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Expression and Characterization of a Thermostable Carrageenase From an Antarctic Polaribacter sp. NJDZO3 Strain
Gui, Yuanyuan1,2,3; Gu, Xiaoqian4; Fu, Liping2,3; Zhang, Qian2,3; Zhang, Peiyu1; Li, Jiang2,3
2021-03-12
发表期刊FRONTIERS IN MICROBIOLOGY
ISSN1664-302X
卷号12页码:11
通讯作者Li, Jiang(lijiang@fio.org.cn)
摘要The complete genome of Polaribacter sp. NJDZ03, which was isolated from the surface of Antarctic macroalgae, was analyzed by next-generation sequencing, and a putative carrageenase gene Car3206 was obtained. Car3206 was cloned and expressed in Escherichia coli BL21(DE3). After purification by Ni-NTA chromatography, the recombinant Car3206 protein was characterized and the antioxidant activity of the degraded product was investigated. The results showed that the recombinant plasmid pet-30a-car3206 was highly efficiently expressed in E. coli BL21(DE3). The purified recombinant Car3206 showed a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular weight of 45 kDa. The optimum temperature of the recombinant Car3206 was 55 degrees C, and it maintain 60-94% of its initial activity for 4-12 h at 55 degrees C. It also kept almost 70% of the initial activity at 30 degrees C, and more than 40% of the initial activity at 10 degrees C. These results show that recombinant Car3206 had good low temperature resistance and thermal stability properties. The optimum pH of recombinant Car3206 was 7.0. Car3206 was activated by Na+, K+, and Ca2+, but was significantly inhibited by Cu2+ and Cr2+. Thin-layer chromatographic analysis indicated that Car3206 degraded carrageenan generating disaccharides as the only products. The antioxidant capacity of the degraded disaccharides in vitro was investigated and the results showed that different concentrations of the disaccharides had similar scavenging effects as vitamin C on O-2(center dot)-, center dot OH, and DPPH center dot. To our knowledge, this is the first report about details of the biochemical characteristics of a carrageenase isolated from an Antarctic Polaribacter strain. The unique characteristics of Car3206, including its low temperature resistance, thermal stability, and product unity, suggest that this enzyme may be an interesting candidate for industrial processes.
关键词carrageenase gene expression enzymatic characterization enzymatic hydrolysis antioxidant capacity
DOI10.3389/fmicb.2021.631039
收录类别SCI
语种英语
资助项目Impact and Response of Antarctic Seas to Climate Change[RFSOCC2020-2022] ; National Key Research and Development Program of China[2018YFC1406704] ; Key Research and Development Program of Shandong Province, China[2018GHY115013]
WOS研究方向Microbiology
WOS类目Microbiology
WOS记录号WOS:000632848400001
出版者FRONTIERS MEDIA SA
引用统计
被引频次:7[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/170115
专题实验海洋生物学重点实验室
通讯作者Li, Jiang
作者单位1.Qingdao Univ, Coll Environm Sci & Engn, Qingdao, Peoples R China
2.Minist Nat Resources, Inst Oceanog 1, Marine Bioresource & Environm Res Ctr, Qingdao, Peoples R China
3.Minist Nat Resources, Inst Oceanog 1, Key Lab Ecol Environm Sci & Technol, Qingdao, Peoples R China
4.Chinese Acad Sci, Inst Oceanol, CAS & Shandong Prov Key Lab Expt Marine Biol, Qingdao, Peoples R China
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Gui, Yuanyuan,Gu, Xiaoqian,Fu, Liping,et al. Expression and Characterization of a Thermostable Carrageenase From an Antarctic Polaribacter sp. NJDZO3 Strain[J]. FRONTIERS IN MICROBIOLOGY,2021,12:11.
APA Gui, Yuanyuan,Gu, Xiaoqian,Fu, Liping,Zhang, Qian,Zhang, Peiyu,&Li, Jiang.(2021).Expression and Characterization of a Thermostable Carrageenase From an Antarctic Polaribacter sp. NJDZO3 Strain.FRONTIERS IN MICROBIOLOGY,12,11.
MLA Gui, Yuanyuan,et al."Expression and Characterization of a Thermostable Carrageenase From an Antarctic Polaribacter sp. NJDZO3 Strain".FRONTIERS IN MICROBIOLOGY 12(2021):11.
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