• Title/Summary/Keyword: Trehalose

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Production of Aminolevulinic Acid by Recombinant Escherichia coli Co-expressing hemA and otsBA Using Crude Glycerol as Carbon Source (폐글리세롤을 탄소원으로 hemA와 otsBA를 공동 발현하는 재조합대장균 배양을 통한 아미노레블린산 생산)

  • Yan, Jingmei;Pham, Diep Ngoc;Kang, Dae-Kyung;Kim, Sung Bae;Kim, Chang-Joon
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.341-348
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    • 2016
  • This study aimed to develop a microbial process for producing aminolevulinic acid (ALA) using crude glycerol. In the culture of ALA-producing cells (Escherichia coli/pH-hemA) in a medium containing crude glycerol, the cell density and production were 1.8-fold and 1.2-fold lower than those obtained from pure glycerol, respectively. However, the cell growth and production were improved by supplementing the medium with trehalose (30 or 100 g/l). Engineered cells (E. coli/pH-hemA/pS-otsBA) were constructed to express otsBA and their culture performance was compared with that of control cells (E. coli/pH-hemA/ pSTV28). The effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and the time of induction were examined to improve the cell growth and ALA production in engineered cells cultured using crude glycerol. When 0.6 mM of IPTG was added at the beginning of the exponential growth phase, the ALA produced by cells was 2,121 mg/l, which was comparable to that from pure glycerol. The results demonstrate that otsBA expression endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.

Transcriptional and Mycolic Acid Profiling in Mycobacterium bovis BCG In Vitro Show an Effect for c-di-GMP and Overlap between Dormancy and Biofilms

  • Cruz, Miguel A. De la;Ares, Miguel A.;Rodriguez-Valverde, Diana;Vallejo-Cardona, Alba Adriana;Flores-Valdez, Mario Alberto;Nunez, Iris Denisse Cota;Aceves-Sanchez, Michel de Jesus;Lira-Chavez, Jonahtan;Rodriguez-Campos, Jacobo;Bravo-Madrigal, Jorge
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.811-821
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    • 2020
  • Mycobacterium tuberculosis produces mycolic acids which are relevant for persistence, recalcitrance to antibiotics and defiance to host immunity. c-di-GMP is a second messenger involved in transition from planktonic cells to biofilms, whose levels are controlled by diguanylate cyclases (DGC) and phosphodiesterases (PDE). The transcriptional regulator dosR, is involved in response to low oxygen, a condition likely happening to a subset of cells within biofilms. Here, we found that in M. bovis BCG, expression of both BCG1416c and BCG1419c genes, which code for a DGC and a PDE, respectively, decreased in both stationary phase and during biofilm production. The kasA, kasB, and fas genes, which are involved in mycolic acid biosynthesis, were induced in biofilm cultures, as was dosR, therefore suggesting an inverse correlation in their expression compared with that of genes involved in c-di-GMP metabolism. The relative abundance within trehalose dimycolate (TDM) of α-mycolates decreased during biofilm maturation, with methoxy mycolates increasing over time, and keto species remaining practically stable. Moreover, addition of synthetic c-di-GMP to mid-log phase BCG cultures reduced methoxy mycolates, increased keto species and practically did not affect α-mycolates, showing a differential effect of c-di-GMP on keto- and methoxy-mycolic acid metabolism.

