• Title/Summary/Keyword: Enzyme Efficiency

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A New Strategy to Improve the Efficiency and Sustainability of Candida parapsilosis Catalyzing Deracemization of (R,S)-1-Phenyl-1,2-Ethanediol Under Non-Growing Conditions: Increase of NADPH Availability

  • Nie, Yao;Xu, Yan;Hu, Qing Sen;Xiao, Rong
    • Journal of Microbiology and Biotechnology
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    • v.19 no.1
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    • pp.65-71
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    • 2009
  • Microbial oxidoreductive systems have been widely used in asymmetric syntheses of optically active alcohols. However, when reused in multi-batch reaction, the catalytic efficiency and sustainability of non-growing cells usually decreased because of continuous consumption of required cofactors during the reaction process. A novel method for NADPH regeneration in cells was proposed by using pentose metabolism in microorganisms. Addition of D-xylose, L-arabinose, or D-ribose to the reaction significantly improved the conversion efficiency of deracemization of racemic 1-phenyl-1,2-ethanediol to (S)-isomer by Candida parapsilosis cells already used once, which afforded the product with high optical purity over 97%e.e. in high yield over 85% under an increased substrate concentration of 15 g/l. Compared with reactions without xylose, xylose added to multi-batch reactions had no influence on the activity of the enzyme catalyzing the key step in deracemization, but performed a promoting effect on the recovery of the metabolic activity of the non-growing cells with its consumption in each batch. The detection of activities of xylose reductase and xylitol dehydrogenase from cell-free extract of C. parapsilosis made xylose metabolism feasible in cells, and the depression of the pentose phosphate pathway inhibitor to this reaction further indicated that xylose facilitated the NADPH-required deracemization through the pentose phosphate pathway in C. parapsilosis. moreover, by investigating the cofactor pool, the xylose addition in reaction batches giving more NADPH, compared with those without xylose, suggested that the higher catalytic efficiency and sustainability of C. parapsilosis non-growing cells had resulted from xylose metabolism recycling NADPH for the deracemization.

알칼리 내성 Bacillus sp. YA-14 유래의 중복 Promotor를 이용한 재조합 Plasmid로부터의 Pectate Iyase의 발현

  • Park, Hee-Kyoung;Hahm, Byoung-Kwon;Yu, Ju-Hyun;Bai, Dong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.571-579
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    • 1997
  • For the overproduction of pectate lyase (PL), the recombinant plasmid pl2BS fl which has strong promoter from alkali-tolerent Bacillus sp. YA-14 was used. In order to overexpress the pectate lyase by the action of overlapping strong promoter in pl2BS$\Delta$fl, 1.6 kb of PL gene was inserted into pl2BS$\Delta$fl to form pl2BS$\delta$f1-PL and the enzyme was expressed. But decreased expression efficiency of the PL gene was observed and it was due to the presence of the transcription terminator region on the upstream of the PL gene. The transcription terminator of the PL gene in pl2BS$\delta$f1-PL was removed and the resulting plasmid p12BS$\Delta$fl$\Delta$PL was formed. Bacillus subtilis 207-25 harboring the recombinant plasmid, p12BS$\Delta$fl$\Delta$PL, revealed increased expression efficiency with chloramphenicol induction when cat-86 was used as a reporter gene.

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Effect of Godulbaegi(Ixeris sonchifolia H.) Powder on Growth, Protein and Lipid Concentration in Rats (고들빼기가 흰쥐의 성장률, 단백질 및 지질농도에 미치는 영향)

