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Authors Yang, EZ; Yi, SZ; Bai, F; Niu, DW; Zhong, JJ; Wu, QH; Chen, SF; Zhou, RC; Wang, F
Author Full Name Yang, Enze; Yi, Shanze; Bai, Fang; Niu, Dewei; Zhong, Junjie; Wu, Qiuhong; Chen, Shufang; Zhou, Renchao; Wang, Feng
Title Cloning, Characterization and Expression Pattern Analysis of a Cytosolic Copper/Zinc Superoxide Dismutase (SaCSD1) in a Highly Salt Tolerant Mangrove (Sonneratia alba)
Source INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Language English
Document Type Article
Author Keywords Sonneratia alba; copper/zinc superoxide dismutase; protein expression; activity and stability; salt stress
Keywords Plus OXIDATIVE STRESS; ANTIOXIDATIVE ENZYMES; BRUGUIERA-PARVIFLORA; AVICENNIA-MARINA; NACL STRESS; SALINITY; RESPONSES; PROTEINS; CDNA; SEEDLINGS
Abstract Mangroves are critical marine resources for their remarkable ability to tolerate seawater. Antioxidant enzymes play an especially significant role in eliminating reactive oxygen species and conferring abiotic stress tolerance. In this study, a cytosolic copper/zinc superoxide dismutase (SaCSD1) cDNA of Sonneratia alba, a mangrove species with high salt tolerance, was successfully cloned and then expressed in Escherichia coli Rosetta-gami (designated as SaCSD1). SaCSD1 comprised a complete open reading frame (ORF) of 459 bp which encoded a protein of 152 amino acids. Its mature protein is predicted to be 15.32 kDa and the deduced isoelectric point is 5.78. SaCSD1 has high sequence similarity (85%-90%) with the superoxide dismutase (CSD) of some other plant species. SaCSD1 was expressed with 30.6% yield regarding total protein content after being introduced into the pET-15b (Sma I) vector for expression in Rosetta-gami and being induced with IPTG. After affinity chromatography on Ni-NTA, recombinant SaCSD1 was obtained with 3.2-fold purification and a specific activity of 2200 U/mg. SaCSD1 showed good activity as well as stability in the ranges of pH between 3 and 7 and temperature between 25 and 55 degrees C. The activity of recombinant SaCSD1 was stable in 0.25 M NaCl, Dimethyl Sulphoxide (DMSO), glycerol, and chloroform, and was reduced to a great extent in beta-mercaptoethanol, sodium dodecyl sulfate (SDS), H2O2, and phenol. Moreover, the SaCSD1 protein was very susceptive to pepsin digestion. Real-time Quantitative Polymerase Chain Reaction (PCR) assay demonstrated that SaCSD1 was expressed in leaf, stem, flower, and fruit organs, with the highest expression in fruits. Under 0.25 M and 0.5 M salt stress, the expression of SaCSD1 was down-regulated in roots, but up-regulated in leaves.
Author Address [Yang, Enze; Yi, Shanze; Niu, Dewei; Zhong, Junjie; Wu, Qiuhong; Wang, Feng] Jinan Univ, Coll Pharm, Guangzhou 510632, Guangdong, Peoples R China; [Yang, Enze; Yi, Shanze; Niu, Dewei; Zhong, Junjie; Wu, Qiuhong; Wang, Feng] Jinan Univ, Guangdong Prov Key Lab Pharmacodynam Constituents, Guangzhou 510632, Guangdong, Peoples R China; [Bai, Fang] Shenzhen Univ, Sch Life Sci, Shenzhen 518060, Peoples R China; [Chen, Shufang; Zhou, Renchao] Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510275, Guangdong, Peoples R China; [Chen, Shufang; Zhou, Renchao] Sun Yat Sen Univ, Sch Life Sci, Guangdong Prov Key Lab Plant Resources, Guangzhou 510275, Guangdong, Peoples R China
Reprint Address Wang, F (corresponding author), Jinan Univ, Coll Pharm, Guangzhou 510632, Guangdong, Peoples R China.; Wang, F (corresponding author), Jinan Univ, Guangdong Prov Key Lab Pharmacodynam Constituents, Guangzhou 510632, Guangdong, Peoples R China.; Zhou, RC (corresponding author), Sun Yat Sen Univ, Sch Life Sci, State Key Lab Biocontrol, Guangzhou 510275, Guangdong, Peoples R China.; Zhou, RC (corresponding author), Sun Yat Sen Univ, Sch Life Sci, Guangdong Prov Key Lab Plant Resources, Guangzhou 510275, Guangdong, Peoples R China.
E-mail Address yangenze@foxmail.com; yishanze317@163.com; 15016733519@163.com; 18739977531@163.com; 18825084106@163.com; qiuhong6506@126.com; chsuf@mail.sysu.edu.cn; zhrench@mail.sysu.edu.cn; jnubiopharm@126.com
Funding Agency and Grant Number National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [31170213, 91231106]; Science and Technology Innovation Project from Department of Education of Guangdong Province [2012KJCX0017]; Administration of Traditional Chinese Medicine of Guangdong Province [20141065]
Funding Text This study was supported by grants from the National Natural Science Foundation of China (31170213 and 91231106), Science and Technology Innovation Project from Department of Education of Guangdong Province (2012KJCX0017) and the research project from Administration of Traditional Chinese Medicine of Guangdong Province (20141065). We thank Maheen Gull at the University of South Florida for critical reading of the manuscript.
Times Cited 8
Total Times Cited Count (WoS, BCI, and CSCD) 8
Publisher MDPI
Publisher City BASEL
Publisher Address ST ALBAN-ANLAGE 66, CH-4052 BASEL, SWITZERLAND
ISSN 1422-0067
29-Character Source Abbreviation INT J MOL SCI
ISO Source Abbreviation Int. J. Mol. Sci.
Publication Date JAN
Year Published 2016
Volume 17
Issue 1
Article Number 4
Digital Object Identifier (DOI) 10.3390/ijms17010004
Page Count 12
Web of Science Category Biochemistry & Molecular Biology; Chemistry, Multidisciplinary
Subject Category Biochemistry & Molecular Biology; Chemistry
Document Delivery Number DK0DW
Unique Article Identifier WOS:000374583800004
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