Mass-Based Metabolomic Analysis of Lactobacillus sakei and Its Growth Media at Different Growth Phases

  • Lee, Sang Bong;Rhee, Young Kyoung;Gu, Eun-Ji;Kim, Dong-Wook;Jang, Gwang-Ju;Song, Seong-Hwa;Lee, Jae-In;Kim, Bo-Min;Lee, Hyeon-Jeong;Hong, Hee-Do;Cho, Chang-Won;Kim, Hyun-Jin
    • Journal of Microbiology and Biotechnology
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    • v.27 no.5
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    • pp.925-932
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    • 2017
  • Changes in the metabolite profiles of Lactobacillus sakei and its growth media, based on different culture times (0, 6, 12, and 24 h), were investigated using gas chromatography-mass spectrometry (MS) and liquid chromatography-MS with partial least squares discriminant analysis, in order to understand the growth characteristics of this organism. Cell and media samples of L. sakei were significantly separated on PLS-DA score plots. Cell and media metabolites, including sugars, amino acids, and organic acids, were identified as major metabolites contributing to the difference among samples. The alteration of cell and media metabolites during cell growth was strongly associated with energy production. Glucose, fructose, carnitine, tryptophan, and malic acid in the growth media were used as primary energy sources during the initial growth stage, but after the exhaustion of these energy sources, L. sakei could utilize other sources such as trehalose, citric acid, and lysine in the cell. The change in the levels of these energy sources was inversely similar to the energy production, especially ATP. Based on these identified metabolites, the metabolomic pathway associated with energy production through lactic acid fermentation was proposed. Although further studies are required, these results suggest that MS-based metabolomic analysis might be a useful tool for understanding the growth characteristics of L. sakei, the most important bacterium associated with meat and vegetable fermentation, during growth.

Comparison of Gene Expression in Larval Fat Body of Helicoverpa assulta in Different Temperature Conditions (온도변화에 따른 담배나방 유충 지방체의 유전자 발현 비교 분석)

  • Cha, Wook Hyun;Kim, Kwang Ho;Lee, Dae-Weon
    • Korean journal of applied entomology
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    • v.57 no.3
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    • pp.165-175
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    • 2018
  • Insects are known to live at wide range of temperature, but can not survive when they are exposed to over $40^{\circ}C$ or below supercooling point. The larvae of Helicoverpa assulta have been reared at high ($35^{\circ}C$), low (3 to $10^{\circ}C$), and room temperature ($25^{\circ}C$; control). To identify stress-related genes, the transcriptomes of fat body have been analyzed. Genes such as cuticular proteins, fatty acyl ${\Delta}9$ desaturase and glycerol 3 phosphate dehydrogenase were up-regulated whereas chitin synthase, catalase, and UDP-glycosyltransferase were down-regulated at low temperature. Superoxide dismutase, metallothionein 2, phosphoenolpyruvate carboxykinase and trehalose transporter have been up-regulated at high temperature. In addition, expressions of heat shock protein and glutathione peroxidase were increased at high temperature, but decreased at low temperature. These temperature-specific expressed genes can be available as markers for climate change of insect pests.

Comparative study on nutritional contents of Auricularia spp. (목이버섯 품종간 영양성분 비교)

  • Kim, Tae-Ho;Jo, Se-Hyun;Kim, Min-Ji;Yu, Young-Bok;Jang, Mi-Hyang;Park, Ki-Moon
    • Journal of Mushroom
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    • v.10 no.1
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    • pp.29-36
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    • 2012
  • We conducted an analysis by comparing the nutritional contents of Auricularia auricula-judae(black), Auricularia polytricha and Auricularia auricula-judae (brown). In nutritional contents of three strains of Auricularia spp., four free sugars, seven organic acids and 24 amino acids were detected. Auricularia auricula-judae (black) was highly contained free sugar, organic acid and amino acid. There was the most prevalent Vitamin $D_2$ content in Auricularia auricula-judae (black) of Auricularia spp. Dietary fiber of three strains showed contents of about 60% but were not significantly different. ${\beta}$-glucan contents of Auricularia auricula-judae (brown) contained the highest contents with $25.21{\pm}0.37%$ and showed significant differences between Auricularia polytricha and Auricularia auricula-judae (black). Total polyphenol contents of Auricularia polytricha showed the highest contents, followed by Auricularia auricula-judae (brown) and Auricularia auricula-judae (black).

