• Journal of Microbiology and Biotechnology
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• v.20 no.12
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• pp.1630-1636
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• 2010

During the fermentation process of Saccharomyces cerevisiae, yeast cells must rapidly respond to a wide variety of external stresses in order to survive the constantly changing environment, including ethanol stress. The accumulation of ethanol can severely inhibit cell growth activity and productivity. Thus, the response to changing ethanol concentrations is one of the most important stress reactions in S. cerevisiae and worthy of thorough investigation. Therefore, this study examined the relationship between ethanol tolerance in S. cerevisiae and a unique protein called alcohol sensitive RING/PHD finger 1 protein (Asr1p). A real-time PCR showed that upon exposure to 8% ethanol, the expression of Asr1 was continuously enhanced, reaching a peak 2 h after stimulation. This result was confirmed by monitoring the fluorescence levels using a strain with a green fluorescent protein tagged to the C-terminal of Asr1p. The fluorescent microscopy also revealed a change in the subcellular localization before and after stimulation. Furthermore, the disruption of the Asr1 gene resulted in hypersensitivity on the medium containing ethanol, when compared with the wild-type strain. Thus, when taken together, the present results suggest that Asr1 is involved in the response to ethanol stress in the yeast S. cerevisiae.

• Journal of Microbiology and Biotechnology
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• v.18 no.5
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• pp.852-858
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• 2008

An extracellular protease (Mc1) was isolated from the nematode-trapping fungus Monacrosporium cystosporium by gel filtration, anion-exchange, and hydrophobic interaction chromatographies. This protease had a molecular mass of approximately 38 kDa and displayed an optimal activity at pH 7-9 and $56^{\circ}C$ (over 30 min). Its proteolytic activity was highly sensitive to the serine protease inhibitor PMSF (phenylmethylsulfonylfluoride, 0.1 mM), indicating that it belonged to the serine-type peptidase group. The Michaelis constant ($K_m$) and $V_max$ for substrate N-Suc-Ala-Ala-Pro-Phe-pNA were $1.67{\times}10^{-4}\;M$ and 0.6071 $OD_{410}$ per 30 s, respectively. This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. Moreover, the enzyme could immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, suggesting that it might playa role in infection against nematodes. The encoding gene of Mc1 was composed of one intron and two exons, coding for a polypeptide of 405 amino acid residues. The deduced amino acid sequence of Mcl showed 61.4-91.9% identity to serine proteases from other nematode-trapping fungi. Our results identified that Mcl possessed biochemical properties including optimal reaction condition and substrate preference that are different from previously identified serine proteases.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.1013-1017
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• 2003

A thermophilic nonspore-forming rod isolated from hay compost in Korea was subjected to a taxonomic study. The microorganism, designated as $SC-1^T$, was identified as a nitrate-reducing and nonmotile bacterium. Although the strain was negatively Gram-stained, a KOH test showed that the strain $SC-1^T$ belonged to a Gram-positive species. Growth was observed between 45 and $70^{\circ}C$. The optimal growth temperature and pH were $60^{\circ}C$ and pH 7.5, respectively. The G+C content of the genomic DNA was 65 mol% and the major quinone types were MK-6 and MK-7. A phylogenetic analysis based on 16S rDNA sequences revealed that the strain $SC-1^T$ was most closely related to Symbiobacterium thermophilum. However, the level of DNA-DNA relatedness between strain $SC-1^T$ and the type strain for Symbiobacterium thermophilum was approximately 30%. Accordingly, on the basis of the phenotypic traits and molecular systematic data, the strain $SC-1^T$ would appear to represent a new species within the genus Symbiobacterium. The type strain for the new species is named $SC-1^T$ ($=KCTC\;0307BP^T;\;DSM15906^T$).

