• 한국축산식품학회지
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• 제42권3호
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• pp.426-440
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• 2022

The bioactive functions of oligosaccharides from human milk have been reported by many studies. Many of oligosaccharides isolated from colostrum and/or milk of dairy animals have been reported to have similar chemical structures with those in human colostrum and/or milk. It has been proved by several studies that the oligosaccharides with similar chemical structure shared common bioactivities. Among domesticated dairy animals, bovine/cattle, caprine/goat, and ovine/sheep are the most commonly used species to isolate oligosaccharides from their colostrum and/or milk. Several studies on the oligosaccharides from goat colostrum and milk have revealed similar properties to that of human milk and possess the highest content of sialyl oligosaccharides (SOS) as compared to other ruminants. Indonesia ranks first in Association of Southeast Asian Nations (ASEAN) for goat milk production. Therefore, goat milk is the second most consumed milk in the country. The most reared dairy goat breed in Indonesia is Etawah Grade. However, oligosaccharides from Indonesia dairy animals including goat, have not been characterized. This is the first study to characterize oligosaccharides from Indonesia dairy animals. The present study was aimed to isolate and characterize oligosaccharides, specifically SOS from the colostrum of Etawah Grade goats by using proton/¹H-nuclear magnetic resonance. The SOS successfully characterized in this study were: Neu5Ac(α2-3)Gal(β1-4)Glc (3'-N-acetylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4)Glc (6'-N-acetylneuraminyllactose), Neu5Gc(α2-3)Gal(β1-4)Glc (3'-N-glycolylneuraminyllactose), Neu5Gc(α2-6)Gal(β1-4)Glc (6'-N-glycolylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4) GlcNAc (6'-N-acetylneuraminyllactosamine) and Neu5Gc(α2-6)Gal(β1-4)GlcNAc (6'-N-glycolylneuraminyllactosamine). This finding shows that Etawah Grade, as a local dairy goat breed in Indonesia, is having significant potential to be natural source of oligosaccharides that can be utilized in the future food and pharmaceutical industries.

• 한국수정란이식학회지
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• 제32권3호
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• pp.227-233
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• 2017

Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

• International Journal of Stem Cells
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• 제16권1호
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• pp.52-65
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• 2023

Background and Objectives: Epithelial-Mesenchymal transition (EMT) is one of the origins of myofibroblasts in renal interstitial fibrosis. Mesenchymal stem cells (MSCs) alleviating EMT has been proved, but the concrete mechanism is unclear. To explore the mechanism, serum-free MSCs conditioned medium (SF-MSCs-CM) was used to treat rat renal tubular epithelial cells (NRK-52E) fibrosis induced by transforming growth factor-β1 (TGF-β1) which ameliorated EMT. Methods and Results: Galectin-3 knockdown (Gal-3 KD) and overexpression (Gal-3 OE) lentiviral vectors were established and transfected into NRK-52E. NRK-52E fibrosis model was induced by TGF-β1 and treated with the SF-MSCs-CM for 24 h after modelling. Fibrosis and autophagy related indexes were detected by western blot and immunocytochemistry. In model group, the expressions of α-smooth muscle actin (α-SMA), fibronectin (FN), Galectin-3, Snail, Kim-1, and the ratios of P-Akt/Akt, P-GSK3β/GSK3β, P-PI3K/PI3K, P-mTOR/mTOR, TIMP1/MMP9, and LC3B-II/I were obviously increased, and E-Cadherin (E-cad) and P62 decreased significantly compared with control group. SF-MSCs-CM showed an opposite trend after treatment compared with model group. Whether in Gal-3 KD or Gal-3 OE NRK-52E cells, SF-MSCs-CM also showed similar trends. However, the effects of anti-fibrosis and enhanced autophagy in Gal-3 KD cells were more obvious than those in Gal-3 OE cells. Conclusions: SF-MSCs-CM probably alleviated the EMT via inhibiting Galectin-3/Akt/GSK3β/Snail pathway. Meanwhile, Gal-3 KD possibly enhanced autophagy via inhibiting Galectin-3/Akt/mTOR pathway, which synergistically ameliorated renal fibrosis. Targeting galectin-3 may be a potential target for the treatment of renal fibrosis.

