• Title/Summary/Keyword: Trehalose

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Control Efficacy of Bacillus velezensis AFB2-2 against Potato Late Blight Caused by Phytophthora infestans in Organic Potato Cultivation

  • Kim, Min Jeong;Shim, Chang Ki;Park, Jong-Ho
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.580-595
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    • 2021
  • Although late blight is an important disease in ecofriendly potato cultivation in Korea, it is highly dependent on the use of eco-friendly agricultural materials and the development of biological control technology is low. It is a necessary to develop an effective biocontrol agent to inactivate late blight in the field. AFB2-2 strain is a gram-positive with peritrichous flagella. It can utilize 20 types of carbon sources, like L-arabinose, and D-trehalose at 35℃. The optimal growth temperature of the strain is 37℃. It can survive at 20-50℃ in tryptic soy broth. The maximum salt concentration tolerated by AFB2-2 strain is 7.5% NaCl. AFB2-2 strain inhibited the mycelial growth of seven plant pathogens by an average inhibitory zone of 10.2 mm or more. Among the concentrations of AFB2-2, 107 cfu/ml showed the highest control value of 85.7% in the greenhouse. Among the three concentrations of AFB2-2, the disease incidence and severity of potato late blight at 107 cfu/ml was lowest at 0.07 and 6.7, respectively. The nucleotide sequences of AFB2-2 strain were searched in the NCBI GenBank; Bacillus siamensis strain KCTC 13613, Bacillus velezensis strain CR-502, and Bacillus amyloliquefaciens strain DSM7 were found to have a genetic similarity of 99.7%, 99.7%, and 99.5%, respectively. The AFB2-2 strain was found to harbor the biosynthetic genes for bacillomycin D, iturin, and surfactin. Obtained data recommended that the B. velezensis AFB2-2 strain could be considered as a promising biocontrol agent for P. infestans in the field.

Effect of Silk Fibroin Biomaterial Coating on Cell Viability and Intestinal Adhesion of Probiotic Bacteria

  • Kwon, Gicheol;Heo, Bohye;Kwon, Mi Jin;Kim, Insu;Chu, Jaeryang;Kim, Byung-Yong;Kim, Byoung-Kook;Park, Sung Sun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.4
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    • pp.592-600
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    • 2021
  • Probiotics can be processed into a powder, tablet, or capsule form for easy intake. They are exposed to frequent stresses not only during complex processing steps, but also in the human body after intake. For this reason, various coating agents that promote probiotic bacterial stability in the intestinal environment have been developed. Silk fibroin (SF) is a material used in a variety of fields from drug delivery systems to enzyme immobilization and has potential as a coating agent for probiotics. In this study, we investigated this potential by coating probiotic strains with 0.1% or 1% water-soluble calcium (WSC), 1% SF, and 10% trehalose. Under simulated gastrointestinal conditions, cell viability, cell surface hydrophobicity, and cell adhesion to intestinal epithelial cells were then measured. The survival ratio after freeze-drying was highest upon addition of 0.1% WSC. The probiotic bacteria coated with SF showed improved survival by more than 10.0% under simulated gastric conditions and 4.8% under simulated intestinal conditions. Moreover, the cell adhesion to intestinal epithelial cells was elevated by 1.0-36.0%. Our results indicate that SF has positive effects on enhancing the survival and adhesion capacity of bacterial strains under environmental stresses, thus demonstrating its potential as a suitable coating agent to stabilize probiotics throughout processing, packaging, storage and consumption.

