• Journal of Chest Surgery
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• 제45권6호
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• pp.368-379
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• 2012

Background: Bioprostheses for cardiovascular surgery have limitations in their use following as calicification. ${\alpha}$-galactosidase epitope is known as a stimulant of immune response and then shows a progressing calcification. The objective of this study was to evaluate histologic characteristics and mechanical properties of decellularization and treated with ${\alpha}$-galactosidase. Materials and Methods: Bovine pericardial tissues were allocated into three groups: fixation only with glutaraldehyde, decellularization with sodium dodesyl sulfate and decellularization plus treatment with ${\alpha}$-galactosidase. We confirmed immunohistological characteristics and mechanical properties as fatigue test, permeability test, compliance test, tensile strength (strain) test and thermal stability test. Results: Decellularization and elimination of ${\alpha}$-gal were confirmed through immunohistologic findings. Decellularization had decreased mechanical properties compared to fixation only group in permeability (before fatigue test p=0.02, after fatigue test p=0.034), compliance (after fatigue test p=0.041), and tensile strength test (p=0.00). The group of decellularization plus treatment with ${\alpha}$-galactosidase had less desirable mechanical properties than the group of decellularization in concerns of permeability (before fatigue test p=0.043) and strain test (p=0.001). Conclusion: Favorable decellularization and elimination of ${\alpha}$-gal were obtained in this study through immunohistologic findings. However, those treatment including decellularization and elimination of ${\alpha}$-gal implied the decreased mechanical properties in specific ways. We need more study to complete appropriate bioprosthesis with decellularization and elimination of ${\alpha}$-gal including favorable mechanical properties too.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1546-1554
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• 2010

A bacterial strain capable of producing extracellular ${\alpha}$-galactosidase was isolated from a sample of sugarcane industrial waste. Microbiological, physiological, and biochemical studies revealed that the isolate belonged to Bacillus sp. Furthermore, based on a 16S rDNA sequence analysis, the new isolate was identified as Bacillus megaterium VHM1. The production of ${\alpha}$-galactosidase was optimized based on various physical culture conditions. Guar gum and yeast extract acted as the best carbon and nitrogen sources, respectively. The optimum pH was 7.5 and the enzyme remained stable over a pH range of 5-9. The enzyme was optimally active at $55^{\circ}C$ and thermostable with a half-life of 120 min, yet lost 90% of its residual activity within 120 min at $60^{\circ}C$. One mM concentrations of $Ag^2$, $Cu^2$, and $Hg^{2+}$ strongly inhibited the ${\alpha}$-galactosidase, whereas the metal ions $Fe^2$, $Mn^{2+}$, and $Mg^{2+}$ had no effect on the ${\alpha}$-galactosidase activity, and $Zn^{2+}$, $Ni^{2+}$, and $Ca^{2+}$ reduced the enzyme activity slightly. When treated with the B. megaterium VHM1 enzyme, the flatulence-causing sugars in soymilk were completely hydrolyzed within 1.5 h.

• 한국축산식품학회지
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• 제27권1호
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• pp.102-109
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• 2007

Lactobacillus salivarius subsp. salivarius CNU27 possessed a high level of ${\alpha}$-galactosidase activity. Purified ${\alpha}$-galactosidase was obtained after sonication of harvested cell pellet followed by DEAE-Sephadex A-50 and Mono Q anion exchange chromatography. The specific activity of the purified enzyme was 8,994 units/mg protein which is 17.09 times higher than that in crude extract. The native enzyme was a monomer with a molecular mass of 56,397.1 dalton. The optimum temperature and pH for the enzyme were $40^{\circ}C$ and 6.0, respectively. The enzyme was stable between 25 and $50^{\circ}C$. However, ${\alpha}$-galactosidase activity was lost rapidly below pH 4.5 and above pH 8.5. The enzyme activity decreased to 6.73% and 4.30% of the original activity by addition of $Cu^{2+}$ and $Hg^{2+}$, respectively. Other metal compounds did not affect the enzyme activity significantly. The enzyme liberated galactose from melibiose, raffinose, and stachyose. The rate of substrates hydrolysis was measured by HPLC. Raffinose, stachyose and melibiose were completely decomposed after 24 hr at $40^{\circ}C$.

• 한국식품영양과학회지
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• 제31권3호
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• pp.465-468
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• 2002

Affinity chromatography법에 의해 해바라기씨 유래 $\alpha$-galactosidase정제를 수행하여 locust bean gum 증점 대체자원의 개발을 위한 기초연구로서 정제효소처리된 guar gum 과 locust bean gum의 구성당인 galactose와 mannose의 구성비율을 비교 검토하였다. 정제효소처리된 guar gum의 중점효과는 galactose와 mannose비율이 1 : 3.2일 때 가장 증가하였다. 이와같은 비율은 효소적 처리시간을 24시간으로 하였을 때 나타내었다. 해바라기씨 유래 $\alpha$-galactosidase처리에 의한 guar gum으로부터의 galactose의 유리가 증점효과에 영향을 미치는 것으로 사료된다. Xanthan gum과 guar gum의 혼합, xanthan gum과 guar gum 및 정제효소처리된 천연 유래 copra meal과의 혼합물에 대한 증점효과도 비교하였다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권3호
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• pp.215-218
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• 2000

