase. Based on this biochemical characterization and existing knowledge, neutral INV is an invertase isoform in plants.

• Polymer(Korea)
• /
• v.30 no.6
• /
• pp.512-518
• /
• 2006

A biodegradable polymer poly((R) -3-hydroxybutyric acid) (PHB) was conjugated with a hydrophilic polymer poly(ethylene glycol) (PEG) by the ttansesterification reaction to form the amphiphilic block copolymer. PHB with low molecular weight ($3000{\sim}30000$) was appropriated for the drug delivery materials. High molecular weight PHB was hydrolyzed by an acid-catalyst to produce the low molecular weight one. Amphiphilic block copolymer was formed the self-assembled polymeric micelle system in the aqueous solution that the hydrophillic PEG was wraped the hydrophobic PHB. Generally, polymeric micelle forms the small particle between $10{\sim}200nm$. These polymeric micelle systems have been widely used for the drug delivery systems because they were biodegradable, biocompatible, non-toxic and patient compliant. The hydroxyl group of PEG was substituted with carboxyl group which has the reactivity to the ester group of PHB. Amphiphilic block copolymer was conjugated between PHB, and modified PEG at $176^{\circ}C$ which was higher than the melting point of PHB. Transesterification reaction was verified with DSC, FTIR, $^1H-NMR$. In the aqueous solution, critical micelle concentration (CMC) of the mPEG-co-PHB copolymer measured by the fluororescence scanning spectrometer was $5{\times}10^{-5}g/L$. The shape and size of the nanoparticle was taken by dynamic light scattering and atomic force microscopy. The size of the nanoparticle was about 130 nm and the shape was spherical. Our polymeric micelle system can be used as the passive targeting drug delivery system.

• Journal of the Korean Applied Science and Technology
• /
• v.36 no.1
• /
• pp.348-354
• /
• 2019

Dipalmitoyl phosphatidyl choline(DPPC), Polyphenol Derivatives, and Hematoxylin-Eosin were directly sonicated in acidic condition for 6 minutes to give clear stock solutions. Absorbtion properties of Polyphenol Derivatives in lecithin vesicle of Diphalmitoyl phosphatidyl choline system at $25^{\circ}C$ have been studied by absorbtion spectroscopy. The equilibrium of Polyphenol Derivatives between monomer and dimer in lecithin vesicles have been existed at low concentration of Polyphenol Derivatives, but oligomer has been formed in vesicle at high concentration of lecithin vesicles. By adding Bacteriorhodopsin(BR) to constant concentration of Polyphenol Derivatives decreased the absorbtion ratio(${\alpha}/{\beta}$) of Polyphenol Derivatives was increased during phase transition of dipalmitoyl phosphatidyl choline. In the presence of column eluted lamella vesicle and mixture of uni- and multilamella aggregates. The differences of rate between column eluted- and mixture were observed, therefore column eluted lamella reaction was represented more catalytic effect. The phase transition temperature of hydrolysis on Dipalmitoyl phosphatidyl choline and Polyphenol Derivatives were measured higher than it of Dipalmitoyl phosphatidyl choline and no Polyphenol Derivatives.

• Journal of the Korean Ceramic Society
• /
• v.41 no.1
• /
• pp.81-85
• /
• 2004

Nanoporous materials with nanometer-sized pores, are of great interest in the various applications such as selective adsorbents, heterogeneous catalysts and catalyst supports because of their high porosity, surface area, and size selective adsorption properties. This study is aimed to prepare nanoporous catalytic materials on the basis of two-dimersional clay by pillaring of $SiO_2$ sol particles. $SiO_2$ Pillared Montmorillonite (Si-PILM) was prepared by ion exchanging the interlayer $Ni^{2+}$ ions of clay with $SiO_2$ nano-sized particles of which the surface was modified with nicked polyhydroxy cations sach as $Ni_4(OH)_4^{4+}$. Nano-sized $SiO_2$ particles were formed by the controlled hydrolysis of tetraethyl orthosilicate (TEOS). Upon pillaring of $Ni^+$-modified $SiO_2$ nano particles between the clay layers, the basal spacing was expanded largely to $45{\AA}$ and the extremely large specific surface area ($S_{BET}$) of $760m^2/g$ was obtained.

