• Journal of Microbiology and Biotechnology
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• 제29권11호
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• pp.1806-1816
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• 2019

Candida albicans is an opportunistic fungus possessing multiple virulence factors controlling pathogenicity. Cell wall proteins are the most important among these factors, being the first elements contacting the host. Ddr48 is a cell wall protein consisting of 212 amino acids. A DDR48 haploinsufficient mutant strain was previously found necessary for proper oxidative stress response and drug resistance. In this study, we aimed to further elucidate the role of Ddr48 by performing additional phenotypic characterization assays. A combinatory proteomic and bioinformatics approach was also undertaken to determine differentially expressed cell wall proteins. Results showed that the mutant strain exhibited a 10% decrease in adhesion mirrored by a 20% decrease in biofilm formation, and slight sensitivity to menadione, diamide, and SDS. Both strains showed similar hyphae formation, virulence, temperature tolerance, and calcofluor white and Congo red sensitivities. Furthermore, a total of 8 and 10 proteins were identified exclusively in the wild-type strain grown under filamentous and non-filamentous conditions respectively. Results included proteins responsible for superoxide stress resistance (Sod4 and Sod6), adhesion (Als3, Hyr4, Pmt1, and Utr2), biofilm formation (Hsp90, Ece1, Rim9, Ipp1, and Pra1) and cell wall integrity (Utr2 and Pga4). The lack of detection of these proteins in the mutant strain correlates with the observed phenotypes.

• Journal of Microbiology and Biotechnology
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• 제31권1호
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• pp.79-91
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• 2021

γ-Glutamylcysteine synthetase (Gcs1) and glutathione reductase (Glr1) activity maintains minimal levels of cellular methylglyoxal in Candida albicans. In glutathione-depleted Δgcs1, we previously saw that NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and alcohol dehydrogenase (Adh1) are the most active methylglyoxal scavengers. With methylglyoxal accumulation, disruptants lacking MGD1 or ADH1 exhibit a poor redox state. However, there is little convincing evidence for a reciprocal relationship between methylglyoxal scavenger genes-disrupted mutants and changes in glutathione-(in)dependent redox regulation. Herein, we attempt to demonstrate a functional role for methylglyoxal scavengers, modeled on a triple disruptant (Δmgd1/Δadh1/Δgcs1), to link between antioxidative enzyme activities and their metabolites in glutathione-depleted conditions. Despite seeing elevated methylglyoxal in all of the disruptants, the result saw a decrease in pyruvate content in Δmgd1/Δadh1/Δgcs1 which was not observed in double gene-disrupted strains such as Δmgd1/Δgcs1 and Δadh1/Δgcs1. Interestingly, Δmgd1/Δadh1/Δgcs1 exhibited a significantly decrease in H2O2 and superoxide which was also unobserved in Δmgd1/Δgcs1 and Δadh1/Δgcs1. The activities of the antioxidative enzymes erythroascorbate peroxidase and cytochrome c peroxidase were noticeably higher in Δmgd1/Δadh1/Δgcs1 than in the other disruptants. Meanwhile, Glr1 activity severely diminished in Δmgd1/Δadh1/Δgcs1. Monitoring complementary gene transcripts between double gene-disrupted Δmgd1/Δgcs1 and Δadh1/Δgcs1 supported the concept of an unbalanced redox state independent of the Glr1 activity for Δmgd1/Δadh1/Δgcs1. Our data demonstrate the reciprocal use of Eapx1 and Ccp1 in the absence of both methylglyoxal scavengers; that being pivotal for viability in non-filamentous budding yeast.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1734-1741
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• 2015

