• Title/Summary/Keyword: Trehalose

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Functions of PUF Family RNA-Binding Proteins in Aspergillus nidulans

  • Son, Sung-Hun;Jang, Seo-Yeong;Park, Hee-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.676-685
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    • 2021
  • RNA-binding proteins are involved in RNA metabolism and posttranscriptional regulation of various fundamental biological processes. The PUF family of RNA-binding proteins is highly conserved in eukaryotes, and its members regulate gene expression, mitochondrial biogenesis, and RNA processing. However, their biological functions in Aspergillus species remain mostly unknown in filamentous fungi. Here we have characterized the puf genes in the model organism Aspergillus nidulans. We generated deletion mutant strains for the five putative puf genes present in the A. nidulans genome and investigated their developmental phenotypes. Deletion of pufA or pufE affected fungal growth and asexual development. pufA mutants exhibited decreased production of asexual spores and reduced mRNA expression of genes regulating asexual development. The pufE deletion reduced colony growth, increased formation of asexual spores, and delayed production of sexual fruiting bodies. In addition, the absence of pufE reduced both sterigmatocystin production and the mRNA levels of genes in the sterigmatocystin cluster. Finally, pufE deletion mutants showed reduced trehalose production and lower resistance to thermal stress. Overall, these results demonstrate that PufA and PufE play roles in the development and sterigmatocystin metabolism in A. nidulans.

Effects of Protein Functionality on Myofibril Protein-Saccharide Graft Reaction

  • Kim, Tae-Kyung;Yong, Hae In;Cha, Ji Yoon;Kim, Yun Jeong;Jung, Samooel;Choi, Yun-Sang
    • Food Science of Animal Resources
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    • v.42 no.5
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    • pp.849-860
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    • 2022
  • The myofibril protein (MP) isolate-saccharide graft reactions was prepared using the Maillard reaction with saccharides. The effects of various saccharides on protein functionality and quality of the Maillard reaction were investigated and compared with those of MP. The grafting degree of the MP isolate-saccharide graft reaction was significantly higher in the reducing sugar-treated groups (lactose, glucose, fructose, and palatinose). The browning intensity of the MP isolate-saccharide graft reaction with fructose, sucrose, and erythitol was higher than that observed in the control reaction (p<0.05). MP that reacted with reducing sugars (glucose, fructose, palatinose, and lactose) had fainter bands than MP that reacted with non-reducing sugars (sucrose, erythitol, trehalose, sorbitol, and xylitol). MPs conjugated with glucose exhibited higher protein solubility. The palatinose and lactose treatments were maximum in water binding capacity, though no significant difference in oil binding capacity among the saccharide treatments was observed. The emulsion stability of the MP isolate-saccharide graft reaction with palatinose and erythitol was higher than that of the control reaction. Therefore, reducing sugars have good protein functionality in the MP isolate-saccharides graft reaction.

UHPLC/TOFHRMS analysis and anti-inflammatory effect of leaf extracts from Zizyphus jujuba in LPS-stimulated RAW264.7 cells

  • Hyun Ji Eo;Sun-Young Lee;Gwang Hun Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.27-33
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    • 2023
  • Zizyphus jujube is a plant in the buckthorn family (Rhamnaceae) that has been the subject of research into antibacterial, antifungal and anti-inflammatory properties of its fruit and seed. However, few studies have investigated its leaves. In this study, the anti-inflammatory activity of ZJL (an extract of Z. jujube leaf) was evaluated to verify its potential as an anti-inflammatory agent and SARS-CoV-2 medicine, using nitric oxide (NO) assay, RT-PCR, SDSPAGE, Western blotting, and UHPLC/TOFHRMS analysis. We found that ZJL suppresed pro-inflammatory mediators such as NO, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor α (TNF-α) in lipopolysaccharide (LPS)-induced RAW264.7 cells. ZJL acted by inhibiting NF-KB and MAPK signaling pathway activity. We also confirmed that ZJL contains a phenol compound and flavonoids with anti-inflammatory activity such as trehalose, maleate, epigallocatechin, hyperoside, catechin, 3-O-coumaroylquinic acid, rhoifolin, gossypin, kaempferol 3-neohesperidoside, rutin, myricitrin, guaiaverin, quercitrin, quercetin, ursolic acid, and pheophorbide a. These findings suggest that ZJL may have great potential for the development of anti-inflammatory drugs and vaccines via inhibition of NF-κB and MAPK signaling in LPS-induced RAW264.7 cells.

