• 한국미생물·생명공학회지
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• 제52권2호
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• pp.152-162
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• 2024

The microbial starter used to produce kefir beverages, kefir grain, contains a microbial exopolysaccharide called kefiran. Kefir grain consisting of water-insoluble polysaccharides, proteins, and fats, which can be applied as a multi-functional biopolymer. The mass of kefir grain can increase in the fermentation process of Kefir, but it is considered very slow. The purpose of this research is to study the impact of ammonium sulfate supplementation and yeast extract on reconstituted skim milk to increase the mass kefir grain and physical properties of kefiran. Results showed that the ammonium sulfate-supplemented substrate increased the mass of kefir grain by 547% in 14 days, with the condition that the substrate must be renewed every 2 days. Refreshing the substrate is considered one of the important factors. Supplementation on substrate did not appear to affect the viability of bacterial and yeast cells. Kefir grain produced from supplemented substrate also yields better thermal stability properties and has more functional groups than without supplementation. Two Lacticaseibacillus rhamnosus (RAL27 and RAL43) and one Limosilactobacillus fermentum (RAL29) were found to produce EPS. The three isolates also showed good skim milk fermentation ability after purification from kefir grain. The kefir grain produced in this study has the potential for wider application. This study also showed that kefir grain can be adjusted in quantity and quality through fermentation substrate engineering.

• Molecules and Cells
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• 제22권3호
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• pp.343-352
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• 2006

Sialic acid is a sugar typically found at the N-glycan termini of glycoproteins in mammalian cells. Lec3 CHO cell mutants are deficient in epimerase activity, due to a defect in the gene that encodes a bifunctional UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE). Sialic acid modification on the cell surface is partially affected in these cells. We have mutagenized Lec3 CHO cells and isolated six mutants (termed C2m) deficient in the cell surface expression of polysialic acid (PSA). Mutant C2m9 was partially defective in expression of cell-surface PSA and wheat germ agglutinin (WGA) binding, while in the other five mutants, both cell-surface PSA and WGA binding were undetectable. PSA expression was restored by complementation with the gene encoding the CMP-sialic acid transporter (CST), indicating that CST mutations were responsible for the phenotypes of the C2m cells. We characterized the CST mutations in these cells by Northern blotting and RT-PCR. C2m9 and C2m45 carried missense mutations resulting in glycine to glutamate substitutions at amino acids 217 (G217E) and 256 (G256E), respectively. C2m13, C2m39 and C2m31 had nonsense mutations that resulted in decreased CST mRNA stability, and C2m34 carried a putative splice site mutation. PSA and CD15s expression in CST-deficient Lec2 cells were partially rescued by G217E CST, but not by G256E CST, although both proteins were expressed at similar levels, and localized to the Golgi. These results indicate that the novel missense mutations isolated in this study affect CST activity.

• 한국균학회지
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• 제23권4호통권75호
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• pp.325-331
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• 1995

상황 (Phellinus linteus) 의 균사체 생장과 다당류 생산 및 다당류에서의 단당류 구성비에 대한 배양 조건의 영향을 조사하였다. 여러 가지 배지를 사용하여 실험한 결과 GP(glucose-peptone)배지에서 균사 생장이 가장 양호하여, 이 배지를 사용하여 탄소원 및 pH 영향을 조사하였다. 탄소원중 glucose에서 최대 균사체 생장 (0.9 g/100 ml)을 보였고, 다당류의 수율은 $5{\sim}14%$ 로서 mannose에서 최대이었다. 최적 pH는 7 이였으며, 산성 pH에서는 비교적 좋은 생장을 보였으나, 알칼리 pH에서는 생장이 급격히 감소하였다. 한편 pH에 따른 다당류 수율은 $5{\sim}8%$로 큰 변화를 보이지 않았다. 또한 탄소원 및 pH에 따른 단당류 구성비의 변화가 관찰되었다. 즉 탄소원에 따라서 다당류에서 glucose, mannose, 및 galactose 함량은 각각 $80{\sim}95%,\;3{\sim}12%$, 그리고 $2{\sim}10%$이었다. 한편 pH 변화에 따라서, glucose는 $83{\sim}92%$, mannose 는 $4{\sim}8%$, 그리고 galactose는$5{\sim}8%$의 구성비를 보였다. 이 결과는 배양 조건에 의한 항암면역활성 다당류의 구조의 변환 및 생산 증강이 가능함을 시사한다.

