• Journal of Microbiology and Biotechnology
• /
• 제19권11호
• /
• pp.1295-1300
• /
• 2009

A 2,172-bp full-length gene (aga-F78), encoding a protease-resistant $\alpha$-galactosidase, was cloned from Rhizopus sp. F78 and expressed in Escherichia coli. The deduced amino acid sequence shared highest identity (45.0%) with an $\alpha$-galactosidase of glycoside hydrolase family 36 from Absidia corymbifera. After one-step purification with a Ni-NTA chelating column, the recombinant Aga-F78 migrated as a single band of ~82 and ~210 kDa on SDS-PAGE and nondenaturing gradient PAGE, respectively, indicating that the native structure of the recombinant Aga-F78 was a trimer. Exhibiting the similar properties as the authentic protein, purified recombinant Aga-F78 was optimally active at $50^{\circ}C$ and pH 4.8, highly pH stable over the pH range 5.0-10.0, more resistant to some cations and proteases, and had wide substrate specificity (pNPG, melidiose, raffinose, and stachyose). The recombinant enzyme also showed good hydrolytic ability to soybean meal, releasing galactose of $415.58\;{\mu}g/g$ soybean meal. When combined with trypsin, the enzyme retained over 90% degradability to soybean meal. These favorable properties make Aga-F78 a potential candidate for applications in the food and feed industries.

• 미생물학회지
• /
• 제52권3호
• /
• pp.344-351
• /
• 2016

대한민국 동해안 울진 앞 바닷물로부터 50-C로 명명한 해양 미생물을 분리하였다. 50-C 균주는 그람-음성, 호기성 세균이며, 노란색 집락을 형성하고, 극성편모를 갖는 박테리아이다. 이 균주는 $20-50^{\circ}C$, pH 5.5-8.5 범위에서 자라며, 비교적 고온인 $40-50^{\circ}C$, pH 6.5-7.5, 2% (w/v) NaCl에서 최적 성장을 보인다. 16S rRNA 유전자 서열 분석결과 50C-3 균주는 Ruegeria 속에 속하는 R. intermedia CC-GIMAT-$2^T$, R. lacuscaerulensis ITI-$1157^T$의 16S rRNA 유전자 서열과 각각 99.4%, 96.98% 상동성을 보였다. 그러나 50C-3 균주는 운동성, 탄소이용능력, 효소생산능력 등의 생리학적 특성에서 두 균주와는 명확히 다른 특성을 보였다. 50C-3 균주의 DNA G+C content는 66.7 mol%이고, 주요한 respiratory quinone은 ubiquinone-10 (Q-10)이었다. 이와 같은 형태학적, 생리학적, 유전학적 특성을 비교하여, 50C-3 균주는 R. intermedia CC-GIMAT-$2^T$와 같은 종에 속하는 새로운 변종으로 판단되며 Ruegeria sp. 50C-3으로 명명하였다(KCTC23890 =DSM25519). 50C-3 균주는 cellulase, agarase 활성은 없었지만, alkaline phosphatase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase를 생산하였고 이들 모두 $50^{\circ}C$ 에서도 활성이 좋은 내열성 효소일 것으로 판단되었다. 특히, ${\beta}$-galactosidase의 경우 $37^{\circ}C$에서 보다 $50^{\circ}C$에서의 활성이 1.9배 증가하여 산업적으로 활용성이 클 것으로 예상된다.

• Journal of Chest Surgery
• /
• 제45권6호
• /
• pp.380-389
• /
• 2012

Background: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and ${\alpha}$-galactosidase (${\alpha}$-gal) to remove major xenoreactive antigens from xenogenic tissues. Materials and Methods: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) ${\alpha}$-gal or decellularization, or both were used to remove ${\alpha}$-gal from bovine pericardium. It was confirmed by ${\alpha}$-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or ${\alpha}$-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000${\times}$ transmission electron microscope (TEM). Results: Recombinant B. thetaiotaomicron ${\alpha}$-gal could effectively remove ${\alpha}$-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human ${\alpha}$-gal removed it recombinant human ${\alpha}$-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant ${\alpha}$-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant ${\alpha}$-gal made no difference in the arrangement of collagen fiber bundles with decellularization. Conclusion: Recombinant B. thetaiotaomicron ${\alpha}$-gal effectively removed ${\alpha}$-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant ${\alpha}$-gal, which may alleviate the harmful xenoreactive immunologic responses of ${\alpha}$-gal. Recombinant ${\alpha}$-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

