戈壁异常球菌Dgl5蛋白生物学功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2017-0875

生物技术通报 ›› 2018, Vol. 34 ›› Issue (3): 177-184.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0875

戈壁异常球菌Dgl5蛋白生物学功能研究

张亨^1,2, 刘盈盈², 陈云², 平淑珍², 王劲^1,2

1. 西南科技大学生命科学与工程学院，绵阳 621000;
2. 中国农业科学院生物技术研究所，北京 100081

收稿日期:2017-10-19 出版日期:2018-03-20 发布日期:2018-04-10
作者简介:张亨，男，硕士研究生，研究方向特殊环境微生物功能基因资源利用；E-mailzhangheng2013@hotmail.com
基金资助:
国家重点基础研究发展计划（“973” 计划）（2013CB733903）

Biological Identification of Dgl5 in Deinococcus gobiensis I-0

ZHANG Heng^1，2, LIU Ying-ying², CHEN Yun², PING Shu-zhen², WANG Jin^1，2

1. College of Life Science and Engineering，Southwest University of Science and Technology，Mianyang 621000;
2. Biotechnology Research Institute，Chinese Academy of Agricultural Sciences，Beijing 100081

Received:2017-10-19 Published:2018-03-20 Online:2018-04-10

摘要/Abstract

摘要： LEA5C（Group 5C Late Embryogenesis Abundant）家族蛋白参与非生物胁迫反应并以分子伴侣的形式保护细胞内生物大分子不受损伤。分离于新疆戈壁沙漠的戈壁异常球菌（Deinococcus gobiensis）I-0具有极端的电离辐射、氧化、干燥等抗性。功能基因组注释表明Dgo_CA1605编码蛋白属于LEA5C家族，命名为Dgl5，对Dgl5生理功能展开研究。采用同源重组的方法构建Dgl5重组表达菌株。非生物胁迫实验结果表明，Dgl5能够显著增强表达菌株BL21盐胁迫抗性，最大限度地保护宿主菌免受反复冻融的损伤；体外生理生化实验结果表明在反复冻融条件下，Dgl5能够有效保护苹果酸脱氢酶（MDH）和乳酸脱氢酶（LDH）酶学活性。因此推测，在冷冻、高盐胁迫条件下，Dgl5蛋白通过保护细胞体内相关酶类的活性，增强宿主菌抵抗外界复杂多变的极端环境。

关键词: LEA5C, 戈壁异常球菌, 非生物胁迫, 分子伴侣

Abstract: Proteins in LEA5C（Group 5C Late Embryogenesis Abundant）family could involve in the responses to abiotic stresses and protect bio-macromolecules in cells from damages under abiotic stresses, due to their functions as molecular chaperones. The Deinococcus gobiensis I-0 isolated from Xinjiang Gobi Desert possessed highly strong resistance to ionizing radiation，oxidation，and desiccation. The genomic annotation of D. gobiensis I-0 indicated that Dgo_CA1605 encoding protein belonged to LEA5C family，named as Dgl5. Accordingly，the function of Dgl5 was investigated in this article. Recombinant expression strain Dgl5 was constructed by homologous recombination. The abiotic stress results revealed that Dgl5 significantly enhanced high-salt resistance of expression strain BL21 and maximally protected the host from the damages in freeze-thaw cycles. Likewise，the physiological and biochemical experiments in vitro demonstrated that the enzymatic activities of malate dehydrogenase and lactate dehydrogenase were effectively maintained due to Dg15 under freeze-thaw cycles. Therefore，we speculated that the improved resistance of the host adapting to harsh environment such as frozen and high-salt was achieved by Dg15 protecting cellular enzyme activities.

Key words: LEA5C, Deinococcus gobiensis I-0, abiotic stress, molecular chaperone

张亨, 刘盈盈, 陈云, 平淑珍, 王劲. 戈壁异常球菌Dgl5蛋白生物学功能研究[J]. 生物技术通报, 2018, 34(3): 177-184.

ZHANG Heng, LIU Ying-ying, CHEN Yun, PING Shu-zhen, WANG Jin. Biological Identification of Dgl5 in Deinococcus gobiensis I-0[J]. Biotechnology Bulletin, 2018, 34(3): 177-184.

