An open reading frame coding for mannanase predicted from the partial genomic sequence of Paenibacillus woosongensis was cloned into Escherichia coli by polymerase chain reaction amplification, and completely sequenced. This mannanase gene, designated man26AT, consisted of 3,162 nucleotides encoding a polypeptide of 1,053 amino acid residues. Based on the deduced amino acid sequence, Man26AT was identified as a modular enzyme, which included a catalytic domain belonging to the glycosyl hydrolase family 26 and two carbohydrate-binding modules, CBM27 and CBM11. The amino acid sequence of Man26AT was homologous to that of several putative mannanases, with identity of 81% for P. ihumii and identity of less than 57% for other strains of Paenibacillus. A cell-free extract of recombinant E. coli carrying the man26AT gene showed maximal mannanase activity at $55^{\circ}C$ and pH 5.5. The enzyme retained above 80% of maximal activity after preincubation for 1 h at $50^{\circ}C$. Man26AT was comparably active on locust bean gum (LBG), galactomanan, and kojac glucomannan, whereas it did not exhibit activity on carboxymethylcellulose, xylan, or para-nitrophenyl-${\beta}$-mannopyranoside. The common end products liberated from mannooligosaccharides, including mannotriose, mannotetraose, mannopentaose, and mannohexaose, or LBG by Man26AT were mannose, mannobiose, and mannotriose. Mannooligosacchrides larger than mannotriose were found in enzymatic hydrolyzates of LBG and guar gum, respectively. However, Man26AT was unable to hydrolyze mannobiose. Man26AT was intracellularly degraded into at least three active proteins with different molecular masses by zymogram.
To increase the fibrinolytic activity and production of mycelium, extracts of 7 plant species were supplemented to the growth media of Armillaria mellea, and mycelial growth and enzymatic activity in the mycelium extracts of A. mellea were estimated. The mycelial production of A. mellea was slightly increased by adding ASH-R, UDVN or RGR extract, whereas KG extract significantly affected the growth. Supplement of ASH-S, UDVN and RGR extracts increased proteolytic activity from 36.8 to 46.1% Fibrinolytic activity was increased to $50{\sim}65%$ by supplement with RVS, ASH-S and RGR extracts, respectively. Enzyme extracts of the fungus grown with RGR extract supplement degraded all chains of fibrinogen within 2 hours, whereas control was required 3 hours. Degradation of fibrin fragments by the enzyme extracts was also observed through microscopy.
The characteristics of enzyme and gene for mannanase B had been reported from Cellulosimicrobium sp. YB-43 producing some kind of mannanase. A gene coding for the enzyme, named mannanase C (ManC), was expected to be located downstream of the manB gene. The manC gene was cloned by polymerase chain reaction and sequenced completely. From this nucleotide sequence, ManC was identified to consist of 448 amino residues and contain a carbohydrate binding domain CBM2 besides a catalytic domain, which was homologous to mannanase belonging to the glycosyl hydrolase family 5. The catalytic domain of ManC showed the highest amino acid sequence similarity of 55% with the mannanases from Streptomyces sp. SirexAA-E (55.8%; 4FK9_A) and S. thermoluteus (57.6%; BAM62868). The His-tagged ManC (HtManC) lacking N-terminal signal peptide with hexahistidine at C-terminus was produced and purified from cell extract of recombinant Escherichia coli. The purified HtManC showed maximal activity at $65^{\circ}C$ and pH 7.5, with no significant change in its activity at pH range from 7.5 to 10. HtManC showed more active on konjac and locust bean gum (LBG) than guar gum and ivory nut mannan (ivory nut). Vmax and Km values of the HtManC for LBG were 68 U/mg and 0.45 mg/ml on the optimal condition, respectively. Mannobiose and mannotriose were observed on TLC as major products resulting from the HtManC hydrolysis of mannooligosacharides. In addition, mannobiose and mannose were commonly detected as the hydrolyzed products of LBG, konjac and ivory nut.
This study was performed to assess the clay impact on alga growth which was a primary producer, in view of food chain in ecosystem. As clay minerals caused turbidity, a low sedimentation, high adsorption capacity with organic matter, adsorption - desorption effect with ionic chemicals, clay minerals were supposed to have a significant effect on the aquatic system. In study we tried to turn out NOAEL (No-observed-adverse-effect-level) of clay materials on the algae growth inhibition using such as kaolinite, sericite and montmorillonite. This study was indicated. (1) In both of kaolinite and sericite, the $72hr-EC_{50}$ of them shows 2,752 mg/L and 2,775 mg/L, respectively. (2) On the other hand, in the case of montmorillonite, the $72hr-EC_{50}$ is not shown a significant difference to that of control samples. (3) It can be explained that is also a very important parameter in an alga growth. Because an alga growth was increased when the permeability of W visible radiation was increased in all clay cases. (4) It is demonstrated alga growth was affected by the characteristics of clay materials. Hence we can assess the $\ulcorner$water environmental risk assessment caused clay materials$\lrcorner$ using the alga growth inhibition level indirectly.