Metabolic profiles of Wolfiporia cocos mycelia cultivated under light and dark conditions

  • Jae-Gu, Han;Sang Suk, Kim;Doo-Ho, Choi;Gi-Hong, An;Kang-Hyo, Lee
    • Journal of Mushroom
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    • v.20 no.4
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    • pp.187-192
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    • 2022
  • Wolfiporia cocos is an edible fungus commercially cultivated in Asia. To investigate metabolic changes of W. cocos mycelia under both light and dark culture conditions, gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) analyses were performed. In terms of the total amount of sugars, alcohols, amino acids, organic acids, fatty acids, and purines, there no significant differences between the W. cocos mycelia cultivated under light (L) or dark (D) conditions (p < 0.05). However, there were some differences with respect to the production of particular sugars and proteins. The levels of trehalose (L: 17.2 ± 0.3% vs. D: 13.9 ± 1.6%), maltose (L: 0.9 ± 0.1% vs. D: 0.3 ± 0.1%), turanose (L: 0.7 ± 0.2% vs. D: 0.1 ± 0.1%), glutamine (L: 1.6 ± 0.3% vs. D: 0.7 ± 0.2%), and proline (L: 0.3 ± 0% vs. D: 0.1 ± 0%) were all significantly higher under light condition (p < 0.05). In contrast, the levels of galactose (L: 13.7 ± 1.2% vs. D: 17.6 ± 2.0%), aspartic acid (L: 0.6 ± 0.1 % vs. D: 0.9 ± 0.1%), cystathionine (L: 0.6 ± 0.1% vs. D: 0.8 ± 0 %), and malic acid (L: 0.7 ± 0.1% vs. D: 1.2 ± 0.1%) were higher under the dark condition. It is worth noting that the amount of pachymic acid, a pharmaceutically active compound of W. cocos, was 1.68 times greater under the light condition (p < 0.05).

Cryopreservation of Siberian tiger (Panthera tigris altaica) epididymal spermatozoa: pilot study of post-thaw sperm characteristics

  • Ibrahim, Saddah;Talha, Nabeel Abdelbagi Hamad;Kim, Jeongho;Jeon, Yubeol;Yu, Iljeoung
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.2
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    • pp.130-135
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    • 2022
  • Epididymal sperm cryopreservation provides a potential method for preserving genetic material from males of endangered species. This pilot study was conducted to develop a freezing method for tiger epididymal sperm. We evaluated post-thaw sperm condition using testes with intact epididymides obtained from a Siberian tiger (Panthera tigris altaica) after castration. The epididymis was chopped in Tyrode's albumin-lactate-pyruvate 1x and incubated at 5% CO2, 95% air for 10 min. The Percoll separation density gradient method was used for selective recovery of motile spermatozoa after sperm collection using a cell strainer. The spermatozoa were diluted with modified Norwegian extender supplemented with 20 mM trehalose (extender 1) and subsequent extender 2 (extender 1 with 10% glycerol) and frozen using LN2 vapor. After thawing at 37℃ for 25 s, Isolate® solution was used for more effective recovery of live sperm. Sperm motility (computerized assisted sperm analysis, CASA), viability (SYBR-14 and Propidium Iodide) and acrosome integrity (Pisum sativum agglutinin with FITC) were evaluated. The motility of tiger epididymal spermatozoa was 40.1 ± 2.0%, and progressively motile sperm comprised 32.7 ± 2.3%. Viability was 56.3 ± 1.6% and acrosome integrity was 62.3 ± 4.4%. Cryopreservation of tiger epididymal sperm using a modified Norwegian extender and density gradient method could be effective to obtain functional spermatozoa for future assisted reproductive practices in endangered species.

Protective Efficacy and Immunogenicity of Rv0351/Rv3628 Subunit Vaccine Formulated in Different Adjuvants Against Mycobacterium tuberculosis Infection

