• 한국미생물·생명공학회지
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• 제23권3호
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• pp.311-315
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• 1995

Assay conditions for screening of $\beta$-1,3-glucan synthase inhibitor were evaluated. Cells in the beginning of mid-log phase showed the highest activity of the $\beta$-1,3-glucan synthase. Cells permeabilized with 1% digitonin treatment could be used as a good crude enzyme source for convenient screening of the $\beta$-1,3-glucan synthase inhibitors. Calcofluor white (0.125% in final) and papulacandin B (25 $\mu$g/ml) inhibit 90% and more than 50% of the $\beta$-1,3-glucan synthase activity, respectively. Cells grown at 37$\circ$C showed higher enzyme activity than those of 25$\circ$C. Catalytic factor of the $\beta$-1,3-glucan synthase was solubilized from particulated membrane preparations, holoenzyme, by extracting with 0.00938% CHAPS.

• Mycobiology
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• 제42권2호
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• pp.167-173
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• 2014

A ${\beta}$-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble ${\beta}$-1,3-glucan (${\beta}$-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a ${\beta}$-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal ${\beta}$-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal ${\beta}$-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa ${\beta}$-glucan synthase was estimated to include catalytically insignificant transmembrane ${\alpha}$-helices and loops. Sequence analysis of 101 fungal ${\beta}$-glucan synthases, obtained from public databases, revealed that the ${\beta}$-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of ${\beta}$-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa ${\beta}$-glucan synthase in this study belonged to Type II family, meaning Type I ${\beta}$-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble ${\beta}$-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa ${\beta}$-glucan synthase will provide better explanations.

• 동의생리병리학회지
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• 제22권5호
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• pp.1196-1201
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• 2008

A examination of the kinetic properties of UDP-glucose : (1,3)-$\beta$-glucan (callose) synthase from mung bean seedings (Sorbus cortex) shows that these enzymes have a complex interaction with UDP-glucose and various effectors. Deoxynojirimycin increased the inhibitory effect of (1,3)-$\beta$-glucan synthase at the concentration-dependent manner by fluorescence assay. The inhibitory effect of Fr. 2-16 (97.15%) showed higher than that of deoxynojirimycin (80.63%). Fr. 2-3 inhibited the growth of the Candida albicans at 1 mm inhibition zone by disk diffusion method. These results suggest that Sorbus cortex extract can be used as a stable antifungal material.

• 한국미생물·생명공학회지
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• 제23권3호
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• pp.316-321
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• 1995

Some properties of $\beta$-1, 3-glucan synthase system in Saccharamyces cerevisiae were investigated. By extraction with detergent and salt, the membrane preparations could be dissociated into two components, one soluble, the other still membrane bound. Both components, in addition to GTP, were necessary for the activity of $\beta$-1, 3-glucan synthase like other fungi. The protective effect of guanosine nucleotides on the soluble factor pointed to the possibility that this fraction contained a GTP-binding protein. Addition of increasing amounts of soluble factor to a constant amount of insoluble catalytic factor, vice versa, gave rise to a saturation curve. These results, including different types of evidence, indicate that the soluble factor and the catalytic factor form a complex.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVII)
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• pp.668-671
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• 2005

In this study, the full gene of the putative ${\beta}-1,3-glucan$ synthase catalytic subunit(gi:40556679) in Agrobacteriujm sp. ATCC31750 was cloned into E. coli BL21(DE). We found that putative ${\beta}-1,3-glucan$ synthase catalytic subunit full gene mutant(E. coli mutant FC) produced soluble glucan.instead of curdlan(insoluble glucan).

• Advances in Traditional Medicine
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• 제10권1호
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• pp.44-49
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• 2010

An examination of the kinetic properties of UDP-glucose, (1,3)-$\beta$-glucans (callose) synthase, from mung bean seedlings (Sorbus commixta cortex) shows that these enzymes have a complex relationship with UDP-glucose and various effectors. Fluorescence assay showed that deoxynojirimycin increased the inhibitory effect of (1,3)-$\beta$-glucan synthase in a concentration-dependent manner. The inhibitory effect of sakuranetin (34.34%) was higher than that of deoxynojirimycin (80.63%). Disk diffusion method revealed that sakuranetin inhibited the growth of Candida albicans to a 1.5 mm inhibition zone. These results suggest that sakuranetin, isolated from Sorbus commixta cortex extract, can be used as stable antifungal material.

