• Journal of Microbiology and Biotechnology
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• v.22 no.7
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• pp.930-938
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• 2012

High levels of xylanase activity (143.98 IU/ml) produced by the newly isolated Paenibacillus campinasensis G1-1 were detected when it was cultivated in a synthetic medium. A thermostable xylanase, designated XynG1-1, from P. campinasensis G1-1 was purified to homogeneity by Octyl-Sepharose hydrophobic-interaction chromatography, Sephadex G75 gel-filter chromatography, and Q-Sepharose ion-exchange chromatography, consecutively. By multistep purification, the specific activity of XynG1-1 was up to 1,865.5 IU/mg with a 9.1-fold purification. The molecular mass of purified XynG1-1 was about 41.3 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Sequence analysis revealed that XynG1-1 containing 377 amino acids encoded by 1,134 bp genomic sequences of P. campinasensis G1-1 shared 96% homology with XylX from Paenibacillus campinasensis BL11 and 77%~78% homology with xylanases from Bacillus sp. YA-335 and Bacillus sp. 41M-1, respectively. The activity of XynG1-1 was stimulated by $Ca^{2+}$, $Ba^{2+}$, DTT, and ${\beta}$-mercaptoethanol, but was inhibited by $Ni^{2+}$, $Fe^{2+}$, $Fe^{3+}$, $Zn^{2+}$, SDS, and EDTA. The purified XynG1-1 displayed a greater affinity for birchwood xylan, with an optimal temperature of $60^{\circ}C$ and an optimal pH of 7.5. The fact that XynG1-1 is cellulose-free, thermostable (stability at high temperature of $70^{\circ}C{\sim}80^{\circ}C$), and active over a wide pH range (pH 5.0~9.0) suggests that the enzyme is potentially valuable for various industrial applications, especially for pulp bleaching pretreatment.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1388-1394
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• 2012

A xylanase-producing thermophilic strain, Geobacillus sp. TC-W7, was isolated from a hot spring in Yongtai (Fuzhou, China). Subsequently, the xylanase gene that encoded 407 amino acids was cloned and expressed. The recombinant xylanase was purified by GST affinity chromatography and exhibited maximum activity at $75^{\circ}C$ and a pH of 8.2. The enzyme was active up to $95^{\circ}C$ and showed activity over a wide pH range of 5.2 to 10.2. Additionally, the recombinant xylanase showed high thermostability and pH stability. More than 85% of the enzyme's activity was retained after incubation at $70^{\circ}C$ for 90 min at a pH of 8.2. The activity of the recombinant xylanase was enhanced by treatment with 10 mM enzyme inhibitors (DDT, Tween-20, 2-Me, or TritonX-100) and was inhibited by EDTA or PMSF. Its functionality was stable in the presence of $Li^+$, $Na^+$, and $K^+$, but inhibited by $Hg^{2+}$, $Ni^{2+}$, $Co^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Pb^{2+}$, $Fe^{3+}$, and $Al^{3+}$. The functionality of the crude xylanase had similar properties to the recombinant xylanase except for when it was treated with $Al^{2+}$ or $Fe^{2+}$. The enzyme might be a promising candidate for various industrial applications such as the biofuel, food, and paper and pulp industries.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.22-28
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• 2013

Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoosusceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAPtagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.15-21
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• 2013

We previously observed that Bacillus subtilis spores from sspE mutants presented a lower germination capacity in media containing high salt concentrations (0.9M NaCl). This deficiency was attributed to the absence of SASP-E (gamma-type small-acid-soluble protein), rich in osmocompatible amino acids released by degradation. Herein we observed that, in addition, this mutant spore presented a reduced capacity to use L-alanine as germinant (L-ala pathway), required longer times to germinate in calcium dipicolinate ($Ca^{2+}$-DPA), but germinated well in asparagine, glucose, fructose, and potassium chloride (AGFK pathway). Moreover, mild sonic treatment of mutant spores partially recovered their germination capacity in L-ala. Spore qualities were also altered, since sporulating colonies from the sspE mutant showed a pale brownish color, a higher adherence to agar plates, and lower autofluorescence, properties related to their spore coat content. Furthermore, biochemical analysis showed a reduced partition in hexadecane and a higher content of $Ca^{2+}$-DPA when compared with its isogenic wild-type control. Coat protein preparations showed a different electrophoretic pattern, in particular when detected with antibodies against CotG and CotE. The complementation with a wild-type sspE gene in a plasmid allowed for recovering the wild-type coat phenotype. This is the first report of a direct involvement of SASP-E in the spore coat assembly during the differentiation program of sporulation.

