Marine Biotechnology Research Center

Jinan, China

Marine Biotechnology Research Center

Jinan, China
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Kim O.-K.,Kyung Hee University | Kim O.-K.,Chonnam National University | Nam D.-E.,Kyung Hee University | Lee M.,Kyung Hee University | And 6 more authors.
Journal of Medicinal Food | Year: 2016

We have provided a protocol for establishing an atopic dermatitis (AD) in vitro model, and evaluated the effects of Costaria costata (CC) extracts on AD in an in vitro model using keratinocytes and splenocytes from AD-induced mice and mast cells. HaCaT cells were each treated with 200 μg/mL of CC water extract (CCW), CC 10% ethanol extract (CCE10%), and CC 70% ethanol extract (CCE70%), immediately followed by stimulation with 20 ng/mL tumor necrosis factor (TNF)-α, and 20 ng/mL interferon (IFN)-γ for inflammation. The splenocytes from AD-induced mice were each treated with 200 μg/mL of CCW, CCE10%, and CCE70%, followed by stimulation with 5 μg/mL ConA or lipopolysaccharide (LPS), to induce T cell or B-cell activation, and 5 μg/mL LPS and 50 ng/mL interleukin-4, to induce immunoglobulin (Ig) E production. We investigated the effects of CCW, CCE10%, and CCE70% on the production of histamine in PMA, and A23187-stimulated MC/9 cells. We found that treatments with CC extracts decreased the production of proinflammatory cytokines in TNF-α and IFN-γ-stimulated HaCaT cells, and the suppression of the imbalance of Th1/Th2 cytokines and IgE production on primary splenocytes. In addition, CC extracts resulted in a decrease in histamine release in the PMA and A23187-simulated MC/9 cells. According to our present results, we can conclude that CC extracts may be effective for the treatment of other allergy diseases, and AD, via the attenuation of allergic reactions. © Copyright Mary Ann Liebert, Inc. and Korean Society of Food Science and Nutrition 2016.

Ran L.-Y.,Shandong University | Ran L.-Y.,Marine Biotechnology Research Center | Su H.-N.,Shandong University | Su H.-N.,Marine Biotechnology Research Center | And 21 more authors.
Journal of Biological Chemistry | Year: 2014

Collagen is an insoluble protein that widely distributes in the extracellular matrix of marine animals. Collagen degradation is an important step in the marine nitrogen cycle. However, the mechanism of marine collagen degradation is still largely unknown. Here, a novel subtilisin-like collagenolytic protease, myroicolsin, which is secreted by the deep sea bacterium Myroides profundi D25, was purified and characterized, and its collagenolytic mechanism was studied. Myroicolsin displays low identity (<30%) to previously characterized subtilisin-like proteases, and it contains a novel domain structure. Protein truncation indicated that the Pro secretion system C-terminal sorting domain in the precursor protein is involved in the cleavage of the N-propeptide, and the linker is required for protein folding during myroicolsin maturation. The C-terminal β-jelly roll domain did not bind insoluble collagen fiber, suggesting that myroicolsin may degrade collagen without the assistance of a collagen-binding domain. Myroicolsin had broad specificity for various collagens, especially fish-insoluble collagen. The favored residue at the P1 site was basic arginine. Scanning electron microscopy and atomic force microscopy, together with biochemical analyses, confirmed that collagen fiber degradation by myroicolsin begins with the hydrolysis of proteoglycans and telopeptides in collagen fibers and fibrils. Myroicolsin showed strikingly different cleavage patterns between native and denatured collagens. A collagen degradation model of myroicolsin was proposed based on our results. Our study provides molecular insight into the collagen degradation mechanism and structural characterization of a subtilisin-like collagenolytic protease secreted by a deep sea bacterium, shedding light on the degradation mechanism of deep sea sedimentary organic nitrogen. © 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

Morin C.,Université de Sherbrooke | Fortin S.,Marine Biotechnology Research Center | Cantin A.M.,Université de Sherbrooke | Rousseau E.,Université de Sherbrooke
American Journal of Respiratory Cell and Molecular Biology | Year: 2011

The effects of a newly synthesized docosahexaenoic acid (DHA) derivative, CRBM-0244, on lung inflammation and airway hyperresponsiveness were determined in an in vitro model of TNF-α-stimulatedhuman bronchi and in an in vivo model of allergic asthma. Mechanical tension measurements revealed that CRBM-0244 prevented bronchial hyperresponsiveness in TNF-α-pretreated human bronchi.Moreover, treatmentwithCRBM-0244 resultedin a decrease in NF-κB activation and cyclooxygenase-2 (COX-2) overexpression triggered by TNF-α. The inhibition of peroxisome proliferator-activated receptor-γ with GW9662 abolished the CRBM-0244-mediated anti-inflammatory effects. CRBM-0244 reduced the Ca2+ sensitivity of bronchial smooth muscle through a decrease in the phosphorylation and expression of the PKC-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase of 17 kDa (CPI-17). Results also revealed an overexpression of CPI-17 protein in lung biopsies derived from patients with asthma. Furthermore, the presence of specialized enzymes such as 5-lipoxygenase and 15- lipoxygenase in the lung may convert CRBM-0244 into active mediators, leading to the resolution of inflammation. The in vivo anti-inflammatory properties of CRBM-0244 were also investigated in a guinea pig model of allergic asthma. After oral administration of CRBM-0244, airway leukocyte recruitment, airwaymucus, ovalbuminspecific IgE, and proinflammatory markers such as TNF-α and COX-2 were markedly reduced. Hence, CRBM-0244 treatment prevents airway hyperresponsiveness, Ca2+ hypersensitivity, and the overexpression of CPI-17 in lung tissue. Together, these findings provide key evidence regarding the mode of action of CRBM-0244 in the lung, and point to new therapeutic strategies for modulating inflammation in patients with asthma.