Effect of 20-hydroxyecdysone on diapausing pupae of the fall webworm, Hyphantria cunea Drury (미국흰불나방(Hyphantria cunea Drury) 휴면번데기에 대한 탈피호르몬의 영향)

  • 박재우;부경생
    • Korean journal of applied entomology
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    • v.32 no.2
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    • pp.139-150
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    • 1993
  • This study was conducted to investigate the effect of 20-hydroxyecdysone on diapausing pupae of Hyphantna cunea. Treatment of 20-HE at the dosage of 5 $\mu$ g or higher/g live weight terminated diapause in about 50% of Hyphantna cunea pupae treated, and some malformed adult emerged at dosages of 10 or 12.5 $\mu$g of 20-hydroxyecdysone/g live wt. The maximum concentration of 20-hydroxyecdysone in the whole body of normally developing pupae was about 3.2 $\mu$g/g on the 6th day after pupation, and in that of diapausing pupae treated with 20 -HE about 4.1 $\mu$g/g on the 6th day after the treatment. But diapausing pupae showed a low level( 1.7 $\mu$g/g) of maximum 20-HE concentration. In diapausing pupae treated with 20- HE, glycongen content was lower than in normally developing pupae, but the changing pattern was similar to that observed in normally developing pupae. Glucose(and/or sorbitol) and trehalose content of 20-HE-injected pupae reached the maximum value on the 6th day after the injection, which were higher than those of normally developing pupae. The amount of soluble haemolymph proteins was lower but the total soluble protein content of haemolymph-rernoved whole body was higher in 20-HE-injected pupae than that in normally developing pupae.

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Development of a Biofungicide Using a Mycoparasitic Fungus Simplicillium lamellicola BCP and Its Control Efficacy against Gray Mold Diseases of Tomato and Ginseng

  • Shin, Teak Soo;Yu, Nan Hee;Lee, Jaeho;Choi, Gyung Ja;Kim, Jin-Cheol;Shin, Chul Soo
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.337-344
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    • 2017
  • To develop a commercial product using the mycoparasitic fungus Simplicillium lamellicola BCP, the scale-up of conidia production from a 5-l jar to a 5,000-l pilot bioreactor, optimization of the freeze-drying of the fermentation broth, and preparation of a wettable powder-type formulation were performed. Then, its disease control efficacy was evaluated against gray mold diseases of tomato and ginseng plants in field conditions. The final conidial yields of S. lamellicola BCP were $3.3{\times}10^9conidia/ml$ for a 5-l jar, $3.5{\times}10^9conidia/ml$ for a 500-l pilot vessel, and $3.1{\times}10^9conidia/ml$ for a 5,000-l pilot bioreactor. The conidial yield in the 5,000-l pilot bioreactor was comparable to that in the 5-l jar and 500-l pilot vessel. On the other hand, the highest conidial viability of 86% was obtained by the freeze-drying method using an additive combination of lactose, trehalose, soybean meal, and glycerin. Using the freeze-dried sample, a wettable powder-type formulation (active ingredient 10%; BCP-WP10) was prepared. A conidial viability of more than 50% was maintained in BCP-WP10 until 22 weeks for storage at $40^{\circ}C$. BCP-WP10 effectively suppressed the development of gray mold disease on tomato with control efficacies of 64.7% and 82.6% at 500- and 250-fold dilutions, respectively. It also reduced the incidence of gray mold on ginseng by 65.6% and 81.3% at 500- and 250-fold dilutions, respectively. The results indicated that the new microbial fungicide BCP-WP10 can be used widely to control gray mold diseases of various crops including tomato and ginseng.

