偏肿革裥菌Lg-mnp2基因在构巢曲霉中的表达

Abstract

Abstract:

The aim of this study was to enhance manganese peroxidase (MnP) gene express by Lenzites gibbosa MnP production for using Aspergillus nidulans. In this study, an auxotrophic strain, TN02A7, of A.nidulans was used as the host of L. gibbosa.The conidiophores of TN02A7 were used to produce the protoplasts by different cell wall lyases, including lywallzyme, cellulase and snailase which were mixed together by 1:1:1 for effectively break down the wall and produce protoplasts. PEG/CaCl₂ were used to mediate transformation of L. gibbosa, containing a gene encoding for manganese peroxidase 2 (Lg-MnP2), into the TN02A7 protoplast. The newly obtained transformant strain, TN02A7-Lg-mnp2 , and TN02A7 were cultured in the same shaking medium containing lignin and the MnP activity was detected. The result showed that TN02A7-Lg-mnp2 could produce MnP activity up to 17 U ·L^-1 in 96 h, while TN02A7 did not produce MnP activity at any time, indicating that the gene Lg-mnp2 had been successfully transformed into TN02A7-Lg-mnp2 and expressed in lignin environment. This study provided a new method to produce MnP.

Key words: Lenzites gibbosa, Lg-mnp2, Aspergillus nidulans, transformation, expression

CLC Number:

S718.81

Hu Meimei, Chi Yujie. Expression of the Gene Lg-mnp2 Encoding for Manganese Peroxidase 2 (MnP2) from White-Rot Fungus Lenzites gibbosa in Aspergillus nidulans[J]. Scientia Silvae Sinicae, 2014, 50(2): 139-143.