Dasania marina gen. nov., sp. nov., of the Order Pseudomonadales, Isolated from Arctic Marine Sediment

  • Lee, Yoo-Kyung;Hong, Soon-Gyu;Cho, Hyun-Hee;Cho, Kyeung-Hee;Lee, Hong-Kum
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.505-509
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    • 2007
  • An obligately aerobic bacterium, strain KOPRI $20902^T$, was isolated from a marine sediment in Ny-${\AA}$lesund, Spitsbergen Islands, Norway. Cells were irregular rods and motile with polar monotrichous flagellum. The optimum growth temperature was $17-22^{\circ}C$. Cells grew best in pH 7.0-10.0 and 3-4% sea salts (corresponding to 2.3-3.1% NaCl). The novel strain required $Ca^{2+}$ or $Mg^{2+}$ in addition to NaCl for growth. Sequence analysis of 16S rRNA gene revealed that the Arctic isolate is distantly related with established species (<92.4% sequence similarity) and formed a monophyletic group with Cellvibrio, which formed a distinct phylogenetic lineage in the order Pseudomonadales. Predominant cellular fatty acids [$C_{16:1}\;{\omega}7c/15:0$ iso 2OH (45.3%), $C_{16:0}$ (18.4%), ECL 11.799 (11.2%), $C_{10:0}$ 3OH (10.4%)]; DNA G+C content (37.0 mol%); nitrate reduction to nitrogen; absence of aesculin hydrolysis, N-acetyl-${\beta}$-glucosaminidase and esterase; no assimilation of arabinose, galactose, glucose, lactose, maltose, and trehalose differentiated the strain from the genus Cellvibrio. Based on the phylogenetic and phenotypic characteristics, Dasania marina gen. nov., sp. nov. is proposed in the order Pseudomonadales. Strain KOPRI $20902^T$ (=KCTC $12566^T$=JCM $13441^T$) is the type strain of Dasania marina.

Efficient plant regeneration from immature embryo cultures of Jatropha curcas, a biodiesel plant

  • Varshney, Alok;Johnson, T. Sudhakar
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.139-148
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    • 2010
  • Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel. In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the establishment of callus. Immature embryos (1.1-1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant) on Murashige and Skoog's (MS) medium supplemented with IBA (0.5 $mg\;l^{-1}$) and BA (1.0 $mg\;l^{-1}$). The above medium when supplemented with growth adjuvants such as 100 $mg\;l^{-1}$ casein hydrolysate + 200 $mg\;l^{-1}$ L-glutamine + 8.0 $mg\;l^{-1}$ $CuSO_4$ resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets (10/explant) was achieved on MS medium supplemented with 500 $mg\;l^{-1}$ polyvinyl pyrrolidone + 30 $mg\;l^{-1}$ citric acid + 1 $mg\;l^{-1}$ BA + 0.5 $mg\;l^{-1}$ Kn + 0.25 $mg\;l^{-1}$ IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-halfstrength MS medium supplemented with 0.5 $mg\;l^{-1}$ IBA and 342 $mg\;l^{-1}$ trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas.

Influence of Physiological and Environmental Factors on Growth and Sporulation of an Antagonistic Strain of Trichoderma viride RSR 7

  • Jayaswal, R.K.;Singh, Rajesh;Lee, Youn-Su
    • Mycobiology
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    • v.31 no.1
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    • pp.36-41
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    • 2003
  • Influence of physiological and environmental factors on an antagonistic strain of Trichoderma viride RSR7 were studied optimize its biocontrol potential. The growth and sporulation of T. viride was greatly influenced by various carbon and nitrogen sources, and the environmental factors such as pH and temperature. The best growth and sporulation of T. viride was observed when sucrose, peptone and trehalose were supplemented in the medium as sole carbon sources. Rhamnose, pyruvic acid and sorbitol also supported a good growth. However, with these carbon sources the sporulation was poor. Growth and sporulation was also affected by various nitrogen sources. Growth and sporulation both were favoured by ammonium forms of nitrogen compared to nitrite or nitrate forms. Urea did not support either growth or sporulation. Among amino acids, glutamic acid, asparagine, leucine, aspartic acid, glutamic acid and alanine supported good growth as well as sporulation. T. viride was able to utilize large number of amino acids as sole nitrogen source. Proline was good for growth, but not for sporulation. Maximum growth and sporulation of T. viride was between pH 4.5 to 5.5. Temperatures between $20^{\circ}C\;and\;37^{\circ}C$ were good for both growth and sporulation of T. viride. At lower temperatures(i.e. below $20^{\circ}C$) growth and sporulation were inhibited. Based on the present study it may be concluded that T. viride RSR7 is capable of growing and sporulating with varied nutritional and environmental conditions and, therefore, this strain of T. viride may be useful as a biocontrol agent under diverse physiological and environmental conditions.