Liu A.,Shandong University | Liu A.,Marine Biotechnology Research Center | Mi Z.-H.,Shandong University | Mi Z.-H.,Marine Biotechnology Research Center | And 16 more authors.
Frontiers in Microbiology | Year: 2016

Most marine bacteria secrete exopolysaccharide (EPS), which is important for bacterial survival in the marine environment. However, it is still unclear whether the self-secreted EPS is involved in marine bacterial motility. Here we studied the role of EPS in the lateral flagella-driven swarming motility of benthic bacterium Pseudoalteromonas sp. SM9913 (SM9913) by a comparison of wild SM9913 and ΔepsT, an EPS synthesis defective mutant. Reduction of EPS production in ΔepsT did not affect the growth rate or the swimming motility, but significantly decreased the swarming motility on a swarming plate, suggesting that the EPS may play a role in SM9913 swarming. However, the expression and assembly of lateral flagella in ΔepsT were not affected. Instead, ΔepsT had a different swarming behavior from wild SM9913. The swarming of ΔepsT did not have an obvious rapid swarming period, and its rate became much lower than that of wild SM9913 after 35 h incubation. An addition of surfactin or SM9913 EPS on the surface of the swarming plate could rescue the swarming level. These results indicate that the self-secreted EPS is required for the swarming of SM9913. This study widens our understanding of the function of the EPS of benthic bacteria. © 2016 Liu, Mi, Zheng, Yu, Su, Chen, Xie, Zhou, Zhang and Qin.

Patra A.K.,Marine Biotechnology Research Center | Patra A.K.,Korean University of Science and Technology | Cho H.H.,Marine Biotechnology Research Center | Kwon Y.M.,Marine Biotechnology Research Center | And 9 more authors.
Ocean Science Journal | Year: 2016

Vestimentiferan tubeworms acquire their symbionts through horizontal transmission from the surrounding environment. In the present study, we constructed a 16S rRNA gene clone library to investigate the phylogenetic relationship between diverse microbes in the sediment and symbiotic bacteria in the trophosome of the tubeworm, Lamellibrachia satsuma, from Kagoshima Bay, Japan. Two symbiotic bacterial phylotypes belonging to the classes γ- and ε-Proteobacteria were found from this tubeworm trophosome. They were very closely related to the symbionts of several other marine invertebrates. The most predominant bacteria in the sediment were ε-Proteobacteria. A broad diversity of bacteria belonged to non-proteobacterial phyla such as Planctomycetes, Acidobacteria, and Chloroflexi was observed. The presence of sulfur oxidizers (i.e., ε-Proteobacteria and γ-Proteobacteria) and sulfur reducers (i.e., δ-Proteobacteria) may play a significant role in the sulfur cycle in these habitats and provide multiple sources of nutrition to the cold-seep communities. Closely related clones of ε-Proteobacteria symbiont in the species level and of γ-Proteobacteria symbiont in the genus level were found in the surrounding sediment. The similarity of symbiont clones of L. satsuma with other symbionts and free-living bacteria suggests the possibility of opportunistic symbiosis in ε-Proteobacteria and the co-evolution of γ-Proteobacteria having occurred after symbiosis with the tubeworms. © 2016, Korea Ocean Research & Development Institute (KORDI) and the Korean Society of Oceanography (KSO) and Springer Science+Business Media Dordrecht.

PubMed | Marine Biotechnology Research Center, Jeju National University, Vietnam Academy of Science and Technology and Chonbuk National University
Type: Journal Article | Journal: Food & function | Year: 2016

We investigated the anti-obesity effects of the potential prebiotic, laminarin, on mice fed a high-fat diet. A metagenomics approach was applied to characterize the ecological and functional differences of gut microbiota among mice fed a normal diet (CTL), a high-fat diet (HFD), and a laminarin-supplemented high-fat diet (HFL). The HFL mice showed a slower weight gain than the HFD mice during the laminarin-feeding period, but the rate of weight gain increased after the termination of laminarin supplementation. Gut microbial community analysis showed clear differences between the CTL and HFD mice, whereas the HFL mice were between the two. A higher abundance of carbohydrate active enzymes was observed in the HFL mice compared to the HFD mice, with especially notable increases in glycoside hydrolase and polysaccharide lyases. A significant decrease in Firmicutes and an increase in the Bacteroidetes phylum, especially the genus Bacteroides, were observed during laminarin ingestion. Laminarin ingestion altered the gut microbiota at the species level, which was re-shifted after termination of laminarin ingestion. Therefore, supplementing laminarin could reduce the adverse effects of a high-fat diet by shifting the gut microbiota towards a higher energy metabolism. Thus, laminarin could be used to develop anti-obesity functional foods. Our results also suggest that laminarin would need to be consumed regularly in order to prevent or manage obesity.

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