• Journal of Microbiology and Biotechnology
• /
• 제15권4호
• /
• pp.800-808
• /
• 2005

Leuconostoc mesenteroides SY1, an isolate from kimchi, was able to ferment $\alpha$-galactosides, such as melibiose and raffinose. $\alpha$-Galactosidase ($\alpha$-Gal) activity was higher in cells grown on melibiose and raffinose than cells grown on galactose, sucrose, and fructose. $\alpha$-Gal activity was not detected in cells grown on glucose, indicating the operation of carbon catabolite repression (CCR). A 6 kb DNA fragment was PCR amplified using a primer set based on the nucleotide sequence of a putative $\alpha$-galactosidase gene (aga) from L. mesenteroides ATCC 8293. Nucleotide sequencing of the 6 kb fragment confirmed the presence of aga and other genes involved in the galactosides utilization, and the gene order was galR (transcriptional regulator)-aga-gaIK (galactokinase)-gaIT (galactose-1-phosphate uridylyltransferase). Northern blotting experiment showed that aga, gaIK, and gaIT constituted the same operon, that the transcription was induced by galactosides, such as melibiose and raffinose, whereas gaIR was independently transcribed as a monocistronic gene, and that the level of transcription was fairly constant. The aga was overexpressed in E. coli BL21 (DE3) using pET26b(+) vector, and $\alpha$-Gal was accumulated in E. coli as an inclusion body.

• Journal of Microbiology and Biotechnology
• /
• 제12권1호
• /
• pp.176-181
• /
• 2002

In order to isolate and identify Bifidobacterium spp. that originated in Korea, feces were sampled from healthy Korean adults and children living in three villages, the first having a history of longevity and the other two where the diet did not include fermented milk or any pharmaceutical preparations. Through the use of Gram staining and microscopic examination for cell morphology, 23 bacterial strains presumed to be the Bifidobacterium genus were isolated from the feces of 13 out of a total of 59 Korean people. To identify the Bifidobacterium strains at the genus level, these bacteria were then analyzed by TLC and the fructose-6-phosphate phosphoketolase (F6PPK) test. The result showed that 22 of the isolated strains were confirmed to be members of the genus Bifidobacterium. All of these bifidobacteria were also identified as Bifidobacterium spp. by the fermentation test. Using a RFLP analysis, an attempt was made to identify the Bifidobacterium spp. that had been isolated from both Korean adults and children. In a genomic Southern blot analysis after digestion with two restriction enzymes (EcoRI, HindIII), all of the 14 randomly selected Korean isolates showed patterns identical to those of three different B. longum species. Another restriction enzyme, CfoI (4-bp recognition enzyme), was then used to identify the strain. Interestingly, all the Korean isolates were identified as B. longum ATCC 15708, indicating that a RFLP analysis was effective for identifying Bifidobacterium spp. at both the strain and species levels.

• Biotechnology and Bioprocess Engineering:BBE
• /
• 제10권6호
• /
• pp.603-606
• /
• 2005

Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

• Animal Bioscience
• /
• 제36권3호
• /
• pp.506-520
• /
• 2023

Objective: Japanese Brown (JBR) cattle, especially the Kochi (Tosa) pedigree (JBRT), is a local breed of moderately marbled beef. Despite the increasing demand, the interbreed differences in muscle metabolites from the highly marbled Japanese Black (JBL) beef remain poorly understood. We aimed to determine flavor-related metabolites and postmortem metabolisms characteristic to JBRT beef in comparison with JBL beef. Methods: Lean portions of the longissimus thoracis (loin) muscle from four JBRT cattle were collected at 0, 1, and 14 d postmortem. The muscle metabolomic profiles were analyzed using capillary electrophoresis time-of-flight mass spectrometry. The difference in post-mortem metabolisms and aged muscle metabolites were analyzed by statistical and bioinformatic analyses between JBRT (n = 12) and JBL cattle (n = 6). Results: A total of 240 metabolite annotations were obtained from the detected signals of the JBRT muscle samples. Principal component analysis separated the beef samples into three different aging point groups. According to metabolite set enrichment analysis, post-mortem metabolic changes were associated with the metabolism of pyrimidine, nicotinate and nicotinamide, purine, pyruvate, thiamine, amino sugar, and fatty acid; citric acid cycle; and pentose phosphate pathway as well as various amino acids and mitochondrial fatty acid metabolism. The aged JBRT beef showed higher ultimate pH and lower lactate content than aged JBL beef, suggesting the lower glycolytic activity in postmortem JBRT muscle. JBRT beef was distinguished from JBL beef by significantly different compounds, including choline, amino acids, uridine monophosphate, inosine 5'-monophosphate, fructose 1,6-diphosphate, and betaine, suggesting interbreed differences in the accumulation of nucleotide monophosphate, glutathione metabolism, and phospholipid metabolism. Conclusion: Glycolysis, purine metabolism, fatty acid catabolism, and protein degradation were the most common pathways in beef during postmortem aging. The differentially expressed metabolites and the relevant metabolisms in JBRT beef may contribute to the development of a characteristic flavor.

