• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.1075-1080
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• 2004

Dextransucrase (DSRB742) from Leuconostoc mesenteroides NRRL B-742CB is a glucosyltransferase that catalyzes the synthesis of dextran using sucrose, or the synthesis of oligosaccharides when acceptor molecules, like maltose, are present. The DSRB742 gene (dsrB742) was cloned and the properties were characterized. In order to identify critical amino acid residues, the DSRB742 amino acid sequence was aligned with glucosyltransferase sequences, and three amino acid residues reported as sucrose binding amino acids in Streptococcus glucosyltransferases were selected for site-directed mutagenesis experiments. Asp-533, Asp-536, and His-643 were independently replaced with Ala or Asn. D533A and D536A dextransucrases showed reduced dextran synthesis activities, 2.3% and 40.8% of DSRB742 dextransucrase, respectively, and D533N, D536N, H643A, end H643N dextransucrases showed complete suppression of dextran synthesis activities altogether. Additionally, D536N dextransucrase showed complete suppression of oligosaccharide synthesis activities. However, modifications at Asp-533 or at His-643 retained acceptor reaction activities in the range of 8.4% to 21.3% of DSRB742 acceptor reaction activity. Thus at least two carboxyl groups of Asp-533 and Asp-536, and His-643 as a proton donor, are essential for the catalysis process.

• Journal of Microbiology and Biotechnology
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• 제11권4호
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• pp.700-703
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• 2001

The influence of process conditions on highly branched glucooligosaccharides production by mixed culture of Leuconostoc mesenteroides ATCC 13146 and Lipomyces starkeyi ATCC 74054 was studied. We divided the batch culture fermentations into two groups according to inoculation method. One-point inoculation was performed by coinoculation of L. mesenteroides and L. starkeyi at the ration of 10 to 1, and two-point inoculation by L. mesenteroides inoculation first and L. starkeyi inoculation after L. mesenteroides grew to the end of the log phase of growth. Two-point inoculation improved the yield of oligosaccharide by 1.5 to 20 fold more than one-point inoculation. In this process, the highest yield of oligosaccharides (48% of theoretical yield) and productivity (0.85 g/l/h) were obtained with starch as an initial substrate for L. starkeyi growth. The estimated composition of the end product consisted of 31.5% oligosaccharides, 17.6% dextran, and 46.5% mannitol.

• Journal of Microbiology and Biotechnology
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• 제8권5호
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• pp.484-489
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• 1998

Galactooligosaccharides (GalOS) were efficiently produced by partially purified $\beta$-galactosidase from the yeast strain Bullera singularis ATCC 24193. Ammonium sulfate precipitation and ultrafiltration methods were used to prepare the enzyme. The enzyme activity decreased at $50^{\circ}C$ and above. A maximum yield of 40% (w/w) GalOS, corresponding to 120 g of GalOS per liter, was obtained from 300 g per liter of lactose solution at $45^{\circ}C$, pH 3.7 when the lactose conversion was 70%. The yield of GalOS did not increase with increasing initial lactose concentration but the total amounts of GalOS did. Volumetric productivity was 4.8 g of GalOS per liter per hour. During this reaction, the by-products, glucose and galactose, were found to inhibit GalOS formation. Reaction products were found to be comprised of disaccharides and trisaccharides according to TLC and HPLC analyses. We propose the structure of the major product, a trisaccharide, to be ο-$\beta$-D-galactopyranosyl-(l-4)-ο-$\beta$-D-galactopyranosyl-(l-4)-$\beta$-D-glucose (4'-galactosyl lactose).

• 동의생리병리학회지
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• 제18권5호
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• pp.1437-1442
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• 2004

In this study, lotus root(Nelumbo nucifera G.) and omija(Schizandrae fructus), which have been used in oriental medicine and folks remedy, were examined to apply to functional foods. We prepared yanggaeng with lotus root, sugar, oligosaccharide, agar, omija extract solution and analyzed the nutritional composition(moisture, protein, fat, ash, crude fiber, carbohydrate, free sugar and minerals). Also we investigated texture profile and evaluated sensory characteristics of developed yanggaengs and 8 commercial ones. Potassium and crude fiber contents of yanggaeng with lotus root were higher than the commercial yanggaengs. Cohesiveness, springiness and gumminess of lotus root yanggaeng were the same levels of commercial yanggaengs. Sensory evaluation with the ones, showed that the lotus yanggaeng was more desirable than the commercial ones. Yanggaeng with lotus had good scores in texture profile and sensory evaluation compared with commercial yanggaengs. These results demonstrated that lotus root hadsufficient values to use a foodstuff for yanggaeng.

