• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.989-998
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• 2016

Acidic endo-polygalacturonases are the major part of pectinase preparations and extensively applied in the clarification of fruits juice, vegetables extracts, and wines. However, most of the reported fungal endo-polygalacturonases are active and stable under narrow pH range and low temperatures. In this study, an acidic endo-polygalacturonase (EPG4) was purified and characterized from a mutant strain of Penicillium oxalicum. The N-terminal amino acid sequence of EPG4 (ATTCTFSGSNGAASASKSQT) was different from those of reported endo-polygalacturonases. EPG4 displayed optimal pH and temperature at 5.0 and 60-70℃ towards polygalacturonic acid (PGA), respectively, and was notably stable at pH 2.2-7.0. When tested against pectins, EPG4 showed enzyme activity over a broad acidic pH range (>15.0% activity at pH 2.2-6.0 towards citrus pectin; and >26.6% activity at pH 2.2-7.0 towards apple pectin). The K_m and V_max values were determined as 1.27 mg/ml and 5,504.6 U/mg, respectively. The enzyme hydrolyzed PGA in endo-manner, releasing oligo-galacturonates from PGA, as determined by TLC. Addition of EPG4 (3.6 U/ml) significantly reduced the viscosity (by 42.4%) and increased the light transmittance (by 29.5%) of the papaya pulp, and increased the recovery (by 24.4%) of the papaya extraction. All of these properties make the enzyme a potential application in the beverage industry.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1636-1643
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• 2012

Enzymatic pre-bleaching by modification of pulp fibers with xylanases is an attractive approach to reduce the consumption of toxic bleaching chemicals in the paper industry. In this study, an alkaliphilic endoxylanase gene was isolated from metagenomic DNA of a structurally stable thermophilic lignocellulose-degrading microbial consortium using amplification with conserved glycosyl hydrolase family 10 primers and subsequent genome walking. The full-length xylanase showed 78% sequence identity to an endo-${\beta}$-1,4-xylanase of Clostridium phytofermentans and was expressed in a mature form with an N-terminal His6 tag fusion in Escherichia coli. The recombinant xylanase Xyn3F was thermotolerant and alkaliphilic, working optimally at $65-70^{\circ}C$ with an optimal pH at 9-10 and retaining >80% activity at pH 9, $60^{\circ}C$ for 1 h. Xyn3F showed a $V_{max}$ of 2,327 IU/mg and $K_m$ of 3.5 mg/ml on birchwood xylan. Pre-bleaching of industrial eucalyptus pulp with no prior pH adjustment (pH 9) using Xyn3F at 50 IU/g dried pulp led to 4.5-5.1% increase in final pulp brightness and 90.4-102.4% increase in whiteness after a single-step hypochlorite bleaching over the untreated pulp, which allowed at least 20% decrease in hypochlorite consumption to achieve the same final bleaching indices. The alkaliphilic xylanase is promising for application in an environmentally friendly bleaching step of kraft and soda pulps with no requirement for pH adjustment, leading to improved economic feasibility of the process.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1701-1710
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• 2017

Classical swine fever virus (CSFV) is the etiologic agent of classical swine fever, a highly contagious disease that causes significant economic losses to the swine industry. The lapinized C-strain, a widely used vaccine strain against CSFV, has low growth efficiency in cell culture, which limits the productivity in the vaccine industry. In this study, a recombinant virus derived from C-strain was constructed and subjected to continuous passaging in PK-15 cells with the goal of acquiring a high progeny virus yield. A cell-adapted virus variant, RecCpp80, had nearly 1,000-fold higher titer than its parent C-strain but lost the ability to induce fever in rabbits. Sequence analysis of cell-adapted RecC variants indicated that at least six nucleotide changes were fixed in RecCpp80. Further adaption of RecCpp80 variant in swine testicle cells led to a higher virus yield without additional mutations. Introduction of each of these residues into the wild-type RecC backbone showed that one mutation, M979R (T3310G), located in the C-terminal region of E2 might be closely related to the cell-adapted phenotype. Rabbit inoculation revealed that $RecCpp40_{+10}$ failed to induce fever in rabbits, whereas $RecCpp80_{+10}$ caused a fever response similar to the commercial C-strain vaccine. In conclusion, the C-strain can be adapted to cell culture by introducing specific mutations in its E2 protein. The mutations in RecCpp80 that led to the loss of fever response in rabbits require further investigation. Continuous passaging of the C-strain-based recombinant viruses in PK-15 cells could enhance its in vitro adaption. The non-synonymous mutations at 3310 and 3531 might play major roles in the enhanced capacity of general virus reproduction. Such findings may help design a modified C-strain for improved productivity of commercial vaccines at reduced production cost.

