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- Identification of Proteolytic Bacteria from the Arctic Chukchi Sea Expedition Cruise and Characterization of Cold-active Proteases
- Ha Ju Park , Yung Mi Lee , Sunghui Kim , Ah Ram Wi , Se Jong Han , Han-Woo Kim , Il-Chan Kim , Joung Han Yim , Dockyu Kim
- J. Microbiol. 2014;52(10):825-833. Published online August 27, 2014
- DOI: https://doi.org/10.1007/s12275-014-4226-6
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Abstract
- Following collection of seawater samples during an Arctic Chukchi Sea expedition cruise of the Korean icebreaker Araon in 2012, a total of 15,696 bacteria were randomly isolated from Marine Broth 2216 agar plates. Of these, 2,526 (16%) showed proteolytic activity and were identified as mainly Alteromonas (31%), Staphylococcus (27%), and Pseudoalteromonas (14%). Among the proteolytic strains, seven were selected based on their significant ability to grow and produce a halo on skim milk plates at low temperatures (<5°C) owing to cold-active proteases. These strains were affiliated with the genus Pseudoalteromonas and were divided into three groups based on phylogenetic analysis of the 16S rRNA genes. Profiling cell membrane fatty acids confirmed the 16S rRNA-based differentiation and revealed the accordance between the two analyses. Seven genes for serine protease precursors were amplified from the corresponding strains, and based on sequence similarities, these genes were divided into three groups that were identical to those identified by the 16S rRNA phylogenetic analysis. Three protease genes from the representative strains of each group were composed of 2,127–2,130 bp, encoding 708–709 amino acids, and these genes yielded products with calculated molecular weights of approximately 72.3–72.8 kDa. Amino acid sequence analysis suggested that the precursors are members of the subtilase serine endo- and exo-peptidase clan and contain four domains (signal peptide, N-terminal prosequence, catalytic domain, and two pre-peptidase C-terminal domains). Upon expression in E. coli, each recombinant protease exhibited proteolytic activity on zymogram gels.
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Citations
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- The Arctic summer microbiome across Fram Strait: Depth, longitude, and substrate concentrations structure microbial diversity in the euphotic zone
Matthias Wietz, Anja Engel, Simon Ramondenc, Matomo Niwano, Wilken‐Jon von Appen, Taylor Priest, Anabel von Jackowski, Katja Metfies, Christina Bienhold, Antje Boetius
Environmental Microbiology.2024;[Epub] CrossRef - Diversity in the utilization of different molecular classes of dissolved organic matter by heterotrophic marine bacteria
Shira Givati, Elena Forchielli, Dikla Aharonovich, Noga Barak, Osnat Weissberg, Natalia Belkin, Eyal Rahav, Daniel Segrè, Daniel Sher, Jennifer F. Biddle
Applied and Environmental Microbiology.2024;[Epub] CrossRef - Benthic bacteria and archaea in the North American Arctic reflect food supply regimes and impacts of coastal and riverine inputs
Alexis M. Walker, Mary Beth Leigh, Sarah L. Mincks
Deep Sea Research Part II: Topical Studies in Oceanography.2023; 207: 105224. CrossRef - Cold-Adapted Proteases: An Efficient and Energy-Saving Biocatalyst
Zhengfeng Yang, Zhendi Huang, Qian Wu, Xianghua Tang, Zunxi Huang
International Journal of Molecular Sciences.2023; 24(10): 8532. CrossRef - Description of Polaribacter batillariae sp. nov., Polaribacter cellanae sp. nov., and Polaribacter pectinis sp. nov., novel bacteria isolated from the gut of three types of South Korean shellfish
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Journal of Microbiology.2022; 60(6): 576. CrossRef - Proteases from the marine bacteria in the genus Pseudoalteromonas: diversity, characteristics, ecological roles, and application potentials
Xiu-Lan Chen, Yan Wang, Peng Wang, Yu-Zhong Zhang
Marine Life Science & Technology.2020; 2(4): 309. CrossRef - Characterization of balofloxacin-stressed proteomics and identification of balofloxacin-binding proteins pre-peptidase and integration host factor in Edwardsiella tarda
Qi Wen, Xian-jie Liu, Wei-cong Zhu, Lu Li, Min-yi Li, Xuna-xian Peng, Hui Li
Journal of Proteomics.2019; 205: 103413. CrossRef - The Place for Enzymes and Biologically Active Peptides from Marine Organisms for Application in Industrial and Pharmaceutical Biotechnology
Jean-Étienne R.L. Morlighem, Gandhi Radis-Baptista
Current Protein & Peptide Science.2019; 20(4): 334. CrossRef - Benefit from decline: the primary transcriptome of Alteromonas macleodii str. Te101 during Trichodesmium demise
Shengwei Hou, Mario López-Pérez, Ulrike Pfreundt, Natalia Belkin, Kurt Stüber, Bruno Huettel, Richard Reinhardt, Ilana Berman-Frank, Francisco Rodriguez-Valera, Wolfgang R Hess
The ISME Journal.2018; 12(4): 981. CrossRef
- The Arctic summer microbiome across Fram Strait: Depth, longitude, and substrate concentrations structure microbial diversity in the euphotic zone
- Cryoprotective Properties of Exopolysaccharide (P-21653) Produced by the Antarctic Bacterium, Pseudoalteromonas arctica KOPRI 21653
- Sung Jin Kim , Joung Han Yim
- J. Microbiol. 2007;45(6):510-514.
- DOI: https://doi.org/2643 [pii]
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Abstract
- Twenty-five bacterial strains that secrete mucous materials were isolated from sediment obtained from King George Island, Antarctica. Seven of these strains proved capable of producing cryoprotective exopolysaccharides. The strain KOPRI 21653 was selected for the further study of an anti-ice-nucleating polysaccharide (ANP), which originated from a polar region. KOPRI 21653 was identified as Pseudoalteromonas arctica as the result of 16S rRNA analysis. The exopolysaccharide, P-21653, was purified completely from the KOPRI 21653 cell culture via column chromatography and protease treatment. The principal sugar components of P-21653 were determined to be galactose and glucose, at a ratio of 1:1.5, via GC-MS analysis. The cryoprotective activity of P-21653 was characterized via an E. coli viability test. In the presence of 0.1% (w/v) P-21653, the survival ratio of E. coli cells was as high as 82.6% over three repeated freeze-thaw cycles. The survival ratio decreased drastically to 71.5 and 48.1%, respectively, in five and seven repeated cycle conditions; however, the survival ratios were greater over three (96.6-92.1%) to seven (100.5-91.6%) freeze-thaw cycles in the presence of 0.5 and 1.0% (w/v) P-21653. In addition, at much lower concentrations (0.1-1.0%), P-21653 resulted in survival ratios (83.1-98.4%) similar to those of two commercially available cryoprotectants (VEG plus X-1000, 92.9% and VM3, 95.3%), which were utilized at the recommended concentrations (90%). The biochemical characteristics of exopolysaccharide P-21653 reflect that this compound may be developed as a useful cryoprotectant for use in medical applications and in the food industry.
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