• Journal of Microbiology and Biotechnology
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• 제21권8호
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• pp.830-837
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• 2011

A genomic DNA fragment encoding a putative maltooligosyltrehalose trehalohydrolase (NfMTH) for trehalose biosynthesis was cloned by the degenerate primer- PCR from cyanobacterium Nostoc flagelliforme. The ORF of NfMTH is 1,848 bp in length and encodes 615 amino acid residues, constituting a 70 kDa protein. The deduced amino acid sequence of NfMTH contains 4 regions highly conserved for MTHs. By expression of NfMTH in E. coli, the function of this protein was demonstrated, where the recombinant protein catalyzed the hydrolysis of maltooligosyl trehalose to trehalose. The expressions of MTH and maltooligosyltrehalose synthase in the filaments of N. flagelliforme were upregulated significantly under dehydration stress, NaCl stress, and high temperature-drought stress. The accumulations of both trehalose and sucrose in the filaments of N. flagelliforme were also improved significantly under the above stresses. Furthermore, trehalose accumulated in smaller quantities than sucrose did when under NaCl stress, but accumulated in higher quantities than sucrose did when under temperature-drought stress, indicating that both trehalose and sucrose were involved in N. flagelliforme adapted to stresses and different strategies conducted in response to various stress conditions.

• 미생물학회지
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• 제13권3호
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• pp.85-90
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• 1975

Crude cellulases freshly prepared from cultures of Aspergillus niger, Prnicillum motatum, Trichoderma vride 16273 and Trichoderma viride 16374 were assayed on 4 different substrates including Na-CMC, cellulose powder, starch and sucrose. Enzyme prepared from A. niger contained highly active hydrolytic enzymes of the 4 substrates assayed. P. notatum [yielded relatively lower amount of cellulase but the extracts were also highly reactive on starch and sucrose. Trichoderma viride 16274 yielded very little cellulase and invertase, but the extracts showed a high degree of amylase activity. Trichoderma viride 16374, however, yielded collulase comparable to that of Penicillium notatum, but lower activities of amylase and invertase were seen. Commercial cellulases prepared from Penicillium notatum (cellulase[K]) and Trichoderma viride(cellulase[J]) indicated enzyme activities closely parallel to the crude enzymes freshly prepared from fungus cultures. The optimum pH's of cellulolytic activities of cellulase[K] and cellulase[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase[K] and cellualse[J] were 4.0 and 5.0 respectively. The optimum temperatures of the cellulolytic activities of cellulase [K] and cellulase [J] were $60{\circ}C$ and $50{\circ}C$ respectively. Assuming the average molecular weight of Na-CMC is about 115,000, the Km values of cellulase [K] and cellulase[J] were found to be $3.3{\times}10^{-5}/nM$ and $3.3{\times}10^{-4}/nM$ respectively.

• Asian-Australasian Journal of Animal Sciences
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• 제15권11호
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• pp.1606-1610
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• 2002

The dynamics of fermentation were studied with Italian ryegrass ensiled in the laboratory silos. The silos were kept in the room set at 25$^{\circ}C$, and then were opened on 0.5, 1, 2, 3, 5, 7 and 14 days after ensiling, respectively. The samples were taken from three silos at each sampling time for chemical analyses. Mono-and disaccharides composition was determined for glucose, fructose and sucrose by high performance liquid chromatography. The Italian ryegrass silage succeeded to achieve lactate type fermentation; high values of lactic acid (85.83 g/kg) and lactic acid/acetic acid at the end of ensiling (14 day), low values of pH (3.74), acetic acid (5.38 g/kg), ethanol (19.20 g/kg) and $NH_3-N/Total\;N$ (75.84 g/kg), no or only small amounts of butyric acid, valeric acid and propionic acid. The fermentation dynamics showed a fast and large pH decrease caused by a fast and large production of lactic acid during the first 5 days. Mono-and disaccharides composition largely decreased within initial 0.5 day (12 h) of ensiling. Sucrose disappeared rapidly within initial 0.5 day of ensiling, and fructose and glucose contents showed an initial rise during ensiling, and then decreased gradually. These indicated that the enzymes of plant tissue were active within 2 days of ensiling, which caused the initial rise in fructose and glucose from the hydrolysis of sucrose and fructans. After 5 days of ensilage, glucose was consumed completely, suggesting that glucose was the first fermentation substrate. After 2 days of ensiling, sum amounts of lactic acid and remaining mono-and disaccharides proved to be larger than the quantity of mono-and disaccharides in the initial grass. From the facts mentioned above, it was suggested that considerable amounts of lactic acid were produced from some other substrate such as fructans than initial mono-and disaccharides.

