红冬孢酵母分子遗传操作技术研究进展
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摘要
红冬孢酵母(Rhodosporidium)是一种天然的微生物油脂和β-类胡萝卜素高产菌,有望被开发为工业平台菌株。近年来,基因组测序完成和各种组学研究的开展,为红冬孢酵母分子遗传操作技术开发奠定了基础。基因元件(如启动子、终止子、报告基因和筛选标记等)的挖掘和引入,基因转化方法开发以及基因操纵技术(如基因失活和过表达)等的完善,加速了红冬孢酵母的代谢工程研究进程。本文中,笔者总结了红冬孢酵母在分子遗传操作技术方面的研究进展,力图勾勒出红冬孢酵母的分子遗传操作技术研究的概貌,为后续研究提供借鉴。
Rhodosporidium is a promising industrial platform strain.It natively produces microbial oil and β-carotenoid with high titer and yield.Recently,great progress has been made in genome sequencing and multi-omics study,providing the basis to develop molecular genetic manipulation technology.Characterization and application of genetic components or elements(such as promoters,terminator,reporter genes and selective markers),development of gene transformation methods,and improvement of genetic manipulation technology(gene inactivation and overexpression) have accelerated the metabolic engineering of Rhodosporidium.Here,we summarize the research progress in molecular genetic manipulation technology of Rhodosporidium.We also outline the research framework of molecular genetic manipulation of Rhodosporidium to provide reference for future research.
Rhodosporidium is a promising industrial platform strain.It natively produces microbial oil and β-carotenoid with high titer and yield.Recently,great progress has been made in genome sequencing and multi-omics study,providing the basis to develop molecular genetic manipulation technology.Characterization and application of genetic components or elements(such as promoters,terminator,reporter genes and selective markers),development of gene transformation methods,and improvement of genetic manipulation technology(gene inactivation and overexpression) have accelerated the metabolic engineering of Rhodosporidium.Here,we summarize the research progress in molecular genetic manipulation technology of Rhodosporidium.We also outline the research framework of molecular genetic manipulation of Rhodosporidium to provide reference for future research.
引文
[1] PARK Y K,NICAUD J M,LEDESMA-AMARO R.The engineering potential of Rhodosporidium toruloides as a workhorse for biotechnological applications[J].Trend Biotechnol,2018,36(3):304-317.
[2] XU J,LIU D.Exploitation of genus Rhodosporidium for microbial lipid production[J].World J Microbiol Biotechnol,2017,33(3):54.
[3] SINGH G,JAWED A,PAUL D,et al.Concomitant production of lipids and carotenoids in Rhodosporidium toruloides under osmotic stress using response surface methodology[J].Front Microbiol,2016,7:1686.
[4] YAEGASHI J,KIRBY J,ITO M,et al.Rhodosporidium toruloides:a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts[J].Biotechnol Biofuel,2017,10:241.
[5] ZHU Z,ZHANG S,LIU H,et al.A multi-omic map of the lipid-producing yeast Rhodosporidium toruloides[J].Nat Commun,2012,3:1112.
[6] WANG Y N,LIN X P,ZHANG S F,et al.Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides[J].Yeast,2016,33(3):99-106.
[7] LIU Y B,KOH C M J,SUN L H,et al.Characterization of glyceraldehyde-3-phosphate dehydrogenase gene RtGPD1 and development of genetic transformation method by dominant selection in oleaginous yeast Rhodosporidium toruloides[J].Appl Microbiol Biotechnol,2013,97(2):719-729.
[8] LIU Y B,YAP S A,KOH C M J,et al.Developing a set of strong intronic promoters for robust metabolic engineering in oleaginous Rhodotorula (Rhodosporidium) yeast species[J].Microb Cell Fact,2016,15(1):200.
[9] LIU Y B,KOH C M J,NGOH S T,et al.Engineering an efficient and tight D-amino acid-inducible gene expression system in Rhodosporidium/Rhodotorula species[J].Microb Cell Fact,2015,14:170.
[10] 马斯佳.圆红冬孢酵母诱导性遗传操作平台的构建[D].大连:大连工业大学,2015.
[11] JOHNS A M B,LOVE J,AVES S J.Four inducible promoters for controlled gene expression in the oleaginous yeast Rhodotorula toruloides[J].Front Microbiol,2016,7:1666.
[12] LIN X,WANG Y,ZHANG S,et al.Functional integration of multiple genes into the genome of the oleaginous yeast Rhodosporidium toruloides[J].FEMS Yeast Res,2014,14(4):547-555.
[13] DIAZ T,FILLET S,CAMPOY S,et al.Combining evolutionary and metabolic engineering in Rhodosporidium toruloides for lipid production with non-detoxified wheat straw hydrolysates[J].Appl Microbiol Biotechnol,2018,102(7):3287-3300.
[14] YANG F,ZHANG S,TANG W,et al.Identification of the orotidine-5′-monophosphate decarboxylase gene of the oleaginous yeast Rhodosporidium toruloides[J].Yeast,2008,25:623-630.
[15] TULLY M,GILBERT H J.Transformation of Rhodosporidium toruloides[J].Gene,1985,36(3):235-240.
[16] 孙文怡.基于农杆菌介导的圆红冬孢酵母遗传重组系统的研究[D].大连:大连理工大学,2017.
[17] LIU H D,JIAO X,WANG Y N,et al.Fast and efficient genetic transformation of oleaginous yeast Rhodosporidium toruloides by using electroporation[J].FEMS Yeast Res,2017,doi:10.1093/femsyr/fox017.
[18] TSAI Y Y,OHASHI T,KANAZAWA T,et al.Development of a sufficient and effective procedure for transformation of an oleaginous yeast,Rhodosporidium toruloides DMKU3-TK16[J].Curr Genet,2017,63(2):359-371.