MicroRNAs (miRNAs) are abundant in bovine milk and milk derived from other livestock, and they have functional roles in infants and in the secretion process of mammary glands. However, few studies have evaluated miRNAs in dairy processes, such as during cheese making and ripening. Thus, we investigated the characteristics of milk-derived miRNAs during the manufacturing and ripening of Camembert cheese as well as the microbiota present using the quantitative reverse transcription polymer chain reaction (RT-qPCR) and 16S rRNA pyrosequencing, respectively. Pyrosequencing showed that the cheese microbiota changed dramatically during cheese processing, including during the pasteurization, starter culture, and ripening stages. Our results indicated that the RNA contents per $200mg/200{\mu}l$ of the sample increased significantly during cheese-making and ripening. The inner cheese fractions had higher RNA contents than the surfaces after 12 and 22 days of ripening in a time-dependent manner (21.9 and 13.2 times higher in the inner and surface fractions than raw milk, respectively). We performed a comparative analysis of the miRNAs in each fraction by RT-qPCR. Large amounts of miRNAs (miR-93, miR-106a, miR-130, miR-155, miR-181a, and miR-223) correlated with immune responses and mammary glands were present in aged cheese, with the exception of miR-223, which was not present on the surface. Considerable amounts of miRNAs were also detected in whey, which is usually disposed of during the cheese-making process. Unexpectedly, there were no significant correlations between immune-related miRNAs and the microbial populations during cheese processing. Taken together, these results show that various functional miRNAs are present in cheese during its manufacture and that they are dramatically increased in amount in ripened Camembert cheese, with differences according to depth.
The ecological health, based on the Index of Biological Integrity (IBI) and Qualitative Habitat Evaluation Index (QHEI) was evaluated in 10 stream sites of Southern Han River. Eleven parameters of 12 parameters (Karr 1981) were modified for the application of regional Korean circumstance. The ecological health, based on IBI grade, was in "good condition" and the IBI score ranged from 33 to 47. Nine parameters of the original 12-parameter metrics in QHEI model (Plafkin et al. 1989) were applied in the habitat assessment. The mean QHEI model values were judged as "partially supporting" and ranged from 75 (non-supporting) to 109 (supporting). Comparative analyses revealed that values of IBI and QHEI models were greater in Gj stream than Ig- and Dn streams. The analysis of fish compositions showed that the proportions of insectivore, omnivore, and carnivore were 61.9%, 19%, and 9.5%, respectively. According to tolerance guild analysis, sensitive species and tolerant species were 76.1% and 4.7%, respectively, indicating a healthy trophic state in terms of food chain. The analysis by habitat guild type indicated that riffle benthic species dominated (57.1%) when compared to water column species (28.5%). The introduced species and individuals with diseases or external abnormality were not observed. Overall, the model values of IBI and QHEI suggested that the ecological health was maintained well in this upstream region.
Production of a high viscosity exoploysaccharide, methylan, by Methylobacterium organophilum from methanol was carried out in fed-batch cultures and the rheological properties of methylan fermentation broth were studied. Bacterial biomass showed little influence on viscosity, but the accumulation of methylan caused the increase of viscosity. With proceeding fermention, the viscosity at the same concentration of methylan was significantly increased and methylan solution showed slightly higher pseudoplasticity. The composition changes of methylan were investigated at various fermentation times. Contents of total sugar, reducing sugar and methylan were decreased but contents of acids(pyruvic acid, uronic acid and acetic acid) were increased with the culture time. It was considered that the increased content of acids resulted in the increase of the hyrodynamic domain in the solution due to charge repulsion. Consequently, the solution viscosity increased in propotion to the acids contents of methylan. Cell growth and methylan production were severely decreased by the limitation of dissolved oxygen. However, the cellular activity for methylan production was almost constant regardless of the level of dissolved oxygen. As a result, the high speed of agitation increased the methylan production, the specific production rate of methylan, and the methylan yield of the cell.
To investigate the spatio-temporal distributions of the mixotrophic dinoflagellate Yihiella yeosuensis in Korean coastal waters and its grazing impact on prey populations, water samples were seasonally collected from 28 stations in the East, West, and South Seas of Korea and Jeju Island from April 2015 to October 2018. The abundances of Y. yeosuensis in the water samples were quantified using quantitative real-time polymerase chain reaction (qPCR). Simultaneously, the physical and chemical properties of water from all sampled stations were determined, and the abundances of the optimal prey species of Y. yeosuensis, the prasinophyte Pyramimonas sp. and the cryptophyte Teleaulax amphioxeia, were quantified using qPCR. Y. yeosuensis has a wide distribution, as is reflected by the detection of Y. yeosuensis cells at 23 sampling stations; however, this distribution has a strong seasonality, which is indicated by its detection at 22 stations in summer but only one station in winter. The abundance of Y. yeosuensis was significantly and positively correlated with those of Pyramimonas sp. and T. amphioxeia, as well as with water temperature. The highest abundance of Y. yeosuensis was 48.5 cells mL-1 in Buan in July 2017, when the abundances of Pyramimonas sp. and T. amphioxeia were 917.6 and 210.4 cells mL-1, respectively. The growth rate of Y. yeosuensis on Pyramimonas sp., calculated by interpolating the growth rates at the same abundance, was 0.49 d-1, which is 37% of the maximum growth rate of Y. yeosuensis on Pyramimonas sp. obtained in the laboratory. Therefore, the field abundance of Pyramimonas sp. obtained in the present study can support a moderate positive growth of Y. yeosuensis. The maximum grazing coefficient for Y. yeosuensis on the co-occurring Pyramimonas sp. was 0.42 d-1, indicating that 35% of the Pyramimonas sp. population were consumed in 1 d. Therefore, the spatio-temporal distribution of Y. yeosuensis in Korean coastal waters may be affected by those of the optimal prey species and water temperature. Moreover, Y. yeosuensis may potentially have considerable grazing impacts on populations of Pyramimonas sp.