• Journal of Microbiology and Biotechnology
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• v.27 no.9
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• pp.1711-1715
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• 2017

In this study, we characterized the $bla_{NDM-5}$-bearing plasmid in a Klebsiella pneumoniae isolate that had lost the plasmid during serial passage. We determined the complete sequences of the plasmid pCC1410-2, which was extracted from a K. pneumoniae ST709 isolate collected at a Korean hospital from which two NDM-5-producing K. pneumoniae isolates were subsequently isolated. As a result, the pCC1410-2 plasmid had a backbone structure that was similar to those of two plasmids previously reported from the same hospital, but lacked some antibiotic resistance genes ($bla_{TEM-1}$, rmtB, mphR(A), mrx(A), and mph(A)). A 9-bp repeating unit encoding three amino acids (Gln-Gln-Pro) was inserted in TraD in pCC1410-2. Thus, the pCC1410-2 plasmid might be transferred from the previously identified carbapenem-resistant K. pneumoniae, but some delections and inversions might have occurred during the process. We compared the transfer frequency and stability of the plasmids. The relative frequency of conjugative transfer and stability in the host were significantly lower in pCC1410-2 than in previously reported $bla_{NDM-5}$-bearing plasmids in Korea. A low transfer frequency and instability in the host may cause underestimation of carbapenemase-producing Enterobacteriaceae in the clinical setting and in surveillance studies.

• Journal of Dairy Science and Biotechnology
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• v.35 no.4
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• pp.262-265
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• 2017

Bombyx mori (silkworm pupae) is a unique and biologically significant insect, which is a recognized source of high quality protein that provides all the essential amino acids required for human health. Recently, many studies have focused on various biomedical applications of B. mori proteins. The purpose of this study was to manufacture protein-fortified kefir containing different concentrations of B. mori powder according to pH and sensory evaluations. The value of the protein-fortified kefir increased but the pH decreased with increasing incubation time, indicating that the amount of B. mori powder did not affect and pH. Addition of B. mori powder also did not affect the sensory properties of overall acceptability, texture, and color compared to control group without addition of B. mori powder. However, flavor and taste were affected by increasing the amounts of B. mori powder, with a significant difference in both flavor and taste between the control and treated groups (both p<0.05). There was no significant difference in overall acceptability, texture, and color. Further studies are needed for producing kefir as a dietary supplement utilizing the functional properties of B. mori.

• Journal of Biosystems Engineering
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• v.41 no.4
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• pp.365-372
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• 2016

Purpose: Tea is the most frequently consumed drink worldwide, next to water. About 75% of the total world tea production includes black tea, and withering is one of the major processing steps critical for the quality of black tea. There are two types of tea withering methods: physical and chemical withering. Withering can be achieved by using tat, tunnel, drum, and trough withering systems. Of these, the trough withering system is the most commonly used. This study focuses on the different types of withering, their effect on the various quality attributes of tea, and other aspects of withering methods that affect superior quality tea. Results: During physical withering, tea shoots loose moisture content that drops from approximately 70-80% to 60-70% (wet basis). This leads to increased sap concentration in tea leaf cells, and turgid leaves become flaccid. It also prevents tea shoots from damage during maceration or rolling. During chemical withering, complex chemical compounds break down into simpler ones volatile flavor compounds, amino acids, and simple sugars are formed. Withering increases enzymatic activities as well as the concentration of caffeine. Research indicates that about 15% of chlorophyll degradation occurs during withering. It is also reported that during withering lipids break down into simpler compounds and catechin levels decrease. Improper withering can cause adverse effects on subsequent manufacturing operations, such as maceration, rolling, fermentation, drying, and tea storage. Conclusion: Freshly harvested leaves are conditioned physically and chemically for subsequent processing. There is no specified withering duration, but 14-18 h is generally considered the optimum period. Proper and even withering of tea shoots greatly depends on the standards of plucking, handling, transportation, environmental conditions, time, and temperature. Thus, to ensure consumption of high quality tea, the withering step must be monitored carefully.