参考文献

［1］Pearce RS. Molecular analysis of acclimation to cold［J］. Plant Growth Regulation, 1999, 29（1-2）：47-76.
［2］Artus NN, Uemura M, Steponkus PL, et al. Constitutive expression of the cold-regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance［J］. Proc Nat Acad Sci, 1996, 93（23）：13404-13409.
［3］Thomashow MF. Plant cold acclimation：freezing tolerance genes and regulatory mechanisms［J］. Annual Review of Plant Biology, 1999, 50（1）：571-599.
［4］Goyal K, Walton LJ, Tunnacliffe A. LEA proteins prevent protein aggregation due to water stress［J］. Biochemical Journal, 2005, 388（1）：151-157.
［5］Reyes JL, Rodrigo MJ, Colmenero-Flores JM, et al. Hydrophilins from distant organisms can protect enzymatic activities from water limitation effects in vitro［J］. Plant, Cell & Environment, 2005, 28（6）：709-718.
［6］Chakrabortee S, Meersman F, Schierle G S K, et al. Catalytic and chaperone-like functions in an intrinsically disordered protein associated with desiccation tolerance［J］. Proc Nat Acad Sci, 2010, 107（37）：16084-16089.
［7］Kanuru M, Aradhyam GK. Chaperone-like activity of calnuc prevents amyloid aggregation［J］. Biochemistry, 2017, 56（1）：149-159.
［8］Battaglia M, Olvera-Carrillo Y, Garciarrubio A, et al. The enigmatic LEA proteins and other hydrophilins［J］. Plant Physiol, 2008, 148（1）：6-24.
［9］He S, Tan L, Hu Z, et al. Molecular characterization and functional analysis by heterologous expression in E. coli under diverse abiotic stresses for OsLEA5, the atypical hydrophobic LEA protein from Oryza sativa L.［J］. Mol Gene Genomics, 2012, 287（1）：39-54.
［10］Jaspard E, Hunault G. Comparison of amino acids physico-chemical properties and usage of late embryogenesis abundant proteins, hydrophilins and WHy domain［J］. PLoS One, 2014, 9（10）：e109570.
［11］Dang NX, Popova AV, Hundertmark M, et al. Functional characterization of selected LEA proteins from Arabidopsis thaliana in yeast and in vitro［J］. Planta, 2014, 240（2）：325-336.
［12］Ciccarelli FD, Bork P. The WHy domain mediates the response to desiccation in plants and bacteria［J］. Bioinformatics, 2004, 21（8）：1304-1307.
［13］Makarova KS, Aravind L, Wolf YI, et al. Genome of the extremely radiation-resistant bacterium Deinococcus radiodurans viewed from the perspective of comparative genomics［J］. Microbiology and Molecular Biology Reviews, 2001, 65（1）：44-79.
［14］Jiang S, Wang J, Liu X, et al. DrwH, a novel WHy domain-containing hydrophobic LEA5C protein from Deinococcus radiodurans, protects enzymatic activity under oxidative stress［J］. Scientific Reports, 2017, 7（1）：9281.
［15］Yuan M, Zhang W, Dai S, et al. Deinococcus gobiensis sp. nov., an extremely radiation-resistant bacterium［J］. Int J Syst Evol Microbiol, 2009, 59（6）：1513-1517.
［16］Dahl JU, Koldewey P, Salmon L, et al. HdeB functions as an acid-protective chaperone in bacteria［J］. J Biol Chem, 2015, 290（1）：65-75.
［17］Liu Y, Wang L, Jiang S, et al. Group 5 LEA protein, ZmLEA5C, enhance tolerance to osmotic and low temperature stresses in transgenic tobacco and yeast［J］. Plant Physiology and Biochemistry, 2014, 84：22-31.
［18］Wu Y, Liu C, Kuang J, et al. Overexpression of SmLEA enhances salt and drought tolerance in Escherichia coli and Salvia miltiorrhiza［J］. Protoplasma, 2014, 251（5）：1191-1199.
［19］Wang H, Wu Y, Yang X, et al. SmLEA2, a gene for late embryogenesis abundant protein isolated from Salvia miltiorrhiza, confers tolerance to drought and salt stress in Escherichia coli and S. miltiorrhiza［J］. Protoplasma, 2017, 254（2）：685-696.
［20］Haaning S, Radutoiu S, Hoffmann S V, et al. An unusual intrinsically disordered protein from the model legume Lotus japonicus stabilizes proteins in vitro［J］. J Biol Chem, 2008, 283（45）：31142-31152.