Kim, Ye-Hwan;Byun, Young Joon;Kim, Won Tae;Jeong, Pildu;Yan, Chunri;Kang, Ho Won;Kim, Yong-June;Lee, Sang-Cheol;Moon, Sung-Kwon;Choi, Yung-Hyun;Yun, Seok Joong;Kim, Wun-Jae
Journal of Korean Medical Science
/
v.33
no.47
/
pp.303.1-303.10
/
2018
Background: Cell division cycle 6 (CDC6) is an essential regulator of DNA replication and plays important roles in the activation and maintenance of the checkpoint mechanisms in the cell cycle. CDC6 has been associated with oncogenic activities in human cancers; however, the clinical significance of CDC6 in prostate cancer (PCa) remains unclear. Therefore, we investigated whether the CDC6 mRNA expression level is a diagnostic and prognostic marker in PCa. Methods: The study subjects included 121 PCa patients and 66 age-matched benign prostatic hyperplasia (BPH) patients. CDC6 expression was evaluated using real-time polymerase chain reaction and immunohistochemical (IH) staining, and then compared according to the clinicopathological characteristics of PCa. Results: CDC6 mRNA expression was significantly higher in PCa tissues than in BPH control tissues (P = 0.005). In addition, CDC6 expression was significantly higher in patients with elevated prostate-specific antigen (PSA) levels (> 20 ng/mL), a high Gleason score, and advanced stage than in those with low PSA levels, a low Gleason score, and earlier stage, respectively. Multivariate logistic regression analysis showed that high expression of CDC6 was significantly associated with advanced stage (${\geq}T3b$) (odds ratio [OR], 3.005; confidence interval [CI], 1.212-7.450; P = 0.018) and metastasis (OR, 4.192; CI, 1.079-16.286; P = 0.038). Intense IH staining for CDC6 was significantly associated with a high Gleason score and advanced tumor stage including lymph node metastasis stage (linear-by-linear association, P = 0.044 and P = 0.003, respectively). Conclusion: CDC6 expression is associated with aggressive clinicopathological characteristics in PCa. CDC6 may be a potential diagnostic and prognostic marker in PCa patients.
Ginseng has been recognized as a lifespan extending medicine which has been regarded as one of the medicines classified as top medicines, as the Boncho (medical herbs) study which is influenced by the idea of guidance's costume and food concept mainly in China is gaining its bona fide form. As the demand for ginseng has been expanded to other levels, the demand for ginseng has been increasing. Ginseng from the nature reached its supply chain limit due to its extinction and difficulty of picking, so it translated into ginseng cultivation of economy rather than harvesting in nature. After the start of ginseng cultivation, the ginseng cultivation was further enhanced by the rapid development of processing methods such as white-ginseng and red-ginseng, and the surge of consumption due to the traditional belief in ginseng drug efficacy and support of scientific research. In the Joseon Dynasty, the name Gasam (cultivated ginseng) had been created as ginseng was cultivated on farmland after the stage of SanYang (wild cultivated ginseng), the purpose of the new name Gasam is to differentiate from natural ginseng, and natural ginseng lost its firm position as the genuine ginseng as the Gasam replaced the genuine ginseng, and the natural ginseng got a new name of SanSam (wild ginseng). Because the real ginseng substance concept dissipated, and as Gasam is being called ginseng, the name Gasam was also disappeared. As a result, it was possible to grow large quantities according to the arrival of the Gasam era, and it was possible to supply the demand for ginseng, and it could become one agricultural industry. In this ginseng cultivation, in Japan where ginseng did not grow naturally, it was difficult to obtain ginseng from Joseon and faced with a shortage of ginseng at all times. Therefore, the shogun cultivated the Gasam systematically at the national level by the inside of the shogunate. However, since the natural ginseng is native to China and Korea, there is a concern about the deterioration of the quality of natural ginseng due to the incorporation of cultivated ginseng (Gasam). To protect the interests, the cultivation of ginseng was subject to control. For this reason, the lack of historical information on Gasam cultivation, which had to be started secretly, would be a natural result. In this paper, althouh not sufficient enough, the historical informations were used to summarize the history of ginseng cultivation in China, Japan and Korea.
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