• Journal of Microbiology and Biotechnology
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• v.22 no.6
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• pp.838-848
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• 2012

Orally administered herbal glycosides are metabolized to their hydrophobic compounds by intestinal microflora in the intestine of animals and human, not liver enzymes, and absorbed from the intestine to the blood. Of these metabolites, some, such as quercetin and kaempherol, are mutagenic. The fecal bacterial enzyme fraction (fecalase) of human or animals has been used for measuring the mutagenicity of dietary glycosides. However, the fecalase activity between individuals is significantly different and its preparation is laborious and odious. Therefore, we developed a fecal microbial enzyme mix (FM) usable in the Ames test to remediate the fluctuated reaction system activating natural glycosides to mutagens. We selected, cultured, and mixed 4 bacteria highly producing glycosidase activities based on a cell-free extract of feces (fecalase) from 100 healthy Korean volunteers. When the mutagenicities of rutin and methanol extract of the flos of Sophora japonica L. (SFME), of which the major constituent is rutin, towards Salmonella typhimurium strains TA 98, 100, 102, 1,535, and 1,537 were tested using FM and/or S9 mix, these agents were potently mutagenic. These mutagenicities using FM were not significantly different compared with those using Korean fecalase. SFME and rutin were potently mutagenic in the test when these were treated with fecalase or FM in the presence of S9 mix, followed by those treated with S9 mix alone and those with fecalase or FM. Freeze-dried FM was more stable in storage than fecalase. Based on these findings, FM could be usable instead of human fecalase in the Ames test.

• The Korean Journal of Mycology
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• v.49 no.4
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• pp.441-454
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• 2021

Fungal diseases including anthracnose, stem rot, blight, wilting, and root rot of crops are caused by phytopathogens such as Colletotrichum species, Sclerotinia sclerotiorum, Phytophthora species, and Fusarium oxysporum and F. solani which threaten the production of chili pepper. In this study, to identify biological control agents (BCAs) of phytopathogenic fungi, potentially useful Bacillus species were isolated from the field soils. We screened out five Bacillus strains with antagonistic capacity that are efficiently inhibiting the growth of phytopathogenic fungi. Bacillus species were characterized by the production of extracellular enzymes, siderophores, and indole-3-acetic acid (IAA). Furthermore, the influence of bacterial strains on the plant growth promoting activity and seedling vigor index were assessed using Brassica juncea as a model plant. Inoculation with Bacillus subtilis SRCM 121379 significantly increased the length of B. juncea shoots and roots by 45.6% and 52.0%, respectively. Among the bacterial isolates, Bacillus subtilis SRCM 121379 showed the superior enzyme activities, antagonistic capacity and plant growth promoting effects. Based on the experimental results, Bacillus subtilis SRCM 121379 (GenBank accession no. NR027552) was finally selected as a BCA candidate.

• Journal of Ginseng Research
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• v.14 no.1
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• pp.21-26
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• 1990

The chemical composition and stabilities of the purified ginseng invertase were investigated. The purified enzyme was found to be a glycoprotein composed of 80.2% protein and 19.7% total sugar. The protein component of the enzyme was composed of acidic amino acid (9.3%), basic amino acid (48.9%), nonpolar amino acid (21.4%), polar amino acid (20.4%) and 6.1% S-containing amino acid. It showed especially high contents of histidine and serine. The enzyme was inactivated almost completely by the treatment with some proteases (papain, pepsin. trypsin, pancreatin and microbial alkaline pretense) and protein denatllrants (8M urea and 6M guanidine-HC1), bolt not with glyrosidase (${\alpha}$-amylase, ${\beta}$-amylase. glcoamylese and cellullase). btonosaccharides sllch as glilrose, fructose, galactose and mannose did not exert any influence on the enzyme activity. The activity of the enzyme was inhibited by Ag+, Mn2+, Hg2+, Zn2+ and Al3+, whereas Ca2+, Mg2+, Ba2+ and Fe3+ gave rather activating effects on the enzyme activity. The enzyme was relatively stable in the VH range of VH 6 and 8, and at the temperatures below 35$^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• v.11 no.3
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• pp.534-538
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• 2001