Detoxification of Aflatoxin B1 Contaminated Maize Using Human CYP3A4

  • Yamada, Marie;Hatsuta, Koji;Niikawa, Mayuko;Imaishi, Hiromasa
    • Journal of Microbiology and Biotechnology
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    • v.30 no.8
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    • pp.1207-1213
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    • 2020
  • Aflatoxin B1 (AFB1) is a mycotoxin produced by Aspergillus flavus (A. flavus). AFB1 is reported to have high thermal stability and is not decomposed by heat treatment during food processing. Therefore, in this study, knowing that AFB1 is metabolized by cytochrome P450 (CYP), our aim was to develop a method to detoxify A. flavus-contaminated maize, under normal temperature and pressure, using Escherichia coli expressing human CYP3A4. First, the metabolic activity of AFB1 by recombinant human CYP3A4 was evaluated. As a result, we confirmed that recombinant human CYP3A4 metabolizes 98% of AFB1. Next, we found that aflatoxin Q1, a metabolite of AFB1 was no longer mutagenic. Furthermore, we revealed that about 50% of the AFB1 metabolic activity can be maintained for 3 months when E. coli expressing human CYP3A4 is freeze-dried in the presence of trehalose. Finally, we found that 80% of AFB1 in A. flavus-contaminated maize was metabolized by E. coli expressing human CYP3A4 in the presence of surfactant triton X-405 at a final concentration of 10% (v/v). From these results, we conclude that AFB1 in A. flavus-contaminated maize can be detoxified under normal temperature and pressure by using E. coli expressing human CYP3A4.

Effect of Preharvest Treatment of $CaCl_2$ and Alkaline-Reduced Water on the Quality of Oyster Mushroom during Storage (수확 전 염화칼슘 및 알카리환원수 처리가 느타리버섯 저장에 미치는 영향)

  • Choi, Yun-Soo;Lee, Kyoung-Min;Seo, Geon-Sik;Kim, Min-Kyung;Hwang, Yong-Soo
    • Korean Journal of Agricultural Science
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    • v.37 no.3
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    • pp.355-360
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    • 2010
  • This experiment was focused to determine the effect of preharvest treatment of alkaine-reduced water (ARW) and $CaCl_2$ on the storage of oyster mushroom. ARW was prepared through electrolysis of water with $CaCl_2$ (0.1%) as salts and $CaCl_2$ (0.1%) was dissolved in deionized water. Mushrooms were harvested after two sprays of each solutions and quality factors including firmness and browning was determined during storage. The severity of browning was more greater when stored 20 days. Unlike $CaCl_2$, ARW containing 139 ug/L calcium was positive on maintaining freshness in terms of delay of firmness loss and respiration drop, keeping soluble carbohydrate content such as trehalose, and inhibiting the occurrence of browning. Browning, however, seemed to be more influenced by the developmental stage of mushroom at harvest. Even no significant difference was found, the level of ${\beta}$-glucan was low in ARW treatment compared to that of control. Further research including application and preparation methods of ARW is required to clarify the potential of ARW on keeping freshness of oyster mushroom through inhibiting tissue browning.

The physiological characteristics of Kuehneromyces mutabilis. (무리우산버섯의 생리적 특성)

  • 채정기;황태익;서승현;김현석;장경수;문형률;강민아;박병인
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.61-61
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    • 2002
  • This study was excuted to decide the physiological characteristics of Kuehneromyces mutabilis. K. mutabilis was tested to select pertinent substract, temperature and pH range for the growth. Mycelial growth of K. mutabilis was mostly supported on MYPA among other tested synthetic or semi-synthetic media. The temperature range for pertinent mycelial growth was about 25~32$^{\circ}C$ and mostly stimulated at $25^{\circ}C$. And the pertinent pH range of MYPA was 5.0~6.0. The required carbon and nitrogen source for mycelial growth of K. mutabilis was tested. The mycelial growth was mostly stimulated by soluble starch at content. The carbon sources for pertinent mycelial growth was mannose or glucose. And the nitrogen source for pertinent mycelial growth was yeast extract or peptone.

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Effect of Monothioglycerol on ROS Inhibition, Mitochondrial Activity, and DNA Integrity in Frozen-thawed Miniature Pig Sperm (Monothiolglycerol이 동결 융해 후 미니돼지 정자의 활성산소 억제, 미토콘드리아 활성 그리고 DNA Integrity에 미치는 영향)

  • Park, Soo-Jung;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.265-271
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    • 2013
  • Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.