• Applied Biological Chemistry
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• 제34권4호
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• pp.299-306
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• 1991

$[U-^{14}C]\;TCAB$를 $MM_2$ 무기배지(無機培地)에 유일(唯一)한 탄소원(炭素源)으로 첨가(添加) 후(後) 분리(分離)한 균주(菌株)들을 순수배양(純粹培養)하였을 때 약간(若干)의 방사성(放射性) 분해산물(分解産物)이 autoradiography에 의(依)하여 검출(檢出)되었다. 또한 유기물(有機物)을 제거한 토양(土壤)에 $^{14}C-TCAB$를 첨가(添加)한 후(後) 각각의 분리균주(分離菌株)들을 접종(接種)하고 $MM_2$ 무기배지(無機培地)를 가(加)하여 일정(一定)한 습도(濕度)를 유지(維持)하면서 $30^{\circ}C$에서 배양(培養)하였을 때 $^{14}CO_2$가 발생(發生)되지 않았다. 이들 분리균주(分離菌株)의 하나인 Achromobacter group VD를 순수배양(純粹培養) 시(時) m/z 250인 분해산물(分解産物)이 GC/MS에 의(依)하여 확인(確認)되었다. 이 분해산물(分解産物)의 가능(可能)한 형성경로(形成經路)는 TCAB의 구조(構造)로부터 dechlorination, hydrorylation, 2개(個) benzene환(環)의 ortho 관열(關裂), 그리고 생성(生成)된 carboxyl group의 환원(還元) 등이 관련(關聯)된다고 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제26권2호
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• pp.266-274
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• 2013

The effect of different phytogenic feed additives on reducing odorous compounds in swine was investigated using in vitro fermentation and analyzed their microbial communities. Soybean meal (1%) added with 0.1% different phytogenic feed additives (FA) were in vitro fermented using swine fecal slurries and anaerobically incubated for 12 and 24 h. The phytogenic FAs used were red ginseng barn powder (Panax ginseng C. A. Meyer, FA1), persimmon leaf powder (Diospyros virginiana L., FA2), ginkgo leaf powder (Ginkgo biloba L., FA3), and oregano lippia seed oil extract (Lippia graveolens Kunth, OL, FA4). Total gas production, pH, ammonianitrogen ($NH_3$-N), hydrogen sulfide ($H_2S$), nitrite-nitrogen ($NO_2{^-}$-N), nitrate-nitrogen ($NO_3{^-}$-N), sulfate (${SO_4}^{--}$), volatile fatty acids (VFA) and other metabolites concentration were determined. Microbial communities were also analyzed using 16S rRNA DGGE. Results showed that the pH values on all treatments increased as incubation time became longer except for FA4 where it decreased. Moreover, FA4 incubated for 12 and 24 h was not detected in $NH_3$-N and $H_2S$. Addition of FAs decreased (p<0.05) propionate production but increased (p<0.05) the total VFA production. Ten 16S rRNA DGGE bands were identified which ranged from 96 to 100% identity which were mostly isolated from the intestine. Similarity index showed three clearly different clusters: I (FA2 and FA3), II (Con and FA1), and III (FA4). Dominant bands which were identified closest to Eubacterium limosum (ATCC 8486T), Uncultured bacterium clone PF6641 and Streptococcus lutetiensis (CIP 106849T) were present only in the FA4 treatment group and were not found in other groups. FA4 had a different bacterial diversity compared to control and other treatments and thus explains having lowest odorous compounds. Addition of FA4 to an enriched protein feed source for growing swine may effectively reduce odorous compounds which are typically associated with swine production.

• Molecules and Cells
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• 제41권4호
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• pp.331-341
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• 2018

L-pipecolic acid is a non-protein amino acid commonly found in plants, animals, and microorganisms. It is a well-known precursor to numerous microbial secondary metabolites and pharmaceuticals, including anticancer agents, immunosuppressants, and several antibiotics. Lysine cyclodeaminase (LCD) catalyzes ${\beta}$-deamination of L-lysine into L-pipecolic acid using ${\beta}$-nicotinamide adenine dinucleotide as a cofactor. Expression of a human homolog of LCD, ${\mu}$-crystallin, is elevated in prostate cancer patients. To understand the structural features and catalytic mechanisms of LCD, we determined the crystal structures of Streptomyces pristinaespiralis LCD (SpLCD) in (i) a binary complex with $NAD^+$, (ii) a ternary complex with $NAD^+$ and L-pipecolic acid, (iii) a ternary complex with $NAD^+$ and L-proline, and (iv) a ternary complex with $NAD^+$ and L-2,4-diamino butyric acid. The overall structure of SpLCD was similar to that of ornithine cyclodeaminase from Pseudomonas putida. In addition, SpLCD recognized L-lysine, L-ornithine, and L-2,4-diamino butyric acid despite differences in the active site, including differences in hydrogen bonding by Asp236, which corresponds with Asp228 from Pseudomonas putida ornithine cyclodeaminase. The substrate binding pocket of SpLCD allowed substrates smaller than lysine to bind, thus enabling binding to ornithine and L-2,4-diamino butyric acid. Our structural and biochemical data facilitate a detailed understanding of substrate and product recognition, thus providing evidence for a reaction mechanism for SpLCD. The proposed mechanism is unusual in that $NAD^+$ is initially converted into NADH and then reverted back into $NAD^+$ at a late stage of the reaction.

