• Journal of Microbiology and Biotechnology
• /
• v.22 no.3
• /
• pp.292-300
• /
• 2012

We report the expression, purification, and characterization of L-asparaginase (AnsA) from Rhizobium etli. The enzyme was purified to homogeneity in a single-step procedure involving affinity chromatography, and the kinetic parameters $K_m$, $V_{max}$, and $k_{cat}$ for L-asparagine were determined. The enzymatic activity in the presence of a number of substrates and metal ions was investigated. The molecular mass of the enzyme was 47 kDa by SDS-PAGE. The enzyme showed a maximal activity at $50^{\circ}C$, but the optimal temperature of activity was $37^{\circ}C$. It also showed maximal and optimal activities at pH 9.0. The values of $K_m$, $V_{max}$, $k_{cat}$, and $k_{cat}/K_m$ were $8.9{\pm}0.967{\times}10^{-3}$ M, $128{\pm}2.8$ U/mg protein, $106{\pm}2s^{-1}$, and $1.2{\pm}0.105{\times}10^4M^{-1}s^{-1}$, respectively. The L-asparaginase activity was reduced in the presence of $Mn^{2+}$, $Zn^{2+}$, $Ca^{2+}$, and $Mg^{2+}$ metal ions for about 52% to 31%. In addition, we found that $NH_4{^+}$, L-Asp, D-Asn, and ${\beta}$-aspartyl-hydroxamate in the reaction buffer reduced the activity of the enzyme, whereas L-Gln did not modify its enzymatic activity. This is the first report on the expression and characterization of the L-asparaginase (AnsA) from R. etli. Phylogenetic analysis of asparaginases reveals an increasing group of known sequences of the Rhizobial-type asparaginase II family.

• Restorative Dentistry and Endodontics
• /
• v.21 no.2
• /
• pp.652-663
• /
• 1996

The purpose of this study was to evaluate the tensile bond strength to tooth structure of composite resin and glass ionomer cement according to filling methods and light curing units. In this study, two class V cavities were prepared on the buccal surface of each tooth of 140 extracted human molars, and they were randomly assigned into 3 experimental groups with 40 teeth and control group with 20 teeth. And then, each experimental groups subdivided into 2 groups(A,B) according to light curing units. The cavities of each group were filled with the CLEARFIL FII self curing resin(Control Group), Z-100 light curing resin(Group 1), Vitremer$^{TM}$ light curing glass ionomer cement(Group 2) and Z-100 light curing resin over the Vitrebond$^{TM}$ liner(Group 3). And subdivided A Group used Argon Laser(SPECTRUM$^{TM}$, U.S.A.), B Group used XL 1,000 curing light (3M, U.S.A.). The specimens underwent temperature changed from $5^{\circ}C$ to $55^{\circ}C$ five hundred times. After thermocycling, specimens were stored in 100% relative humidity at $37^{\circ}C$ for 24 hours. And then, the tensile bond strength of specimens were calculated with Universal Testing Machine(AGS-100A, Japan). The results were as follows : 1. Among the experimental groups, the group 2-B showed the highest tensile bond strength ($18.89{\pm}7.80$) and the group 1-A showed the lowest tensile bond strength ($11.68{\pm}2.28$). There was significant difference between group 2-B and group 1-A(p<0.01). 2. Between the light curing units, the XL 1,000 unit showed higher tensile bond strength ($16.63{\pm}3.20$) than that of the Argon Laser unit ($13.73{\pm}2.30$). There was significant difference between XL 1,000 and Argon Laser(p<0.01). 3. About filling methods and materials, the group 2 showed the highest tensile bond strength ($17.56{\pm}1.89$) and the group 1 showed the lowest tensile bond strength($13.03{\pm}1.90$). There was significant difference between group 2 and group 1,3(p<0.01). In conclusion, the results showed that the glass-ionomer cement that cured by XL 1,000 light curing unit demonstrated significantly higher tensile bond strength than other curing unit and filling methods.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.5
• /
• pp.621-628
• /
• 2004