A Study on the Quality Characteristics of Rice Beer Using Brewing Yeast isolated from Nuruk (누룩에서 분리한 양조용 효모를 이용한 쌀맥주의 품질특성 연구)

  • Lee, Young Bog;Ko, Dong Jun;Cheong, Chul
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.12
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    • pp.340-347
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    • 2019
  • This study investigated the brewing properties of rice beer made with yeast isolated from traditional whole wheat nuruk. The experimental method was followed by alcohol fermentation and quality analysis for six months after separating the yeast from nuruk. The yeasts isolated from traditional nuruk showed normal fermentation characteristics, like those of commercial yeast, in terms of alcohol production capability, sugar content, and pH reducing power. Especially, the yeast (KCCM 301) isolated from whole wheat nuruk showed higher contents of glycogen and trehalose than that of commercial yeast, and so KCCM 301 yeast has excellent yeast vitality. Meanwhile, the yeast (KCCM 90301) isolated from traditional Nuruk produced significantly higher alcohol and ester contents than that of commercial yeast. This has a positive effect for supplementing the taste and aroma of rice beer. In conclusion, the yeasts isolated from whole wheat nuruk showed the general alcohol fermentation pattern and aroma content of rice beer. These yeasts seem to be effective in strengthening the flavor of rice beer.

Enhanced drought and oxidative stress tolerance in transgenic sweetpotato expressing a codA gene (CodA 고발현 형질전환 고구마의 산화 및 건조 스트레스 내성 증가)

  • Park, Sung-Chul;Kim, Myoung Duck;Kim, Sun Ha;Kim, Yun-Hee;Jeong, Jae Cheol;Lee, Haeng-Soon;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.42 no.1
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    • pp.19-24
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    • 2015
  • Glycine betaine (GB) is one of the compatible solutes that accumulate in the chloroplasts of certain halotolerant plants under salt or cold stress. The codA gene for choline oxidase, the enzyme that converts choline into GB, has been cloned from a soil bacterium Arthrobacter globiformis. We generated transgenic sweetpotato plants [Ipomoea batatas (L.) Lam] expressing codA gene in chloroplasts under the control of the SWPA2 promoter (referred to as SC plants) and evaluated SC plants under oxidative and drought stresses. SC plants showed enhanced tolerance to methyl viologen (MV)-mediated oxidative stress and drought stress due to induced expression of codA. At $5{\mu}M$ of MV treatment, all SC plants showed enhanced tolerance to MV-mediated oxidative stress through maintaining low ion leakage and increased GB levels compared to wild type plants. When plants were subjected to drought conditions, SC plants showed enhanced tolerance to drought stress through maintaining high relative water contents and increased codA expression compared to wild type plants. These results suggest that the SC plants generated in this study will be useful for enhanced biomass production on global marginal lands.

Gene Silencing Induced by Cytosine Methylation in Transgenic Tomato (형질전환 토마토에서 Cytosine Methylation에 의한 유전자발현 억제)

  • Jung, Seo-Hee;Min, Sung-Ran;Lee, Soo-Young;Park, Ji-Young;Davarpanah, S Javad;Chung, Hwa-Jee;Jeon, Jae-Heung;Liu, Jang-Ryol;Jeong, Won-Joong
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.323-329
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    • 2007
  • Transgene expression was analyzed in tomato plants. Four lines of neomycin phosphotransferase II gene (NPTII) and the trehalose biosynthetic fusion gene (TPSP) transformed $T_0$ plants showed kanamycin resistance on selection medium. However, the analysis of phenotype (kanamycin resistance) and mRNA expression in $T_1$ plants indicated that the expression of the NPTII and TPSP transgenes was down-regulated to an undetectable level in two independent lines 1 and 11. Southern analysis demonstrated that the lines 1 and 11 had multicopies of the transgenes, whereas the typical transgenic lines 2 and 10 had 1 or 2 copies. DNA methylation analysis using methylation sensitive enzyme detected accumulated CpG DNA methylation on TPSP coding region and CaMV35S promoter region in the line 11, but not the typical transgenic line 2. These results suggest that multicopy transgene in plants is attributed to down-regulation of the transgene expression via transcriptional gene silencing.