• Journal of the Korean institute of surface engineering
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• v.34 no.2
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• pp.105-114
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• 2001

$Al_2$$O_3$ coatings were deposited on M2 high speed steels by the plasma enhanced chemical vapor deposition (PECVD) process, using a gas mixture of AlC1$_3$, $H_2$, $CO_2$ and Ar $Al_2$$O_3$ coatings had interference color and showed amorphous phase. $A1_2$X$A1_3$/($Ti_{0.5}$ /$Al_{0.5}$ )N double layer coatings were produced in the sequence of substrate $NH_3$ plasma pretreatment, ($Ti_{0.5}$$Al_{0.5}$)N depoition process, $Al_2$$O_3$ deposition process. $Al_2$ $O_3$/( $Ti_{0.5}$A $l_{0.5}$)N double layer coatings showed NaCl structure in ( $Ti_{0.5}$A $l_{0.5}$)N layer and amorphous phase in A1$_2$ $O_3$ layer. It was shown that $Al_2$ $O_3$ columns continuously grew onto ( $Ti_{0.5}$A $l_{0.5}$)N columns. ( $Ti_{0.5}$A $l_{0.5}$)N single coating and $Al_2$ $O_3$/( $Ti_{0.5}$A $l_{0.5}$)N double layer coating were oxidized at $700^{\circ}C$, 80$0^{\circ}C$, 90$0^{\circ}C$ for 1hr, 3hr in atmosphere. At 80$0^{\circ}C$, single layer coatings were oxidized, which were examined substrate oxide particle. But $Al_2$ $O_3$/ ( $Ti_{0.5}$A $l_{0.5}$)N double layer coatings maintained the asdeposited state. Therefore, $Al_2$ $O_3$/ ( $Ti_{0.5}$A $l_{0.5}$)N double layer coatings have moreexcellent oxidation resistance than ( $Ti_{0.5}$A $l_{0.5}$)N single layer coatings.X> 0.5/)N single layer coatings.s.

• The Plant Pathology Journal
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• v.26 no.4
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• pp.313-320
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• 2010

Genetic instability of the rice blast fungus Magnaporthe oryzae has been suggested as a major factor underlying the rapid breakdown of host resistance in the field. However, little information is available on the mechanism of genetic instability. In this study, we assessed the stability of repetitive DNA elements and several key phenotypic traits important for pathogenesis after serially transferring two isolates though rice plants and an artificial medium. Using isolate 70-15, we obtained a total of 176 single-spore isolates from 10 successive rounds of culturing on artificial medium. Another 20 isolates were obtained from germ tubes formed at the basal and apical cells of 10 three-celled conidia. Additionally, 60 isolates were obtained from isolate KJ201 after serial transfers through rice plants and an artificial medium. No apparent differences in phenotypes, including mycelial growth, conidial morphologies, conidiation, conidial germination, appressorium formation, and virulence, or in DNA fingerprints using MGR586, MAGGY, Pot2, LINE, MG-SINE and PWL2 as probes were observed among isolates from the same parent isolate. Southern hybridization and sequence analysis of two avirulence genes, AVR-Pita1 and AVR-Pikm, showed that both genes were also maintained stably during 10 successive generations on medium and plants. However, one reversible loss of restriction fragments was found in the telomere-linked helicase gene (TLH1) family, suggesting some telomere regions may be more unstable than the rest of the genome. Taken together, our results suggest that phenotype and genotype of M. oryzae isolates do not noticeably change, at least up to 10 successive generations on a cultural medium and in host plants.

• The Plant Pathology Journal
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• v.36 no.6
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• pp.536-557
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• 2020

Sugarcane yellow leaf virus (SCYLV) is a distinct member of the Polerovirus genus of the Luteoviridae family. SCYLV is the major limitation to sugarcane production worldwide and presently occurring in most of the sugarcane growing countries. SCYLV having high genetic diversity within the species and presently ten genotypes are known to occur based on the complete genome sequence information. SCYLV is present in almost all the states of India where sugarcane is grown. Virion comprises of 180 coat protein units and are 24-29 nm in diameter. The genome of SCYLV is a monopartite and comprised of single-stranded (ss) positive-sense (+) linear RNA of about 6 kb in size. Virus genome consists of six open reading frames (ORFs) that are expressed by sub-genomic RNAs. The SCYLV is phloem-limited and transmitted by sugarcane aphid Melanaphis sacchari in a circulative and non-propagative manner. The other aphid species namely, Ceratovacuna lanigera, Rhopalosiphum rufiabdominalis, and R. maidis also been reported to transmit the virus. The virus is not transmitted mechanically, therefore, its transmission by M. sacchari has been studied in different countries. SCYLV has a limited natural host range and mainly infect sugarcane (Sachharum hybrid), grain sorghum (Sorghum bicolor), and Columbus grass (Sorghum almum). Recent insights in the protein-protein interactions of Polerovirus through protein interaction reporter (PIR) technology enable us to understand viral encoded proteins during virus replication, assembly, plant defence mechanism, short and long-distance travel of the virus. This review presents the recent understandings on virus biology, diagnosis, genetic diversity, virus-vector and host-virus interactions and conventional and next generation management approaches.