Twenty male buffalo calves of 6-9 months of age (average body weight, 97 kg) were randomly allocated into two main groups of four (control) and sixteen (supplemented) calves. The supplemented group was further divided in to four equal sub-groups, with the two groups supplemented with a liquid preparation of urea-molasses, UML1, containing fish meal and UML2, containing formaldehyde treated deoiled mustard cake (FDMC) and the other two, with a semi-solid preparation, UMC1 with FDMC and deoiled rice bran (DORB) contributing similar level of CP as in UML2 and UMC2 with double the level of FDMC to that in UMC1. The control group was fed with DORB along with ad libitum wheat straw at 40:60 ratios. The rest of the groups were fed on the above diet supplemented with 500 g (as fed basis) of urea-molasses preparations. The experimental feeding was carried out for 24 weeks including a metabolism trial towards the end of experimental feeding. Daily feed intake and fortnightly change in live weight were also recorded during the study. Catalytic supplementation of 500 g urea-molasses induced 8-25% higher voluntary feed intake of wheat straw, resulting in 15-25% higher DM and OM intake. The digestibility of DM, OM, total carbohydrate, NDF, ADF, hemicellulose and cellulose in all the dietary groups were comparable. The CP digestibility of calves in supplemented groups were higher (p<0.05) than the control group. The balance of nutrients, viz. N, Ca and P, was also higher in the supplemented groups. Significantly higher intake of digestible CP coupled with other digestible nutrients attributed to higher TDN (1.67-1.78 vs. 1.37 kg) and ME (5.94-6.31 vs. 4.87 Mcal) intake in urea-molasses supplemented groups which resulted in higher live weight gain compared to that in control group (p<0.01). Between the supplements, UML2 and UMC2 faired non-significantly, indicating formalin treated mustard cake as a suitable replacement to fishmeal in the supplement. The overall ranking based on intake and digestibility of nutrients, live weight gain, economic evaluation and input-output relationship revealed that the rations with UML2 and UMC1 to be of greater value compared to other types. From the study it can be concluded that young ruminants can be reared successfully on a basal diet of deoiled rice bran and wheat straw supplemented with cheaper urea-molasses-mineral mix.

• Proceedings of the Korean Vacuum Society Conference
• /
• 2011.08a
• /
• pp.293-294
• /
• 2011

It is known that semiconductor quantum-dot (QD) heterostructures have superior zero-dimensional quantum confinement, and they have been successfully applied to semiconductor laser diodes (QDLDs) for optical communication and infrared photodetectors (QDIPs) for thermal images [1]. The self-assembled QDs are normally formed at Stranski-Krastanov (S-K) growth mode utilizing the accumulated strain due to lattice-mismatch existing at heterointerfaces between QDs and cap layers. In order to increase the areal density and the number of stacks of QDs, recently, sub-monolayer (SML)-thick QDs (SQDs) with reduced strain were tried by equivalent thicknesses thinner than a wetting layer (WL) existing in conventional QDs (CQDs) by S-K mode. Despite that it is very different from CQDs with a well-defined WL, the SQD structure has been successfully applied to QDIP[2]. In this study, optical characteristics are investigated by using photoluminescence (PL) spectra taken from self-assembled InAs/GaAs QDs whose coverage are changing from submonolayer to a few monolayers. The QD structures were grown by using molecular beam epitaxy (MBE) on semi-insulating GaAs (100) substrates, and formed at a substrate temperature of 480$^{\circ}C$ followed by covering GaAs cap layer at 590$^{\circ}C$. We prepared six 10-period-stacked QD samples with different InAs coverages and thicknesses of GaAs spacer layers. In the QD coverage below WL thickness (~1.7 ML), the majority of SQDs with no WL coexisted with a small amount of CQDs with a WL, and multi-peak spectra changed to a single peak profile. A transition from SQDs to CQDs was found before and after a WL formation, and the sublevel of SQDs peaking at (1.32${\pm}$0.1) eV was much closer to the GaAs bandedge than that of CQDs (~1.2 eV). These revealed that QDs with no WL could be formed by near-ML coverage in InAs/GaAs system, and single-mode SQDs could be achieved by 1.5 ML just below WL that a strain field was entirely uniform.