[19] TAKAHASHI S,OKADA H,ABE K,et al.Genetic Transformation of the yeast Rhodotorula gracilis ATCC 26217 by electroporation[J].Appl Biochem Microbiol,2014,50(6):624-628.
[20] 田李,陈捷胤,汪佳妮,等.高效大丽轮枝菌(Verticillium dahliae)基因敲除体系的构建[J].微生物学报,2011,51(7):906-913.
[21] FILLET S C,SUáREZ GONZáLEZ B,RONCHEL BARRENO M D C,et al.Production of microbial oils with an elevated oleic acid content:WO2016185073(A1)[P].2016-11-24.
[22] SUN W Y,YANG X B,WANG X Y,et al.Homologous gene targeting of a carotenoids biosynthetic gene in Rhodosporidium toruloides by Agrobacterium-mediated transformation[J].Biotechnol Lett,2017,39(7):1001-1007.
[23] KOH C M J,LIU Y B,MOEHNINSI,et al.Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides[J].BMC Microbiol,2014,14(1):50.
[2] XU J,LIU D.Exploitation of genus Rhodosporidium for microbial lipid production[J].World J Microbiol Biotechnol,2017,33(3):54.
[3] SINGH G,JAWED A,PAUL D,et al.Concomitant production of lipids and carotenoids in Rhodosporidium toruloides under osmotic stress using response surface methodology[J].Front Microbiol,2016,7:1686.
[4] YAEGASHI J,KIRBY J,ITO M,et al.Rhodosporidium toruloides:a new platform organism for conversion of lignocellulose into terpene biofuels and bioproducts[J].Biotechnol Biofuel,2017,10:241.
[5] ZHU Z,ZHANG S,LIU H,et al.A multi-omic map of the lipid-producing yeast Rhodosporidium toruloides[J].Nat Commun,2012,3:1112.
[6] WANG Y N,LIN X P,ZHANG S F,et al.Cloning and evaluation of different constitutive promoters in the oleaginous yeast Rhodosporidium toruloides[J].Yeast,2016,33(3):99-106.
[7] LIU Y B,KOH C M J,SUN L H,et al.Characterization of glyceraldehyde-3-phosphate dehydrogenase gene RtGPD1 and development of genetic transformation method by dominant selection in oleaginous yeast Rhodosporidium toruloides[J].Appl Microbiol Biotechnol,2013,97(2):719-729.
[8] LIU Y B,YAP S A,KOH C M J,et al.Developing a set of strong intronic promoters for robust metabolic engineering in oleaginous Rhodotorula (Rhodosporidium) yeast species[J].Microb Cell Fact,2016,15(1):200.
[9] LIU Y B,KOH C M J,NGOH S T,et al.Engineering an efficient and tight D-amino acid-inducible gene expression system in Rhodosporidium/Rhodotorula species[J].Microb Cell Fact,2015,14:170.
[10] 马斯佳.圆红冬孢酵母诱导性遗传操作平台的构建[D].大连:大连工业大学,2015.
[11] JOHNS A M B,LOVE J,AVES S J.Four inducible promoters for controlled gene expression in the oleaginous yeast Rhodotorula toruloides[J].Front Microbiol,2016,7:1666.
[12] LIN X,WANG Y,ZHANG S,et al.Functional integration of multiple genes into the genome of the oleaginous yeast Rhodosporidium toruloides[J].FEMS Yeast Res,2014,14(4):547-555.
[13] DIAZ T,FILLET S,CAMPOY S,et al.Combining evolutionary and metabolic engineering in Rhodosporidium toruloides for lipid production with non-detoxified wheat straw hydrolysates[J].Appl Microbiol Biotechnol,2018,102(7):3287-3300.
[14] YANG F,ZHANG S,TANG W,et al.Identification of the orotidine-5′-monophosphate decarboxylase gene of the oleaginous yeast Rhodosporidium toruloides[J].Yeast,2008,25:623-630.
[15] TULLY M,GILBERT H J.Transformation of Rhodosporidium toruloides[J].Gene,1985,36(3):235-240.
[16] 孙文怡.基于农杆菌介导的圆红冬孢酵母遗传重组系统的研究[D].大连:大连理工大学,2017.
[17] LIU H D,JIAO X,WANG Y N,et al.Fast and efficient genetic transformation of oleaginous yeast Rhodosporidium toruloides by using electroporation[J].FEMS Yeast Res,2017,doi:10.1093/femsyr/fox017.
[18] TSAI Y Y,OHASHI T,KANAZAWA T,et al.Development of a sufficient and effective procedure for transformation of an oleaginous yeast,Rhodosporidium toruloides DMKU3-TK16[J].Curr Genet,2017,63(2):359-371.
[19] TAKAHASHI S,OKADA H,ABE K,et al.Genetic Transformation of the yeast Rhodotorula gracilis ATCC 26217 by electroporation[J].Appl Biochem Microbiol,2014,50(6):624-628.
[20] 田李,陈捷胤,汪佳妮,等.高效大丽轮枝菌(Verticillium dahliae)基因敲除体系的构建[J].微生物学报,2011,51(7):906-913.
[21] FILLET S C,SUáREZ GONZáLEZ B,RONCHEL BARRENO M D C,et al.Production of microbial oils with an elevated oleic acid content:WO2016185073(A1)[P].2016-11-24.
[22] SUN W Y,YANG X B,WANG X Y,et al.Homologous gene targeting of a carotenoids biosynthetic gene in Rhodosporidium toruloides by Agrobacterium-mediated transformation[J].Biotechnol Lett,2017,39(7):1001-1007.
[23] KOH C M J,LIU Y B,MOEHNINSI,et al.Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides[J].BMC Microbiol,2014,14(1):50.
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