Sialytrisaccharides based on $\beta$-galactosyldisaccharides were synthesized using $\beta$-galactosidase and trans-sialidase in one pot. Using $\beta$-galactosidase from Bacillus Ciculans and trans-sialidase from Trypanosoma cruzi simulaneously, 6mM sialyltrisaccharides composed of about 95% NeuAc$\alpha$(2,3)Gal$\beta$(1,4)GlcNAc and 5% NeuAc$\alpha$(2,3)Gal$\beta$(1,6)GlcNAc were produced from a reaction mixture containing 25mM o-nitropheny1-$\beta$-D-galsctolneuraminic acid. One beauty of this reaction was that a secondary hydrolysis of the disaccharide intermediate occurring between the activated galactopyranoside and N-acetylgucosamine was prevented. Using $\beta$-galactosidase from Escherichia cloi and the same trans-sialidase, 15mM sialyltrisaccharides composed of about 90% NeuAc$\alpha$(2,3)Gal$\beta$(1,6)GlcNac and 10% NeuAc$\alpha$(2,3)Gal$\beta$(1,4)GlcNAc were produced from a reaction misture containing 400nM galactose, 800nM N-acetylglucosylation rection between galactose and N-actylgucosamine was diminant since the disaccharide intermediate mainly resulted sreulted in the silylated product.

• 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.

• 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.

• Food Science and Biotechnology
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• 제17권5호
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• pp.1115-1118
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• 2008

${\alpha}$-Galactosidase gene (aga) from Leuconostoc mesenteroides SY1 was expressed in a heterologous host, Lactobacillus brevis 2.14 using an Escherichia coli-Leuconostoc shuttle vector, pSJE. pSJEaga (pSJE carrying aga) was introduced into Lactobacillus brevis 2.14 by electroporation and transformation efficiency was $1.1{\times}10^3$ per ${\mu}g$ DNA. L. brevis transformants (TFs) showed higher ${\alpha}$-galactosidase (${\alpha}$-Gal) activities than cells containing pSJE. Transcription levels of aga in L. brevis 2.14 grown on different carbon sources (1%, w/v) were examined by slot blot analysis. Aga transcript levels and ${\alpha}$-Gal activities were higher in cells grown on melibiose, raffinose, and galactose than cells on glucose, sucrose, and fructose. Western blot result showed that L. brevis 2.14 harboring pSJEaga produced much more ${\alpha}$-Gal when grown on melibiose than on glucose.

• 한국식품과학회지
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• 제20권1호
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• pp.79-84
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• 1988

대두(大豆)중에 존제하는 장내(腸內) 가스 발생인자인 raffinose와 stachyose의 제거에 이용될 수 있는 Aspergillus niger의 ${\alpha}$-galactosidase는 Czapeck-Dox 액체배지와 밀기울 배지에서 5일간 배양하였을 때 효소생성이 제일 높았다. Czapeck-Dox 액체배양에서 raffinose와 $NaNO_3$가 효소생산에 가장 효과적인 탄소원과 질소원으로 나타났으며 밀기울 배양에서는 질소원 또는 탄소원의 첨가효과가 없었다. 생성된 조효소(組酵素)용액의 작용 최적 pH는 4.0-5.0, 안정도 최적 pH는 3.5-6.5, 최적온도는 $40-50^{\circ}C$ 였다. 합성기질인 p-nitrophenyl-${\alpha}$-D-galactoside에 대한 Michaelis상수는 0.42mM, 최대반응속도는 152${\mu}moles$ substrate/min/kg (고체배지)이었다. $HgCl_2$는 강력한 비경쟁 저해제로 작용하였고 p-chloromercuribenzoate는 낮은 농도에서 경쟁 저해제로 작용하였다. 조효소 용액은 raffinose와 stachyose를 완전 분해하여 단당류 만을 생성하였다.

• 한국식품과학회지
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• 제18권6호
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• pp.450-457
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• 1986

대두(大豆)중에 존재하는 장내(腸內) 가스발생인자(發生因子)(flatulence factor)인 raffinose와 stachyose의 제거(除去)에 효소제(酵素劑)를 이용하기 위한 기초연구로서, 시판효소제(市販酵素劑)에 대한 ${\alpha}-Galactosidase$의 활성도(活性度)를 비교하고 그의 효소적(酵素的) 특성(特性)을 조사하였다. 시판(市販)되는 6종의 효소제(酵素劑)중 Aspergillus niger에서 얻은 pectinase 제제가 가장 강력하였다. 선정된 효소제(酵素劑)의 특성(特性)을 조사한 결과 작용최적(作用最適) pH는 4.0-4.5, 안정도최적(安定度最適) pH는 4-5, 작용최적(作用最適) 온도는 $45^{\circ}C$였다. 합성기질(合成基質)인 $p{\cdot}nitropheyl-{\alpha}-D{\cdot}galactoside$에 대한 Michaelis constant(Km)는 2.08mM, 최대반응속도(最大反應速度)(Vmax)는 435 ${\mid}{\mu}{\mid}$moles substrate/minute/g enzyme preparation 이었으며 조해제(阻害劑)의 실험 결과 SH기(基)를 필수로 하는 효소(酵素)로 밝혀졌다. 본(本) 효소제(酵素劑)는 3가지 기질(基質)인 raffinose, sucrose, $p-nitrophenyl-{\alpha}-D{\cdot}galactoside$를 거의 완전하게 가수분해(加水分解)하였으며, raffinose와 stachyose의 분해(分解)과정을 thin-layer chromatography로 조사한 결과 단당류의 Spot만이 나타났다. 따라서 대두식품(大豆食品)중의 장내(腸內) 가스발생인자(發生因子)는 ${\alpha}-Galactosidase$ 역가(力價)를 가지는 효소제(酵素劑)의 이용에 의하여 쉽게 제거될 수 있을 것이다.