• Journal of Dairy Science and Biotechnology
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• 제37권3호
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• pp.167-176
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• 2019

Milk contains essential nutrients and functional compounds, such as calcium, fat-soluble vitamins A, D, E, and K, carotenoids, bioactive peptides, and sphingolipids. The bioactive molecules from milk are not expensive and have an added advantage of being derived from food. Therefore, they are more stable and have a broader spectrum than that of other chemicals. Bioactive milk components are useful for treating non-digestive tract disorders, such as cancer, cognitive decline, and hypertension. However, the clinical application of certain breast milk ingredients is limited due to the lack of a large-scale production technology. Once the scaled-up production of lactoferrin became possible, clinical applications were devised and evaluated. Similarly, human alpha-lactalbumin made lethal to tumor cells (HAMLET) can be produced on a large scale as a recombinant protein in microorganisms or in transgenic cattle using suitable separation systems. HAMLET can be used to treat human skin papilloma and cancer. Studies on breast milk that explored the clinical applications of the bioactive components of breast milk have spurred the development of translational medicine and breast milk-derived therapeutics. Some breast-milk derived therapeutic agents are already available to clinicians. Many components of breast milk have shown efficacy in pre-clinical studies and have valid clinical evaluations.

• Journal of Microbiology
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• 제45권1호
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• pp.69-74
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• 2007

Escherichia coli is a clonal species, and occurs as both commensal and pathogenic strains, which are normally classified on the basis of their O, H, and K antigens. The O-antigen (O-specific polysaccharide), which consists of a series of oligosaccharide (O-unit) repeats, contributes major antigenic variability to the cell surface. The O-antigen gene cluster of E. coli O66 was sequenced in this study. The genes putatively responsible for the biosynthesis of dTDP-6-deoxy-L-talose and GDP-mannose, as well as those responsible for the transfer of sugars and for O-unit processing were identified based on their homology. The function of the wzy gene was confirmed by the results of a mutation test. Genes specific for E. coli O66 were identified via PCR screening against representatives of 186 E. coli and Shigella O type strains. The comparison of intergenic sequences located between galF and the O-antigen gene cluster in a range of E. coli and Shigella showed that this region may perform an important function in the homologous recombination of the O-antigen gene clusters.

• Food Science and Biotechnology
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• 제14권3호
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• pp.380-386
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• 2005

Optimal conditions of ultrafiltration (UF) and nanofiltration (NF) were investigated for separation and concentration of isoflavones and oligosaccharides from Sunmul. Levels of COD, BOD, and suspended solids (SS) in UF and NF permeates were also determined to evaluate effectiveness of these processes for reducing water pollution. Optimal UF operation conditions to achieve minimal fouling and maximal flux were $33-34^{\circ}C$ operating temperature and 2.3-2.4 bar trans-membrane pressure. Recovery yields of isoflavones and oligosaccharides in UF retentate were 11.49-28.16% and 12.77-27.57%, respectively. Increase in volumetric concentration factor (VCF) resulted in more functional compounds of isoflavones and oligosaccharides passing through UF membrane. Total isoflavone and oligosaccharide yields decreased by 3% as VCF increased from 6.0 to 8.0 and from 8.0 to 10.0, while decreased significantly by 10% as VCF decreased from 4.0 to 6.0. Optimal NF operating conditions were 192-195 psig operating pressure at $30-33^{\circ}C$. Total yields of isoflavones and oligosaccharides significantly decreased at VCF 8.0, whereas did not decrease up to VCF 6.0 during NF operation. Therefore, VCF 6.0 was recommended for economical process. COD and BOD decreased by more than 98% after NF process, and SS were not detected after UF process. These results indicated sequential filtration process was useful for separation of isoflavones and oligosaccharides from Sunmul and for reducing water contaminants.