Few research had investigated the secretion of phospholipase and aspartyl proteinase from Candida spp. causing infection in females with type 2 diabetes mellitus. This research aimed to investigate the prevalence of vulvovaginal candidiasis (VVC) in diabetic versus non-diabetic women and compare the ability of identified Candida isolates to secrete phospholipases and aspartyl proteinases with characterization of their genetic profile. The study included 80 females with type 2 diabetes mellitus and 100 non-diabetic females within the child-bearing period. Candida strains were isolated and identified by conventional microbiological methods and by API Candida. The isolates were screened for their extracellular phospholipase and proteinase activities by culturing them on egg yolk and bovine serum albumin media, respectively. Detection of aspartyl proteinase genes (SAP1 to SAP8) and phospholipase genes (PLB1, PLB2) were performed by multiplex polymerase chain reaction. Our results indicated that vaginal candidiasis was significantly higher among the diabetic group versus nondiabetic group (50% versus 20%, respectively) (p = 0.004). C. albicans was the most prevalent species followed by C. glabrata in both groups. No significant association between diabetes mellitus and phospholipase activities was detected (p = 0.262), whereas high significant proteinase activities exhibited by Candida isolated from diabetic females were found (82.5%) (p = 0.000). Non-significant associations between any of the tested proteinase or phospholipase genes and diabetes mellitus were detected (p > 0.05). In conclusion, it is noticed that the incidence of C. glabrata causing VVC is increased. The higher prevalence of vaginal candidiasis among diabetics could be related to the increased aspartyl proteinase production in this group of patients.

• 한국안광학회지
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• 제21권3호
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• pp.215-225
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• 2016

목적: 본 연구에서는 소프트콘택트렌즈의 재질 특성과 착색 여부가 눈물 성분이 침착된 소프트콘택트렌즈에 칸디다균 흡착에 미치는 영향을 알아보았다. 방법: Etafilcon A, hilafilcon B, nelfilcon A 재질의 투명 소프트콘택트렌즈와 써클 소프트콘택트렌즈를 대상으로 하여 인공눈물에 침착시키기 전과 후의 흡착 균 수를 측정하였다. 또한, 주사전자현미경을 통해 렌즈에 흡착된 균을 관찰하였다. 결과: 칸디다균의 흡착은 콘택트렌즈 재질에 따라 통계적으로 유의하게 차이가 있었다. Etafilcon A 재질에서는 투명 소프트콘택트렌즈와 써클 콘택트렌즈간 균 흡착양 차이가 없었으며, hilafilcon B 및 nelfilcon A 재질에서는 써클 소프트콘택트렌즈에 흡착된 균 수가 다소 많았다. 투명 중심부와 착색부를 비교하였을 때 투명 중심부에 칸디다균 흡착이 더 많았으며, 투명 중심부와 투명 주변부를 비교하였을 때 투명 주변부에서의 균 흡착양이 더 많았다. 눈물 단백질이 침착되었을 경우 흡착된 칸디다균의 수가 감소하였으며 눈물 단백질의 칸디다균에 대한 항균력 유지는 재질에 따라 차이가 있어 etafilcon A 재질 렌즈에서 유지 기간이 가장 길었다. 결론: 본 연구에서는 소프트콘택트렌즈의 재질 특성, 착색여부 및 부위에 따라 흡착되는 칸디다균수가 달라진다는 것을 밝혔으며, 칸디다균의 흡착 특성에 따라 관리 방법을 달리해야 할 것을 제안한다.

• 대한의생명과학회지
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• 제25권4호
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• pp.407-416
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• 2019

Recent years have seen an increase in the incidence of candidiasis caused by non-albicans Candida (NAC) species. In fact, C. glabrata is now second only to C. albicans as the most common cause of invasive candidiasis. Therefore, the rapid genotyping specifically for C. glabrata is required for early diagnosis and treatment of candidiasis. A number of genotyping assays have been developed to differentiate C. glabrata sequence types (STs), but they have several limitations. In the previous study, multi-locus sequence typing (MLST) has performed with a total of 101 C. glabrata clinical isolates to analyze the prevalent C. glabrata STs in Korea. A total of 11 different C. glabrata STs were identified and, among them, ST-138 was the most commonly classified. Thus, a novel probe-based quantitative PCR (qPCR) assay was developed and evaluated for rapid and accurate identification of the predominant C. glabrata ST-138 in Korea. Two primer pairs and hybridization probe sets were designed for the amplification of internal transcribed spacer 1 (ITS1) region and TRP1 gene. Analytical sensitivity of the probe-based qPCR assay was 100 ng to 10 pg and 100 ng to 100 pg (per 1 μL), which target ITS1 region and TRP1 gene, respectively. This assay did not react with any other Candida species and bacteria except C. glabrata. Of the 101 clinical isolates, 99 cases (98%) were concordant with MLST results. This novel probe-based qPCR assay proved to be rapid, sensitive, highly specific, reproducible, and cost-effective than other genotyping assay for C. glabrata ST-138 identification.