• 한국식품영양과학회지
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• 제41권5호
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• pp.687-694
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• 2012

본 연구는 저식염 오징어 젓갈에 대한 과학적 자료를 제공함과 동시에 전통수산발효식품 소비의 확산에 기여하기 위하여 염 농도에 따른 발효기간별 품질특성 측정, 발효최적조건, 유통기한 설정 및 발효최적조건에서 제조한 저식염 오징어 젓갈의 생리활성 등을 연구하였으며, 그 결과를 요약하면 다음과 같다. 숙성온도가 증가할수록 pH는 급격히 감소하였으며, 기간에 따라 대조구인 10% 젓갈의 pH 변화는 거의 없는 반면 3 및 5% 첨가군은 급격히 감소하였으나 pH 5 이하에서는 서서히 감소하였다. 아미노질소와 휘발성염기 질소 또한 발효온도가 증가할수록 각각의 함량이 급격히 증가하였으며 식염의 농도가 낮을수록 휘발성 염기질소의 함량은 증가하였다. 아미노질소의 함량이 최고치가 되는 시점을 최적발효조건 지표로 산출하였으며 그 결과 $-1^{\circ}C$에서 저장한 염 농도 3% 젓갈일 때 41일의 최적발효기한을 나타내었다. 저식염 오징어 젓갈의 냉수 및 에탄올 추출물의 항산화 활성, 항당뇨 및 항대장암은 뚜렷한 활성을 보이지 않았으며, 일반성분 함량은 식염과 부재료 첨가로 수분과 회분의 함량에 변화를 보였지만 그 이후 저온저장 하는 기간 동안에는 일정하게 유지되다. 구성아미노산 및 유리아미노산은 glycine, arginine, alanine 및 glutamic acid 등의 함량이 절반 이상을 차지함으로써 맛에서 우수하였고, 관능검사결과는 시중에서 판매하고 있는 젓갈과 비교했을 때 큰 차이를 보이지 않았다. 이상의 결과를 고려할 때 염 농도 3% 젓갈을 $-1^{\circ}C$에서 제조, 숙성시킨다면 최근 소비자의 건강상 기호패턴의 변화에 부응시킴과 동시에 전통 수산발효식품의 보급 및 소비 확산에 기여할 수 있을 것이라 판단된다.

• 한국지반신소재학회논문집
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• 제18권4호
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• pp.107-114
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• 2019

본 논문에서는 진원지 주변에서 빠르고 정확한 지진 조기 경보를 수행하기 위한 선착 P파 다중 탐지 시스템과 이를 구동하기 위한 지진파 초동 탐지 및 경보 알고리즘을 개발하였다. 공용 중인 5개소(포항지역 4개소)의 건축물을 선정하여 개발한 계측 시스템을 설치하였고, 지반 진동을 실시간 모니터링하며 실증시험을 진행하였다. 실증 모니터링 중 2019년 9월 26일 포항 지역에서 규모 2.3의 지진이 발생하였다. 포항지역에 설치된 총 4개소의 시스템 중 3개소에서 P파 초동 탐지 알고리즘이 작동되어 지진동 이벤트로 기록되었다. 진원지로부터 5.5 km로 가장 가까운 계측소는 지진 발생 후 1.2초 후 P파 초동이 감지되었으며, P파 도달 후 약 1.02초 후 S파가 도달하여 다소의 경보시간을 제공해주었다. P파가 탐지된 3곳의 최대 가속도는 각각 6.28gal, 6.1gal, 5.3gal로 기록되었으며, 이벤트 경보 발령을 위한 최대 지반 가속도의 임계값(25.1gal)을 초과하지 않아 경보 알고리즘이 작동하지 않았다. 향후 지속적인 모니터링 및 분석을 통해 추가 검증이 이루어진다면 국내 실정에 맞는 실효성 높은 지진 경보 시스템으로 활용 될 수 있을 것으로 판단된다.

• Applied Biological Chemistry
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• 제36권3호
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• pp.191-196
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• 1993

본 연구는 합성기질인 $X-{\alpha}-Gal$를 이용하여 발효유 및 유제품내의 Bifidobacteria 생균수를 측정할 목적으로 하였다. 젖산균과 Bifidobacteria의 ${\alpha}-galactosidase$ specific activity를 측정한 결과 Bifidobacteria 균주에서는 높은 ${\alpha}-galactosidase$ activity를 가지고 있었으며, 그 중 Bif. longum KCTC 3215의 specific activity는 8.57 unit/mg protein으로 가장 높게 나타났다. Lactobacillus, Streptococcus, Pediococcus와 Leuconostoc 균주에서는 활성이 미약하거나 없었다. 합성기질인 $X-{\alpha}-Gal$을 MRS agar 배지에 $100{\;}{\mu}M$ 첨가한 결과 Bifidobacteria는 blue colony로, Lac. bulgaricus, Lac. casei와 Leu. mescenteroides 균주는 light blue colony로, 그 외 젖산균에서는 white colony로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제32권8호
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• pp.1064-1071
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• 2022