References

曹进玲. 2011. 钙离子通道蛋白MidA在构巢曲霉中的功能特征研究. 南京:南京师范大学硕士学位论文.
陈凤. 2012. 米曲霉niaD300转化系统的构建及其RNA干扰效应的研究. 广州: 华南理工大学硕士学位论文.
(Chen F. 2012. Study on the transformation system and RNA interference of niaD300. Guangzhou: MS thesis of South China Uniersity. [in Chinese] )
胡美美,池玉杰. 2013. 偏肿革裥菌 MnP 基因在构巢曲霉中转化方法的建立. 东北林业大学学报, 41(7): 129-133.
(Hu M M, Chi Y J. 2013. Construction of the transformation system in aspergillus nidulans for manganese peroxidases gene of white-rot fungus lenzites gibbosa. Journal of Northeast Forestry University, 41(7): 129-133. [in Chinese] )
黄卫, 汪天虹, 吴志红, 等. 2000. 丝状真菌遗传转化系统研究进展. 微生物学杂志, 20 (3): 40-44.
(Huang W, Wang T H, Wu Z H, et al. 2000. Advance in genetic transformation system of filamentous fungi. Journal Of Microbiology, 20 (3): 40-44. [in Chinese] )
李娟, 杨金奎, 梁连铭, 等. 2006. 丝状真菌遗传转化系统研究进展. 江西农业大学学报, 28 (4): 516-520 .
(Li J, Yang J K, Liang L M, et al. 2006. Advance in the research on genetic transformation system of filamentous fungi. Acta Agriculturae Universitatis Jiangxiensis, 28 (4): 516-520. [in Chinese] )
钱科, 胡昌华. 2012. 根癌农杆菌介导丝状真菌遗传转化的影响因子研究进展. 中国医药指南, 10 (9): 371-373.
王金良, 陈宏文.2010. 米曲霉pyrG基因克隆及其同源转化系统的建立. 食品科学, 31(11): 202-205.
(Wang J L, Chen H W. 2010. pyrG gene cloning and establishment of homologous transformation system for Aspergillus oryzae. Food Science, 31(11): 202-205. [in Chinese] )
闫洪波. 2009. 偏肿拟栓菌锰过氧化物酶cDNA基因克隆及在毕赤酵母中的表达. 哈尔滨:东北林业大学博士学位论文.
(Yan H B. 2009. Molecular cloning of cDNA gene encoding a manganese peroxidase from pseudotrametes gibbosa and heterologous expression of this gene in Pichia pastoris. Haerbin: MS thesis of Northeast Forestry University. [in Chinese] )
Abuodeh R O, Orbach M J, Mandel M A, et al. 2000. Genetic transformation of Coccidioides immitis facilitated by Agrobacterium tumefaciens. Infect Dis, 181(6): 2106-2110.
Gu L, Kelly C, Lajoie C. 2000. Cloning of the white rot fungus Phanerochaete chrysosporium manganese peroxidase gene (MnP2) into the methylotrophic yeast Pichia pastoris // Smith L C. Wood,Cellulose.Building Blocks for Chemicals, Fuels and Advanced Materials. Second Annual Partnerships for Environmental Improvement and Economic Development Conference, Syracuse, Syracuse University, 9-11.
Herzog R W, Daniell H, Singh N K, et al.1996. A comparative study on the transformation of Aspergillus nidulans by microprojectile bombardment of conidia and a more conventional procedure using protoplasts treated with polyethyleneglycol. Appl Microbiol Biotechnol, 45: 333-337.
Koukaki M, Giannoutsou E, Karagouni A, et al. 2003. A novel improved method for Aspergillus nidulans transformation. Journal of Microbiological Methods, 55: 687-695.
Li D M,Youngs H L, Gold M H. 2001. Heterologous expression of a thermostable manganese peroxidase from Dichomitus squalens in Phanerochaete chrysosporium. Archives of Biochemistry and Biophysics, 385 (2): 348-356.
Larrondo L F,Lobos S,Stewart P J, et al. 2001.Isoenzyme multiplicity and characterization of recombinant manganese peroxidases from Ceriporiopsis subvermispora and Phanerochaete chrysosporium. Applied and Environmental Microbiology, 67(5): 2070-2075.
Michielse C B, Salim K, Ragas P, et al. 2004. Development of a system for integrative and stable transformation of the zygomycete Rhizopus oryzae by Agrobacterium-mediated DNA transfer. Mol Genet Genom, 271(4): 499-510.
Ruiz-Dueńas F J, Martínez M J, Martínez Á T.1999. Heterologous expression of Pleurotus eryngii peroxidase confirms its ability to oxidize Mn²⁺ and different aromatic substrates. Applied and Environmental Microbiology, 65(10): 4705-4707.
Stewart P,Whitwam R E, Kersten P J, et al.1996. Efficient expression of a Phanerochaete chrysosporium manganese peroxidase gene in Aspergillus oryzae. Applied and Environmental Microbiology, 62(3): 860-864.
Tilburn J, Scazzocchio C, Taylor G, et al. 1983. Transformation by intergration in Aspergillus nidulans. Gene, 26(2/3): 205-221.
Whitwam R E, Gazarian I G, Tien M. 1995. Expression of fungal Mn peroxidase in E.coli and refolding to yield active enzyme. Biochemical and Biophysical Research Communications, 216(3): 1013-1017.
Whitwam R, Tien M.1996. Heterologous expression and reconstitution of fungal Mn peroxidase. Archives of Biochemistry and Biophysics, 333(2): 439-446.