• 한국약용작물학회지
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• 제11권5호
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• pp.358-363
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• 2003

미치광이풀의 모상근 뱌양에 의해 의학적으로 중요한 물질인 scopolamine과 hyoscyamine 생산을 증진시키기 위해 3종의 곰팡이와 1종의 효모 유래의 생물학적 elicitor를 supernatant와 homogenate의 형태로 처리하였다. Elicitor 중 Candida albicans는 supernatant와 homogenate 모두 scopolamine 생산성을 증가시켰다. Fusarium solani의 homogenate와 C. albicans의 supernatant 처리로 hyoscyamine 함량이 증가되었다. 그러나 나머지 fungus strain도 대조구에 비해 tropane alkaloids의 생산을 다소 증가시켰다. 본 연구에서는 C. albicans가 tropane alkaloids 생산에 적합한 elicitor임으로 구명되었다. 본 연구 결과는 미치광이풀 모상근 대량배양에 의한 tropane alklaoids 대량생산에 기여할 것이다.

• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.689-699
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• 2020

Brevibacillus brevis GZDF3 is a gram-positive, plant growth-promoting rhizosphere bacterium (PGPR) isolated from the rhizosphere soil of Pinellia ternata (an important herb in traditional Chinese medicine). The GZDF3 strain produces certain active compounds, such as siderophores, which are the final metabolite products of non-ribosomal peptide synthetase (NRPS) and independent non-ribosomal peptide synthetase (NIS) activity. With the present study, we attempted to investigate the siderophore production characteristics and conditions of Bacillus sp. GZDF3. The antibacterial activity of the siderophores on pathogenic fungi was also investigated. Optimal conditions for the synthesis of siderophores were determined by single factor method, using sucrose 15 g/l, asparagine 2 g/l, 32℃, and 48 h. The optimized sucrose asparagine medium significantly increased the production of siderophores, from 27.09% to 54.99%. Moreover, the effects of different kinds of metal ions on siderophore production were explored here. We found that Fe³⁺ and Cu²⁺ significantly inhibited the synthesis of siderophores. The preliminary separation and purification of siderophores by immobilized-metal affinity chromatography (IMAC) provides strong antibacterial activity against Candida albicans. The synergistic effect of siderophores and amphotericin B was also demonstrated. Our results have shown that the GZDF3 strain could produce a large amount of siderophores with strong antagonistic activity, which is helpful in the development of new biological control agents.

• 한국생물정보학회:학술대회논문집
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• 한국생물정보시스템생물학회 2005년도 BIOINFO 2005
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• pp.139-143
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• 2005

Cytochrome P450 14${\alpha}$-sterol demethylase enzyme (CYP51) is the target a of azole type antifungals. The azole blocks the ergosterol synthesis and thereby inhibits fungal growth. A three-dimensional (3D) homology model of CYP51 from Candida albicans was constructed based on the X-ray crystal structure of CYP51 from Mycobacterium tuberculosis. Using this model, the binding modes for the substrate (24-methylene-24, 25-dihydrolanosterol) and the known inhibitors (fluconazole, voriconazole, oxiconazole, miconazole) were predicted from docking. Virtual screening was performed employing Structure Based Focusing (SBF). In this procedure, the pharmacophore models for database search were generated from the protein-ligands interactions each other. The initial structure-based virtual screening selected 15 compounds from a commercial available 3D database of approximately 50,000 molecule library, Being evaluated by a cell-based assay, 5 compounds were further identified as the potent inhibitors of Candida albicans CYP51 (CACYP51) with low minimal inhibitory concentration (MIC) range. BMD-09-01${\sim}$BMD-09-04 MIC range was 0.5 ${\mu}$g/ml and BMD-09-05 was 1 ${\mu}$g/ml. These new inhibitors provide a basis for some non-azole antifungal rational design of new, and more efficacious antifungal agents.

