• Journal of Microbiology and Biotechnology
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• 제1권2호
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• pp.90-95
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• 1991

A ${\alpha}-galactosidase{\;}({\alpha}-D-galactoside$ galactohydrolase; EC 3.2.1.22) was purified from the culture filtrate of Penicillium purpurogenum by DEAE-cellulose column chromatography, gel filtration of Bio gel p-l00, and subsequent SP-Sephadex C-25 chromatography. The final preparation thus obtained showed a single band on polyacrylamide disc-gel and SDS-polyacrylamide gel electrophoresis. The molecular weight and isoelectric point were determined to be 63,000 and pH 4.0 by SDS-polyacrylamide gel electrophoresis and isoelectric focusing, respectively. The galactosidase exhibited maximum activity at pH 4.5 and $55^{\circ}C$, and was stable between pH 2 and 5, and also stable up to $40^{\circ}C$. The enzyme activity was not affected considerably by treatment with other metal compounds except mercuric chloride and silver nitrate. Copra galactomannan was finally hydrolyzed to galactose, mannose and mannobiose through the sequential actions of the purified galactosidase and mannanase from the same strain. The enzyme hydrolyzed melibiose and raffinose, but not lactose.

• Asian-Australasian Journal of Animal Sciences
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• 제23권10호
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• pp.1340-1347
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• 2010

To study the effects of ${\alpha}$-galactosidase (${\alpha}$-Gal) supplementation on performance and energy metabolism, 216 Arbor Acres male broilers were placed in 36 cages of 6 birds each and allotted to 4 diets for 42 d, with 0-21 d as starter period and 22-42 d as grower period. The 4 diets were based on corn non-dehulled soybean meal in a $2{\times}2$ factorial arrangement, with 2 levels of ${\alpha}$-Gal (0 vs. 60 U/kg feed) and 2 levels of ME (normal metabolizable energy (NME) and low metabolizable energy (LME)). Bird performance was obtained at 21 and 42 d of age with samples of feces collected for nutrient digestibility from 19-21 d and 40-42 d. At 21 and 42 d, 1 bird from 6 cages of each treatment was killed to determine liver weight, intestinal pH and chyme viscosity. With the addition of ${\alpha}$-Gal the 42 d body weight (BW) and 0-42 d average daily gain (ADG) were significantly improved (p<0.05). Average daily feed intake (ADFI) of birds fed the LME diet was significantly increased compared to those fed the NME diet during starter (p<0.01) and grower (p<0.05) periods and overall (p<0.01). There was an interaction of ${\alpha}-Gal{\times}ME$ on 0-21 d ADFI (p<0.01). Supplementation of ${\alpha}$-Gal significantly improved (p<0.01) feed efficiency during the grower period and overall. Feed efficiency of birds fed the LME diet was significantly decreased (p<0.05) compared to those fed the NME diet during the starter period and overall. With the addition of ${\alpha}$-Gal apparent metabolizable energy (AME) was improved (p<0.01) by 2.1% and 1.8% during starter and grower periods, respectively. There was a main effect (p<0.05) of ${\alpha}$-Gal on the digestion of neutral detergent fiber (NDF) during the starter period and crude protein (CP), NDF and acid detergent fiber (ADF) during the grower period. With the addition of ${\alpha}$-Gal, the relative weight of liver was reduced (p<0.01) during the two phases. The duodenal and jejunal pH were significantly decreased (p<0.01) with the supplementation of ${\alpha}$at the two phases. ${\alpha}$-Gal addition reduced (p<0.01) chyme viscosity of the ileum during the starter and grower periods. Overall, ${\alpha}$-Gal showed a major effect on nutrient efficiency, improved ADG and feed efficiency, whereas LME decreased feed efficiency. The incorporation of ${\alpha}$-Gal into a LME diet could at least partially offset ME deficiency of non-dehulled soybean meal.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.852-860
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• 2012