• Asian-Australasian Journal of Animal Sciences
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• 제16권5호
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• pp.748-763
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• 2003

Anaerobic rumen microorganisms mainly bacteria, protozoa and fungi degrade ligno-cellulosic feeds consumed by the ruminants. The ruminants in developing countries are predominantly maintained on low grade roughage and grazing on degraded range land resulting in their poor nutrient utilization and productivity. Hence, manipulation of rumen fermentation was tried during last two decades to optimize ruminal fermentation for improving nutrient utilization and productivity of the animals. Modification of rumen microbial composition and their activity was attempted by using chemical additives those selectively effect rumen microbes, introduction of naturally occurring or genetically modified foreign microbes into the rumen and genetically manipulation of existing microbes in the rumen ecosystem. Accordingly, rumen protozoa were eliminated by defaunation for reducing ruminal methane production and increasing protein outflow in the intestine, resulting in improve growth and feed conversion efficiency of the animals. Further, Interspecies trans-inoculation of rumen microbes was also successfully used for annulment of dietary toxic factor. Additionally, probiotics of bacterial and yeast origin have been used in animal feeding to stabilize rumen fermentation, reduced incidence of diarrhoea and thus improving growth and feed conversion efficiency of young stalk. It is envisaged that genetic manipulation of rumen microorganisms has enormous research potential in developing countries. In view of feed resource availability more emphasis has to be given for manipulating rumen fermentation to increase cellulolytic activity for efficient utilization of low grade roughage.

• Journal of Microbiology and Biotechnology
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• 제30권4호
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• pp.477-481
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• 2020

Viability plays an important role in the beneficial microbes (probiotics) to produce health benefits. However, this idea has been changed after the invention of the term "paraprobiotics," indicating that non-viable microbes could produce health benefits similar to those produced by live probiotics. Occasionally, it might be dangerous to administer live probiotics to people with weak immunity. In such cases, ingestion of paraprobiotics could be a potential alternative. The definition of paraprobiotics refers to the use of inactivated (non-viable) microbial cells or cell fractions to provide health benefits to the consumer. Paraprobiotics have attracted much attention because of their long shelf life, safety, and beneficial effects, such as modulation of immunity, modification of biological responses, reduction of cholesterol, anti-inflammatory, and antiproliferative properties. These features indicate that paraprobiotics may play a vital role in improving the health of the consumer by enhancing particular physiological functions, even though the exact underlying mechanisms have not yet been completely elucidated. In this mini-review, we briefly discuss the historical backgrounds of paraprobiotics and evidence of their health-promoting effects, prophylactic, and therapeutic properties.

• Plant Biotechnology Reports
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• 제5권3호
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• pp.289-295
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• 2011

The SpoU family of proteins catalyzes the methylation of transfer RNAs (tRNAs) and ribosomal RNAs (rRNAs). We characterized a putative tRNA/rRNA methyltransferase, VaSpoU1 of the SpoU family, from Viscum album (mistletoe). VaSpoU1 and other plant SpoU1s exhibit motifs of the SpoU methylase domain that are conserved with bacterial and yeast SpoU methyltransferases. VaSpoU1 transcripts were detected in the leaves and stems of V. album. VaSpoU1-GFP fusion proteins localized to both chloroplasts and mitochondria in Arabidopsis protoplasts. Sequence analysis similarly predicted that the plant SpoU1 proteins would localize to chloroplasts and mitochondria. Interestingly, mitochondrial localization of VaSpoU1 was inhibited by the deletion of a putative N-terminal presequence in Arabidopsis protoplasts. Therefore, VaSpoU1 may be involved in tRNA and/or rRNA methylation in both chloroplasts and mitochondria.

