Key Laboratory of Marine Bioactive Substances

Qingdao, China

Key Laboratory of Marine Bioactive Substances

Qingdao, China
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Che S.,First Institute of Oceanography | Che S.,Key Laboratory of Marine Bioactive Substances | Song W.,First Institute of Oceanography | Song W.,Key Laboratory of Marine Bioactive Substances | And 2 more authors.
Current Microbiology | Year: 2013

Temperature and salinity fluctuations are two of the most important factors affecting the growth of polar bacteria. In an attempt to better understand the function of heat-shock proteins (HSPs) in the adaptive mechanisms of the Antarctic psychrotrophic bacterium Psychrobacter sp. G to such conditions, genes Hsp845, Hsp2538, Hsp2666, and Hsp2667 were cloned on the basis of the draft genome. The expression characteristics of these HSP genes under different stress conditions were analyzed by the qRT-PCR method. Expression of Hsp845 and Hsp2667 was inhibited significantly by low temperature (0 and 10 C, respectively). There was no difference of expression when Hsp2538 and Hsp2666 were exposed to 0 C but the expression of Hsp2666 was inhibited when exposed to 10 C. Expression of Hsp2538 and Hsp2667 was not sensitive but expression of Hsp845 and Hsp2666 was increased at low salinity (0 and 15, respectively). Expression of the four HSP genes was enhanced at high salinity (90 and 120) and at high temperature independent of salinity. By contrast, low temperature had no significant effect independent of salinity. © 2013 Springer Science+Business Media New York.

He Y.-Y.,State Oceanic Administration | He Y.-Y.,Key Laboratory of Marine Bioactive Substances | Wang Y.-B.,State Oceanic Administration | Wang Y.-B.,Key Laboratory of Marine Bioactive Substances | And 18 more authors.
Current Microbiology | Year: 2017

Calmodulin (CaM) is a Ca2+-binding protein that plays a role in several Ca2+ signaling pathways, which dynamically regulates the activities of hundreds of proteins. The ice alga Chlamydomonas sp. ICE-L, which has the ability to adapt to extreme polar conditions, is a crucial primary producer in Antarctic ecosystem. This study hypothesized that Cam helps the ICE-L to adapt to the fluctuating conditions in the polar environment. It first verified the overall length of Cam, through RT-PCR and RACE-PCR, based on partial Cam transcriptome library of ICE-L. Then, the nucleotide and predicted amino acid sequences were, respectively, analyzed by various bioinformatics approaches to gain more insights into the computed physicochemical properties of the CaM. Potential involvements of Cam in responding to certain stimuli (i.e., UVB radiation, high salinity, and temperature) were investigated by differential expression, measuring its transcription levels by means of quantitative RT-PCR. Results showed that CaM was indeed inducible and regulated by high UVB radiation, high salinity, and nonoptimal temperature conditions. Different conditions had different expression tendencies, which provided an important basis for investigating the adaptation mechanism of Cam in ICE-L. © 2017, Springer Science+Business Media New York.

Wang Y.-B.,First Institute of Oceanography | Wang Y.-B.,Key Laboratory of Marine Bioactive Substances | Liu F.-M.,First Institute of Oceanography | Liu F.-M.,Key Laboratory of Marine Bioactive Substances | And 5 more authors.
Journal of Pure and Applied Microbiology | Year: 2014

Laser tweezers Raman spectroscopy (LTRS) can help with observing and studying individual cells or organelles in a natural state for a relatively long period. In this study, LTRS and GC-MS were used to report physiological metabolism of active psychrophilic petroleum hydrocarbon-degrading strains isolated from the Antarctic Ocean. Based on GC-MS analysis and analysis of the Raman spectrum of degradation productions, the results showed Planococcus sp.NJ41 and Shewanella sp.NJ49 degraded diesel, n-hexadecane, polycyclic aromatic hydrocarbons, and other petroleum hydrocarbons with high efficiency at low temperature (0°C-10°C). GC-MS showed the long straight-chain hydrocarbons were decomposed into short straight-chain hydrocarbons. The Raman spectrum showed there were more proteins and carbohydrate than lipids were produced during the Planococcus sp.NJ41 and Shewanella sp.NJ49 growth and degradation. Finally, the Raman intensity at 1512 cm-1 to 1514cm -1 represented β-carotene that was involved in the regulation of membrane fluidity in Antarctic bacteria, allowing it to adapt to the extreme low-temperature environment of the Antarctic.