[1]	Liu Daofeng, Wang Xia, Dai Yin, Yang Jianfeng, Ma Jing, Li Mingyang, Sui Shunzhao. Cloning and Function Analysis of CpTAF10 from Wintersweet (Chimonanthus praecox) [J]. Scientia Silvae Sinicae, 2019, 55(6): 176-183.
[2]	Lu Huijun, Li Ziyi, Liang Hanyu, Yue Yuanzhi, Zhou Tianchang, Yang Yuzhang, Wang Yucheng, Ji Xiaoyu. Expression and Stress Tolerance Analysis of NAC24 from Tamarix hispida [J]. Scientia Silvae Sinicae, 2019, 55(3): 54-63.
[3]	Zhang Jiaqi, Hu Hengkang, Xu Chuanmei, Hu Yuanyuan, Huang Youjun, Xia Guohua, Huang Jianqin, Chang Yingying, Ye Lei, Lou Heqiang, Zhang Qixiang. Cloning, Subcellular Localization and Function Verification of Gibberellin 2-Oxidase Gene in Walnut (Juglans regia) [J]. Scientia Silvae Sinicae, 2019, 55(2): 50-60.
[4]	Fan Weijian, Xiang Weifang, Wang Jing, Bai Penghua, Pan Lina, Yang Yixin, Zhu Gengping, Li Min. Effects of Insecticides and Ultraviolet on the Expression of sHSP12.2 Gene in Chouioia cunea [J]. Scientia Silvae Sinicae, 2019, 55(2): 128-136.
[5]	Huang Ren, Zhang Yun, Zhang Qixiang, Wang Zhengjia, Xia Guohua, Huang Jianqin, Hu Yuanyuan. Transcriptome Analysis of Photosynthetic Capacity of Exocarp of Heterogeneously Pollinated Carya cathayensis [J]. Scientia Silvae Sinicae, 2019, 55(1): 128-137.
[6]	Zhang Yang, Du Lin, Tang Xianfeng, Liu Huanhuan, Zhou Gongke, Chai Guohua. Identification and Expression Pattern Analyses of Populus BAG Genes [J]. Scientia Silvae Sinicae, 2019, 55(1): 138-145.
[7]	Wei Mingke, Yu Jinjian, Huang Xiaolong, Liu Qiongyao, Huang Huahong, Lin Erpei, Tong Zaikang. Cloning, Expression and Single Nucleotide Polymorphisms Analysis of NAC Transcription Factor Gene ClNAC1 in Cunninghamia lanceolata [J]. Scientia Silvae Sinicae, 2018, 54(9): 49-59.
[8]	Wang Haibo, Gong Ming, Guo Junyun, Xin Hu, Tang Lizhou, Liu Chao, Gao Yong, Dai Dongqin. Molecular Cloning and Prokaryotic Expression of Orphan Gene Soloist of AP2/ERF Gene Family in Jatropha curcas [J]. Scientia Silvae Sinicae, 2018, 54(9): 60-69.
[9]	Wang Juan, Chen Hui. Identification and Expression of Cold Shock Domain-containing Protein Gene in Dendroctonus armandi [J]. Scientia Silvae Sinicae, 2018, 54(4): 58-66.
[10]	Li Hui, Gu Tianzi, Chen Changyu, Huang Kairu, Tian Shuo, Zhao Xudong, Hao Dejun. cDNA Cloning, Sequence Analysis and Expression Profile of a Chemosensory Protein from the Clostera restitura (Lepidoptera: Notodontidae) [J]. Scientia Silvae Sinicae, 2018, 54(4): 67-75.
[11]	Zhang Hehua, Liu Jiaxin, Luo Chaobing, Zhang Lingyun. Cloning and Analysis of a Transcription Factor PwERF8 and the Promoter Sequences in Picea wilsonii [J]. Scientia Silvae Sinicae, 2018, 54(3): 48-60.
[12]	Li Jianbo, Jia Huixia, Zhang Jin, Liu Bobin, Hu Jianjun, Wang Lijuan, Lu Mengzhu. Effect of Overexpression of Populus tomentosa WUSCHEL-related homeobox 4 (PtoWOX4a) on the Secondary Growth of Poplar [J]. Scientia Silvae Sinicae, 2018, 54(2): 52-59.
[13]	Ni Fei, Li Wenhao, Lin Erpei, Tong Zaikang, Huang Huahong. Cloning, Expression and Regulation of MYB Genes in Betula luminifera [J]. Scientia Silvae Sinicae, 2018, 54(12): 70-81.
[14]	Gu Zhanying, Yang Ruonan, Chen Hao. The Establishment of Isolation and Transient Transformation Methods of Protoplasts of Vernicia fordii Mesophyll Cells [J]. Scientia Silvae Sinicae, 2018, 54(1): 46-53.
[15]	Xu Jiajuan, Li Huogen. Cloning and Primary Functional Analysis of LcPAT8 Gene from Liriodendron chinense [J]. Scientia Silvae Sinicae, 2017, 53(9): 45-54.

Expression of the Gene Lg-mnp2 Encoding for Manganese Peroxidase 2 (MnP2) from White-Rot Fungus Lenzites gibbosa in Aspergillus nidulans

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments