• 생명과학회지
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• 제24권1호
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• pp.54-60
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• 2014

방향족 화합물은 독성 환경오염물질로 생태계와 인간의 건강에 해로운 영향을 미친다. 그중 chlorotoluene과 nitrotoluene 화합물은 수생생물에 독성을 나타내며 인간의 피부, 눈, 호흡기를 자극한다. 본 연구에서는 폐수의 chlorotoluene과 nitrotoluene 화합물을 저렴하고 간단하게 검출하고자 재조합 미생물 바이오센서를 개발하였다. BTEX (benzene, toluene, ethylbenzene, xylene) 분해 조절 유전자 xylR를 Po' (upstream activating sequences를 제거한 DmpR 조절단백질 promoter Po) 또는 Pu (XylR 고유의 프로모터)::lacZ 유전자(${\beta}$-galactosidase 유전자)의 upstream에 연결한 플라스미드를 제작한 후, E. coli $DH5{\alpha}$에 형질 전환하였다. 유도 화합물 존재 하에서, 아가로스에 고정된 이 재조합 바이오센서 세포는 유도 화합물에 의해 ${\beta}$-galactosidase를 발현하고 기질인 chlorophenol red ${\beta}$-D-galactopyranoside (CPRG)를 분해하여 1~2시간에 붉은색을 나타내었다. BTEX 화합물 중, 특이적으로 o-, m-, p-chlorotoluene ($0.1{\mu}M-100 mM$) 그리고 o-, m-, p-nitrotoluene (0.1 mM-100 mM)에서 높은 반응을 나타내었으며 Po'가 Pu보다 높은 반응성을 보여주었다. 아가로스에 고정된 바이오센서는 $4^{\circ}C$에서 21일간 보존 후에도 활성의 큰 변화 없이 안정하였으며, chlorotoluene과 nitrotoluene 화합물들로 spike된 전처리 하지 않은 폐수 시료 중에서도 좋은 반응을 보여 주어 폐수 중 chlorotoluene과 nitrotoluene 화합물의 간단한 초기 검출에 활용될 수 있음을 제시하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권12호
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• pp.1761-1768
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• 2005

Effects of alpha-galactosidase (GAL) on broiler corn-soybean meal diet was investigated. In experiment 1, sixty cockerels were allocated to five groups, including three enzyme treatments (GAL added at 0, 500, and 1,000 mg/kg diet), a nitrogen-free diet group and a fast group. The true nitrogen-corrected ME (TME$_n$) and true amino acid availability were determined. In experiment 2, 324 day-old chicks were used in a 2${\times}$3 factorial design consisting of two energy contents (high and low) and three GAL levels (0, 250, and 500 mg/kg). Three feeding phases, comprising 0-21 d, 22-35 d and 36-48 d, were involved. GAL addition improved TME$_n$ and the availability of methionine and cystine (p<0.05). The apparent ME (AME) or nitrogen-corrected AME (AME$_n$) and digestibility of dry matter, organic matter, calcium, and phosphorus were improved significantly on d 21, so was crude protein and an interaction of energy and GAL on AME$_n$ (p<0.05) was found on d 35. However, daily intake and daily gain were significantly improved with GAL addition (p<0.05) during 21 d. The small intestine relative weight decreased at 250 mg/kg GAL (p<0.05) on d 35, whereas presented an interaction between GAL and energy on d 21 (p<0.05). Likewise, this treatment increased breast muscle ratio (p<0.05). On d 21, triglycerides level of broilers showed interaction between energy and enzyme levels (p<0.05). Uric acid level in 500 mg/kg GAL declined linearly (p<0.05). On d 35, quadratic effects (p<0.05) were observed in total protein, albumin, globulin and cholesterol content for enzyme supplementation. And the interactive effects of energy and GAL on serum values showed more obviously. The study implies that GAL improved energy and nutrient availability of corn-soybean meal diet in broiler. The GAL supplementation to corn-soybean meal based diet can improve performance of broilers in early stages of growth.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1997년도 한국생물과학협회 학술발표대회
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• pp.261.1-261
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• 1997

No Abstract, See Full Text

• Food Science and Biotechnology
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• 제16권1호
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• pp.127-132
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• 2007

[ ${\alpha},\;{\alpha}$ ]-Trehalose was efficiently modified by a transgalactosylation reaction of Escherichia coli ${\beta}-galactosidase$ using lactose as a donor to yield two galactosyl trehalose trisaccharides. The reaction products of trehalose by the enzyme were observed by thin layer chromatography (TLC) and high performance anion exchange chromatography (HPAEC) and were purified by BioGel P2 gel permeation chromatography and recycling preparative HPLC. Liquid chromatography-mass spectrometry (LC-MS) and ^{13}C$ nuclear magnetic resonance (NMR) analyses revealed that the structures of the main products were $6^2-{\beta}-D-galactosyl$ trehalose (1) and $4^2-{\beta}-D-galactosyl$ trehalose (2). A reaction of 30%(w/v) trehalose and 15%(w/v) lactose at pH 7.5 and $45^{\circ}C$ resulted in a total yield of approximately 27-30% based on the amount of trehalose used. The galactosyl trehalose products were not hydrolyzed by trehalose. In addition the mixture of transfer products (9:1 ratio of 1 to 2) showed higher thermal stability than glucose, lactose, and maltose, but less than trehalose, against heat treatment over $100^{\circ}C$ at pH 4 and 7. It also exhibited better thermal stability than sucrose at pH 4 alone.

• Journal of Microbiology and Biotechnology
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• 제22권12호
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• pp.1621-1628
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• 2012

The agar-degrading bacterium GNUM-1 was isolated from the brown algal species Sargassum serratifolium, which was obtained from the West Sea of Korea, by using the selective artificial seawater agar plate. The cells were Gram-negative, $0.5-0.6{\mu}m$ wide and $2.0-2.5{\mu}m$ long curved rods with a single polar flagellum, forming nonpigmented, circular, smooth colonies. Cells grew at $20^{\circ}C-37^{\circ}C$, between pH 5.0 and 9.0, and at 1-10% (w/v) NaCl. The DNA G+C content of the GNUM-1 strain was 45.5 mol%. The 16S rRNA sequence of the GNUM-1 was very similar to those of Alteromonas stellipolaris LMG 21861 (99.86% sequence homology) and Alteromonas addita $R10SW13^T$(99.64% sequence homology), which led us to assign it to the genus Alteromonas. It showed positive activities for agarase, amylase, gelatinase, alkaline phosphatase, esterase (C8), lipase (C14), leucine arylamidase, valine arylamidase, ${\alpha}$-chymotrypsin, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\beta}$-glucosidase, catalase, and urease. It can utilize citrate, malic acid, and trisodium citrate. The major fatty acids were summed feature 3 (21.5%, comprising $C_{16:1}{\omega}7c/iso-C_{15:0}$ 2-OH) and C16:0 (15.04%). On the basis of the variations in many biochemical characteristics, GNUM-1 was considered as unique and thus was named Alteromonas sp. GNUM-1. It produced the highest agarase activity in modified ASW medium containing 0.4% sucrose, but lower activity in rich media despite superior growth, implying that agarase production is tightly regulated and repressed in a rich nutrient condition. The 30 kDa protein with agarase activity was identified by zymography, and this report serves as the very first account of such a protein in the genus Alteromonas.

• 한국식품조리과학회지
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• 제14권5호
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• pp.589-596
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• 1998

본 연구는 대두단백질의 풍미와 기능성의 향상을 목적으로 효소처리한 분리대두단백에 fructooligosaccaride, galactooligosaccaride , isomaltooligosaccaride를 첨가하여 3종류의 비피더스균으로 각기 배양시킨 soy yogurts의 pH, 산도, 생균수, $\alpha$-galactosidase의 활성 , 유기산과 n-hexanal 및 acetaldehyde의 함량을 연구하였다. pH에 있어서 올리고당의 종류에 따라서는 유의차가 없었으나 균주의 종류에 따라서는 유의차가 있었다. 따라서 B. bifidum 배양군의 pH가 5.26~5.30로 가장 높았고 B. breve 배양군은 4.76~4.86이었으며, B. infantis 배양군은 4.38~4.40로서 가장 낮았다. 적정산도는 pH와 유사한 경향을 나타내어 올리고당의 종류에 따라서는 유의차가 없었으나 균주의 종류에 따라서는 유의차가 있었다. 따라서 B. bifidum 배양군의 산도가 0.81~0.84%로 가장 낮았고 B. breve 배양군은 0.97-1.04% 였으며, B. infantis 배양군은 1.39~1.41%로 가장 높았다. 생균수는 올리고당의 종류에 따라서는 대체로 근사한 수치를 나타내었으나 균주의 종류에 따라서는 경시적인 차이가 있었으며 모든 시료들이 $10^{9}$ CFU/ml 이상의 생균수를 포함하는 것으로 나타났다. 한편 B. infantis로 배양한 soy yogurts를 4$^{\circ}C$에서 7일간 저장한 후 생균수를 측정한 결과 $10^{9}$ CFU/ml를 나타내어 초기생균수를 거의 유지하였다. $\alpha$-Galactosidase 활성은 B. infantis, B. bifidum, B. breve 순으로 높았으며 활성이 가장 높았던 B. infantis의 활성을 100%로 하였을 경우 B. bifidum B. breve의 활성은 각각 32.7, 26.0% 이었다. 유기산 중에서 lactic acid와 acetic acid은 적정산도의 결과와 유사한 경향을 나타내어 B. infantis 배양군, B. breve 배양군, B. bifidum 배양군 순으로 높았으며 propionic acid는 반대경향을 나타내었고 올리고당 종류에 따라서는 모든 시료에서 유의차가 없었다. 휘발성 향기성분 중에서 콩비린내를 발생시키는 성분인 n-hexanal은 B. breve배양군이 B. bifidum과 B. infantis 배양군 보다 농도가 높았으며 acetaldehyde는 0.02~0.52 ppm으로 극미량이 검출되었고 올리고당 종류에 따라서는 모든 시료에서 유의차가 없었다.

• 한국식품과학회지
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• 제20권3호
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• pp.426-432
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• 1988

${\alpha}-galactosidase$ 및 protease활 을 가지는 효소제 제를 두유(豆乳) 제조시에 처리하여 장내(腸內) 가스발생인자(flatulence factor)를 제거 하는 동시에 두유의 수율(收率)과 소화율(消化率)을 향상시키고자 실험한 결과는 다음과 같다. 두유의 생산량 및 단백질 수율은 효소제(酵素劑) 처리에 의하여 증가하지 않았다. 두유 중 고형분의 양은 pH 6에서 pH10으로 높아질수록, 효소제 처리온도가 $30^{\circ}C$에서 $60^{\circ}C$로 높아질수록 증가하였다. 두유중 TCA가용성 질소화합물의 비율은 효소제의 농도가 높을 수록, pH 4에서 pH10으로 증가 할수록, 처리시간이 길어질수록 증가하였고 처리온도 $50^{\circ}C$까지 증가하는 경향을 보였다. 장내 가스 발생인자로 알려진 raffinose와 stachyose의 함량도 효소제의 농도가 증가할수록, 처리 온도가 $30^{\circ}C$에서 $60^{\circ}C$로 증가함에 따라 많이 감소하였으며 최적 pH는 4-5사이였다.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1659-1669
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• 2020