• KSBB Journal
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• 제9권2호
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• pp.191-199
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• 1994

김치 발효에 주관여 미생물로 작용하는 Lactoba-cillus plantarum, Leuconostoc mesenteroides 균주에 세포 융합법을 적용시켜 우수한 형질의 재조합체를 얻기 위한 균주 개량의 방법으로 이용하고자 하여 그 기초가 되는 원형질 형성의 최적 조건을 조사하였다. Protoplast를 생성하기 위하여 세포벽 용균 효소로 lysozyme $40{\mu}$g/ml와 protoplast 형성률이 가장 좋은 생육시기인 15hr으로 결정하였다. $30^{\circ}C$, pH 7.5에서 현저한 흡광도 감소 경향과 protoplast 형성률도 우수하였다. 15% sucrose, 20mM $MgCl_2$, 6mM $CaCl_2$에서는 흡광도 감소 경향을 보이지 않았으나 protoplast 형성률은 다른 농도에서보다 증가하는 경향을 나타냈다. Plasma expander로 알려진 gela tin을 첨가하였을 경우에는 660nm에서 유의적인 흡광도 감소 경향을 보이지 않았으며 단지 proto-plast를 형성시키는데 있어서 중요한 보조적 역할을 수행하는 것으로 여겨진다. 균주 발육을 하기 위한 최소 저지 농도를 조사한 결과 marker로 이용할 수 있는 항생제는 erythromycin($15{\mu}$g/ml), ampicillin($50{\mu}$/ml), tetracyclin($30{\mu}$g/ml), chloramphe-nicol($30{\mu}$g/ml)이었다. Leu. mesenteroides는 모든 조건에서 L.plantarum보다 용균 작용과 protoplast 형성 효율이 낮아 L. plantarum에 준하여 protoplast 생성조건을 결정하였다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.192-192
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• 2022

Sweetpotato is rich in starch, which is converted to sugar during storage due to enzymatic hydrolysis. The sugar content of sweetpotato is a component related to taste and storability. In this study, the sugar content (fructose, glucose, maltose, sucrose and total sugar content) of 94 genotypes was evaluated and the GWAS (Genome-Wide Association Study) was conducted to search for candidate genes for sugar content. The fructose and glucose content were 0.2 ~ 8.8 and 0.2 ~ 9.4 g/100g, respectively. The maltose, sucrose and total sugar content were 0.2 ~ 9.1,3.2 - 30.0 and 7.9 ~ 40.2 g/100g, respectively. The fructose and glucose showed a positive correlation (0.98). The 94 genotypes were genotyped with genotyping-by-sequencing (GBS) and aligned against the reference genome sequences of sweetpotato. The GBS libraries from 94 genotypes were sequenced on an Illumina HiSeqXten system, and 1,339,892 SNPs (Single Nucleotide Polymorphism) were generated. Filtering for < 60% missing rate and > 0.05 minor allele frequency resulted in a total of 44,255 SNPs used in GWAS. The GAPIT (Genome Association and Prediction Integrated Tool) was used to conduct based on the mean of sugar content with a Bonferroni-corrected chromosome-wide significance threshold with a -logio(P) of 5.95. The significant SNPs were obtained with fructose (seven), glucose (six), maltose (four) and sucrose (nine). There were several genes related to sugar content around the significant SNPs such as sugar transport protein 8-like, probable galactose-1 -phosphate uridyltransferase-like and beta-amylase. These results will contribute to understanding of sugar content and conversion in sweetpotato.

• 한국식품영양과학회지
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• 제40권11호
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• pp.1575-1581
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• 2011

본 연구에서는 유채박 섬유질을 고부가가치로 활용하기에 앞서 적합한 화학적 전처리 조건을 찾고자, 산 및 알칼리 촉매로 다양한 조건 하에 처리한 후, 각 인자들에 따른 유채박의 분해율 및 유리당 함량을 측정하였다. 유채박은 $H_2SO_4$, HCl, NaOH 촉매 중 HCl 촉매 하에서 가장 효과적으로 분해되었으며, 특히 1%의 촉매 농도에서 높은 분해율을 보였다. 반면 $H_2SO_4$, HCl 촉매 하에서 유채박 투입량이 낮을수록 분해율이 증가하였으나, NaOH 촉매 하에서는 유채박 투입량에 따른 분해율 차이가 거의 관찰되지 않았다. 분해 후 측정된 총 유리당 함량은 HCl를 처리한 경우가 가장 높았으며, 유채박 투입량이 증가함에 따라 총 유리당 함량이 감소하였으나 이에 따른 영향은 미미했다. 또한 2% 촉매에서 총 유리당 함량이 높았으나 HCl 촉매의 경우 1%일 때가 다른 처리 조건들보다 총 유리당 함량이 높았다. 각각의 유리당으로는 $H_2SO_4$, HCl 촉매 하에서 glucose, galactose, arabinose, fructose가 주로 분리되었으며 NaOH 촉매 하에서는 대부분 sucrose가 분리되었다. 촉매 농도가 증가함에 따라 glucose, galactose 함량은 증가하였고, fructose 함량은 감소하였으며, 거의 모든 유리당에서 유채박 투입량이 증가함에 따라 그 함량이 감소하였다. 상기 결과를 종합해 보면 총 유리당은 유채박을 1% HCl를 사용하여 2 g/100 mL의 비율로 화학적 처리하는 것이 본 연구 범위 내에서 총 당을 분리해내는데 가장 우수한 것으로 조사되었으며 glucose, galactose는 2% HCl-0.5 g/100 mL, fructose는 0.5% $H_2SO_4$-0.5 g/100 mL 조건에서 효율적인 분리가 이루어짐을 확인하였다. 따라서 목표하는 당에 따라 적합한 촉매, 촉매 농도, 시료 투입량을 조절하여 분리해낼 수 있을 것으로 생각한다. 현재 분리되어 나온 당을 바탕으로 기능성 당 및 수용성 식이섬유를 얻는 연구를 수행하고 있으며, 실질적으로 대량의 당들을 순도 높게 얻기 위해서는 처리 규모(시료양, 촉매)를 늘리거나 각각의 당들을 효과적으로 정제하는 연구도 필요할 것으로 생각한다.