• 한국지하수토양환경학회지:지하수토양환경
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• 제20권6호
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• pp.85-94
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• 2015

RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) is the most important explosive contaminant, both in concentration and in frequency, at military shooting ranges in which green technologies such as phytoremediation or constructed wetlands are the best option for mitigation of explosive compounds discharge to the environment. A study was conducted with two identical lab-scale plug flow constructed wetlands planted with Amur silver grass to treat water artificially contaminated with 40 mg/L of toxic explosive compound, RDX. The reactor was inoculated with or without RDX degrading mixed culture to evaluate plant-microorganism interactions in RDX removal, transformation products distribution, and kinetic constants. RDX and its metabolites in water, plant, and sediment were analyzed by HPLC to determine mass balance and kinetic constants. After 30 days of operation, the reactor reached steady-state at which more than 99% of RDX was removed with or without the mixed culture inoculation. The major transformation product was TNX (Trinitroso-RDX) that comprised approximately 50% in the mass balance of both reactors. It was also the major compound in the plant root and shoot system. Acute toxicity analysis of the water samples showed more than 30% of toxicity reduction in the effluent than that of influent containing 40 mg/L of RDX. In the Amur silver grass mesocosm seeded with the mixed culture, the specific RDX removal rate, that is 1st order removal rate normalized to plant fresh weight, was estimated to be 0.84 kg⁻¹ day⁻¹ which is 16.7% higher than that in the planted only mesocosm. Therefore, the results of this study proved that Amur silver grass is an effective plant for RDX removal in constructed wetlands and the efficiency can be increased even more when applied with RDX degrading microbial consortia.

• 한국미생물·생명공학회지
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• 제25권6호
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• pp.592-597
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• 1997

During the screening of inhibitors against phospholipase C (PLC) and the formation of inositol phosphates (IP$_{t}$) at NIH3T3${\gamma}$1 cells from microbial secondary metabolites, we selected a fungal strain MT60109 which was capable of producing an inhibitor. By the taxonomic studies, this fungus was identified as Pseudallescheria sp. MT60109 and an inhibitor of PLC was purified by BuOH extraction and chromatographic techniques from the culture broth of Pseudallescheria sp. MT60109. The inhibitor was identified as thielavin B by the physico-chemical properties and spectroscopic analysis of UV, FAB-MS, $^{1}$H, $^{13}$C-NMR, $^{1}$H-$^{1}$H COSY and HMBC. Thielavin B showed potent inhibitory activity against PLC purified from bovine brain with an IC$_{50}$ of 20 $\mu$M. And it also inhibited the formation of inositol phosphates in platelet-derived growth factor (PDGF) -stimulated NIH3T3${\gamma}$1 cells with an IC$_{50}$ of 20 $\mu$M.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.900-907
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• 2007

Three male Gayal (Bos frontalis) and three male Yunnan Yellow cattle (Bos taurus) were fed pelleted lucerne and measurements made of digestibility, nitrogen utilisation, rumen fermentation and microbial population and key plasma metabolites. Total actual dry matter intake was similar but when expressed in terms of live weight or metabolic live weight feed intakes were significantly higher (p<0.05) for Gayal than cattle. Apparent digestibilities of dry matter, organic matter, fibre and dietary nitrogen were similar for both Gayal and cattle. Rumen ammonia nitrogen and total volatile fatty acids were significantly higher (p<0.05) for Gayal than cattle and total numbers of viable rumen bacteria, cellulolytic and amylolytic bacteria, but not proteolytic bacteria nor protozoa, were significantly greater (p<0.05) for Gayal than cattle. Although Gayal have a different rumen ecology to cattle, similar digestive parameters were exhibited. Further research is required to establish relationship between rumen ecology and digestive parameters.

• KSBB Journal
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• 제14권6호
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• pp.682-687
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• 1999