Li J.,First Institute of Oceanography | Li J.,Key Laboratory of Marine Bioactive Substances | Sha Y.,First Institute of Oceanography | Sha Y.,Key Laboratory of Marine Bioactive Substances | And 5 more authors.
Journal of Microbiology and Biotechnology | Year: 2014

An extracellular agarase was purified from Bacillus sp. BI-3, a thermophilic agar-degrading bacterium isolated from a hot spring in Indonesia. The purified agarase revealed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular mass of 58 kDa. The optimum pH and temperature of the agarase were 6.4 and 70°C, respectively. The activity of the agarase was stable at high temperatures, and more than 50% activity was retained at 80°C for 15 min. Furthermore, the enzyme was stable in the pH range of 5.8-8.0, and more than 60% of the residual activity was retained. Significant activation of the agarase was observed in the presence of K+, Na+, Ca2+, Mg2+, and Sr2+; on the other hand, Ba2+, Zn2+, Cu2+, Mn2+, Co2+, Fe2+, and EDTA inhibited or inactivated the enzyme activity. The components of the hydrolytic product analyzed by thin-layer chromatography showed that the agarase mainly produced neoagarobiose. This study is the first to present evidence of agarolytic activity in aerobic thermophilic bacteria. © 2014 by The Korean Society for Microbiology and Biotechnology.

Shuo-Shuo C.,First Institute of Oceanography | Shuo-Shuo C.,Key Laboratory of Marine Bioactive Substances | Xue-Zheng L.,First Institute of Oceanography | Xue-Zheng L.,Key Laboratory of Marine Bioactive Substances | And 2 more authors.
Protein Expression and Purification | Year: 2011

The cold-active lipase gene Lip-948, cloned from Antarctic psychrotrophic bacterium Psychrobacter sp. G, was ligated into plasmid pColdI. The recombinant plasmid pColdI + Lip-948 was then transformed into Escherichia coli BL21. SDS-PAGE analysis showed that there was substantive expression of lipase LIP-948 in E. coli with a yield of about 39% of total protein, most of which was present in the inclusion body. The soluble protein LIP-948 only consisted of 1.7% of total LIP-948 with a specific activity of 66.51 U/mg. Co-expression of molecular chaperones with the pColdI + Lip-948 were also carried out. The results showed that co-expression of different chaperones led to an increase or decrease in the formation of soluble LIP-948 in varying degrees. Co-expression of pColdI + Lip-948 with chaperone pTf16 and pGro7 decreased the amount of soluble LIP-948, while the soluble expression was enhanced when pColdI + Lip-948 was co-expressed with "chaperone team" plasmids (pKJE7, pG-Tf2, pG-KJE8), respectively. LIP-948 was most efficiently expressed in soluble form when it was co-expressed with pG-KJE8, which was up to 19.8% of intracellular soluble proteins and with a specific activity of 108.77 U/mg. The soluble LIP-948 was purified with amylase affinity chromatography and its enzymatic characters were studied. The optimal temperature and pH of LIP-948 was 35 °C and 8, respectively. The activity of LIP-948 dropped dramatically after incubation at 50 °C for 15 min and was enhanced by Sr2+, Ca 2+. It preferentially hydrolyzed 4-nitrophenyl esters with the shorter carbon chain. © 2011 Elsevier Inc. All rights reserved.

Song W.,First Institute of Oceanography | Song W.,Key Laboratory of Marine Bioactive Substances | Lin X.,First Institute of Oceanography | Lin X.,Key Laboratory of Marine Bioactive Substances | And 2 more authors.
Polar Biology | Year: 2012

Low temperature is one of the major environmental challenges that Antarctic bacteria must face. Detailed studies of cold shock responses of cold-adapted microorganisms are still insufficient. Here, we cloned three cold shock protein (CSP) genes (Csp1137, Csp2039, and Csp2531) in the Antarctic bacterium Psychrobacter sp. G and their regulatory sequences were identified. The three CSPs were highly conserved with other known CspAs. qRT-PCR was performed to evaluate their expression characteristics under stress conditions, and the potential influence of regulatory sequences also was analyzed. The expression of Csp1137 was enhanced both by low (0, 10 °C) and high temperature (30 °C). The expression of Csp2039 was enhanced by low temperature (0 °C), but was lower than that of Csp1137. This can be explained by the absence in Csp2039 of the AT-rich UP element. Different from Csp1137, the expression of Csp2531 was inhibited by low temperature (0 °C), even with the presence of AT-rich UP element, and it was not sensitive to high temperature (30 °C). The expression of Csp1137 was enhanced by high salinity (90, 120), whereas that of Csp2531was enhanced by low salinity (0, 15). At 0 °C and a salinity of 15, the expression of Csp1137 was repressed initially, but then it increased greatly during the next 10 h. The expressions of Csp2039 and Csp2531 were repressed significantly under four different combinations of stress conditions. Our results showed that the role of the upstream regulation sequences were much more complex than previously thought. Also, gene expressions were also affected by the environmental salinity. These are helpful in further clarification of the adaptation mechanism of Psychrobacter sp. G. © 2012 Springer-Verlag.

Yi-Bin W.,State Oceanic Administration | Yi-Bin W.,Key Laboratory of Marine Bioactive Substances | Fang-Ming L.,State Oceanic Administration | Fang-Ming L.,Key Laboratory of Marine Bioactive Substances | And 10 more authors.
Journal of the Marine Biological Association of the United Kingdom | Year: 2016

Ice algae have successfully adapted to the extreme environmental conditions in the Antarctic, however the underlying mechanisms involved in the regulation and response of thylakoid membranes and chloroplast to low-temperature stress are still not well understood. In this study, changes in pigment concentrations, lipids, fatty acids and pigment protein complexes in thylakoid membranes and chloroplast after exposure to low temperature conditions were investigated using the Antarctic ice algae Chlamydomonas sp. ICE-L. Results showed that the chloroplasts of Chlamydomonas sp. ICE-L are distributed throughout the cell except in the nuclear region in the form of thylakoid lamellas which exists in the gap between organelles and the starch granules. Also, the structure of mitochondria has no obvious change after cold stress. Concentrations of Chl a, Chl b, monogalactosyl diacylglycerol, digalactosyl diacylglycerol and fatty acids were also observed to exhibit changes with temperature, suggesting possible adaptations to cold environments. The light harvesting complex, lutein and β-carotene played an important role for adaptation of ICE-L, and increasing of monogalactosyl diacylglycerol and digalactosyl diacylglycerol improved the overall degree of unsaturation of thylakoid membranes, thereby maintaining liquidity of thylakoid membranes. The pigments, lipids, fatty acids and pigment-protein complexes maintained the stability of the thylakoid membranes and the normal physiological function of Chlamydomonas sp. ICE-L. Copyright © Marine Biological Association of the United Kingdom 2016

Wang F.,Henan Agricultural University | Xu B.,Henan Agricultural University | Sun Y.,Henan Agricultural University | Zang J.,Key Laboratory of Marine Bioactive Substances | And 2 more authors.
Journal of Chinese Institute of Food Science and Technology | Year: 2013

A fungus named WNF-15A which secreted a special amaranthine pigment was isolated from soil collected from Antarctic. It was determined as a member of Geomyces based on colony morphologic characteristics, microscopic morphology and the phylogenetic analysis of ITS. And it was a psychrophiles which has optimum growth temperature between 15~20°C. The hydro soluble amaranthine pigment produced by WNF-15A has the maximum absorption at 533 nm. Its thermal degradation dynamics corresponded with Arrhenius empirical formula and the activation energy was 65.4~81.2 kJ/mol, and this pigment has a nearly higher thermostability than monacolin K and betacyanin. Changes of pH had little effect on the stability of the pigment that was most stable under pH3~9 condition. Although the pigment was faded quickly when it exposed to sunlight, scattered light was almost ineffective on its stability. This pigment was nontoxic determined by LD50>15000 mg/kg observed from mice toxicity test. This pigment could be largely exploited as a natural pigment.

Wang Y.B.,State Oceanic Administration | Wang Y.B.,Key Laboratory of Marine Bioactive Substances | Gao C.,Key Laboratory of Marine Bioactive Substances | Gao C.,Qingdao University of Science and Technology | And 7 more authors.
BioResources | Year: 2015

Immobilization is an effective way to solve the problem associated with the application of cold-active cellulase in industrial processes. In this study, a cold-active cellulase from the Antarctic psychrophilic bacterium Pseudoalteromonas sp. NJ64 was obtained, immobilized, and analyzed for optimal immobilization conditions. Then it was used in kelp cellulose ethanol fermentation, achieving a higher purity level of kelp cellulose ethanol. The enzymatic activity of this cold-active cellulase was 49.7 U/mL. The optimal immobilization process conditions were as follows: sodium alginate, 30 g/L; calcium chloride, 5 g/L; glutaraldehyde, 0.4%; and cross-linking time, 5 h. Under these conditions, the activity recovery rate was 51.58%. The optimum reaction temperature was at 40 °C, the optimum initial pH was 9.0, and the relative enzyme activity was 58.37% after being recovered seven times. A higher purity level of kelp cellulose ethanol has reached (37.37%). Immobilized cold-active cellulase can effectively hydrolyze the cellulose of kelp residue, which is a valuable component of cellulose bio-ethanol production and will have broad implications in the development of the ethanol industry in China.

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