• 한국미생물·생명공학회지
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• 제20권6호
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• pp.642-646
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• 1992

본인 등은 베타-1,3-글루칸 합성효소능이 낮은 Saccharomyces cerevisiae의 조건성 돌연변이주들을 분리하여 그 특성을 조사하였다. 이 돌연변이주들은 비허용온도인 37'C에서 삼투감수성을 나타나며, 세포벽 성분 중 알칼리 비용해성 글루칸의 함량이 낮았다. 베타-1,3-글루칸의 합성효소능의 결함원인을 조사한 결과, 효모의 베타-1,3-글루칸 합성효소를 구성하는 촉매성분(cataytic component)과 GTP-결합성분 두가지 중 촉매 성분의 결함이 이 돌연변이주들의 베타-1,3-글루칸 합성효소능의 손상원인인 것으로 추정되었다.

• 한국균학회지
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• 제23권2호통권73호
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• pp.129-138
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• 1995

비허용온도인 $37^{\circ}C$에서 삼투감수성을 보이며 베타-1,3-글루칸 합성능이 현저히 손상된 Saccharomyces cerevisiae mutant(LP353)를 YCp50으로 제조한 yeast genomic library로 형질전환시킨 후, 콜로니 자기방사법으로 형질전환체의 선별을 시도한 결과, LP353의 베타-1,3-글루칸 합성능을 부분적으로 회복시켜 주는 약 8.5-kb 크기의 DNA 절편을 클로닝하는데 성공하였다. 클로닝된 8.5-kb의 DNA 절편은 copy 수에 무관하게 LP353의 또 다른 표현형질인 온도의존적 삼투감수성은 회복시켜 주지 못하였으나, 세포벽의 베타-1,3-글루칸 함량과 베타-1,3-글루칸 분해효소인 ${\beta}-glucanase$에 대한 내성은 copy수에 무관하게 증가시켜 주었다. 한편, 8.5-kb의 DNA 절편은 $37^{\circ}C$의 삼투안정제가 첨가된 액체배지에서 잘 자라지 못하는 LP353의 돌연변이 형질을 회복시켜 야생형의 수준에 근접하는 생장양상을 보여 주었다. 이상의 결과로 클로닝된 8.5-kb 크기의 DNA 절편은 S. cerevisiae의 베타-1,3-글루칸 생합성에 관여하는 유전자의 하나인 BGS2를 포함하고 있는 것으로 보여지며, subcloning을 통한 기능부위 분석 결과, 4.8-kb 크기의 BglII-KpnI DNA 절편에 BGS2가 존재하는 것으로 추정되었다.

• Mycobiology
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• 제38권2호
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• pp.102-107
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• 2010

We identified a gene for $\beta$-1,3-glucan synthesis (GBG1), a nonessential gene whose disruption alters cell wall synthesis enzyme activities and cell wall composition. This gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. Disruption of the GBG1 gene did not affect cell viability or growth rate, but did cause alterations in cell wall synthesis enzyme activities: reduction of $\beta$-1,3-glucan synthase and chitin synthase III activities as well as increased chitin synthase I and II activities. GBG1 disruption also showed altered cell wall composition as well as susceptibility toward cell wall inhibitors such as Zymolyase, Calcofluor white, and Nikkomycin Z. These results indicate that GBG1 plays a role in cell wall biogenesis in S. cerevisiae.

• 동의생리병리학회지
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• 제17권6호
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• pp.1509-1513
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• 2003

Antifungal activities of the extracts from 26 medicinal plants were investigated utilizing paper-disk diffusion method and (1,3)β-glucan synthase inhibitory assay. (1,3)β-glucan synthase is considered as valuable target in the development of antifungal agents. Among the screened extracts, the ethyl acetate extract of Equisetum arvense, the ethyl acetate extract of Polygonum aviculare, the butanol extract of Crataegus pinnatifida and the n-hexane extract of Saussurea lappa showed significant antifungal activities on Candida albacans in both disk diffusion and enzyme assays.