Time filter

Source Type

Zhou G.,Third Institute of Oceanography | Jin M.,Third Institute of Oceanography | Cai Y.,Inspection and Quarantine Technology Center | Zeng R.,Third Institute of Oceanography | Zeng R.,Utilization Collaborative Innovation Center
International Journal of Biological Macromolecules

A thermostable α-amylase (designated as Amy16) has been previously identified in Flammeovirga pacifica isolated from deep-sea sediments. The DNA sequence of Amy16 exhibited no significant similarity with those of any known protein, including the glycoside hydrolases. Amino acid sequence analysis revealed that Amy16 belonged to GH13 family and possessed a conserved DXEXD motif, which was essential for its hydrolysis activities. The recombinant Amy16 purified with Ni+ affinity column after its heterologous expression in Escherichia coli cells was most active at 50°C and retained more than 81% of its initial activity after incubation at 60°C for 20min. The optimal pH for Amy16 was determined to be 6.5, and a good tolerance to alkaline environment was observed. Low concentration of Mg2+, Sr2+, Na+ and K+ slightly increased the activity of Amy16. Results of thin layer chromatography experiments revealed that Amy16 was able to hydrolyse starch into maltose in a time-dependent manner, suggesting that Amy16 is a liquid-type endoenzyme with starch hydrolysis activities. Therefore, our study presented thermostable and alkali-stable Amy16, which may be suitable for use as an additive in detergents. © 2015 Elsevier B.V. Source

Xu F.,CAS South China Sea Institute of Oceanology | Xu F.,University of Chinese Academy of Sciences | Xu F.,Guangdong Laboratory Animals Monitoring Institute | Li J.,CAS South China Sea Institute of Oceanology | And 8 more authors.
Fish and Shellfish Immunology

Inhibitor of NF-κB (IκB), the important regulator of NF-κB/Rel signaling pathway, plays the crucial role in immune response of both vertebrates and invertebrates. Here, a novel homologue of IκB was cloned from Crassostrea gigas, and designated as CgIκB3. The complete CgIκB3 cDNA was 1282 bp in length, including a 942 bp open reading frame (ORF), a 51 bp 5' UTR and a 289 bp 3' UTR. The ORF encodes a putative protein of 313 amino acids with a predicted molecular weight of approximately 34.7 kDa. Sequence analysis reveals that CgIκB3 contains a conserved degradation motif but with only five ankyrin repeats. Neither a PEST domain nor a C-terminal casein kinase II phosphorylation site was identified through either alignment or bioinformatic prediction. Phylogenetic analysis suggested that CgIκB3 shares common ancestor with CgIκB1 rather CgIκB2, and theoretically it may originate from one duplication event prior to divergence of CgIκB1 and CgIκB2. Tissue expression analyses demonstrated that CgIκB3 mRNA is the most abundant in gills and heart. The expression following PAMP infection showed that CgIκB3 was significantly up-regulated in a similar pattern when challenged with LPS, HKLM or HKVA, respectively. Moreover, similar to CgIκB1 and CgIκB2, CgIκB3 can also inhibit Rel dependent NF-κB activation in HEK293 cells in a dose-dependent manner. In summary, these findings suggest that CgIκB3 can be as the functional inhibitor of NF-κB/Rel and involved in the host defense of C. gigas. The discovery of the third IκB emphasizes the complexity and importance of the regulation on NF-κB activation. © 2015 Elsevier Ltd. Source

Li Q.,Ningbo University | Li Q.,State Oceanic Administration | Li Q.,Utilization Collaborative Innovation Center | Ao J.,State Oceanic Administration | And 11 more authors.
Fish and Shellfish Immunology

Two cysteine proteases, cathepsin S (CatS) and cathepsin L (CatL), have been identified as the key enzymes involved in the processing of invariant chain (Ii chain) in mammals. However, little is known about the roles of fish cathepsins in the Ii chain processing. In this study, large yellow croaker cathepsin S (LycCatS) and L (LycCatL) were identified and characterized. Based on the sequence comparison and phylogenetic analysis, both LycCatS and LycCatL are highly conserved to their counterparts in teleost. These two cathepsins were constitutively expressed in all tissues and immune-related cells tested, although at different levels. Both recombinant LycCatS (rLycCatS) and LycCatL (rLycCatL) possess the typical cysteine protease activity. Like other mammalian endopeptidase cathepsins, rLycCatS and rLycCatL could be autocatalytically activated to remove propeptides and release active mature peptides. On the other hand, the autocatalytic activation of rLycCatL could be inhibited by recombinant large yellow croaker Ii chain (rLyc-TR-Ii), but the autocatalytic activation of rLycCatS was not affected by rLyc-TR-Ii. Furthermore, the activated rLycCatS can efficiently process rLyc-TR-Ii in a stepwise manner in vitro, while the activated rLycCatL can not. These data indicate that cathepsin S may be the main cathepsin involved in the Ii chain processing in bony fish. © 2015 Elsevier Ltd. Source

Discover hidden collaborations