• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.98-104
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• 1988

A bacterial isolate FRI-33 which produces hydrophillic polysaccharide was identified and its cultural condition was investigated. FRI-33 was identified as Enterobacter agglomerans. The optimum cultural conditions for polysaccharide production were 3$0^{\circ}C$, pH 5.7, using medium composed of glucose 25 g/$\ell$, peptone 2.0 g/$\ell$, yeast extract 0.5 g/$\ell$, KH$_2$PO$_4$ 1.0g/$\ell$, MgSO$_4$.7H$_2$ O 1.0g/$\ell$, CaCO$_3$ 2.5g/$\ell$. The polysaccharide production after 72 hours was 8.41 g/$\ell$. The polysaccharide was composed of galactose (1.0 mole), xylose (1.5 mole), gluconodeltalactone (1.9 mole) and ribose (0.03 mole). The apparent viscosity of 1% polysaccharide solution was 504 mPa.s at 60 rpm and intrinsic viscosity was 45.80 d$\ell$/ g.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1773-1784
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• 2008

Effects of various industrially important carbon sources (glucose, sucrose, xylose, gluconate, and glycerol) on shikimic acid (SA) biosynthesis in Escherichia coli were investigated to gain new insight into the metabolic capability for overproducing SA. At the outset, constraints-based flux analysis using the genome-scale in silico model of E. coli was conducted to quantify the theoretical maximum SA yield. The corresponding flux distributions fueled by different carbon sources under investigation were compared with respect to theoretical yield and energy utilization, thereby identifying the indispensable pathways for achieving optimal SA production on each carbon source. Subsequently, a shikimate-kinase-deficient E. coli mutant was developed by blocking the aromatic amino acid pathway, and the production of SA on various carbon sources was experimentally examined during 51 batch culture. As a result, the highest production rate, 1.92 mmol SA/h, was obtained when glucose was utilized as a carbon source, whereas the efficient SA production from glycerol was obtained with the highest yield, 0.21 mol SA formed per mol carbon atom of carbon source consumed. The current strain can be further improved to satisfy the theoretically achievable SA production that was predicted by in silico analysis.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1559-1565
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• 2014

Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

• Journal of Microbiology and Biotechnology
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• v.23 no.3
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• pp.397-404
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• 2013

Paenibacillus xylanilyticus KJ-03 was isolated from soil samples obtained from a field with Amorphophallus konjac plants. A gene encoding xylanase was isolated from KJ-03 and cloned using a fosmid library. The xynA gene encodes xylanase; it consists of 1,035 bp and encodes 345 amino acids. The amino acid sequence deduced from the P. xylanilyticus KJ-03 xylanase showed 81% and 69% identities with those deduced from the P. polymyxa E681 and Paenibacillus sp. HPL-001 xylanases, respectively. The xynA gene comprises a single domain, consisting of a catalytic domain of the glycosyl hydrolase (GH) 10 family. The xynA gene was expressed in Escherichia coli BL21 (trxB), and the recombinant xylanase was purified by Niaffinity chromatography. The purified xylanase showed optimum activity with birchwood xylan as a substrate at $40^{\circ}C$ and pH 7.4. Treatment with $Mg^{2+}$ and $Li^+$ showed a slight decrease in XynA activity; however, treatment with 5 mM $Cu^{2+}$ completely inhibited its activity. The results of the thin layer chromatography analysis indicated that the major hydrolysis product was xylobiose and small amounts of xylose and xylotriose. XynA showed increased activity with oat spelt xylan and birchwood xylan, but showed only slight activity with locust bean gum.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1891-1907
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• 2016

Yeasts that are present in marine environments have evolved to survive hostile environments that are characterized by high exogenous salt content, high concentrations of inhibitory compounds, and low soluble carbon and nitrogen levels. Therefore, yeasts isolated from marine environments could have interesting characteristics for industrial applications. However, the application of marine yeast in research or industry is currently very limited owing to the lack of a suitable isolation method. Current methods for isolation suffer from fungal interference and/or low number of yeast isolates. In this paper, an efficient and non-laborious isolation method has been developed and successfully isolated large numbers of yeasts without bacterial or fungal growth. The new method includes a three-cycle enrichment step followed by an isolation step and a confirmation step. Using this method, 116 marine yeast strains were isolated from 14 marine samples collected in the UK, Egypt, and the USA. These strains were further evaluated for the utilization of fermentable sugars (glucose, xylose, mannitol, and galactose) using a phenotypic microarray assay. Seventeen strains with higher sugar utilization capacity than the reference terrestrial yeast Saccharomyces cerevisiae NCYC 2592 were selected for identification by sequencing of the ITS and D1/D2 domains. These strains belonged to six species: S. cerevisiae, Candida tropicalis, Candida viswanathii, Wickerhamomyces anomalus, Candida glabrata, and Pichia kudriavzevii. The ability of these strains for improved sugar utilization using seawater-based media was confirmed and, therefore, they could potentially be utilized in fermentations using marine biomass in seawater media, particularly for the production of bioethanol and other biochemical products.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.10a
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• pp.143.2-144
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• 2003