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.912-920
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• 2021

SOS response is a conserved response to DNA damage in prokaryotes and is negatively regulated by LexA protein, which recognizes specifically an "SOS-box" motif present in the promoter region of SOS genes. Myxococcus xanthus DK1622 possesses a lexA gene, and while the deletion of lexA had no significant effect on either bacterial morphology, UV-C resistance, or sporulation, it did delay growth. UV-C radiation resulted in 651 upregulated genes in M. xanthus, including the typical SOS genes lexA, recA, uvrA, recN and so on, mostly enriched in the pathways of DNA replication and repair, secondary metabolism, and signal transduction. The UV-irradiated lexA mutant also showed the induced expression of SOS genes and these SOS genes enriched into a similar pathway profile to that of wild-type strain. Without irradiation treatment, the absence of LexA enhanced the expression of 122 genes that were not enriched in any pathway. Further analysis of the promoter sequence revealed that in the 122 genes, only the promoters of recA2, lexA and an operon composed of three genes (pafB, pafC and cyaA) had SOS box sequence to which the LexA protein is bound directly. These results update our current understanding of SOS response in M. xanthus and show that UV induces more genes involved in secondary metabolism and signal transduction in addition to DNA replication and repair; and while the canonical LexA-dependent regulation on SOS response has shrunk, only 5 SOS genes are directly repressed by LexA.

• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1577-1585
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• 2013

Bioethanol production from lignocellulose is considered as a sustainable biofuel supply. However, the low cellulose hydrolysis efficiency limits the cellulosic ethanol production. The cellulase is strongly inhibited by the major end product cellobiose, which can be relieved by the addition of ${\beta}$-glucosidase. In this study, three ${\beta}$-glucosidases from different organisms were respectively expressed in Saccharomyces cerevisiae and the ${\beta}$-glucosidase from Saccharomycopsis fibuligera showed the best activity (5.2 U/ml). The recombinant strain with S. fibuligera ${\beta}$-glucosidase could metabolize cellobiose with a specific growth rate similar to the control strain in glucose. This recombinant strain showed higher hydrolysis efficiency in the cellulose simultaneous saccharification and fermentation, when using the Trichoderma reesei cellulase, which is short of the ${\beta}$-glucosidase activity. The final ethanol concentration was 110% (using Avicel) and 89% (using acid-pretreated corncob) higher than the control strain. These results demonstrated the effect of ${\beta}$-glucosidase secretion in the recombinant S. cerevisiae for enhancing cellulosic ethanol conversion.

• 한국환경농학회:학술대회논문집
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• 2009.07a
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• pp.119-126
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• 2009

In recent years, research at the Environmental Microbial and Food Safety Laboratory (EMFSL), Agricultural Research Service (ARS) has focused on the development of novel image-based sensing technologies to address agro-food safety concerns, and transformation of these novel technologies into practical instrumentation for industrial implementations. The line-scan-based hyperspectral imaging techniques have often served as a research tool to develop rapid multispectral methods based on only a few spectral bands for rapid online applications. We developed a newer line-scan hyperspectral imaging platform for high-speed inspection on high-throughput processing lines, capable of simultaneous multiple inspection algorithms for different agro-food safety problems such as poultry carcass inspection for wholesomeness and apple inspection for fecal contamination and defect detection. In addition, portable imaging devices were developed for in situ identification of contamination sites and for use by agrofood producer and processor operations for cleaning and sanitation inspection of food processing surfaces. The aim of this presentation is to illustrate recent advances in the above agro.food safety sensing technologies.

• YAKHAK HOEJI
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• v.42 no.3
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• pp.302-306
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• 1998

In the previous study we described the antitumor activity of GLB, a protein-polysaccharide fraction of the growing tips of Ganoderma lucidum, against sarcoma 180 solid tu mor in ICR mice. In this study we investigated the stimulatory activity of GLB on macrophages. When analyzed using a flow cytometer, GLB ($100{\mu}g/ml$) was found to increase the phagocytic activity of the BALB/c mouse peritoneal macrophages as well as chicken macrophage BM2CL cells against FITC-labeled C.albicans by 55.2% and 21.2%, respectively. GLB also increased the spreading and the expression of MHC class II molecules of BM2CL cells as well as the mouse peritoneal macrophages. From these results, it is clear that GLB is a strong stimulator to the macrophages.

