• Journal of Microbiology and Biotechnology
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• 제22권6호
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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.

• 한국균학회지
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• 제49권4호
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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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• 제14권1호
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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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• 제11권3호
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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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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.441-444
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• 2002

본 연구의 목적은 기존의 단일 균주보다 두 종의 서로 다른 균주를 복합시켰을 때 두 균 주간의 상호 보완적인 면을 이용하여 효소생산을 극대화하기 위함이며, 다음과 같은 결론을 얻었다. 실험에서 사용된 T. viride와 FB01는 최적 pH와 온도가 비슷하여 혼합배양이 가능 하였으며, 이들의 복합균주를 개발하는데 성공하였다. 두 균주간의 상호작용으로 인해 단일 균주일 때보다 CMCase, ${\beta}-glucosidase$ 및 avicelase의 활성이 우수했으며 또한 계대배양 및 pH 조절을 통해 ${\beta}-glucosidase$이 활성이 최고 3.2배까지 증가함을 알 수 있었다. 탄소원으로 섬유소 폐기물중 특히 볏짚을 이용했을 때 복합균주에 의한 효소생산이 효과적이었으므로 앞으로 이 조건에서 복합균주를 장기적으로 계대배양하면서 효소의 생산성 향상과 복합균주의 안정성을 관찰할 필요가 있다고 사료된다.

• 한국토양비료학회지
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• 제42권6호
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• pp.500-512
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• 2009

유기물원을 항온 배양했을 때 온도가 토양미생물체량과 효소활성 및 PLFA함량에 미치는 영향을 토양 특성별로 평가하고자 입상과 분상 혼합유기질비료, 돈분퇴비, 음식물퇴비를 화산회토양과 비화산회토양 30g에 2g을 잘 혼합 후 10, 20, $30^{\circ}C$에서 항온배양을 하면서 pH, 토양질소, 유기물 함량, Microbial biomass C와 N, 토양효소활성, 인지질지방산 함량을 분석하였다. 돈분과 음식물퇴비는 토양종류와 온도에 상관없이 토양 pH의 변화가 크지 않았으나 입상유기질비료는 온도가 높을수록 낮아졌으며, 비화산회토양에서 변화폭이 컸다. 미생물체량 C는 화산회토양의 경우 배양 온도가 높고 유기질비료를 처리할 때 높아지는 경향이었지만 토양종류에 상관없이 점차 감소하였다. 미생물체량 N은 비화산회토양에서 유기질비료, 화산회토양에서는 돈분과 음식물퇴비를 처리할 때 높게 나타났다. 인지질 지방산함량은 항온배양 75일후 비화 산회토양이 화산회토양보다 높았고, 270일후 화산회토양에서 높게 나타났으나 점차 낮아지는 경향이었다. Urease활성은 150일에 비화산회토양의 입상유기질비료처리에서 $10^{\circ}C$(75.0)>$20^{\circ}C$(16.3)>$30^{\circ}C$($4.6ug\;NH{_4-}N\;g^{-1}\;2h^{-1}$)순으로 $10^{\circ}C$에서 가장 높게 나타났으며 온도가 높고, 시간이 경과할수록 낮아졌으며 화산회토양의 Urease활성은 $10^{\circ}C$에서 높게 나타났다. ${\beta}-glucosidase$ 활성은 비화산회토양이 화산회토양보다 높았고 시간이 경과할수록 낮아졌다. 토양미생물체량 C와 미생물활성지표간의 상관계수는 비화산회토양에서 PLFA ($r^2=0.91$), 화산회토양에서 ${\beta}-glucosidase$ ($r^2=0.83$)가 높았으며, 미생물활성지표는 토양종류와 온도에 따라 상대적으로 민감도가 다르게 나타났다.

• Journal of Microbiology and Biotechnology
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• 제22권4호
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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.

• 한국미생물·생명공학회지
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• 제9권3호
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• pp.129-137
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• 1981

인삼엽 및 인삼피를 식물조직부양효소 (macerating enzyme)로 분리시킴으로써 인삼의 유효성분인 saponin을 효율적으로 추출하고자 인삼부패균중 조직부양력이 강력한 1종의 G-211 균주를 선정하여 조제한 조효소로써 실험하였다. 조직부양작용의 최적 pH 는 인삼엽, 인삼피 공히 pH 5.0이었으며 가용성물질추출율 및 saponin추출율은 인삼엽이 pH4.5, 인삼피가 pH5.5에서 가장 효과가 컸다. 조직부양효소와 조 cellulase을 qudydd처리하므로써 인삼엽, 인삼피 각각 3.45%, 3.90%의 saponin을 추출할 수 있었는데 이 추출율은 전체 saponin 함량의 39.8%, 39.3%에 해당되는 수율이었다. 또한 추출된 saponin을 HPLC를 이용해서 각 ginsenoside별 pattern도 조사하였다.

• 미생물학회지
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• 제47권4호
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• pp.302-307
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• 2011

A. nidulans chitin deacetylase를 자가분해 용액으로부터 소수성 상호작용 컬럼 크로마토그래피와 이온 교환 컬럼 크로마토그래피를 통해 순수 분리하였다. 효소 활성에 관여하는 아미노산을 분석하기 위해 효소 단백질과 특정 아미노산 잔기에 작용하는 화학 수식제를 반응시켜 효소를 화학 수식하였다. histidine 잔기가 화학 수식된 효소는 효소활성을 100% 상실하였으며, arginine의 잔기 혹은 tyrosine 잔기는 100 ${\mu}M$보다 높은 농도의 수식제로 화학수식 되었을 때 효소활성이 감소하였다. Aspartic acid 혹은 glutamic acid의 carboxyl group 잔기의 화학수식은 효소활성의 상대적으로 작은감소를 나타냈다. 이것은 산성 아미노산의 잔기가 화학 촉매 반응에 직접 관여하지 않았거나 혹은 산성 아미노산 잔기는 효소단백질의 전반적인 구조에 영향을 미친다는 것을 추론할 수 있다. 이러한 결과는 효소 단백질의 촉매활성에 histidine, tyrosine 및 arginine 잔기가 중요한 역할을 담당하는 것을 의미한다.

• Archives of Pharmacal Research
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• 제20권6호
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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.