[1]	梅欢, 李玥, 刘可蒙, 刘吉华. 小檗碱桥酶高效原核表达及生物合成l-SLR的研究[J]. 生物技术通报, 2023, 39(7): 277-287.
[2]	赵雪婷, 高利燕, 王俊刚, 沈庆庆, 张树珍, 李富生. 甘蔗AP2/ERF转录因子基因ShERF3的克隆、表达及其编码蛋白的定位[J]. 生物技术通报, 2023, 39(6): 208-216.
[3]	董聪, 高庆华, 王玥, 罗同阳, 王庆庆. 基于联合策略提高FAD依赖的葡萄糖脱氢酶的酵母表达[J]. 生物技术通报, 2023, 39(6): 316-324.
[4]	李苑虹, 郭昱昊, 曹燕, 祝振洲, 王飞飞. 外源植物激素调控微藻生长及目标产物积累研究进展[J]. 生物技术通报, 2023, 39(6): 61-72.
[5]	冯珊珊, 王璐, 周益, 王幼平, 方玉洁. WOX家族基因调控植物生长发育和非生物胁迫响应的研究进展[J]. 生物技术通报, 2023, 39(5): 1-13.
[6]	翟莹, 李铭杨, 张军, 赵旭, 于海伟, 李珊珊, 赵艳, 张梅娟, 孙天国. 异源表达大豆转录因子GmNF-YA19提高转基因烟草抗旱性[J]. 生物技术通报, 2023, 39(5): 224-232.
[7]	姚姿婷, 曹雪颖, 肖雪, 李瑞芳, 韦小妹, 邹承武, 朱桂宁. 火龙果溃疡病菌实时荧光定量PCR内参基因的筛选[J]. 生物技术通报, 2023, 39(5): 92-102.
[8]	杨春洪, 董璐, 陈林, 宋丽. 大豆VAS1基因家族的鉴定及参与侧根发育的研究[J]. 生物技术通报, 2023, 39(3): 133-142.
[9]	苗淑楠, 高宇, 李昕儒, 蔡桂萍, 张飞, 薛金爱, 季春丽, 李润植. 大豆GmPDAT1参与油脂合成和非生物胁迫应答的功能分析[J]. 生物技术通报, 2023, 39(2): 96-106.
[10]	许睿, 祝英方. 中介体复合物在植物非生物胁迫应答中的功能[J]. 生物技术通报, 2023, 39(11): 54-60.
[11]	孙雨桐, 刘德帅, 齐迅, 冯美, 黄栩筝, 姚文孔. 茉莉酸调控植物生长发育和胁迫的研究进展[J]. 生物技术通报, 2023, 39(11): 99-109.
[12]	杨旭妍, 赵爽, 马天意, 白玉, 王玉书. 三个甘蓝WRKY基因的克隆及其对非生物胁迫的表达[J]. 生物技术通报, 2023, 39(11): 261-269.
[13]	安昌, 陆琳, 沈梦千, 陈盛圳, 叶康卓, 秦源, 郑平. 植物bHLH基因家族研究进展及在药用植物中的应用前景[J]. 生物技术通报, 2023, 39(10): 1-16.
[14]	位欣欣, 兰海燕. 植物MYB转录因子调控次生代谢及逆境响应的研究进展[J]. 生物技术通报, 2022, 38(8): 12-23.
[15]	王慧, 马艺文, 乔正浩, 常彦彩, 术琨, 丁海萍, 聂永心, 潘光堂. AOX基因家族的结构和功能特征分析[J]. 生物技术通报, 2022, 38(7): 160-170.

戈壁异常球菌Dgl5蛋白生物学功能研究

Biological Identification of Dgl5 in Deinococcus gobiensis I-0

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价