An Antarctic bacterial isolate displaying extracellular ${\alpha}$-galactosidic activity was named Paenibacillus sp. LX-20 based on 16S rRNA gene sequence analysis. Optimal activity for the LX-20 ${\alpha}$-galactosidase occurred at pH 6.0-6.5 and $45^{\circ}C$. The enzyme immobilized on the smart polymer Eudragit L-100 retained 70% of its original activity after incubation for 30 min at $50^{\circ}C$, while the free enzyme retained 58% of activity. The enzyme had relatively high specificity for ${\alpha}$-D-galactosides such as p-nitrophenyl-${\alpha}$-galactopyranoside, melibiose, raffinose and stachyose, and was resistant to some proteases such as trypsin, pancreatin and pronase. Enzyme activity was almost completely inhibited by $Ag^+$, $Hg^{2+}$, $Cu^{2+}$, and sodium dodecyl sulfate, but activity was not affected by ${\beta}$-mercaptoethanol or EDTA. LX-20 ${\alpha}$-galactosidase may be potentially useful as an additive for soybean processing in the feed industry.

• 생명과학회지
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• 제28권11호
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• pp.1347-1353
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• 2018

본 연구는 한국전통약용식물(구기자, 오미자 잎)의 발효물로부터 분리한 Bacillus coagulanse NRR1207의 발효 특성을 파악하고 Bacillus coagulans NRR1207의 ${\alpha}$-galactosidase의 활성과 이를 통한 대두박의 비소화성분의 분해를 확인하였다. Bacillus coagulans NRR1207이 생산하는 효소 중 ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase가 가장 높은 40 nmol 이상의 활성을 나타내었다. Bacillus coagulans NRR1207의 발효 특성은 10% skim milk에서 배양했을 때, pH는 급속히 감소했고 적정 산도는 1.9%까지 증가했고 생균수도 발효 24시간에 8.8 log CFU/ml로 증가했다. 유당은 배양 72시간째 완전히 고갈되었고 유산 생산 능력도 탁월했다. Bacillus coagulans NRR1207을 대두박에 접종 후 배양시간에 따른 생균수의 변화는 배양 시작 시 7.6 log CFU/ml, 배양 16시간에 최고에 도달하여 9.0 log CFU/ml이었고 배양 72시간에 8.3 log CFU/ml로 Bacillus coagulans NRR1207이 왕성하게 잘 성장하였다. 대두박의 비소화성분 분해는 Bacillus coagulans NRR1207 접종 후 발효 24, 48, 72시간이 경과하면서 이 균이 생산한 ${\alpha}$-galactosidase에 의해 비소화성분인 stachyose와 raffinose가 대부분 분해되고 galactose가 생성되었다. 따라서 Bacillus coagulans NRR1207은 대두박의 비소화성분을 분해하는 생균제(Probiotics)로써 이용하여 식품 및 가축 사료 이용성 증대에 활용이 가능할 것으로 사료된다.

• Applied Biological Chemistry
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• 제41권7호
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• pp.489-495
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• 1998

한국 재래 간장으로부터 분리한 Scopulariopsis brevicaulis가 생성하는 ${\alpha}-galactosidase$의 최적 생산조건은 tryptone 1.5%, $NH_4NO_3$ 0.2%, raffinose 2.5%, $KH_2PO_4$ 0.5%, yeast extract 0.5%, pH 7.0, $27^{\circ}C$에서 3일간 배양 했을 때였다. ${\alpha}-galactosidase$의 최적 pH는 7.0, 최적온도는 $27^{\circ}C$였으며, $pH\;6.0{\sim}8.0$범위와 $40^{\circ}C$이하에서 안정하였다. $Ag^{2+},\;Hg^{2+},\;Cu^{2+}$ 등의 금속이온 및 p-chloromercuribenzoic acid, Iodine에 의해 강하게 저해되어 본 효소의 catalytic부위에 -SH기가 있음이 추정되었다. Km값은 1.9 mM, Vmax값은 $9.66{\times}10^2\;{\mu}M/min$이었다. 배양기간에 따른 raffinose의 분해양상을 high performance liquid chromatography로 살펴본 결과 배양초기에 raffinose가 분해되어 sucrose, glucose, fructose가 관찰되었다. 배양기간이 경과함에 따라 raffinose가 감소하였고 sucrose, glucose, fructose도 소실됨을 확인하였다.