• The Korean Journal of Physiology and Pharmacology
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• v.5 no.1
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• pp.19-25
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• 2001

We have studied the effects of excitatory amino acids on the expression of the c-fos and c-jun mRNA in rat C6 glioma cells. The glutamate, $N-methyl-_D-aspartate$ (NMDA), and kainic acid (KA) increased c-fos mRNA level in a concentration-dependent manner. However, they did not affect c-jun mRNA level. In addition, forskolin and phorbol 12-myristate 13-acetate (PMA) increased c-fos mRNA level. Furthermore, PMA increased c-jun mRNA level whereas forskolin downregulated c-jun mRNA level. The glutamate, NMDA and KA, at a concentration of 0.25 mM, did not affect the basal c-fos and c-jun mRNA levels, and also did not affect forskolin- and PMA-induced responses. Furthermore, both forskolin and PMA itself increased the phosphorylation of ERK (extracellular signal regulated kinase) and CREB (cyclicAMP responsible element binding protein) proteins. The KA, NMDA, and glutamate did not affect forskolin- induced increase of ERK and CREB phosphorylation. The KA decreased PMA-induced increase of phosphorylation of ERK and CREB proteins, whereas glutamate and NMDA did not affect the phosphorylation of ERK and CREB proteins induced by PMA. These findings suggest that, in C6 glioma cells, c-fos mRNA induction induced by EAAs is not mediated by phosphorylation of ERK and CREB proteins.

• Journal of Applied Biological Chemistry
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• v.59 no.2
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• pp.137-143
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• 2016

Hair is a dermal adjunctive organ that protects the body from external physical and chemical stimuli; hair undergoes anagen, catagen, and telogen phases, with hair-loss occurring during the telogen phase. Alopecia is a condition wherein a person undergoes hair-loss far exceeding the normal amount, owing to diverse external factors. Wild beans are rich in isoflavone and amino acids known to prevent hair-loss; compared to cultivated beans, many wild bean species have higher protein content. This study aimed to develop a hair growth promoting solution, with superior hair growth promoting effects and fewer side effects, using naturally obtained Glycine soja Siebold et Zucc (GSSZ) extracts. Seven-week-old C57BL/6N male mice were classified into different experimental groups. Hair growth was observed in GSSZ-treated mice, and compared against that seen in 3 % minoxidil (MXD, positive control)-treated mice. Visual observations revealed a greater reduction in hair-loss in MXD and GSSZ application groups, compared to that in TXN group (hair loss induction using 1 % testosterone). Evaluation using an image analysis software revealed that compared to the positive control, TXN + GSSZ group showed the highest hair growth. TXN + MXD and control groups exhibited similar follicular cell growth, while the hair growth promotion patterns were similar in the negative control (normal), TXN + GSSZ, and TXN groups, as observed via histological analysis. GSSZ did not induce cytotoxicity (even at 2 mg/mL) in keratinocytes and dermal papilla cells; alternately, dermal papilla cell proliferation was activated in a (GSSZ) concentration-dependent manner. Therefore, the GSSZ extract promoted hair growth and increased hair growth-related cell activity, and could therefore be utilized in alopecia treatment.

• Microbiology and Biotechnology Letters
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• v.13 no.4
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• pp.339-344
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• 1985

Cephalosporin C(CPC) inhibited the growth of Cephalosporium acremonium M-113, a potent CPC producer derived from C acremonium ATCC 20339. Similar inhibitory effects of CPC were also observed in growth of C. acremonium ATCC 20339 and ATCC 14553. Minimum inhibitory concentrations (MIC) of CPC on the growth of conidia and hyphae of C. acremonium M-113 were 200-500 and 3000-4000$\mu\textrm{g}$/$m\ell$ respectively in synthetic medium. MIC values were increased in complex media. The inhibitory effect of CPC was due to CPC-exerted inhibition of amino acids uptake by the cells. 3'-Group of CPC might be important in its inhibitory action. In audition, CPC itself could be utilized by the cells as a nitrogen source under nitrogen limited condition.