  • Kee Woong Kwon;Tae Gun Kang;Ara Lee;Seung Mo Jin;Yong Taik Lim;Sung Jae Shin;Sang-Jun Ha
    • IMMUNE NETWORK
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    • v.23 no.2
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    • pp.16.1-16.19
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    • 2023
  • Bacillus Calmette-Guerin (BCG) vaccine is the only licensed vaccine for tuberculosis (TB) prevention. Previously, our group demonstrated the vaccine potential of Rv0351 and Rv3628 against Mycobacterium tuberculosis (Mtb) infection by directing Th1-biased CD4+ T cells co-expressing IFN-γ, TNF-α, and IL-2 in the lungs. Here, we assessed immunogenicity and vaccine potential of the combined Ags (Rv0351/Rv3628) formulated in different adjuvants as subunit booster in BCG-primed mice against hypervirulent clinical Mtb strain K (Mtb K). Compared to BCG-only or subunit-only vaccine, BCG prime and subunit boost regimen exhibited significantly enhanced Th1 response. Next, we evaluated the immunogenicity to the combined Ags when formulated with four different types of monophosphoryl lipid A (MPL)-based adjuvants: 1) dimethyldioctadecylammonium bromide (DDA), MPL, and trehalose dicorynomycolate (TDM) in liposome form (DMT), 2) MPL and Poly I:C in liposome form (MP), 3) MPL, Poly I:C, and QS21 in liposome form (MPQ), and 4) MPL and Poly I:C in squalene emulsion form (MPS). MPQ and MPS displayed greater adjuvancity in Th1 induction than DMT or MP did. Especially, BCG prime and subunit-MPS boost regimen significantly reduced the bacterial loads and pulmonary inflammation against Mtb K infection when compared to BCG-only vaccine at a chronic stage of TB disease. Collectively, our findings highlighted the importance of adjuvant components and formulation to induce the enhanced protection with an optimal Th1 response.

The Forkhead Gene fkhB is Necessary for Proper Development in Aspergillus nidulans

  • Seo-Yeong Jang;Ye-Eun Son;Dong-Soon Oh;Kap-Hoon Han;Jae-Hyuk Yu;Hee-Soo Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.11
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    • pp.1420-1427
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    • 2023
  • The forkhead domain genes are important for development and morphogenesis in fungi. Six forkhead genes fkhA-fkhF have been found in the genome of the model filamentous Ascomycete Aspergillus nidulans. To identify the fkh gene(s) associated with fungal development, we examined mRNA levels of these six genes and found that the level of fkhB and fkhD mRNA was significantly elevated during asexual development and in conidia. To investigate the roles of FkhB and FkhD, we generated fkhB and fkhD deletion mutants and complemented strains and investigated their phenotypes. The deletion of fkhB, but not fkhD, affected fungal growth and both sexual and asexual development. The fkhB deletion mutant exhibited decreased colony size with distinctly pigmented (reddish) asexual spores and a significantly lower number of conidia compared with these features in the wild type (WT), although the level of sterigmatocystin was unaffected by the absence of fkhB. Furthermore, the fkhB deletion mutant produced sexual fruiting bodies (cleistothecia) smaller than those of WT, implying that the fkhB gene is involved in both asexual and sexual development. In addition, fkhB deletion reduced fungal tolerance to heat stress and decreased trehalose accumulation in conidia. Overall, these results suggest that fkhB plays a key role in proper fungal growth, development, and conidial stress tolerance in A. nidulans.

Improving antistress capacity and lipid productivity in the green alga Chlorella pyrenoidosa by adding abscisic acid under salt stress conditions

  • Ke Ding;Zheng Chen;Qiwu Wan;Xiaolin Hu;Xiaohui Wang;Heng Li;Yongzhong Wang;Yang Luo;Debing Xiang
    • ALGAE
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    • v.39 no.3
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    • pp.207-222
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    • 2024
  • In this study, the regulation of abscisic acid (ABA) on cell growth and lipid biosynthesis was investigated under salt-induced stress in Chlorella pyrenoidosa. It is found that as suffering from only salt stress, although the lipid content of single cell was improved, the inhibitory effects of stress on cell proliferation was visible. When the algal cells were exposed to salt stress and ABA conditions, lipid productivity was increased (45.35 mg L-1 d-1) by 1.17-fold compared to that of control cells (20.91 mg L-1 d-1), and the inhibition to cell growth was relieved. Transcriptomic analysis revealed that after adding ABA, these genes involved in antioxidant activity, jasmonic acid (JA) biosynthesis, and lipid biosynthesis were upregulated. Subsequently, we observed that the levels of glutathione, total antioxidant capacity, trehalose, and JA were elevated and the levels of reactive oxygen species were reduced. This study presents an effective approach to improve lipid production in algal cells, a new mechanism on that ABA alleviates intracellular oxidative stress through JA signaling pathway was elaborated.

Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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Comparative analysis of total sugar and sugar alcohol contents of the fruiting bodies in edible and medicinal mushrooms (식용 및 약용버섯류의 단당류, 이당류, 당알코올 함량 비교 분석)

  • Cho, Jae-Han;Park, Hye-Sung;Han, Jae-Gu;Lee, Gang-Hyo;Sung, Gi-Ho;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.12 no.4
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    • pp.316-323
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    • 2014
  • We analyzed saccharide by dividing and comparing Monosaccharide, Disaccharide and sugar Alcohol. At first, Glucose had outstanding contained quantity of ASI 7114 with 81.11 g/l even comparing with other mushrooms for medical use and edibility. And 119.98 g/l of Fructose was observed at Hericium erinaceum that was more contained quantity than Flammulina velutipes and Lentinus edodes. But, the most contained quantity observed in Ganoderma lucidum was ASI 7015 with 15.70 g/l that was the level of 1/8 approximately against Hericium erinaceum. Ribose was found at low level generally that was hardly contained. Xylose was also observed low level. ASI 7004 was detected at 0.96g/l that was the most content with imperceptible difference by comparing with other mushrooms for medical use and edibility. Next, 35.21 g/l of Trehalose, disaccharide was observed at Agaricus bisporus that was around 11 times of content than ASI 3.09 g/l that was the most content of Ganoderma lucidum. For ${\alpha}$-Lactose, Sparassis crispa has the most amount of 3.38 g/l that was around 12.5 times of ASI 7060 0.27 g/l that was the most content of Ganoderma lucidum. For Glycerol, sugar alcohol, 64.74 g/l was observed at Pleurotus eryngii. We knew it was around 8 times of ASI 7004 8.61 g/l that was the most content of Ganoderma lucidum. 0.72 g/l of Solbitol was observed at Flammulina velutipes. We knew it was around 2times of ASI 7003 0.31 g/l that was the most content of Ganoderma lucidum. Moreover most of Ganoderma lucidum didn't contain Solbitol. 2.96 g/l of Mannitol was observed at Agaricus bisporus. that was the most content among other mushrooms. Also Mannitol was contained in Lentinus edodes and leurotus cornucopiae only. Even Ganoderma lucidum didn't have Mannitol. At last, as a result of myo-Inosito content analysis, it was seemed not to be involved in any of mushrooms.

Analysis of Nutritional Components of Lepista nuda (민자주방망이버섯(L. nuda)의 영양성분 분석)

  • Lee Yang-Suk;Kim Jong-Bong;Shin Seung-Ryeul;Kim Nam-Woo
    • Food Science and Preservation
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    • v.13 no.3
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    • pp.375-381
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    • 2006
  • This study was carried out to analyze the nutritional component of Lepista nuda in order to estimate its nutritional and functional values. The content of moisture were $90.90{\pm}0.09%$ carbohydrate $4.34{\pm}0.07%$, crude protein $3.70{\pm}0.03%$, ash $0.55{\pm}0.04%$, and crude fat $0.47{\pm}0.01%$, respectively The reducing sugar was $636.17{\pm}7.50mg$. Total amount of free sugar was 97.32 mg, which were mainly composed of galactose (51.98 mg) and trehalose (22.48 mg). In the results of mined analysis, the content of K was the highest as $103.10{\pm}0.18mg$, followed by $56.01{\pm}0.20mg$ of Mg. The total content of hydrolyzed amino acid was 310.39 mg which consist of essential amino acids including valine (25.40 mg), leucine (22.52 mg), lysine (21.34 mg) and 198.89 mg of non-essential amino acids including glutamic acid (60.72 mg), aspartic acid (30.17 mg), and alanine (29.72 mg), as the major amino acids. free amino acids were 220.86 mg which were composed of essential amino acids (36.21 mg) inclusive of isoleucine (21.57 mg) and threonine (11.20 mg) and non-essential amino acids (184.65 mg) including glutamic acid (42.78 mg) and aspartic acid (32.44 mg) as major amino acids. Total content of amino acid derivatives was 187.85 me and that of hydroxyproline was highest as 121.50 mg.