• Biomolecules & Therapeutics
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• 제6권4호
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• pp.364-370
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• 1998

Glycosidase inhibitors from urine of Bombyx mori were isolated and their inhibitory activities on glycosidases were evaluated. Six compounds were isolated by using several ion exchange columns, and their chemical structures were identified by the physicochemical and spectral data. Compound IV, V and Ⅵ were identified as 1-deoxynojirimycin, fagomine and 1,4-dideoxy-1,4-imino-D-arabinitol, respectively. Among six compounds isolated,1-deoxynojirimycin(IV) was the most potent inhibitor on $\alpha$-glucosidase and $\beta$-galactosidase of rat intestine, and its inhibitory activities for trehalase and almond $\beta$-glucosidase were relatively weak. Compound V and Ⅵl retained a little inhibitory potency toward $\alpha$-glucosidase and $\beta$-galactosidase. Compound II and III, however, have been found to have no effect on all glycosidases tested in this study.

• Journal of Microbiology and Biotechnology
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• 제22권11호
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• pp.1532-1539
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• 2012

The ${\alpha}$-galactosidase-coding gene agaAJB13 was cloned from Sphingomonas sp. JB13 showing 16S rDNA (1,343 bp) identities of ${\leq}97.2%$ with other identified Sphingomonas strains. agaAJB13 (2,217 bp; 64.9% GC content) encodes a 738-residue polypeptide (AgaAJB13) with a calculated mass of 82.3 kDa. AgaAJB13 showed the highest identity of 61.4% with the putative glycosyl hydrolase family 36 ${\alpha}$-galactosidase from Granulicella mallensis MP5ACTX8 (EFI56085). AgaAJB13 also showed <37% identities with reported protease-resistant or Sphingomonas ${\alpha}$-galactosidases. A sequence analysis revealed different catalytic motifs between reported Sphingomonas ${\alpha}$-galactosidases (KXD and RXXXD) and AgaAJB13 (KWD and SDXXDXXXR). Recombinant AgaAJB13 (rAgaAJB13) was expressed in Escherichia coli BL21 (DE3). The purified rAgaAJB13 was characterized using p-nitrophenyl-${\alpha}$-D-galactopyranoside as the substrate and showed an apparent optimum at pH 5.0 and $60^{\circ}C$ and strong resistance to trypsin and proteinase K digestion. Compared with reported proteaseresistant ${\alpha}$-galactosidases showing thermolability at $50^{\circ}C$ or $60^{\circ}C$ and specific activities of <71 U/mg with or without protease treatments, rAgaAJB13 exhibited a better thermal stability (half-life of >60 min at $60^{\circ}C$) and higher specific activities (225.0-256.5 U/mg). These sequence and enzymatic properties suggest AgaAJB13 is the first identified and characterized Sphingomonas ${\alpha}$-galactosidase, and shows novel protease resistance with a potential value for basic research and industrial applications.

• Journal of Animal Science and Technology
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• 제45권2호
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• pp.211-218
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• 2003