1,3-α-3,6-anhydro-L-galactosidase (α-neoagarooligosaccharide hydrolase) catalyzes the last step of agar degradation by hydrolyzing neoagarobiose into monomers, D-galactose, and 3,6-anhydro-L-galactose, which is important for the bioindustrial application of algal biomass. Ahg943, from the agarolytic marine bacterium Gayadomonas joobiniege G7, is composed of 423 amino acids (47.96 kDa), including a 22-amino acid signal peptide. It was found to have 67% identity with the α-neoagarooligosaccharide hydrolase ZgAhgA, from Zobellia galactanivorans, but low identity (< 40%) with the other α-neoagarooligosaccharide hydrolases reported. The recombinant Ahg943 (rAhg943, 47.89 kDa), purified from Escherichia coli, was estimated to be a monomer upon gel filtration chromatography, making it quite distinct from other α-neoagarooligosaccharide hydrolases. The rAhg943 hydrolyzed neoagarobiose, neoagarotetraose, and neoagarohexaose into D-galactose, neoagarotriose, and neoagaropentaose, respectively, with a common product, 3,6-anhydro-L-galactose, indicating that it is an exo-acting α-neoagarooligosaccharide hydrolase that releases 3,6-anhydro-L-galactose by hydrolyzing α-1,3 glycosidic bonds from the nonreducing ends of neoagarooligosaccharides. The optimum pH and temperature of Ahg943 activity were 6.0 and 20℃, respectively. In particular, rAhg943 could maintain enzyme activity at 10℃ (71% of the maximum). Complete inhibition of rAhg943 activity by 0.5 mM EDTA was restored and even, remarkably, enhanced by Ca²⁺ ions. rAhg943 activity was at maximum at 0.5 M NaCl and maintained above 73% of the maximum at 3M NaCl. K_m and V_max of rAhg943 toward neoagarobiose were 9.7 mg/ml and 250 μM/min (3 U/mg), respectively. Therefore, Ahg943 is a unique α-neoagarooligosaccharide hydrolase that has cold- and high-salt-adapted features, and possibly exists as a monomer.

• Fisheries and Aquatic Sciences
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• 제7권2호
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• pp.70-75
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• 2004

The CMV promoter driven lacZ reporter gene (pcDNA-lacZ) was constructed and used for DNA immunization study. The expression of the lacZ gene was confirmed in vitro using RTG-2 cell line before using for in vivo study in olive flounder (Paralichthys olivaceus). In the dose response study, the maximum expression of the lacZ gene was found in the group injected with 5 ${\mu} g$ of the plasmid DNA. Kinetic study showed a significantly increased expression of $\beta-galactosidase$ gene at 7 days after injection. Effects of DNA vaccine on specific and nonspecific immune responses such as antibody and NO production were studied and the significant effect was found in olive flounder injected with 10 and 15 ${\mu} g$ DNA (sub optimal dose for lacZ gene expression). Two pro inflammatory cytokine genes, $IL-l\beta$ and $TNF-\alpha$, were also found to be up regulated in the muscle injected with the plasmid, suggesting an induction of local inflammatory response.

• 약학회지
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• 제55권1호
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• pp.56-63
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• 2011

Fabry disease (FD) is an X-linked inborn error of glycoshpingolipid metabolism resulting from mutation in the enzyme ${\alpha}$-galactosidase A gene. The disease is an X-linked lipid storage disorder and the lack of ${\alpha}$-Gal A causes an intracellular accumulation of glycosphingolipids, mainly globotriaosylceramide (Gb-3). Measurement of Gb-3 in plasma has clinical importance for monitoring after enzyme replacement therapy for confirmed FD patients. Using electrospray ionization MS/MS we had developed, a simple, rapid, and highly sensitive analytical method for Gb-3 in plasma was used for the purpose of screening FD among high risk groups in Korean population. To date, no comprehensive results for FD screening have been performed and reported in Korea. We screened 1,100 outpatients from 13 hospitals (including clinics) to assess the incidence of FD among patients in high risk groups. For patients with borderline level amount of Gb-3, we repeated Gb-3 or performing complementary or confirmative assay with ${\alpha}$-Gal A activity and DNA mutaion analysis for confirmation diagnosis. Of 1,100 we diagnosed 3 FD with 2 classical type and 1 carrier (0.27%).