품종별 감잎의 주요성분과 항산화 활성을 비교하여 고기능성 감잎차의 우수성을 알리는 기초자료로 활용하고자 이 실험을 수행하였다. 상주둥시, 갑주백목, 월하시, 청도반시, 녹차(대조구)의 생엽을 7월 초순에 채엽 후 건조시켜 시료로 사용하였고, 비타민C, 카페인, 카테친, Gallic acid, 향기성분, 유리당, 유리아미노산, 칼슘, 항산화 활성을 조사하였다. 비타민C는 월하시 0.089%, 청도반시 0.079%, 갑주배목 0.032%, 상주둥시 0.015%, 녹차 0.01%로 비타민C 함량이 가장 많은 월하시가 녹차에 비해 8배정도 높았고, 카페인은 감잎 4품종 모두 발견되지 않았고, 녹차는 6.63%이었다. 카테친은 청도반시 0.35%, 갑주백목 0.34%, 월하시 0.24%, 상주둥시 0.18%, 녹차(대조구) 0.07%이였고, Gallic acid는 상주 둥시 0.32%, 갑주백옥 0.2%, 월하시 0.05%, 청도반시 0.03%, 녹차(대조구) 1.41%이었다. 칼슘은 청도반시 9516.1PPM, 영동월하시 6863.5PPM, 봉옥 6563.5PPM, 상주둥시 5420.1PPM, 녹차(대조구) 2349.7PPM이였고, 유리당은 감품종간에는 큰 차이는 없었으나 녹차에 비해 Xylose, fructose, Glucose, Sucrose의 함량이 높았고, Maltose의 함량에 일어서는 녹차가 높았다. 유리아미노산은 31항목을 조사하였고, 전체 함량은 상주둥시 60.40, 봉옥 53.21, 월하시 52.29, 청도반시 47.58, 녹차(대조구) 114.72nmo1/${\mu}\ell$이었다. 시료의 향기성분은 생엽을 건조시켜 전자코를 이용하여 분석하였고, 감잎 품종간에는 향기 패턴이 비슷한 경향이였으나, 녹차의 패턴과는 차이가 있었다. 감잎의 DPPH radical 소거활성은 상주둥시 RC$_{50}$($\mu\textrm{g}$)=64.5 청도반시 64.0, 월하시 42.0, 갑주백목 47.0, 녹차(대조구) 19.0으로 김잎품종중 월하시의 항산화 활성이 높았고, 녹차의 항산화 활성이 감잎에 비해 높았다.

• Food Industry And Nutrition
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• v.2 no.1
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• pp.7-9
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• 1997

내열설 미생물, Thermus caldophilus CK24에 대한 탄수화물 생합성을 연구하는 과정에서 다양한 탄수화물 관련효소를 탐색하고 그레 대한 생화학적 및 분자생물학적 연구를 수행하고 있다. 일차로 내열성 미생물내 1) 당핵산염 합성효소와 당전이 효소, 2) 탄수화물 대사효소. 3)탄수화물 분해 및 전환효소의 존재를 HPLC/Bio-LC분석을 통하여 확인하고 이들에 대한 연구를 진행하고 있다. 본 연구발표에서는 포도당을 과당으로 전환하는 이성화효소(xylose isomerase), 그리고 맥아당을 트레할로스로 전환하는 트레할로스 합성효소(trehalose synthase)를 소개하고저 한다. 이성화효소는 이미 산업적 과당 생산에서 대규모적으로 사용되고 있는 식품산업효소이다. 본 연구에서는 Thermus caldophilus GK24, Thermus thermophilus HB8, Thermus flavus AT62 3종의 내열성 미생물에 대한 이성화효소 유전자를 클로닝 하고, 각 재조합하고 이성화효소를 대량생산하였다. 이 내열성 이성화효소는 최적 반응 온도가 8$0^{\circ}C$이고, 포도당을 과당으로 전환하는 수유른 55%이었다. 이러한 과당전환률은 이미 산업적으로 사용되고 있는 이성화효소의 과당전환률(43%)보다 훨씬 높은 것으로 과당 생산공정의 단순화의 생산성 향상에 결정적인 요인이라 할 수 있다. 한편 본 이성화효소의 산업적 특성을 증대하기 위하여 구조-기능관계 연구를 착수하였다. 우선 내열성 이산화 효소의 입체 구조를 결정하였고, 구조조정에 따른 기능적 특성을 조사하기 위하여 특정 위치의 선택적 변이 연구를 진행하고 있다. 끝으로 포도당 전이 효소를 추적하던 과정에서 맥아당을 트레할로스로 전환하는 새로운 효소를 Thermus caldo-philus GK24에서 발견하였다. 그 트레할로스 합성효소는 분자량이 약 110kDa이고 최적 반응온도가 75$^{\circ}C$이면, 조효소없이 맥아당을 트레할로스로 80%이상 전환해 주는 가역효소이었다. 본 연구에서는 효소반응의 조건과 특성을 조사하였고, 효소 아미노-밀단의 서열결정정보를 통하여 효소의 유전자를 클로닝 하고 그 유전자의 구조와 발현연구를 진행하고 있다.