본 연구는 양돈사료내 대두박 항영양인자인 $\alpha$-galactosides와 galatomannan의 분해를 유도하는 $\alpha$-1,6-galactosidase와 $\beta$-1,4-mannanase의 사료내 첨가가 자돈 및 육성돈의 성장과 영양소 소화율에 미치는 영향을 조사하기 위하여 실시하였다. 시험 1은 개시시 체중 10.57$\pm$0.30kg의 3원 교잡종 자돈 60두를 공시하였으며, 시험설계는 옥수수－건조유청－대두박 위주의 사료에 NRC (1998)의 영양소 요구량에 따라 처리한 대조구 (CON), 대조구 사료내 복합효소제를 0.1% 첨가한 처리구로 하였다. 사양시험기간동안, 일당증체량에 있어서는 대조구와 비교하여 EC0.1 처리구가 높은 것으로 평가되었으나 유의적인 차이는 보이지 않았다. 그러나 사료효율에 있어서는 대조구와 비교하여 EC0.1 처리구가 유의적으로 높게 평가되었다 (P<0.05). 건물과 질소 소화율에 있어서 대조구와 비교하여 처리구가 향상된 것으로 조사되었다 (P<0.05). 시험 2는 개시시 체중 22.30$\pm$0.45kg의 3원 교잡종 육성돈 36두를 공시하였으며, 시험설계는 옥수수－대두박 위주의 사료에 NRC (1998)의 영양소 요구량에 따라 처리한 대조구 (AME, adequate ME diet), 대조구 사료내 복합효소제를 0.1% 첨가한 처리구 (AME+EC0.1, Adequate ME diet+ 0.1% 복합효소제), 대조구 사료에서 대사에너지 함량을 4% 낮춘 사료에 복합효소제를 0.1% 첨가한 처리구 (LME+EC0.1, Low ME diet + 0.1% 복합효소제)로 하였다. 총 30일간의 사양시험 기간동안, 일당증체량에 있어서는 AME 처리구와 비교하여 복합효소제 처리구가 유의적인 성장율이 높은 것으로 조사되었다 (P<0.05). 건물 및 질소 소화율에 있어서는 AME 처리구와 비교하여 복합효소제 첨가구가 유의적으로 높게 평가되었다 (P<0.05). 결론적으로, 자돈 및 육성돈 사료에 복합효소제의 첨가는 성장능력 및 영양소 소화율을 향상시키는 것으로 사료된다.

• 대한유전성대사질환학회지
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• 제14권2호
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• pp.135-141
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• 2014

목적: 파브리병(Fabry disease)은 alpha-galactosidase A의 결핍으로 인하여 리소좀에 globotriaosylceramide(Gb3)가 축적되어 여러 장기에 이상을 일으키는 질병이다. 본 연구에서는 파브리병의 만성 합병증 중 심장 질환을 주로 보이는 환자들, 그 중에서도 좌심실 비대증을 보이는 한국인 환자들을 대상으로 파브리병의 빈도를 알아보고자 하였다. 방법: 좌심실비대증을 진단받은 환자 257명을 연구대상으로 선정하였고, 남성이 172명(평균 56세, 범위 30-81세), 여성이 84명(평균 66세, 범위 45-85세)이었다. 파브리병 선별을 위하여 고성능액체크로마토그래피-탠덤질량분석기를 이용하여 소변 Gb3 농도를 측정하였다. 확진은 형광분석법에 의한 말초혈액의 alpha-galactosidase A 활성도와 염기서열분석법에 의한 GLA 유전자 돌연변이 유무를 검사하여 이루어졌다. 결과: 소변 Gb3 검사에서 cutoff (25 ug/mmoL creatinine)를 초과하는 환자는 4명이었지만, 최종적으로 추가 검사를 통해 진단된 파브리병 환자는 여성 환자 한 명이었다(1/257명, 0.4%). 확진된 환자는 54.3 ug/mmoL creatinine의 Gb3 농도와 15.5 nmole/hr/mg protein (참고범위, $55.2{\pm}12.7nmole/hr/mg$ protein)의 alpha-galactosidase A 활성도를 보였다. GLA 유전자에서는 c.796G>A (p.D266N) 돌연변이가 이형접합체로 관찰되었다. 추가로 시행한 가족검사에서 환자의 딸은 아직 파브리병의 증상을 보이지 않았지만, 엄마와 같은 GLA 돌연변이(c.796G>A)를 가지고 있었으며, alpha-galactosidaseA 활성도는 42.5 nmole/hr/mg protein, 소변 Gb3 농도는 25.5 ug/mmoL creatinine을 나타냈다. 결론: 한국인 좌심실 비대증을 가진 환자들에서 파브리병의 유병율은 0.4%였다. 유병율이 낮아 보임에도 불구하고, 파브리병 진단 전 환자와 가족 구성원을 발견할 수 있는 장점 덕분에 선별검사의 의의가 있는 것으로 사료된다.

• 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.