• 생명과학회지
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• 제25권9호
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• pp.1021-1026
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• 2015

Invertase (β-D-fructosfuranosidase, EC 3.2.1.26)는 설탕을 포도당과 과당으로 가수분해하는 반응을 촉매한다. 3종류의 invertases [액포(수용성 산), 세포질(수용성 알칼리) 및 세포벽 결합]가 식물에서 연구되어 왔다. 우리는 순차적인 ammonium sulfate 침전, 이온교환크로마토그래피, 흡수크로마토그래피, Green-19 친화크로마토그래피 과정을 통해 완두콩(Pisum sativum L.) 발아체로부터 중성 invertase의 세포막 연결 isoform을 430배 순수 분리하였다. 분리된 세포막과 결합 된insoluble invertase (IN-INV)는 최적 pH는 중성에서 알칼리 사이(pH 6.8-7.5)로 나타났다. 이 효소는 Tris 뿐만 아니라 Hg²⁺ and Cu²⁺와 같은 중금속에 의해 저해되었다. IN-INV 의 K_m과 V_max 값은 각각 12.95 mM과 2.98 U/min으로 측정되었다. IN-INV는 기질로써 과당뿐만 아니라 라피노오스와 반응하기 때문에 진정한 β-fructofuranosidase로 판명되었다. IN-INV의 분자량 20 kDa이었다. 위 결과로 볼 때 GA 영향으로 급속히 자라는 발아체에서 단백질이 분리되었는데 특징적으로 invertase였다.

• Journal of Ginseng Research
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• 제4권2호
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• pp.133-145
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• 1980

In order to find out the inhibitors of acetic acid fermentation in Korean ginseng (Panax Sin son C. A. Meyer), total aglycone, panaxadiol, panaxadiol, oleanolic acid and ${\beta}$ -sitosterol were added to the basal medium, respectively, and a surface culture was carried out at 30$^{\circ}C$. The results were as follows: 1 . Saponins lost their activity to inhibit the acetic acid fermentation by hydrolysis. 2 Panaxadiol inhibited slightly, and the degree of inhibition was about 1/300 of that of free saponins. 3. Panaxadiol and oleanolic acid inhibited silighly similar to total aglycone. 4. Acetic acid fermentation was stimulated at the early stage when ${\beta}$-sitosterol was added to the media below the level of 0.000815%. But the fermentation was inhibited when media contained it more than that media 5. An over-oxidation of acetic acid was observed when the media contained total aglycone. panaxadiol, panaxatriol, oleanolic acid and ${\beta}$-sitosterol, respectively, while the media which contained sucrose, ginseng extracts ginseng saponins was shown not to be over-oxidized.

• 한국식품영양과학회지
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• 제26권5호
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• pp.794-799
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• 1997

The effective fermentation methods of Kimchi juice for utilization of outer layer of Chinese cabbage, an waste of Kimchi industry were studied. The Kimchi juice prepared with brining and grinding the waste of Chinese cabbage and addition of spices was fermented at $25^{\circ}C$. Addition of 5$^{\circ}C$15% fermented Kimchi juice of pH 5.4 at initial stage and pH 4.4 at middle state resulted in a significant increase in fermentation rate and solid content after 12 hours of fermentation. The combined method of enzymatic hydrolysis(0.1% viscozyme) of the brined and ground cabbage and addition of 2.0% NaCl, 1.0% sucrose and 10% fermented juice of pH 5.4 first and 4.4 during fermentation, respectively resulted in more rapid fermentation. The solid concentration was 5 times higher than control at maximum point and acidic and total flavor intensity were also significantly high.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.188-191
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• 2002

A glucan and a fructan producing enzymes from Leuconostoc mesenteroides NRRL B-1149 were prepared and concentrated from the cu1ture of 1.5% sucrose using polysulfone ultrafiltration hollow fiber in the presence of 0.1% (w/v) Tween 80, 1 mM $CaCl_2$, and 0.02% $NaN_3$. The molecular masses of the enzymes were estimated to be about 213.6 kDa and 180 kDa, respectively, based on the PAS staining for the glucosyltransferase and Mukasa method for fructosyltransferase. Polymers produced by the enzymes showed different solubility; an insoluble glucan and a soluble fructan. The linkages of polymers were determined by methylation using Hakomori reagent and following acid hydrolysis. The glucan was composed of ${\alpha}$-1,6 and 1,3 linkages and the fructan showed similar linkage data of levan.