• 대한유전성대사질환학회지
• /
• 제17권3호
• /
• pp.92-95
• /
• 2017

Fabry 병은 성염색체 연관 유전성 리소좀 대사 질환으로 ${\alpha}$-galactosidase A 를 코딩하는 GLA 유전자의 변이로 인한 ${\alpha}$-galactosidase A 효소의 결핍에 의해 발생한다. 이 질환은 globotriaosylceramide (GL-3) 및 관련된 글리코스핑고리피드(glycophospholipids)가 신장 사구체, 심근, 후근 신경절 및 자율 신경계, 혈관 내피 세포 및 평활근 등에 축적되어 사지 말단 동통, 신부전, 심부전 등의 다양한 임상양상을 보이게 된다. 대증적 요법으로만 치료하던 Fabry 병은 효소대체요법의 발전으로 신부전을 포함하여, 심각한 합병증의 예방 및 호전과 함께 질환의 예후를 향상시키고 있다. 또한 사지 말단 동통은 Fabry 병 환자들의 삶의 질을 특히 떨어뜨리며, 적절한 효소대체요법에 효과가 있는 것으로 알려져 있다. 저자들은 40대 후반에 Fabry 병을 진단받고 효소대체요법을 시작하여 사지 말단 동통이 호전된 중국인 남자 환자에 대해 보고하는 바이다.

• Journal of Animal Science and Technology
• /
• 제55권4호
• /
• pp.289-293
• /
• 2013

The objective of this study was to assess the effects of dietary carbohydrases on growth performance, nutrient digestibility and blood characteristics in finishing pigs. A total of 90 pigs [(Landrace ${\times}$ Yorkshire) ${\times}$ Duroc] (initial BW = $56.15{\pm}1.26kg$) were used for a 35 d feeding trial. The dietary treatments included: 1) CON (control diet), 2) MIX (CON + mixture with ${\alpha}$-galactosidase and ${\beta}$-mannanase 0.05%) and 3) MAN (CON + ${\beta}$-mannanase 0.05%). There were six replications per treatment with five pigs per pen. The average daily gain (ADG) in MIX was higher than in CON (p<0.05). No significant differences were noted in the average daily feed intake (ADFI) and feed efficiency (G:F) among dietary treatments (p>0.05). Apparent total tract digestibility (ATTD) of dry matter (DM) and energy (E) in MIX increased (p<0.05) relative to CON and MAN. The ATTD of nitrogen (N) in MIX was higher (p<0.05) than in CON. No differences in red blood cells (RBC), white blood cells (WBC), lymphocytes and IgG concentrations were observed among dietary treatments (p>0.05). In conclusion, the addition of the mixture of carbohydrases (${\alpha}$-galactosidase and ${\beta}$-mannanase 0.05%) increased ADG and nutrient digestibility in finishing pigs.

• Childhood Kidney Diseases
• /
• 제20권2호
• /
• pp.79-82
• /
• 2016

Fabry disease is an X-linked lysosomal storage disorder caused by a deficiency of the enzyme ${\alpha}-galactosidase$ A, resulting in the accumulation of glycosphingolipids within the lysosomes of various cell types. It has a wide spectrum of clinical phenotypes, and renal failure is a serious complication. Fabry disease is confirmed either by measurement of ${\alpha}-galactosidase$ A activity or by genetic testing for GLA mutations. Renal biopsy findings on light microscopy, specifically enlarged podocytes with foamy cytoplasm, and osmiophilic inclusion bodies in the cytoplasm in all types of renal cells on electron microscopy, are characteristic of this disease. The predominant differential diagnosis is iatrogenic phospholipidosis in association with certain drugs that can cause cellular injuries indistinguishable from Fabry disease. Here, we report the case of a 10-year-old boy with microscopic hematuria who underwent a renal biopsy that showed morphological findings consistent with Fabry disease, although the patient had neither a GLA mutation nor a history of drug consumption. Six years later, spontaneous regression of this renal pathology was observed in a second renal biopsy examination.