• Bulletin of the Korean Mathematical Society
• /
• v.25 no.2
• /
• pp.203-209
• /
• 1988

In this paper, we define a g-regular semigroup which is a generalization of a regular semigroup. And we want to find some properties of g-regular semigroup. G-regular semigroups contains the variety of all regular semigroup and the variety of all periodic semigroup. If a is an element of a semigroup S, the smallest left ideal containing a is Sa.cup.{a}, which we may conveniently write as $S^{I}$a, and which we shall call the principal left ideal generated by a. An equivalence relation l on S is then defined by the rule alb if and only if a and b generate the same principal left ideal, i.e. if and only if $S^{I}$a= $S^{I}$b. Similarly, we can define the relation R. The equivalence relation D is R.L and the principal two sided ideal generated by an element a of S is $S^{1}$a $S^{1}$. We write aqb if $S^{1}$a $S^{1}$= $S^{1}$b $S^{1}$, i.e. if there exist x,y,u,v in $S^{1}$ for which xay=b, ubv=a. It is immediate that D.contnd.q. A semigroup S is called periodic if all its elements are of finite order. A finite semigroup is necessarily periodic semigroup. It is well known that in a periodic semigroup, D=q. An element a of a semigroup S is called regular if there exists x in S such that axa=a. The semigroup S is called regular if all its elements are regular. The following is the property of D-classes of regular semigroup.group.

• Proceedings of the Korean Vacuum Society Conference
• /
• 1998.02a
• /
• pp.17-17
• /
• 1998

Synchrotron radiation is produced when charged particles moving with relativistic velocities a are accelerated - for example, deflected by the bending magnets which guide the electron or p positrons in circular accelerators or storage rings. By using special focusing magnetic lattices i in the particle accelerators it is possible to make the dimensions of the particle beam very small with a hi맹 charge density which results in a light source with high b디lIiance. Synchrotron light h has important properties which make it ideal for a wide range of investigations in surface s science. The fact that the spectrum of electromagnetic radiation emitted in a bending magnet e extends in a continuum from the 얹r infra red region to hard x-rays means that it is id않I for a v variety of spectroscopic studies. Since there are no convenient lasers, or other really bright l light sources, in the vacuum ultraviolet and soft x-ray re.밍ons the development of synchrotron r radiation has enabled enormous advances to be made in this di펌C비t spectr따 re밍on. P Polarization-dependent measurements, for ex없nple ellipsometry or circular dichroism studies a are possible because the radiation has a well-defined polarization - linear in the plane of orbit w with additional right-circular, or left-circular, components for emission an생es above, or below, t the horizontal, respectively. Since the synchrotron light is emitted from a bunch of charge c circulating in a ring the light is emitted with a well-defined time structure with a short flash of l light every time a bunch passes an exit port. The time structure depends on the size of the ring a and the number and sequence of filling of the bunches. A pulsed light source enables time¬r resolved studies to be performed which provide direct information on the lifetimes and decay m modes of excited states and in addition opens up the possibility of using time of flight t techniques for spectroscopic studies. The fact that synchrotron radiation is produced in a clean u ultrahi야 vacuum environment is of gr않t importance for surce science studies. The current t비rd generation synchrotron light sources provide exceptionally high baliance and stability a and open up possibilities for experiments which would have been inconceivable only a short time ago.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.11
• /
• pp.1944-1953
• /
• 2015

A novel alkaline protease from Streptomyces sp. M30, SapHM, was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and DEAE-Sepharose chromatography, with a yield of 15.5% and a specific activity of 29,070 U/mg. Tryptic fragments of the purified SapHM were obtained by electrospray ionization quadrupole time-of-flight mass spectrometry. Nucleotide sequence analysis revealed that the gene sapHM contained 1,179 bp, corresponding to 392 amino acids with conserved Asp156, His187, and Ser339 residues of alkaline protease. The first 24 amino acid residues were predicted to be a signal peptide, and the molecular mass of the mature peptide was 37.1 kDa based on amino acid sequences and mass spectrometry. Pure SapHM was optimally active at 80℃ in 50 mM glycine-NaOH buffer (pH 9.0), and was broadly stable at 0-50℃ and pH 4.0-9.0. The protease relative activity was increased in the presence of Ni²⁺, Mn²⁺, and Cu²⁺ to 112%, 113%, and 147% of control, respectively. Pure SapHM was also activated by dimethylformamide, dimethyl sulfoxide, Tween 80, and urea. The activity of the purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. The K_m and V_max values were estimated to be 35.7 mg/ml, and 5 × 10⁴ U/mg for casein. Substrate specificity analysis showed that SapH was active on casein, bovine serum albumin, and bovine serum fibrin.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
• /
• 2009.11a
• /
• pp.5-5
• /
• 2009