  • 고진복;오세원;김재영
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.3
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    • pp.525-530
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    • 1998
  • The effects of godulbaegi(Ixeris sonchifolia H.) powder on body weight gain, lipid and protein concentrations, and enzyme activities were studied in growing male rats for 4 and 8 week. The fats (Sprague-Dawley) were fed diets containing 5% godulbaegi or without. The body weght gain, food efficiency ratio, protein efficiency ratio, hemoglobin concentration and hematocrit value of rats fed godulbaegi diet were similar to the control diet. The ratio of spleen weight to body weight in godulbaegi group for 8 weeks was greater than in that of control group, but they were similar both feeding groups in liver and kidney. The concentrations of the total lipid and triglyceride in serum were increased in rats fed godulbaegi diet for 4 week than in those fed control diet, but those were decreased in rats fed godulbaegi diet for 8 week than in those fed the control diet. The concentrations of the phospholipid, total cholestrol and HDL-cholestrol in serum were decreased in rats fed godulbaegi diet for 8 week than in those fed control diet. The HDL-cholestrol/total cholestrol ratio and athergenic index were similar to both diet groups. There were no differences in growth, serum total protein, albumin concentrations, aspartate aminotransferase and alanine aminotransferase activities between rats fed godulbaegi diet and those fed the control diet.

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Establishment of Optimum Extraction Conditions and Wrinkle Improvement Evaluation of Glycosaminoglycans in Styela plicata (오만둥이(Styela plicata)에서 글리코스아미노글리칸의 최적 추출조건 설정 및 주름개선 효능)

  • Neri, Therese Ariane N.;Choi, Byeong-Dae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.5
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    • pp.717-724
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    • 2020
  • Styela plicata are naturally-occurring marine resources easily found along the coastlines that have established their niche as functional food and nutraceuticals ingredient along with their increasing consumer demand. Ascidian contain a large amount of dietary fiber but only the meat has been utilized and consumed while the rest of its parts are discarded. Also, various studies have been conducted on the meat of ascidians while studies on the functionality of the ascidian tunics, which were mostly undervalued, were scarce. In this study, we investigated and explored the glycosaminoglycans (GAGs) contents in the tunics of S. plicata, and their potential use as functional ingredient in pharmaceutical and nutraceutical applications. Sulfated GAGs and uronic acids contents were 8.9-10.7 g/100 g and 9.4-11.3 g/100 g, respectively. Highest GAGs content was extracted with optimum Brix at 7-9. Extraction efficiency using hot water at 121℃ was 4.22% while enzyme extraction using Protamex was more efficient at 5.91%. GAGs extracted from S. plicata tunics exhibited collagenase inhibitory activity of 75.2% at 100 ㎍/mL and procollagen synthesis activity of 80.1% at 100 ㎍/mL.

Digestion efficiency differences of restriction enzymes frequently used for genotype-by-sequencing technology

  • Chung, Yong Suk;Jun, Taehwan;Kim, Changsoo
    • Korean Journal of Agricultural Science
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    • v.44 no.3
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    • pp.318-324
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    • 2017
  • With the development of next-generation sequencing (NGS), a cutting-edge technology, genotype-by-sequencing (GBS) became available at a low cost per sample. GBS makes it possible to customize the process of library preparation to obtain high-quality single nucleotide polymorphisms (SNPs) in the most efficient way. However, a GBS library is hard to construct due to fine-tuning of concentration of each reagent and set-up. The major reason for this is the presence of undigested genomic DNA (gDNA) owing to the efficiency of different restriction enzymes for different species with unknown reasons. Therefore, this proof-concept study is to demonstrate the unpredictable patterns of enzyme digestion from various plants in order to make the reader aware of the caution needed when choosing restriction enzymes for their GBS library preparations. Indeed, no pattern was found for the digestibility of gDNA samples and restriction enzymes in the current study. We suggest that more data should be accumulated on this matter to help researchers who want to apply GBS technologies in a variety of genetic approaches.