• Microbiology and Biotechnology Letters
• /
• v.38 no.4
• /
• pp.428-433
• /
• 2010

Enzymatic hydrolysis of sulfate groups in agaropectin or agar simplifies the production process of high-quality or low sulfate-content agarose. This study was investigated that cell surface displayed arylsulfatase can be applied to desulfatation of agar for production of agarose. Sulfate content of agarose prepared by treatment of yeast surface-displayed arylsulfatase was decreased in a enzyme dosedependent manner. Especially, 35 unit/mL of yeast surface arylsulfatase attenuated sulfate content of agarose up to 0.2%. In the 0.6% agar(Junsei), 35 unit/mL enzyme treated at $40^{\circ}C$ for 3 h showed the lowest content of sulfate. Therefore, this result was determined to be the optimal condition to desulfatation of agar for production of agarose. In addition, the gel strength of yeast surface arylsulfatase treated agar and commercial agarose were compared. Agarose prepared by treatment of yeast surface arylsulfatase showed $559.8{\pm}0.12$ of gel strength, and it is a similar compared to the commercial agarose.

• Korean Chemical Engineering Research
• /
• v.48 no.3
• /
• pp.322-326
• /
• 2010

Stability of sodium borohydride solution during storage was studied. In order to enhance the $NaBH_4$ stability, NaOH and KOH were added to the $NaBH_4$ solution. The effect of concentration of the borohydride and alkaline solution, temperature and materials of storage vessels on the rate of borohydride hydrolysis was investigated. The rate of hydrogen evolution decreased as the concentration of alkaline increased due to increase of $NaBH_4$ stability in the solution. The stability of $NaBH_4$ solution decreased when the borohydride concentration raised from 10 to 15 wt% and then increased when the $NaBH_4$ concentration increased above 15 wt% due to increase in the pH of the concentrated solution. The activity coefficient of hydrolysis of $NaBH_4$ solution(NaOH 3.0 wt%, $NaBH_4$ 25 wt%) was 115.1 kJ/mol and this value was 1.5~4.0 times higher than that of hydrolysis of $NaBH_4$ solution with catalyst. The borohydride solutions in glass and stainless-steel vessel were more stable than the solution in plastic(PE) vessel.

• Journal of Life Science
• /
• v.27 no.7
• /
• pp.849-856
• /
• 2017

The ${\beta}$-D-fructofuranosidase (EC 3.2.1.26) is an important enzyme from a historical point of view, discovered by French biologist Berthelot in 1860 and was first used to study enzymology. ${\beta}$-D-fructosfuranosidase catalyzes the hydrolysis of sucrose into D-glucose and D-fructose. Four biochemical subgroups of ${\beta}$-D-fructofuranosidase have been investigated in plants. There are vacuolar (soluble acid), cytoplasmic (soluble alkaline), membrane-bound (insoluble alkaline), and cell wall-bound (insoluble acid) ${\beta}$-D-fructofuranosidase by purification. Their biochemical characteristics are distinct. It suggested that those enzymes might be different gene products. The contribution of each of these enzymes to sucrose management in the plant is likely to be correlated with their localization. Common localization in developing cells in tissues from a range of developmental stages and plant parts suggests that all of the isoforms may be closely involved in nutrient transport. The ${\beta}$-D-fructofuranosidases were most commonly found associated with maturing tissues in developing fruits, leaves, and roots. The ${\beta}$-D-fructofuranosidase activity varies in the relationship between growth and expansion through cell division, development of storage organs and tissues, and the relationship of plant defense responses. It is necessary to summarize more researches in order to know the definite physiological function.