• Journal of Microbiology and Biotechnology
• /
• v.28 no.12
• /
• pp.2036-2045
• /
• 2018

An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

• Applied Microscopy
• /
• v.51
• /
• pp.17.1-17.10
• /
• 2021

Our previous study on the binding activity between Cel5H and clay minerals showed highest binding efficiency among other cellulase enzymes cloned. Here, based on previous studies, we hypothesized that the positive amino acids on the surface of Cel5H protein may play an important role in binding to clay surfaces. To examine this, protein sequences of Bacillus licheniformis Cel5H (BlCel5H) and Paenibacillus polymyxa Cel5A (PpCel5A) were analyzed and then selected amino acids were mutated. These mutated proteins were investigated for binding activity and force measurement via atomic force microscopy (AFM). A total of seven amino acids which are only present in BlCel5H but not in PpCel5A were selected for mutational studies and the positive residues which are present in both were omitted. Of the seven selected surface lysine residues, only three mutants K196A(M2), K54A(M3) and K157T(M4) showed 12%, 7% and 8% less clay mineral binding ability, respectively compared with wild-type. The probable reason why other mutants did not show altered binding efficiency might be due to relative location of amino acids on the protein surface. Meanwhile, measurement of adhesion forces on mica sheets showed a well-defined maximum at 69±19 pN for wild-type, 58±19 pN for M2, 53±19 pN for M3, and 49±19 pN for M4 proteins. Hence, our results demonstrated that relative location of surface amino acids of Cel5H protein especially positive charged amino acids are important in the process of clay mineral-protein binding interaction through electrostatic exchange of charges.

• KSCE Journal of Civil and Environmental Engineering Research
• /
• v.34 no.3
• /
• pp.895-906
• /
• 2014

A fundamental study of the dynamically penetrating anchor (DPA - colloquially known as torpedo anchor) embedded into deep seabed was conducted using measurement data and numerical approaches. Numerical simulation of such a structure penetration was often suffered by severe mesh distortion arising from very large soil deformation, complex contact condition and nonlinear soil behavior. In recent years, a Coupled Eulerian-Lagrangian method (CEL) has been used to solve geomechanical boundary value problems involving large deformations. In this study, 3D finite element analyses using the CEL formulation are carried out to simulate the construction process of dynamic anchors. Through comparisons with results of field measurements, the CEL method in the present study is in good agreement with the general trend observed by in-situ measurements and thus, predicts a realistic large deformation movement for the dynamic anchors by free-fall dropping, which the conventional FE method cannot. Additionally, the appropriate parametric studies needed for verifying the characteristic of dynamic anchor are also discussed.

• Journal of Microbiology and Biotechnology
• /
• v.15 no.2
• /
• pp.302-309
• /
• 2005

Phytopathogenic Pectobacterium carotovorum subsp. carotovorum (Pcc) LY34 secretes multiple isozymes of the plant cell wall degrading enzyme endoglucanases. We have cloned a third cel gene encoding CMCase from Pcc LY34. The structural organization of the celC gene (AY188753) consisted of an open reading frame (ORP) of 1,116 bp encoding 371 amino acid residues with a signal peptide of 22 amino acids within the NH$_2$-terminal region of pre-CelC. The predicted amino acid sequence of CelC was similar to that of Peetobaeterium ehrysanthemi Cel8Y (AF282321). The CelC has the conserved region of the glycoside hydrolase family 8. The apparent molecular mass of CelC was calculated to be 39 kDa by CMC-SDS-PAGE. The cellulase­minus mutant of Pee LY34 was as virulent as the wild-type in pathogenicity tests on tubers of potato. The results suggest that the CelC of Pce LY34 is a minor factor for the pathogenesis of soft-rot.

• Journal of Life Science
• /
• v.22 no.4
• /
• pp.437-446
• /
• 2012

A metagenomic library of cow rumen in the pCC1FOS phage vector was screened in $E.$ $coli$ EPI300 for cellulase activity on carboxymethyl cellulose agar plates. One clone was partially digested with $Sau$3AI, ligated into the $Bam$HI site of the pBluescript II SK+ vector, and transformed into $E.$ $coli$ $DH5{\alpha}$. We obtained a 1.5 kb insert DNA, designated $cel$5C, which hydrolyzes carboxymethyl cellulose. The cel5C gene has an open reading frame (ORF) of 1,125 bp encoding 374 amino acids. It belongs to the glycosyl hydrolase family 5 with the conserved domain LIMEGFNEIN. The molecular mass of the Cel5C protein induced from $E.$ $coli$ $DH5{\alpha}$, as analyzed by CMC SDS-PAGE, appeared to be approximately 42 kDa. The enzyme showed optimum cellulase activity at pH 4.0, and $50^{\circ}C$. We examined whether the $cel$5C gene comes from the 49 identified cow rumen bacteria using PCR. No PCR bands were identified, suggesting that the $cel$5C gene came from the unidentified cow rumen bacteria.