Streptomyces virginiae 유래의 virginiamycin 생합성 유도인자인 VB-C를 이용하여 다른 방선균에의 항생물질 생산 유도 기능을 검토하였다. Erythromycin 생산균인 Streptomyces erythraeus를 공시균주로 사용하였으며, VB-C 결합 receptor 유전자가 포함된 plasmid(pARS 701, 9 kb)를 공시균주에 도입하여 transformant로 사용하였다. Parent와 transformant 모두 유도인자인 VB류를 생산하였으며, VB-C의 signal 전달에 관여하는 결합 단백질의 존재가 확인되었다. Parent는 본배양 0, 20, 44시간째에 합성 VB-C 300 ng/ml를 첨가시 초기 항생물질 생산시기가 약 8시간 이상 단축되었다. Transformant는 본배양 44시간에 첨가시 6시간 이상 항생물질 생산시기가 단축되었고 항생물질 생산량도 증가되었으며, parent에 비해 B. subtilis에도 강한 항균력을 나타내었다. 또한 항균 spectrum은 parent보다는 transformant가 생산한 항생물질의 항균호과가 더 큰 것으로 나타났으며, VB-C를 첨가한 경우 미첨가 경우보다 넓은 항균 spectrum을 보였다. 이들 결과에서 공시균인 S. erythraeus에서 VB signal 전달에 따른 항생물질 생산유도가 입증되었으며, VB-C receptor gene의 도입에 따른 발현과 함께 VBs의 유도, 항생물질의 생산시기의 단축 및 생산량의 증가가 예상됨에 따라 다양한 Streptomyces sp. 균주를 대상으로 항생물질의 대량생산을 비롯한 새로운 2차 대사산물 생산에의 응용이 예상되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권4호
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• pp.479-482
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• 2005

The nutritional worth of Sarson Saag Waste (SSW), a cannery waste, was assessed in comparison with conventional complete diet as a total mixed ration (TMR), and a conventional green fodder, Avena sativa. Each diet was offered ad libitum, supplemented with mineral mixture and common salt, to 4 male murrah buffaloes. The control TMR was made iso-nitrogenous to SSW. Simultaneously, each diet was offered to 3 rumen fistulated male buffaloes for assessing the biochemical changes in the rumen. The nutrient digestibility of unconventional SSW was comparable to that of conventional green fodder-A. sativa but significantly (p<0.05) higher than that of control TMR. The tri-chloro acetic acid (TCA) precipitable-N in the strained rumen liquor of animals fed SSW was considerably higher than that of animals fed A.sativa. The urinary excretion of total purine derivatives was comparable in animals fed SSW and conventional green fodder but significantly (p<0.05) higher than those fed conventional control TMR. The significantly (p<0.05) lower purine nitrogen index (PNI) in animals fed control TMR resulted in significantly (p<0.05) lower microbial protein synthesis than that in animals fed SSW and conventional green fodder. The N-excretion as per cent of nitrogen intake was significantly (p<0.05) lower in animals fed SSW as compared to either of the conventional feeds tested, resulting in significantly (p<0.05) higher Nretention and apparent biological value. SSW supplemented with mineral mixture could serve as an excellent source of nutrients for ruminants.

• Journal of Microbiology and Biotechnology
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• 제28권10호
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• pp.1581-1588
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• 2018

The growth of lactic acid bacteria (LAB) generates a high number of metabolites related to aromas and flavors in fermented dairy foods. These microbial proteases are involved in protein hydrolysis that produces necessary peptides for their growth and releases different molecules of interest, like bioactive peptides, during their activity. Each genus in particular has its own proteolytic system to hydrolyze the necessary proteins to meet its requirements. This review aims to highlight the differences between the proteolytic systems of Streptococcus thermophilus and other lactic acid bacteria (Lactococcus and Lactobacillus) since they are microorganisms that are frequently used in combination with other LAB in the elaboration of fermented dairy products. Based on genetic studies and in vitro and in vivo tests, the proteolytic system of Streptococcus thermophilus has been divided into three parts: 1) a serine proteinase linked to the cellular wall that is activated in the absence of glutamine and methionine; 2) the transport of peptides and oligopeptides, which are integrated in both the Dpp system and the Ami system, respectively; according to this, it is worth mentioning that the Ami system is able to transport peptides with up to 23 amino acids while the Opp system of Lactococcus or Lactobacillus transports chains with less than 13 amino acids; and finally, 3) peptide hydrolysis by intracellular peptidases, including a group of three exclusive of S. thermophilus capable of releasing either aromatic amino acids or peptides with aromatic amino acids.

• Applied Biological Chemistry
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• 제39권2호
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• pp.153-158
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• 1996

광독립 영양세포를 이용하여 미생물유래 제초활성 물질을 탐색하던 중 강한 제초활성을 갖는 균주를 선발하고, 그 균주가 생산하는 활성 물질들을 분리 정제한 후 구조를 결정하였으며, 이들의 활성을 조사하였다. 선발된 ME-13 균주의 미생물학적 특성을 조사한 결과 Streptomyces로 동정되어 본 균주를 Streptomyces sp. ME-13으로 명명하였다. 균배양 상등액으로부터 활성탄소흡착, sillca gel, MCI gel등의 column chromatography 및 HPLC를 통하여 2 가지 활성 물질을 순수 분리하였다. 이들 화합물은 기기분석결과, anisomycin계의 AB3217-A와 B로 각각 동정되었다. AB3217-A와 B는 25ppm 농도에서 무와 피의 종자 발아를 완전히 억제하였고, 종자 발아후 50% 생육저해농도는 약 6 ppm 정도로 서로 비슷하였으며 anisomycin보다 6배 정도 강한 제초활성을 보였다. 이와 같은 제초활성은 본 연구에 의하여 처음으로 밝혀졌다.