• 미생물학회지
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• 제39권4호
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• pp.230-234
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• 2003

Bacillus amyloliquefaciens psychrotrophic strain이 분비하는 세포 외 단백질 가수분해효소를 정제하여, endopeptidase 활성에 관한 특성을 분석하였다. Protease SE910로 명명된 효소는 단백질 내부의 leucine에 연결된 peptide 결합만을 가수분해하는 endopeptidase로 작용한다. 효소를 특이한 아미노산 잔기에 작용하는 화학수식제와 반응하였을 때 효소의 활성부위에 관여하는 아미노산 잔기가 수식되었을 때는 효소활성이 저해를 받는다. 본 효소는 serine을 수식하는 PMSF에의해 endopeptidase 활성이 완전히 저해되었으며, 카르복실 기능기를 수식하는 화학수식제에 의해 저해되었고, lysine을 화학수식하는 PLP에 의해서는 큰 영향을 받지 않았다. 이것은 본 효소의 endopeptidase 활성에 serine과 aspartic acid 잔기가 관여하는 것을 의미한다. 구조적으로 leucine을 포함하는 유도체인 bestatin은 효소의 endopeptidase 활성을 경쟁적으로 저해하였다.

• Journal of Plant Biotechnology
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• 제45권4호
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• pp.315-321
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• 2018

Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

• Mycobiology
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• 제50권5호
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• pp.357-365
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• 2022

Schizophyllum commune has emerged as the most promising model mushroom to study developmental stages (mycelium, primordium), which are two primary processes of fruit body development. Long non-coding RNA (lncRNA) has been proved to participate in fruit development and sex differentiation in fungi. However, potential lncRNAs have not been identified in S. commune from mycelium to primordium developmental stages. In this study, lncRNA-seq was performed in S. commune and 61.56 Gb clean data were generated from mycelium and primordium developmental stages. Furthermore, 191 lncRNAs had been obtained and a total of 49 lncRNAs were classified as differently expressed lncRNAs. Additionally, 26 up-regulated differently expressed lncRNAs and 23 down-regulated between mycelium and primordia libraries were detected. Further, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that differentially expressed lncRNAs target genes from the MAPK pathway, phosphatidylinositol signal, ubiquitin-mediated proteolysis, autophagy, and cell cycle. This study provides a new resource for further research on the relationship between lncRNA and two developmental stages (mycelium, primordium) in S. commune.

• 동아시아식생활학회지
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• 제21권3호
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• pp.392-400
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• 2011

본 연구는 선선편이 농산물 및 가공환경 중에 발생하는 미생물학적 위해 요소를 분석하여 공정 중 오염을 유발하는 지점을 재분석하고, 현재 사용되는 HACCP 계획서의 CCP 검증을 통하여 새로운 HACCP 가이드라인을 제시하였다. 본 연구의 대상인 A업체의 공장 내의 환경 오염도를 나타내는 공중 낙하균의 구획 분리 등의 관리로 인하여 $10^1$ log CFU/plate/15 min 이하의 대체적으로 낮은 오염도를 보였다. 기구 및 설비에 대한 표면 오염도를 검사 한 결과, E. coli는 검출되지 않았으며, 기구 및 설비의 총균수와 대장균군 오염도는 1~2 log CFU/100 $cm^2$ 수준을 보였다. 그러나 재료가 직접적으로 닿는 기구 중 원재료를 다듬는 도마($4.20{\pm}2.12$ log CFU/$cm^2$)와 세척 후 이용하는 탈수기 의 표면($4.57{\pm}0.92$ log CFU/$cm^2$)의 총균수의 오염도가 높게 나와 도마와 탈수기를 통해 교차 오염의 위험성이 높은 것을 확인할 수 있었다. A업체의 주요 생산 품목인 양상추와 양배추의 공정별 미생물 감소 효과를 본 결과, 양상추와 양배추 원재료의 총균수는 5~6 log CFU/g, 대장균군은 2.5 log CFU/g 정도의 오염도를 보였으며, 박피 및 절단, 1차 세척, 2차 세척, 건조의 과정을 거친 후의 최종 완제품에 대하여 양상추와 양배추의 총균수는 3~4 log CFU/g, 대장균군은 1~2 log CFU/g의 오염도를 보여 공정 과정을 거치는 동안 미생물이 감소하는 것을 확인할 수 있었다. 그러나 2차 세척과 탈수의 과정에서 미생물의 오염도가 유지 또는 증가하여 2차 세척과 탈수 공정에 대한 관리가 필요하였다. 공정 환경과 공정 효과에 대한 미생물학적 위해 요소를 분석한 결과, 기존의 HACCP 계획서에서 지정한 CCP(1차 세척공정, 금속 검출 공정)외에 세 개의 CP(2차 세척 공정, 탈수공정, 완제품의 저장)를 추가 설정한 새로운 HACCP 가이드라인을 제안하였다.