Thin-film-transistors (TFTs) that can be prepared at low temperatures have attracted much attention because of the great potential for transparent and flexible electronics. One of the mainstreams in this field is the use of organic semiconductors such as pentacene. But device performance of the organic TFTs is still limited due to low field-effect mobility and rapid degradation after exposing to air. Alternative approach is the use of amorphous oxide semiconductors as a channel. Amorphous oxide semiconductors (AOSs) based TFTs showed the fast technological development, because AOS films can be fabricated at room temperature and exhibit the possibility in application like flexible display, electronic paper, and larges solar cells. Among the various AOSs, a-IGZO has lots of advantages because it has high channel mobility, uniform surface roughness and good transparency. [1] The high mobility is attributed to the overlap of spherical s-orbital of the heavy post-transition metal cations. This study demonstrated the effect of the variation in channel thickness from 30nm to 200nm on the TFT device performance. When the thickness was increased, turn-on voltage and subthreshold swing was decreased. The a-IGZO channels and source/drain metals were deposited with shadow mask. The a-IGZO channel layer was deposited on $SiO_2$/p-Si substrates by RF magnetron sputtering, where RF power is 150W. And working pressure is 3m Torr, at $O_2/Ar$ (2/28 sccm) atmosphere. The electrodes were formed with electron-beam evaporated Ti (30 nm) and Au (70 nm) bilayer. Finally, Al (150nm) as a gate metal was thermal-evaporated. TFT devices were heat-treated in a furnace at 250 $^{\circ}C$ and nitrogen atmosphere for 1hour. The electrical properties of the TFTs were measured using a probe-station. The TFT with channel thickness of 150nm exhibits a good subthreshold swing (SS) of 0.72 V/decade and on-off ratio of $1{\times}10^8$. The field effect mobility and threshold voltage were evaluated as 7.2 and 8 V, respectively.

• BMB Reports
• /
• v.40 no.4
• /
• pp.588-594
• /
• 2007

A novel hot spring thermophile, Anoxybacillus gonensis A4 (A. gonensis A4) was investigated in terms of capability of tributyrin degradation and characterization of its thermostable esterase activity by the hydrolysis of p-nitrophenyl butyrate (PNPB). It was observed that A. gonensis A4 has an esterase with a molecular weight of 62 kDa. The extracellular crude preparation was characterized in terms of substrate specificity, pH and temperature optima and stability, kinetic parameters and inhibition/activation behaviour towards some chemicals and metal ions. Tributyrin agar assay showed that A. gonensis A4 secreted an esterase and $V_{max}$ and $K_m$ values of its activity were found to be 800 U/L and 176.5 ${\mu}M$, respectively in the presence of PNPB substrate. The optimum temperature and pH, for A. gonensis A4 esterase was $60-80^{\circ}C$ and 5.5, respectively. Although the enzyme activity was not significantly changed by incubating crude extract solution at $30-70^{\circ}C$ for 1 h, the enzyme activity was fully lost at $80^{\circ}C$ for same incubation period. The pH-stability profile showed that original crude esterase activity increased nearly 2-fold at pH 6.0. The effect of some chemicals on crude esterase activity indicated that A. gonensis A4 produce an esterase having serine residue in active site and -SH groups were essential for its activity.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.2
• /
• pp.347-355
• /
• 2016

An exo-β-D-glucosaminidase (AorCsxA) from Aspergillus oryzae FL402 was heterologously expressed and purified. The deduced amino acid sequence indicated that AorCsxA belonged to glycoside hydrolase family 2. AorCsxA digested colloid chitosan into glucosamine but not into chitosan oligosaccharides, demonstrating exo-β-D-glucosaminidase (CsxA) activity. AorCsxA exhibited optimal activity at pH 5.5 and 50℃; however, the enzyme expressed in Pichia pastoris (PpAorCsxA) showed much stronger thermostability at 50℃ than that expressed in Escherichia coli (EcAorCsxA), which may be related to glycosylation. AorCsxA activity was inhibited by EDTA and most of the tested metal ions. A single amino acid mutation (F769W) in AorCsxA significantly enhanced the specific activity and hydrolysis velocity as revealed by comparison of V_max and k_cat values with those of the wild-type enzyme. The three-dimensional structure suggested the tightened pocket at the active site of F769W enabled efficient substrate binding. The AorCsxA gene was heterologously expressed in P. pastoris, and one transformant was found to produce 222 U/ml activity during the high-cell-density fermentation. This AorCsxA-overexpressing P. pastoris strain is feasible for large-scale production of AorCsxA.