• Journal of Ginseng Research
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• v.40 no.2
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• pp.127-134
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• 2016

Background: Rhizospheric fungi play an essential role in the plantesoil ecosystem, affecting plant growth and health. In this study, we evaluated the fungal diversity in the rhizosphere soil of 2-yr-old healthy Panax notoginseng cultivated in Wenshan, China. Methods: Culture-independent Illumina MiSeq and culture-dependent techniques, combining molecular and morphological characteristics, were used to analyze the rhizospheric fungal diversity. A diffusion test was used to challenge the phytopathogens of P. notoginseng. Results: A total of 16,130 paired-end reads of the nuclear ribosomal internal transcribed spacer 2 were generated and clustered into 860 operational taxonomic units at 97% sequence similarity. All the operational taxonomic units were assigned to five phyla and 79 genera. Zygomycota (46.2%) and Ascomycota (37.8%) were the dominant taxa; Mortierella and unclassified Mortierellales accounted for a large proportion (44.9%) at genus level. The relative abundance of Fusarium and Phoma sequenceswas high, accounting for 12.9% and 5.5%, respectively. In total,113 fungal isolates were isolated from rhizosphere soil. They were assigned to five classes, eight orders (except for an Incertae sedis), 26 genera, and 43 species based on morphological characteristics and phylogenetic analysis of the internal transcribed spacer. Fusarium was the most isolated genus with six species (24 isolates, 21.2%). The abundance of Phoma was also relatively high (8.0%). Thirteen isolates displayed antimicrobial activity against at least one test fungus. Conclusion: Our results suggest that diverse fungi including potential pathogenic ones exist in the rhizosphere soil of 2-yr-old P. notoginseng and that antagonistic isolates may be useful for biological control of pathogens.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.96-103
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• 2007

Plant growth-promoting rhizobacteria (PGPR) have the potential to be used as microbial inoculants to reduce disease incidence and severity and to increase crop yield. Some of the PGPR have been reported to be able to enter plant tissues and establish endophytic populations. Here, we demonstrated an approach to screen bacterial endophytes that have the capacity to promote the growth of pepper seedlings and protect pepper plants against a bacterial pathogen. Initially, out of 150 bacterial isolates collected from healthy stems of peppers cultivated in the Chungcheong and Gyeongsang provinces of Korea, 23 putative endophytic isolates that were considered to be predominating and representative of each pepper sample were selected. By phenotypic characterization and partial 16S rDNA sequence analysis, the isolates were identified as species of Ochrobacterium, Pantoea, Pseudomonas, Sphingomonas, Janthinobacterium, Ralstonia, Arthrobacter, Clavibacter, Sporosarcina, Acidovorax, and Brevundimonas. Among them, two isolates, PS4 and PS27, were selected because they showed consistent colonizing capacity in pepper stems at the levels of $10^6-10^7CFU/g$ tissue, and were found to be most closely related to Pseudomonas rhodesiae and Pantoea ananatis, respectively, by additional analyses of their entire 16S rDNA sequences. Drenching application of the two strains on the pepper seedlings promoted significant growth of peppers, enhancing their root fresh weight by 73.9% and 41.5%, respectively. The two strains also elicited induced systemic resistance of plants against Xanthomonas axonopodis pv. vesicatoria.

• Journal of Microbiology and Biotechnology
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• v.2 no.4
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• pp.278-283
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• 1992

Aldehyde production by cells of a methanol utilizing yeast, Hansenula nonfermentans KYP-1 was improved when they were grown in a methanol-limited continuous culture, in comparison with cells grown in a batch culture. A higher cell yield was also obtained in continuous culture than in batch culture. This could be due to the fact that a lower methanol concentration was maintained in the jar fermentor to minimize growth inhibition by methanol. A maximum cell productivity of 0.219 g.$liter^{-1}.hr^{-l}$ and a cell yield of 47% were obtained at dilution rates of 0.1 $hr{-1}$ and 0.06 hr{-1}, respectively. The greatest amount of aldehyde was measured at a dilution rate of 0.08 $hr{-1}$. Under optimum reaction conditions, 915.7 mM of acetaldehyde was produced from 1.5 M ethanol after 21 hours reaction, with a conversion rate of 61%. Propionaldehyde and acrolein were produced with conversion rates of 32.7% and 44%, respectively.