• Mycobiology
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• v.42 no.2
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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.

• Microbiology and Biotechnology Letters
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• v.23 no.3
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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.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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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).

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.5
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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.

• Microbiology and Biotechnology Letters
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• v.23 no.3
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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.

• Microbiology and Biotechnology Letters
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• v.20 no.6
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• pp.642-646
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• 1992

We have isolated conditional lethal mutants of Saccharomyces cerevisiae which are low in (1 ~3)-~-D-glucan synthase activity. These mutants were osmotic sensitive at nonpermissive temperature (37$^{\circ}$C) and showed a decreased level of alkali-insoluble cell wall glucan. The decrease in (1 ~3)-~-D-glucan synthase activity of the mutants appeared to be mainly due to the defect in catalytic component rather than in GTP-binding component.

• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.758-764
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• 2019

${\beta}$-Glucan is a chief structural polymer in the cell wall of yeast. ${\beta}$-Glucan has attracted intensive attention because of its wide applications in health protection and cosmetic areas. In the present study, the ${\beta}$-glucan biosynthesis pathway in S. Cerevisiae was engineered to enhance ${\beta}$-glucan accumulation. A newly identified bacterial ${\beta}-1$, 6-glucan synthase GsmA from Mycoplasma agalactiae was expressed, and increased ${\beta}$-glucan content by 43%. In addition, other pathway enzymes were investigated to direct more metabolic flux towards the building of ${\beta}$-glucan chains. We found that overexpression of Pgm2 (phosphoglucomutase) and Rho1 (a GTPase for activating glucan synthesis) significantly increased ${\beta}$-glucan accumulation. After further optimization of culture conditions, the ${\beta}$-glucan content was increased by 53.1%. This study provides a new approach to enhance ${\beta}$-glucan biosynthesis in Saccharomyces cerevisiae.

• Advances in Traditional Medicine
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• v.10 no.1
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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.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.6
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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.

• Mycobiology
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• v.38 no.2
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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.