• 원예과학기술지
• /
• 제30권4호
• /
• pp.463-470
• /
• 2012

최근 원예산업의 발달로 인해 모종의 연중 생산량이 증가되고 플러그 트레이를 이용한 양질의 모종을 편리하게 키울수 있는 육묘기술이 보급되면서 상토의 사용량이 크게 증가되었다. 본 연구에서는 4가지 점토광물 일라이트, 필라이트, 제올라이트, 벤토나이트의 상토 첨가 시 고추의 초기 성장에 대한 효과를 알아보았다. 또한 일라이트 처리 시 고추 잎과 줄기의 시료에 대한 프로테옴 분석을 수행하였다. 포트에 있는 상토에 4가지 점토광물을 각각 처리한 이후에 트레이서 재배된 모종 중에서 건강하고 규칙적인 크기를 가진 모종을 선별하여 포트에서 재배하였다. 본 연구는 6주 동안 충북대학교 유리 온실에서 수행되었다. 4가지 점토광물 처리에 따른 고추의 초장, 뿌리와 줄기의 생중량과 건중량은 무처리와 비교해서 모든 점토광물 처리구에서 높게 나타났다. 그리고 고추의 뿌리, 줄기, 잎에서 양이온 $K^+$, $Ca^{2+}$, $Mg^{2+}$의 흡수량은 무처리구보다 모든 점토광물 처리구에서 높게 나타났다. 고추 시료에 대한 프로테옴 분석을 실시한 결과 잎 조직에서는 에너지 대사에 관련 단백질인 plastid fructose-1, 6-bisphosphat aldolase class 1, aldolase 및 glyceraldehydes 3-phosphate dehydrogenase가 대조구보다 더 많이 발현되는 것을 보여주었다. 줄기 조직에서는 에너지 대사에 관련된 NAD-dependent formate dehydrogenase, potassium(K) 운송 단백질, 지베렐린 반응 조절에 관련된 GIA/RGA 단백질 등이 대조구보다 더 많이 발현되는 것을 보여주었다. 프로테옴 분석으로 얻어진 결과를 보면 일라이트 처리 시 특이하게 차별적으로 발현된 단백질이 고추의 초기 생장 증진에 관련된 것으로 사료된다. 그러므로 점토광물의 처리에 대한 채소작물의 반응에 관련된 몇몇 단백질의 동정은 그들의 분자적인 기작 구명을 이해하는 데 새로이 기여할 것으로 사료된다.

• 생명과학회지
• /
• 제34권7호
• /
• pp.465-475
• /
• 2024

Lactiplantibacillus plantarum K9은 굼벵이에서 분리된 다양한 생리활성물질에 기인하여 프로바이오틱스 균주로 활용 가능한 유산균이다. L. plantarum K9 유전체 분석결과로써 박테리아 염색체와 3 plasmid가 존재하는 것으로 나타났다. L. plantarum K9의 핵심 대사경로 분석 결과 해당과정, 오탄당대사(pentose phosphate pathway)는 정상적으로 수행되는 것으로 나타났다. 그러나 포도당신생합성과 ED pathway의 핵심 효소인 fructose-1,6-bisphosphatase (EC: 3.1.3.11)와 6-phosphogluconate dehydratase (EC: 4.2.1.12) / 2-keto-de- oxy-6-phosphogluconate (KDPG) aldolase (EC: 4.2.1.55)가 각각 결여되어 있기 때문에 포도당신생합성과 ED pathway는 수행하지 못하는 것으로 제의된다. 또한, TCA 회로에서 fumarate 및 malate를 형성하는 일부 효소만 존재하는 반면에 나머지 TCA 회로에 연관되는 효소들이 모두 결여되어 있었기 때문에 TCA 회로는 진행되지 못하는 것으로 추정되었다. 산화적 전자전달계는 NADH dehydrogenase complex I과 cytochrome reductase complex IV에 해당하는 요소들을 보유하고 있기 때문에 class IIB 타입(bd-type)의 전자전달시스템을 수행할 것으로 예측되었다. 종합적으로, L. plantarum K9은 lactic acid 동형발효를 수행하며, 포도당신생합성 및 오탄당대사가 가능하며, class IIB 타입(bd-type) 산화적 전자전달시스템에 의해 에너지 대사를 수행하는 것으로 제의된다. 따라서, L. plantarum K9은 다른 유산균주에 비교하여 lactic acid 생성량이 비교적 높아 생리활성도가 우수할 것으로 제의된다. 다른 한편으로, L. plantarum K9은 산화적 전자전달이 가능한 것으로 추정되어 산소에 대한 내성이 높아서 배양 특성이 양호하여 프로바이틱스로써 활용가능성이 높은 것으로 제의된다.

• Journal of Microbiology and Biotechnology
• /
• 제18권4호
• /
• pp.746-753
• /
• 2008

The hprK gene encoding bifunctional HPrK/P (kinase/phosphorylase) was cloned from L. mesenteroides SY1, a strain isolated from kimchi. hprK was transcribed as a monocistronic gene. His-tagged HPrH16A and HPrK/P were produced in E. coli BL21 (DE3) using pET26b(+) and purified. HPrK/P phosphorylation assay with purified proteins showed that the kinase activity of HPrK/P increased at slightly acidic pHs. Divalent cations such as $Mg^{2+}$ and $Mn^{2+}$ and glycolytic intermediates such as fructose-1, 6-bisphosphate (FBP) and phosphoenolpyruvate (PEP) increased the kinase activity of HPrK/P, but inorganic phosphate strongly inhibited it. Kinetic studies for the kinase activity of HPrK/P showed that the apparent $K_m$ values were 0.18 and $14.57{\mu}M$ for ATP and HPr, respectively. The $K_m$ value for the phosphorylase activity of HPrK/P was $14.16{\mu}M$ for P-Ser-HPr (HPr phosphorylated at the serine residue).

• 한국축산식품학회지
• /
• 제38권4호
• /
• pp.806-815
• /
• 2018

This study was performed to isolate some strains of Bifidobacterium breve from fecal materials of neonates and to screen them for the biotransformation activity of converting linoleic acid into conjugated linoleic acid (CLA). Fecal samples were collected from twenty healthy neonates between 14 and 100 days old, and four hundred colonies were randomly selected from a Bifidobacterium selective transoligosaccharide medium. A duplex polymerase chain reaction technique was developed for the rapid and accurate molecular characterization of the B. breve strains that have been reported to show the species-specific characteristic of CLA production. They are identified by 16S ribosomal DNA, fructose-6-phosphate phosphoketolase encoding genes (xfp), and rapid pulsed field gel electrophoresis. Thirty-six isolates were identified as B. breve, and just two of the 12 neonates were harboring B. breve strains. Each isolate showed different CLA-producing ability in the spectrophotometric assay. All of the positive strains from the primary spectrophotometric assay were confirmed for their CLA-producing activities using gas-chromatographic analysis, and their conversion rates were different, depending on the strain isolated in this study. Some strains of B. breve were successfully isolated and characterized based on the CLA-producing activity, and further studies are necessary to characterize the enzyme and the gene responsible for the enzyme activity.

• Molecular & Cellular Toxicology
• /
• 제4권2호
• /
• pp.164-169
• /
• 2008

Methylmercury (MeHg) is known to have devastating effects on the mammalian nervous system. In order to characterize the mechanism of MeHg-induced neurotoxicity, we investigated the analysis of transcriptional profiles on human 8k cDNA microarray by treatment of $1.4{\mu}M$ MeHg at 3, 12, 24 and 48h in human neuroblastoma SH-SY5Y cell line. Some of the identified genes by MeHg treatment were significant at early time points (3h), while that of others was at late time points (48h). The early response genes that may represent those involved directly in the MeHg response included pantothenate kinase 3, a kinase (PRKA) anchor protein (yotiao) 9, neurotrophic tyrosine kinase, receptor, type 2 gene, associated with NMDA receptor activity regulation or perturbations of central nervous system homeostasis. Also, when SH-SY5Y cells were subjected to a longer exposure (48h), a relative increase was noted in a gene, glutamine-fructose-6-phosphate transaminase 1, reported that overexpression of this gene may lead to the increased resistance to MeHg. To confirm the alteration of these genes in cultured neurons, we then applied real time-RT PCR with SYBR green. Thus, this result suggests that a neurotoxic effect of the MeHg might be ascribed that MeHg alters neuronal receptor regulation or homeostasis of neuronal cells in the early phase. However, in the late phase, it protects cells from neurotoxic effects of MeHg.

• 한국토양비료학회지
• /
• 제49권1호
• /
• pp.36-43
• /
• 2016

With transient flooding followed by poor or slow drainage plant roots may become reduction conditions because the root zone was fully filled with water. This study was examined the effects of calcium treatment in the early growth stage on biochemical changes in leaves and roots of melon (Cucumis melo L.) seedlings kept under flooding condition for 72 h. The activities of lactate dehydrogenase more gradually enhanced in the roots than those of leaves of melon seedlings treated with calcium. The activities of alcohol dehydrogenase associated with alcohol fermentation under low oxygen conditions continuously increased in the leaves and roots of seedlings untreated with calcium under flooding at least 72 h but those was constant within at least 12 h in treated with calcium. These results showed that calcium supplying in the early growth stage mitigated alcohol fermentation of melon seedlings kept under flooding condition for 72 h. Activities of nitrate reductase and acid phosphatase in the leaves and roots of seedlings in treated with calcium somewhat higher than those of non-treated with calcium. The activities of sucrose phosphate synthase and fructose-1,6-bisphosphatase of leaves of seedlings in treated with calcium more higher than those of non-treated with calcium. These results indicated that calcium nutrition mitigate the reduction of activities of some enzymes of melon seedling kept under flooding condition for 72 h.