The xynA gene encoding an alikali-tolerant endo-1,4-${\beta}$-xylanase (XYN) was cloned from the alkalophilic Bacillus pumilus A-30. The nucleotide sequence of a 974-bp DNA fragment containing the xynA was determined. An ORF of 684 nucleotides that encoded a protein of 228 amino aicds was detected. Asparagine-71 of XYN from B. Pumilus A-30 showed to be highly conservative in alkaline xylanases of family G/11, upon comparing the amino acid sequences of 17 family G/11 xylanases. Site-directed mutation of N71D of the xynA gene resulted in a decrease of 12.4% in the specific acitivity and a significant decline in the enzyme activity in the alkaline pH range.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.441-444
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• 2002

This study was targeted to develope a microbial consortium having high cellulase production. A filamentous fungus, strain FB01, isolated from a compost showed high ${\beta}-glucosidase$ activity especially. The strain FBOl was co-cultured with Trichoderma viride to enhance the productivity of ${\beta}-glucosidase$, changing inoculation time of one strain (FB01). The microbial consortium prepared showed the higher cellulytic enzyme production than T. viride well-known. The maximal enzyme production was obtained when the microbial consortium was cultured at $30^{\circ}C$ and pH 6.0 for 10days and the activities of CMCase, ${\beta}-glucosidase$, and avicelase were 2.0, 0.8, and 0.2 U/mL, respectively. These enzyme activities were 2, 4, and 2 times as high as those of CMCase, ${\beta}-glucosidase$, avicelase from T. viride, respectively, indicating that a synergistic interaction appeared between T viride and strain FB01. The serial subcultures by pH control increased ${\beta}-glucosidase$ production about 3.2 times. Also, enzyme production using rice-straw as a carbon source showed that the activities of CMCase, ${\beta}-glucosidase$, and avicelase were 3.69, 0.76, 0.17 U/mL, respectively, and ${\beta}-glucosidase$ activity was 1.5 times higher than that of T. viride. Consequently, microbial consortium showed the considerabely enhanced production of the cellullolytic enzymes, such as CMCase, ${\beta}-glucosidase$, and avicelase compared those of T. viride, and a favorable stability for the enzyme production even in the serial subcultures.

• Korean Journal of Soil Science and Fertilizer
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• v.42 no.6
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• pp.500-512
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• 2009

This study was carried out to evaluate the effect of temperature on soil microbial biomass, enzyme activities, and PLFA content in the volcanic(VAS) and the non-volcanic ash soil(NVAS). The soils were treated with organic materials such as organic fertilizer pelleted(OFPL), organic fertilizer powdered(OFPD), pig manure compost(PMC), and food waste compost(FWC). Two grams of organic materials were well mixed with 30g of dried volcanic and non-volcanic ash soil(< 2 mm) with 50% of soil moisture content. And the soils were incubated at 10, 20, $30^{\circ}C$ in incubator. Soils were analysed on the incubation times as followed; soil pH, total nitrogen, organic matter(at 75, 150, 270 days), microbial biomass C and PLFA (at 75, 270 days), microbial biomass N and soil enzyme(at 150, 270 days). pH values of soils treated with PMC and FWC had no changes on soil type, and incubation temperature. However, the pH was increased with temperature in the soils treated with OFPL. The changes in NVAS was higher than in VAS. Soil microbial biomass C content were high in the condition of high temperature and organic fertilizers treatment in VAS. But the contents were gradually decreased with incubation period in both NVAS and VAS. Soil microbial biomass N was high in NVAS treated with organic fertilizers and in VBS treated with PMC and FWC. PLFA content was higher in NVBS than in VBS at 75 days but showed high in VBS at 270 days. Urease activity of NVBS treated with OFPL showed $10^{\circ}C$ (75.0)> $20^{\circ}C$ (16.3)>$30^{\circ}C$ ($4.6ug\;NH{_4-}N\;g^{-1}\;2h^{-1}$) at 150 days. It were decreased gradually high temperature and time passes. And it showed high at $10^{\circ}C$ in VBS. Glucosidase activity was higher in NVBS than in VBS. Correlation coefficient of between soil microbial biomass C and microbial activity indicators showed that PLFA was high significantly at $r^2=0.91$ in NVBS and ${\beta}-glucosidase$ was $r^2=0.83$ in VBS. Soil microbial activities showed differences in the relative sensitivities of soil type and soil temperature.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.462-469
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• 2012