Arabinogalactans have diverse biological properties and can be used as pharmaceutical agents. Most arabinogalactans are composed of β-(1→3)-galactan, so it is particularly important to identify β-1,3-galactanases that can selectively degrade them. In this study, a novel exo-β-1,3-galactanase, named PoGal3, was screened from Penicillium oxalicum sp. 68, and hetero-expressed in P. pastoris GS115 as a soluble protein. PoGal3 belongs to glycoside hydrolase family 43 (GH43) and has a 1,356-bp gene length that encodes 451 amino acids residues. To study the enzymatic properties and substrate selectivity of PoGal3, β-1,3-galactan (AG-P-I) from larch wood arabinogalactan (LWAG) was prepared and characterized by HPLC and NMR. Using AG-P-I as substrate, purified PoGal3 exhibited an optimal pH of 5.0 and temperature of 40℃. We also discovered that Zn²⁺ had the strongest promoting effect on enzyme activity, increasing it by 28.6%. Substrate specificity suggests that PoGal3 functions as an exo-β-1,3-galactanase, with its greatest catalytic activity observed on AG-P-I. Hydrolytic products of AG-P-I are mainly composed of galactose and β-1,6-galactobiose. In addition, PoGal3 can catalyze hydrolysis of LWAG to produce galacto-oligomers. PoGal3 is the first enzyme identified as an exo-β-1,3-galactanase that can be used in building glycan blocks of crucial glycoconjugates to assess their biological functions.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2002년도 제38회 학술심포지움
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• pp.13-19
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• 2002

To elucidate the regulatory mechanism for expression of sialyl-glycoconjugates and their biological functions, ninetheen sialyltransferase cDNAs including eleven by our group or co-works have been cloned and characterized so far. The cloned sialyltransferases are classified into four families according to the carbohydrate linkages they synthesize: ${\alpha}2,3-sialyltransferase$ (ST3Gal I-VI), ${\alpha}$ 2,6-sialyltransferase (ST6Gal I), GalNAc ${\alpha}$ 2,6-sialyltransferase (ST6GalNAc I-VI), and ${\alpha}2,8-sialyltransferase$ (ST8Sia I-VI). Each of the sialyltransferase genes is differentially expressed in a tissue-, cell type-, and stage-specific manner. These enzymes differ in their substrate specificity and various biochemical parameters. However, enzymatic analysis conducted in vitro with recombinant enzyme revealed that one linkage can be synthesized by multiple enzymes. We present here an overview of structure and function of sialyltransferases performed by our group and co-works. Genomic structures and transcriptional regulation of two kinds of human sialyltransferase gene are also presented.

• Journal of Microbiology and Biotechnology
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• 제26권7호
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• pp.1267-1277
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• 2016

Currently, enzymatic synthesis of lactulose, a synthetic prebiotic disaccharide, is commonly performed with glycosyl hydrolases. In this work, a new type of lactulose-producing biocatalyst was developed by displaying β-galactosidase from Bacillus stearothermophilus IAM11001 (Bs-β-Gal) on the surface of Bacillus subtilis 168 spores. Localization of β-Gal on the spore surface as a fusion to CotX was verified by western blot analysis, immunofluorescence microscopy, and flow cytometry. The optimum pH and temperature for the resulting spore-displayed β-Gal was 6.0 and 75℃, respectively. Under optimal conditions, it showed maximum activity of 0.42 U/mg spores (dry weight). Moreover, the spore-displayed CotX-β-Gal was employed as a whole cell biocatalyst to produce lactulose, yielding 8.8 g/l from 200 g/l lactose and 100 g/l fructose. Reusability tests showed that the spore-displayed CotX-β-Gal retained around 30.3% of its initial activity after eight successive conversion cycles. These results suggest that the CotX-mediated spore-displayed β-Gal may provide a promising strategy for lactulose production.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권5호
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• pp.337-340
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• 2001

A galactooligosaccharide(GalOS)-producing yeast, OE-20 was selected from forty seven strains of yeast growing in Korean traditional Meju (cooked soybean) and the yeast was tentatively identified as Kluyveromyces maxianus var lactis by its morphology and fermentation profile. A maximum yield of 25.1%(w/w) GalOS, which corresponds to 25.1 g of GalOS per liter, was obtained from the reaction of 100 g per liter of lactose solution at 3$0^{\circ}C$, pH 7.0 for 18 h with an intracellular crude $\beta$-galactosidase. Glucose and galactosidase were found to inhibit GalOS formation. The GalOS that were purified by active carbon and celite 545 column chromatography were supplemented in MRS media and a stimulated growth was observed of some intestinal bacteria. In particular the growth rate of Bifidobacterium infantis in the GalOS containing MRS broth increased up to 12.5% compared to that of the MRS-glucose broth during a 48h incubation period.