Ant-Obesity Effect of Coriandrum sativum L. Ethanol Extract in High Fat-Induced Obesity Animal Model (고수 에탄올 추출물의 고지방식이 비만 동물모델에서의 항비만효과)

  • Rak Won Lee;Soon Ah Kang
    • The Korean Journal of Food And Nutrition
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    • v.36 no.4
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    • pp.296-308
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    • 2023
  • This study investigated the anti-obesity effects of Coriandrum sativum L. ethanol extracts in a high fat diet-induced obesity model (DIO). We confirmed the anti-obesity effects by analysing the expression of the related proteins, weight gain, dietary intake, dietary efficiency, blood biochemistry, histological analysis and western blot analysis. After oral administration of Coriandrum sativumL. ethanol extracts at concentrations of 250 and 500 mg/kg, a significant improvement in dietary efficiency, reduction in weight gain, triglycerides, total cholesterol and LDL-cholesterol in blood lipid was observed for 8 weeks. In addition, improvement in blood glucose and metabolism confirmed through glucose tolerance test was observed. Further, the concentration of alanine transaminase (ALT) in blood was significantly decreased, which improved the fatty liver caused by high-fat diet intake as confirmed by liver tissue analysis. This phenomenon was confirmed to decrease the expression of fat accumulation-related PPARγ and FAS protein in the liver tissue. Especially, it is believed that FAS, a liposynthetic enzyme, has a stronger inhibitory effect than PPARγ. Therefore, Coriandrum sativum L. ethanol extract is thought to improve obesity by reducing blood lipids levels, improving glucose metabolism and inhibiting synthesis of the fat that accumulates in the liver in high-fat diet-induced obesity animal models.

Dynamics of Functional Genes and Bacterial Community during Bioremediation of Diesel-Contaminated Soil Amended with Compost

  • Hyoju Yang;Jiho Lee;Kyung-Suk Cho
    • Journal of Microbiology and Biotechnology
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    • v.33 no.4
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    • pp.471-484
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    • 2023
  • Compost is widely used as an organic additive to improve the bioremediation of diesel-contaminated soil. In this study, the effects of compost amendment on the remediation performance, functional genes, and bacterial community are evaluated during the bioremediation of diesel-contaminated soils with various ratios of compost (0-20%, w/w). The study reveals that the diesel removal efficiency, soil enzyme (dehydrogenase and urease) activity, soil CH4 oxidation potential, and soil N2O reduction potential have a positive correlation with the compost amendment (p < 0.05). The ratios of denitrifying genes (nosZI, cnorB and qnorB) to 16S rRNA genes each show a positive correlation with compost amendment, whereas the ratio of the CH4-oxidizing gene (pmoA) to the 16S rRNA genes shows a negative correlation. Interestingly, the genera Acidibacter, Blastochloris, Erythrobacter, Hyphomicrobium, Marinobacter, Parvibaculum, Pseudoxanthomonas, and Terrimonas are strongly associated with diesel degradation, and have a strong positive correlation with soil CH4 oxidation potential. Meanwhile, the genera Atopostipes, Bacillus, Halomonas, Oblitimonas, Pusillimonas, Truepera, and Wenahouziangella are found to be strongly associated with soil N2O reduction potential. These results provide useful data for developing technologies that improve diesel removal efficiency while minimizing greenhouse gas emissions in the bioremediation process of diesel-contaminated soil.

Determination of Ozone Tolerance on Environmental Tree Species Using Standard Index (표준화 지수를 이용한 환경수목의 오존 내성 결정)

  • Han, Sim-Hee;Kim, Du-Hyun
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.11 no.1
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    • pp.3-12
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    • 2009
  • Ozone tolerance of tree species was determined by standard index of physiological damages and biochemical defense responses under short-term ozone exposure. At the end of 150ppb $O_3$ fumigation, photosynthetic characteristics and antioxidative enzyme activities were analyzed in the leaves of five species(Koelreuteria paniculata, Firmiana simplex, Styrax japonica, Fraxinus rhynchophylla, Viburnum sargentii). Injury index was determined by the effect of ozone on photosynthetic parameters and malondialdehyde(MDA) content, and tolerance index was calculated using the rate of increase in superoxide dismutase(SOD), ascorbate-peroxidase(APX), glutathione reductase(GR) and catalase(CAT) activities. Apparent quantum yield(AQY), carboxylation efficiency(Ce) and photo-respiration rate(PR) decreased in the leaves of five species with increasing ozone exposure time. These parameters were considered as an appropriate indicator for stress evaluation. Antioxidative enzyme activities showed various results depending on the tree species, exposure time, and enzyme types. SOD activity of K. paniculata increased with ozone exposure time, and that of F. rhynchophylla increased only after 6 hours of ozone exposure. CAT activity of $O_3$-exposed F. simplex was lower than the control. Based on standard index, ozone tolerance ability of five species was determined as two tolerant species(F. rhynchophylla > K. paniculata) and three sensitive species(S. japonica > F. simplex > V. sargentii).