A metagenomic fosmid library was constructed from genomic DNA isolated from the microbial community residing in hindguts of a wood-feeding higher termite (Microcerotermes sp.) collected in Thailand. The library was screened for clones expressing lignocellulolytic activities. Fourteen independent active clones (2 cellulases and 12 xylanases) were obtained by functional screening at pH 10.0. Analysis of shotgun-cloning and pyrosequencing data revealed six ORFs, which shared less than 59% identity and 73% similarity of their amino acid sequences with known cellulases and xylanases. Conserved domain analysis of these ORFs revealed a cellulase belonging to the glycoside hydrolase family 5, whereas the other five xylanases showed significant identity to diverse families including families 8, 10, and 11. Interestingly, one fosmid clone was isolated carrying three contiguous xylanase genes that may comprise a xylanosome operon. The enzymes with the highest activities at alkaline pH from the initial activity screening were characterized biochemically. These enzymes showed a broad range of enzyme activities from pH 5.0 to 10.0, with pH optimal of 8.0 retaining more than 70% of their respective activities at pH 9.0. The optimal temperatures of these enzymes ranged from $50^{\circ}C$ to $55^{\circ}C$. This study provides evidence for the diversity and function of lignocellulose-degrading enzymes in the termite gut microbial community, which could be of potential use for industrial processes such as pulp biobleaching and denim biostoning.

• Microbiology and Biotechnology Letters
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• v.9 no.3
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• pp.129-137
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• 1981

The purpose of this study was to extract saponin efficiently from ginseng leaves and peelings by macerating them with microbial enzyme. To begin with, we selected G-211 strain having the highest macerating activity among several rotting molds of fresh ginseng. Crude macerating enzyme was prepared from this G-2l1 strain by ammonium sulfate precipitation, and was applied to macerating leaves and peelings of ginseng. The optimal pH of the enzyme for maceration was 5.0 in both leaves and peelings of ginsen g. The optimal pH for the extraction of soluble matters and saponins was 4.5 and 5.5 in ginseng leaves and ginseng peelings, respectively. When this enzyme was treated together with crude cellulase from Trichoderma viride (To4), the extract content of saponin was 3.45% for ginseng leaves and 3.90% for ginseng peelings. Their yields were 39.8 % and 39.3 % of total saponin amounts in ginseng leaves and ginseng peelings, respectively. The ginsenoside patterns of saponins extracted with the treatment of enzymes were also studied by HPLC technics.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.302-307
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• 2011

Native chitin deacetylase of Aspergillus nidulans was purified to apparent homogeneity by a combination of phenyl-Sepharose and Q-Sepharose column chromatography. In order to analyze the amino acid residues involved in the enzyme activity, the enzyme was chemically modified with chemical agent, which selectively reacted with the specific amino acid residue on the protein. When the enzyme was chemically modified with diethylpyrocarbonate, which specifically reacted with histidine residues on the protein, the activity was eliminated. The chitin deacetylase, chemically modified with 100 ${\mu}M$ modifier at the residue of arginine or tyrosine, has shown to have decreased activities. It was shown that the modification at aspartic acid or glutamic acid did not affect the enzyme activity to a greater extent, which would not implicate that acid amino residues were directly involved in catalytic reaction and would affect on the global structures of the proteins. This results demonstrated that histidine and tyrosine residues of enzyme would participate in an important function of the chitin deacetylase activity.

• Archives of Pharmacal Research
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• v.20 no.6
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• pp.519-524
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• 1997

Steroid $9{\alpha}$-hydroxylase is an enzyme found in nocardioform microorganisms which can utilize steroids as a sole carbon source. After fractional centrifugation of the cell homogenates, the enzyme activity in Nocardia and Rhodococcus was found in cytoplasmic membrane fraction. On the contrary, Mycobacterium had its 9.alpha.-hydroxylation activity in cytosolic fraction. To characterize the enzyme in these microorganisms, several potential inhibitors of 9.alpha.-hydroxylase were tested and the cofactor requirement for the same enzyme was also examined. The inhibitory effect of ferrous ion chelators indicated involvement of iron containing proteins in the 9.alpha.-hydroxylase system. On the other hand, metyrapone, an inhibitor known to be specific for cytochrome P450 interfered with the enzyme in Mycobacterium, but didn't inhibit the enzyme activity in Nocardia and Rhodococcus. While the $9{\alpha}$-hydroxylase system in Nocardia and Rhodococcus required NADPH, NADH was required as an election donor in Mycobacterium.