• Journal of Ginseng Research
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• v.28 no.3
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• pp.143-148
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• 2004

Brown color intensity has been a major factor to estimate the quality of red ginseng and its products. This study deals with the relationship between the browning reaction of ginseng root and two compounds, arginyl-fructosyl-glucose(Arg-fru-glc) and arginyl-fructose (Arg-fru), in the model system of steaming and heat-drying processes for the preparation of red ginseng. During the steaming process, a marked decrease of starch and a considerable formation of maltose occurred in main roots of raw ginseng, but the formation of glucose was scarcely observed. After the heat-drying process, the brown color intensity of the powdered preparation of steamed main roots was 3 to 4 times higher than that of the powdered preparation of raw main roots. Also, when the heat- drying process was done with the addition of L-arginine, brown color intensity of the powdered preparation of steamed main roots was 12 to 13 times higher than that of the powdered preparation of raw main roots. The amount ratios of browning reaction products formed from sugar compounds and amino acids in the model system of steaming and heat-drying treatments in vitro were in order of xylose > glucose > fructose > maltose > dextrin (DE 9) > sucrose > dextrin (DE 8) and soluble starch. Each solution of Arg-fru-glc and Arg-fru that were synthesized chemically from maltose plus L-arginine and glucose plus L-arginine, respectively, changed from colorless to brown color during the heat-drying treatment. Amino acids or sugars were effective on the acceleration of each browning reaction of Arg-fru-gIc and Arg-fru during the heat-drying treatment.

• Korean journal of applied entomology
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• v.46 no.2
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• pp.303-311
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• 2007

From the course of screening of useful xylanase producing microorganism from a phytophagous longicorn beetle, we isolated an extra-cellular xylanase producing strain, Paenibacillus sp. HY-8 from the intestine of Moechotypa diphysis adult. On the basis of morphological, biochemical and phylogenetic studies of the new isolate was identified as a Paenibacillus species. Production of xylanase in this strain was strongly induced by adding xylan to the growth medium and repressed by glucose or xylose. The highest xylanase production was attained in the M9 media containing 1% yeast extract and 0.5% birchwood xylan when cultured at $25^{\circ}C$ for 24 hrs. HY-8 producing xylanase showed superior hydrolytic activities against various plant source feedstuff than control xylanase produced by Tricoderma sp. at pH 6.0.

• Journal of the Korean Society of Food Culture
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• v.29 no.3
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• pp.278-285
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• 2014

This study investigated the quality characteristics of fermented wines using a selected strain to obtain basic data on sugar-free tomato wine production. Alcohol content of the tomato and commercial wine was 8.2~12.9%, whereas tomato wine showed a relatively low alcohol content of 8.2%. For total phenol content, Chile wine showed the highest value of 162.89 mg/100 g, followed by persimmon wine at 122.33 mg/100 g. Tomato wine showed a relatively high total phenol content of 96.57 mg/100 g. In all sections, xylose was not detected, although there were differences in free sugars depending on each product. Acetic and citric acid contents were highest in tomato wine, and the six kinds of commercial wine and tomato wine showed acetic acid content of 800.6 mg/100 g and citric acid content of 1,064.4 mg/100 g. The total free amino acid content was 100.63 mg/100 g in tomato wine, which was 2~3 times higher compared to that in commercial wine ($26.33{\pm}52.15mg/100g$).