Evaluation of the Estrogenic Activity by Yeast Two-hybrid Assay and Enzyme-linked Immunosorbent Assay in Sewage Treatment Plant (하수처리장의 내분비계장애물질에 대한 Yeast Two-hybrid Assay와 Enzyme-linked Immunosorbent Assay에 의한 에스트로겐활성도 평가)

  • Lee, Byoung-Cheun;Ra, Jin-Sung;Kim, Sang-Don;Kawai, Hukiko;Lee, Chul-Hee
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.771-777
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    • 2007
  • Several endocrine disrupting chemicals(EDCs) were monitored to evaluate the estrogenic activities and the concentrations by yeast two-hybrid assay and enzyme-linked immunosorbent assay(ELISA) in sewage treatment plant(STP) which consist of industrial and domestic line. In the influent of domestic line, estrone, 17$\beta$-estradiol, 17$\alpha$-ethinylestradiol and alkylphenolethoxylate(APE) were detected up to 167.1, 39.7, 7.3 and 145.4 ng/L, respectively. The average removal efficiency of 17$\beta$-estradiol after the activated sludge process was 77.5% and further removed to 80.8% after the sand filtration-ozonation step. These results suggests that the activated sludge process has limited potential to remove the estrogenic activity effectively. The contributions of the estrogenic chemicals to the estrogenic activities were 70.7, 23.3, 3.7 and 2.3% for estrone, 17$\beta$-estradiol 17$\alpha$-ethinylestradiol and APE, respectively, in the domestic line effluents. Therefore, 17$\beta$-estradiol and estrone contributed most of the estrogenic activity in the domestic line effluents.

Improvement of Transglycosylation Efficiency using a Glycosynthase Mutant derived from Thermoplasma acidophilum ${\alpha}$-Glucosidase (Thermoplasma acidophilum 유래 ${\alpha}$-glucosidase로 부터 생산된 glycosynthase 돌연변이 단백질의 개선된 당전이 효율)

  • Hwang, Sung-Min;Seo, Seong-Hwa;Park, In-Myoung;Choi, Kyoung-Hwa;Kim, Do-Man;Cha, Jae-Ho
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.104-110
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    • 2012
  • Glycosynthase is an active site nucleophile mutant enzyme, prepared from glycosidase, which is capable of synthesizing oligosaccharide derivatives without the hydrolysis of the product. Thermoacidophilic ${\alpha}$-glucosidase of Thermoplasma acidophilum (AglA) exhibits a transglycosylating activity yielding various glycosides. AglA was converted to glycosynthase by the substitution of the catalytic nucleophile Asp-408 residue into non-nucleophile glycine in order to increase its ability to synthesize various glycosides by transglycosylation. The glycosynthase mutant was purified by Ni-NTA chromatography and its glycoside-synthesizing activity was measured by using an external nucleophile, sodium formate buffer, providing maltose as a donor and p-nitrophenyl-${\alpha}$-D-glucopyranoside ($pNP{\alpha}G$) as an acceptor, respectively. In addition, $pNP{\alpha}G$ was examined for its feasibility to act as both a donor and an acceptor, and products were compared with those of the wildtype enzyme. The mutant enzyme was found to catalyze the formation of a specific product from $pNP{\alpha}G$ with a yield of 42.5% without further hydrolysis, while the wild-type enzyme produced two $pNP{\alpha}G$ products at low yields. The results demonstrate the possibility of satisfactory yields for the reactions in the presence of small amounts of acceptor, and demonstrate that the high activity of the mutant, at pHs below neutrality, was applicable in the transfer of glucose from the natural donor.