• Proceedings of the Korean Vacuum Society Conference
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• 2013.08a
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• pp.91-91
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• 2013

Imagine a world where we could biomanufacture hybrid nanomaterials having atomic-scale resolution over functionality and architecture. Toward this vision, a fundamental challenge in materials science is how to design and synthesize protein-like material that can be fully self-assembled and exhibit information-specific process. In an ongoing effort to extend the fundamental understanding of protein structure to non-natural systems, we have designed a class of short peptides to fold like proteins and assemble into defined nanostructures. In this talk, I will talk about new strategies to drive the self-assembled structures designing sequence of peptide. I will also discuss about the specific interaction between proteins and inorganics that can be used for the development of new hybrid solar energy devices. Splitting water into hydrogen and oxygen is one of the promising pathways for solar to energy convertsion and storage system. The oxygen evolution reaction (OER) has been regarded as a major bottleneck in the overall water splitting process due to the slow transfer rate of four electrons and the high activation energy barrier for O-O bond formation. In nature, there is a water oxidation complex (WOC) in photosystem II (PSII) comprised of the earthabundant elements Mn and Ca. The WOC in photosystem II, in the form of a cubical CaMn4O5 cluster, efficiently catalyzes water oxidation under neutral conditions with extremely low overpotential (~160 mV) and a high TOF number. The cluster is stabilized by a surrounding redox-active peptide ligand, and undergo successive changes in oxidation state by PCET (proton-coupled electron transfer) reaction with the peptide ligand. It is fundamental challenge to achieve a level of structural complexity and functionality that rivals that seen in the cubane Mn4CaO5 cluster and surrounding peptide in nature. In this presentation, I will present a new strategy to mimic the natural photosystem. The approach is based on the atomically defined assembly based on the short redox-active peptide sequences. Additionally, I will show a newly identified manganese based compound that is very close to manganese clusters in photosystem II.

• Journal of Life Science
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• v.18 no.4
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• pp.482-487
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• 2008

In this study, we screened and identified the bacterial strain showing high alkaline pretense inhibitor activity from marine environment. Nine bacterial strains with alkaline pretense inhibitor activity were isolated from marine sediments. Among them, strain C12 had the highest alkaline pretense inhibitor activity and was selected for further taxonomical study. On the basis of morphological and physiological characteristics, strain C12 was identified as the genus Streptomyces. A phylogenetic analysis of the 165 rDNA showed that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces thermocarboxydus. Morphological characteristics showed cylindrical spore chain and smooth spore surface by scanning electron microscope. Strain C12 was grown on all media except for ISP 9 agar. This strain could be grown in the medium containing up to 9% NaCl.

• Molecules and Cells
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• v.27 no.4
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• pp.423-428
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• 2009

Superoxide dismutases (SODs) constitute the first line of cellular defense against oxidative stress in plants. SODs generally occur in three different forms with Cu/Zn, Fe, or Mn as prosthetic metals. We cloned the full-length cDNA of the Thellungiella halophila Cu/Zn-SOD gene ThCSD using degenerate RT-PCR and rapid amplification of cDNA ends (RACE). Sequence analysis indicated that the ThCSD gene (GenBank accession number EF405867) had an open reading frame of 456 bp. The deduced 152-amino acid polypeptide had a predicted molecular weight of 15.1 kDa, an estimated pI of 5.4, and a putative Cu/Zn-binding site. Recombinant ThCSD protein was expressed in Escherichia coli and assayed for SOD enzymatic activity in a native polyacrylamide gel. The SOD activity of ThCSD was inactivated by potassium cyanide and hydrogen peroxide but not by sodium azide, confirming that ThCSD is a Cu/Zn-SOD. Northern blotting demonstrated that ThCSD is expressed in roots, stems, and leaves. ThCSD mRNA levels increased by about 30-fold when plants were treated with sodium chloride (NaCl), abscisic acid (ABA), and indole-acetic acid (IAA) and by about 50-fold when treated with UVB light. These results indicate that ThCSD is involved in physiological pathways activated by a variety of environmental conditions.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.4
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• pp.495-500
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• 1999

The carrageenans are linear, sulfated Polysaccharides extracted from various species of the Rhodophyta (marine red algae). The carrageenan backbone is based on a repeating disaccharide sequence of $\beta$-D-galactopyranose residues linked glycosidically through position 1 and 3, and $\alpha$-D-galactopyranose residues linked glycosidically through position 1 and 4. Carrageenans are typical food polysaccharides in that food applications overwhelmingly dominate their end uses. Other applications, hewer, including cosmetics, pharmaceuticals, industrial suspensions and paints are also of importance But because of its high degree of gelling and viscosity with low solubility, carrageenan is limited to use beyond $0.03\%$ as food additives. Response Surface Methodology was applied for optimizing the processing parameters of ultrasound treatment in order to produce low-molecular-weight carrageenan. The use of ultrasound significantly reduced viscosity of $\lambda$-carrageenan solutions. Optimal parameters for ultrasound reduction of carrageenan molecular weight were: temperature, $10^{\circ}C$; ultrasound intensity, 121.64 $W/cm^2$ ; tarrageenan concentration, $2\%$; treatment time, 40 min. As the gel permeation chromatogram of dextran standards (M.W.= 500,000 ; 260,000 ; 167,000 ; 71,400 ; 42,000) and ultrsound treated carrageenan, the molecular weight of ultrasound treated carrageenan were approximately 200,000 (peak 1) and 60,000 (peak 2), respectively.