• Journal of the Korean Geotechnical Society
• /
• v.31 no.8
• /
• pp.29-38
• /
• 2015

This paper presents the application of the Coupled Eulerian-Lagrangian (CEL) numerical technique to simulate the driving of open-ended piles into sandy soil. The main objective of this study was to investigate the applicability of CEL technique to the behavior of the driven pile penetration. Comprehensive studies to verify the behavior of driven pile penetration are presented in this paper. Through comparison with results of field load tests, the CEL methodology was found to be in good agreement with the general trend observed by in situ measurement, and the CEL approach accurately simulated the behavior of driven pipe piles.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.10
• /
• pp.1446-1454
• /
• 2021

Herein, we cloned and expressed an endo-β-1,4-glucanase gene (celA1805) from Bacillus subtilis B111 in Escherichia coli. The recombinant celA1805 contains a glycosyl hydrolase (GH) family 8 domain and shared 76.8% identity with endo-1,4-β-glucanase from Bacillus sp. KSM-330. Results showed that the optimal pH and temperature of celA1805 were 6.0 and 50℃, respectively, and it was stable at pH 3-9 and temperature ≤50℃. Metal ions slightly affected enzyme activity, but chemical agents generally inhibited enzyme activity. Moreover, celA1805 showed a wide substrate specificity to CMC, barley β-glucan, lichenin, chitosan, PASC and avicel. The K_m and V_max values of celA1805 were 1.78 mg/ml and 50.09 µmol/min/mg. When incubated with cellooligosaccharides ranging from cellotriose to cellopentose, celA1805 mainly hydrolyzed cellotetrose (G4) and cellopentose (G5) to cellose (G2) and cellotriose (G3), but hardly hydrolyzed cellotriose. The concentrations of reducing sugars saccharified by celA1805 from wheat straw, rape straw, rice straw, peanut straw, and corn straw were increased by 0.21, 0.51, 0.26, 0.36, and 0.66 mg/ml, respectively. The results obtained in this study suggest potential applications of celA1805 in biomass saccharification.

• Journal of information and communication convergence engineering
• /
• v.1 no.3
• /
• pp.129-132
• /
• 2003

In this paper, cobalt (Co)-electrode FBAR devices fabricated on seven-layered Bragg Reflectors are presented along with their resonance characteristics. ZnO films are used as the resonating material in FBAR devices where the Co electrode is 3000${\AA}$ thick. All processes are preformed in an RF magnetron sputtering system. As a result of characterization, the resonance characteristics are observed to depend strongly on the quality of ZnO film and Bragg Reflectors. In addition, the FBAR devices with W/$SiO_2$ reflectors show good resonance characteristics in term of return loss and quality-factor (Q-factor).

• Journal of Radiation Industry
• /
• v.11 no.3
• /
• pp.145-149
• /
• 2017

The cel7A sequence variation was analyzed between the wild type (Lentinula edodes KACC42378) and its cellulase activity enhanced mutant LER277. LER277 was induced by using gamma ray radiation ($^{60}Co$) at the $LD_{99}$ dose (0.94 kGy). Cloning and sequencing results showed that the cel7A coding DNA sequence (CDS) of LER277 had five nucleotide substitutions ($T{\rightarrow}C$, 201, 285 and 744 nt; $A{\rightarrow}G$, 525 nt; $C{\rightarrow}T$, 540 nt) and one hexanucleotide repeat insertion (GGCACC, within 1375-1392 nt) compared to that of the wild type. The Five nucleotide substitutions did not change the deduced amino acids and the hexanucleotide insertion elongated the GT repeat in a serine/threonine/glycine-rich linker. These results suggest that the enhancement of the cellulase activity in LER277 partly stemmed from cel7A changes by which the GT repeat of the linker is elongated.

• Journal of information and communication convergence engineering
• /
• v.2 no.2
• /
• pp.71-75
• /
• 2004

In non-selective frequency environment, it is difficult to take the advantage of path diversity. In the literature, some methods have been proposed to solve the issue. This paper presents a new method to obtain the resolvable path in Indoor Multi-carrier Code Division Multiple Access (MC-CDMA) system by using downlink beam forming. With the aid of downlink beamforming, the most reliable path is found and chosen for the communication link. The new approach is evaluated in term of bit error rate (BER) and power consumption. The simulation results show that the new approach can get better BER performance. However, the cost of BER improvement is a small degradation in power reservation.