• 대한약침학회지
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• 제15권1호
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• pp.23-28
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• 2012

Objective: This experimental study was designed to investigate the safety and the effectiveness of Samjeong pharmacopuncture solution (SPS) manufactured by using a the lowtemperature extract on process. Methods: To identify the safety and the effectiveness of using SPS as eye drops, we performed applied eye irritation tests on rabbits and antibacterial tests for Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, and Candida albicans. The eye irritation test was performed according to the toxicity testing regulation of the Korea Food & Drug Administration (2009. 8. 24, KFDA 2009-116). After SPS had been applied on the left eye of the rabbits, eye irritation in the cornea, iris and conjunctiva was observed on the 1st, 2nd, 3rd, 4th & 7th day. After SPS had been dropped on bacterial species that cause keratitis, the minimum inhibition concentration and the size of the inhibition zone were measured. The anti-bacterial potency was also measured by taking the size of inhibition zone. Results: After SPS had been administered on the left eye of the rabbits, none of nine rabbits were found to show abnormal signs or weight changes. After SPS had been administered on the left eye of the rabbits, no eye irritation in the cornea, iris and conjunctiva was observed on the 1st, 2nd, 3rd, 4th & 7th day. No specific response was detected in MIC for bacterial species Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Aspergillus niger, Fusarium oxysporum, and Candida albicans after SPS had been applied. Conclusions: This study suggests that SPS is a non-toxic and non-irritant medicine that does not cause any of eye irritation in rabbits, but it has no antibacterial effects on bacterial species that are well known to cause keratitis. These results suggest that more research is required on extracts from herbal medicines for treating keratitis.

• Journal of Ginseng Research
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• 제30권3호
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• pp.128-131
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• 2006

본 연구는 홍삼엑스로부터 분리된 홍삼사포닌이 일부 병원성 미생물 Staphylococcus aureus 및 Candida albicans의 성장에 미치는 영향을 조사하였다. 사포닌의 농도를 달리하여 첨가하고 5 일 동안 배양한 후 측정된 S. aureus의 pH 는 사포닌 무첨가군 pH 8.55에 비해 0.013, 0.125, 0.5, 1.0% 첨가군에서 각각 8.35, 7.80, 6.87, 6.34로 사포닌 농도가 증가할수록 pH는 감소하는 경향을 나타내었다. S. aureus의 성장에 미치는 사포닌의 영향을 조사한 결과 사포닌 무첨가군의 성장률을 100%로 놓았을 때 상대적으로 계산된 균의 성장률은 0.013, 0.125, 0.5, 1.0% 첨가군에서는 각각 55.56, 52.22, 0.001, 0%로 나타나서 사포닌 농도가 증가할수록 농도 의존적으로 균의 성장이 강하게 억제되는 경향을 나타내었다. 특히 조사포닌 0.5% 이상의 농도에서는 균의 생육은 완전히 억제되었다. 한편 병원성 효모인 C. albicans의 pH도 사포닌 무첨가군 pH 5.97에서 사포닌 첨가시 pH 5.83-5.60으로 약간씩 감소하는 경향이었다. 각 첨가군과 성장률을 비교하였을 경우 사포닌 0.013, 0.125, 0.5, 1.0% 첨가군은 각각 50.0, 50.0, 37.5, 25.0%의 성장율을 나타내어 S. aureus와 마찬가지로 사포닌 첨가농도가 증가할수록 성장률이 감소하는 경향을 보였다. 그러나 C. albicans의 생균수는 무첨가군 $8.0{\times}10^7\;CFU/ml$에 비해 사포닌 첨가군 모두에서 $2.0-4.0{\times}10^7\;CFU/ml$으로 무첨가군에 비해서는 낮은 수준이었지만 유의적인 변화는 관찰되지 않았다. 따라서 사포닌은 0.13% 이상의 농도에서 병원성효모 C. albicans 보다는 세균인 S. aureus의 성장을 크게 억제하였으며, 이러한 결과로부터 홍삼사포닌은 진균류인 효모보다는 세균에 대해 더욱 큰 생육저해효과를 나타낸다는 사설을 알 수 있었다.