• The Korean Journal of Pain
• /
• 제23권3호
• /
• pp.207-210
• /
• 2010

Fabry disease is an X-linked lysosomal disease caused by deficiency of ${\alpha}$-galactosidase, in which early diagnosis may be missed due to the wide variety of clinical symptoms presenting during disease progression. A 13 year-old boy visited our pain clinic complaining of pricking and burning pain in the toe tips of both feet. Continuous epidural infusion for pain management was performed because of oral analgesics ineffectiveness. The patient underwent ${\alpha}$-galactosidase A (GLA) enzyme analysis based on the clinical impression of Fabry disease from pain with a peripheral neuropathic component and history of anhidrosis. He was diagnosed with Fabry disease after confirming mutation of the GLA gene through a screening test of GLA activity. Enzyme replacement therapy was initiated and pain was tolerated with oral analgesics.

• Clinical and Experimental Pediatrics
• /
• 제53권2호
• /
• pp.235-238
• /
• 2010

파브리병은 알파-갈락토시다아제(alpha galactosidase) A 효소에 위치하는 유전자 변이에 의한 글라이코스핑고리피드(glycospingolipid) 대사이상에 의한 질환으로 X-염색체와 연관된다. 말단 지각이상을 호소하던 12세 남자 환자와 증상이 미미한 그의 형에서 알파-갈락토시다아제 A 효소 활성도 측정과 유전자 변이 확인을 통하여 파브리병을 확진한 2례를 보고하고자 한다.

• Journal of Microbiology and Biotechnology
• /
• 제18권2호
• /
• pp.338-342
• /
• 2008

This study was performed to screen and select Lactobacillus strains from chicken feces for probiotic use in animals. Of these strains, strain AU had the highest immunostimulatory effect. Therefore, strain A12 was characterized as a potential probiotic. Strain A12 was tentatively identified as Lactobacillus acidophilus A12, using the API 50 CHL kit based on a 99.9% homology. L. acidophilus A12 was highly resistant to artificial gastric juice (pH 2.5) and bile acid (oxgall). Based on results from the API ZYM kit, leucine arylamidase, crystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, ${\alpha}$-galactosidase, ${\beta}$-galactosidase, ${\alpha}$-glucosidase, ${\beta}$-glucosidase, and N-acetyl-${\beta}$-glucosamidase were produced by strain A12. L. acidophilus A12 showed resistance to several antibiotics (nisin, gentamicin, and erythromycin). The amount of interleukin $(IL)-1{\alpha}$ in $20{\times}$ concentrated supernatant from L. acidophilus A12 was approximately 156pg/ml. With regard to antioxidant activity, L. acidophilus A12 supernatant showed 60.6% DPPH radical scavenging activity. These results demonstrate the potential use of L. acidophilus A12 as health-promoting probiotics.

• 한국미생물·생명공학회지
• /
• 제33권4호
• /
• pp.241-247
• /
• 2005

선별마커로써 Lactococcus lactis ssp. lactis ATCC 7962 유래의 $\beta$-galactosidase 유전자를 포함하는 Lactococcus용 셔틀/발현 벡터 pWgall3T를 제조하여 Escherichia coli DH5$\alpha$와 고. Lactis MG1363내로 도입하였다. 이들 형질 전환체들은 X-gal을 포함하는 배지에서 파란색의 표현형을 보임으로써 쉽게 확인할 수 있었다. 또한, L. lactis MG1363 형질전환체로부터 $\beta$-galactosidase 활성을 측정한 결과 기존에 $\beta$-galactosidase를 활성을 지닌 L. lactis ATCC 7962에 비해 glucose를 포함하는 M17배지에서 4배정도 높은 활성을 보임으로써 선별마커로써의 효율성을 나타내었다. pWgal13T는 $\beta$-galactosidase 유전자 외에 L. lactis Wg2유래의 replicon과 외래 유전자의 발현을 위한 L. lactis ssp. cremoris LM0230의 promoter P13C, terminator를 포함하고 있다. 이 벡터의 이용가능성을 확인하기 위하여 외래 유전자 EGFP유전자를 P13C 아래에 삽입하여 E. coli와 L. iactis에서 발현을 확인하였다. 이 연구에서 제조된 Lactococcus용 발현 벡터 pWgal13T는 E. coli와 L. lactis에서 외래 유용 유전자를 생산을 위해 이용 할 수 있을 것이다.