• 한국축산식품학회지
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• 제39권5호
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• pp.725-741
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• 2019

In the current study, we first investigated a method for directly transforming lactose into galacto-oligosaccharides (GOS) for manufacturing low-lactose and GOS-enriched skim milk (GSM) and then evaluated its prebiotic potential by inoculating five strains of Bifidobacterium spp. In addition, fermented GSM (FGSM) was prepared using a potentially probiotic Lactobacillus strain and its fermentation characteristics and antioxidant capacities were determined. We found that GOS in GSM were metabolized by all five Bifidobacterium strains after incubation and promoted their growth. The levels of antioxidant activities including radical scavenging activities and 3-hydroxy-3-methylglutaryl-CoA reductase inhibition rate in GSM were significantly increased by fermentation with the probiotic Lactobacillus strain. Moreover, thirty-nine featured peptides in FGSM was detected. In particular, six peptides derived from ${\beta}$-casein, two peptides originated from ${\alpha}s_1$-casein and ${\kappa}$-casein were newly identified, respectively. Our findings indicate that GSM can potentially be used as a prebiotic substrate and FGSM can potentially prevent oxidative stress during the production of synbiotic fermented milk in the food industry.

• 한국식품저장유통학회지
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• 제15권3호
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• pp.361-366
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• 2008

본 연구는 두유의 기능성을 향상시키고자 효소 가수분해 방법으로 저분자화에 따른 기능적 특성변화를 조사하였다. 두유액(SM), 두유액 가수분해물(HSM), 전두유액(WSM) 및 전두유 가수분해물(HWSM)의 일반성분은 비지 제거의 유무에 따라 성분함량 차이를 보였으나 저분자화 효소처리에 따른 변하는 거의 없었다. 색도는 대두의 비지 제거 유무에 차이를 나타내었으나 가수분해 처리에 의한 차이는 없었다. 유리당과 올리고당의 총함량은 SM이 1,389.88 mg%, HSM 1,013.51 mg%, WSM 1,539.51 mg%, HWSM 1,331.13 mg%로 효소 가수분해 처리 후 감소하였다. HSM과 HWSM의 DPPH free radical 소거활성은 각각 49.26%, 45.34%로 SM과 WSM에 비해 높은 활성을 보였으며, ACE 저해활성도 약 1.6배 높게 나타났다. Superoxide radical 소거활성은 HSM과 HWSM에서 높은 활성을 보였구, 원료 콩에서 비지제거에 따른 차이는 없었다. 이상의 결과 두유액 가수분해물(HSM) 및 전두유 가수분해물(HWSM)은 영양성분이나 기능적 특성이 두유액(SM)과 전두유액(WSM)에 비하여 우수한 것으로 나타나 향후 다양한 기능성 강화 소재로 활용이 기대된다.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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• pp.278-281
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• 2004

The $\alpha$1,6-mannosyltransferase encoded by Saccharomyces cerevisiae OCH1 plays a key role for the outer chain initiation of the N-linked oligosaccharides. A search for Hansenula polymorpha genes homologous to S. cerevisiae OCHI (ScOCH1) has revealed seven open reading frames (ORF100, ORF142, ORF168, ORF288, ORF379, ORF576, ORF580). All of the seven ORFs are predicted to be a type II integral membrane protein containing a transmembrane domain near the amino-terminal region and has a DXD motif, which has been found in the active site of many glycosyltransferases. Among this seven-membered OCH1 gene family of H. polymorpha, we have carried out a functional analysis of H. polymorpha ORF168 (HpOCH2) showing the highest identity to ScOCH1. Inactivation of this protein by disruption of corresponding gene resulted in several phenotypes suggestive of cell wall defects, including hypersensitivity to hygromycin B and sodium deoxycholate. The structural analysis of N-glycans synthesized in HpOCH2-disrupted strain (Hpoch2Δ) and the in vitro $\alpha$1,6-mannosyltransferase activity assay strongly indicate that HpOch2p is a key enzyme adding the first $\alpha$1,6-mannose residue on the core glycan Man$_{8}$GlcNAc$_2$. The Hpoch2Δ was further genetically engineered to synthesize a recombinant glycoprotein with the human compatible N-linked oligosaccharide, Man$_{5}$GlcNAc$_2$, by overexpression of the Aspergillus saitoi $\alpha$1,2-mannosidase with the 'HDEL” ER retention signal.gnal.