干旱区生态环境知识资源中心(Arid): Characterization of a Mn-SOD from the desert beetle Microdera punctipennis and its increased resistance to cold stress in E. coli cells

Arid

DOI	10.7717/peerj.8507
	Characterization of a Mn-SOD from the desert beetle Microdera punctipennis and its increased resistance to cold stress in E. coli cells
	Xikeranmu, Zilajiguli; Ma, Ji; Liu, Xiaoning
通讯作者	Liu, Xiaoning
来源期刊	PEERJ
ISSN	2167-8359
出版年	2020
卷号	8
英文摘要	Insects have developed a complex network of enzymatic antioxidant systems for handling reactive oxygen species (ROS) generated during stress. Superoxide dismutases (SODs) play a determinant role in balancing ROS in insect. However, studies devoted to SODs functions in insects under cold stress are limited. In the present study, we attempted to identify and characterize a mitochondrial manganese SOD (mMn-SOD) from the desert beetle Micordera punctipennis (denoted as MpmMn-SOD) and explore its protective effects on bacteria cells under cold stress. MpmMn-SOD is composed of 202 amino acids with conserved domains required for metal ions binding and enzyme activity. RT-qPCR experiments revealed that the expression of MpmMn-SOD was ubiquitous but tissue-specific and was induced by cold stress. An E. coli (BL21) system was applied to study the function of MpmMn-SOD. The MpmMn-SOD gene was cloned into the prokaryotic expression vector pET-32a to generate a recombinant plasmid pET-32a(MpmMn-SOD). After transformation of the plasmid into E. coli BL21, the fusion protein Trx-His-MpmMn-SOD was overexpressed and identified by SDS-PAGE and Western blotting. Antioxidant activity assay showed that the death zones of the transformed bacteria BL21 (pET32a-mMn-SOD) were smaller in diameter than the control bacteria BL21 (pET32a). Survival curves under -4 degrees C showed that BL21 (pET32a-mMn-SOD) had significant enhanced cold resistance compared to BL21 (pET32a). Its SOD activity under -4 degrees C had a significant negative correlation (r = -0.995) with superoxide anion O-2(center dot-) content. Accordingly, under cold stress BL21 (pET32a-mMn-SOD) had lower electric conductivity and malondialdehyde (MDA) content than BL21 (pET32a). Taken together, our results showed that cold stress stimulated the expression of MpmMn-SOD in M. punctipennis. The E. coli cells that overexpress MpmMn-SOD increase their resistance to cold stress by scavenging ROS, and mitigate potential cell damage caused by ROS under cold conditions.
英文关键词	Microdera punctipennis mMn-SOD Cold stress Superoxide radicals Oxidative stress E. coli Recombinant enzyme Antioxidant defense
类型	Article
语种	英语
国家	Peoples R China
开放获取类型	Green Published, gold
收录类别	SCI-E
WOS记录号	WOS:000513590700006
WOS关键词	MANGANESE SUPEROXIDE-DISMUTASE ; COLEOPTERA-TENEBRIONIDAE ; MESSENGER-RNA ; BOMBYX-MORI ; GENE ; EXPRESSION ; SILKWORM ; CATALASE ; TRANSFERASE ; EVOLUTION
WOS类目	Multidisciplinary Sciences
WOS研究方向	Science & Technology - Other Topics
来源机构	新疆大学
资源类型	期刊论文
条目标识符	http://119.78.100.177/qdio/handle/2XILL650/315289
作者单位	Xinjiang Univ, Coll Life Sci & Technol, Xinjiang Key Lab Biol Resources & Genet Engn, Urumqi, Peoples R China
推荐引用方式 GB/T 7714	Xikeranmu, Zilajiguli,Ma, Ji,Liu, Xiaoning. Characterization of a Mn-SOD from the desert beetle Microdera punctipennis and its increased resistance to cold stress in E. coli cells[J]. 新疆大学,2020,8.
APA	Xikeranmu, Zilajiguli,Ma, Ji,&Liu, Xiaoning.(2020).Characterization of a Mn-SOD from the desert beetle Microdera punctipennis and its increased resistance to cold stress in E. coli cells.PEERJ,8.
MLA	Xikeranmu, Zilajiguli,et al."Characterization of a Mn-SOD from the desert beetle Microdera punctipennis and its increased resistance to cold stress in E. coli cells".PEERJ 8(2020).