益生芽孢杆菌表达系统的研究进展
|
摘要
益生芽孢杆菌尤其是枯草芽孢杆菌是目前分泌外源蛋白的理想宿主,将益生菌作为表达外源基因的宿主菌,除了必须有适合基因操作的菌株外,还必须建立合适的表达质粒以及解决外源基因表达的相关问题。本文详细阐述了益生芽孢杆菌宿主菌、启动子和信号肽的改造优化及不同类型载体的优缺点,为益生芽孢杆菌表达系统的完善奠定了基础。同时结合本实验室研究提出以芽孢表面展示技术投递抗原,口服后除了保持芽孢杆菌益生菌原有的功能外,还同时发挥疫苗黏膜免疫作用。
Probiotic Bacillus,especially B.subtilis is an attractive host for the production of heterologous proteins.Probiotics as host bacteria expressing foreign genes have to construct appropriate expression vectors and gene expression regulation in addition to the host bacteria suitable for gene manipulation.This paper not only reviewed the improving of host bacteria,promoters and signal peptide,but also explained the advantages and disadvantages of various vectors,which provided a basic information for improving probiotic Bacillus expression system.At the same time,the method for delivering antigens that was displayed on probiotic Bacillus sp.spore surface proposed in our previous studies,which not only keeps the original function of Bacillus probiotics,but also plays the role of mucosal immunity.
Probiotic Bacillus,especially B.subtilis is an attractive host for the production of heterologous proteins.Probiotics as host bacteria expressing foreign genes have to construct appropriate expression vectors and gene expression regulation in addition to the host bacteria suitable for gene manipulation.This paper not only reviewed the improving of host bacteria,promoters and signal peptide,but also explained the advantages and disadvantages of various vectors,which provided a basic information for improving probiotic Bacillus expression system.At the same time,the method for delivering antigens that was displayed on probiotic Bacillus sp.spore surface proposed in our previous studies,which not only keeps the original function of Bacillus probiotics,but also plays the role of mucosal immunity.
引文
[1]李育阳.基因表达技术[M].北京:科学出版社,2002.
[2]卢争辉,周玉玲,张晓舟,等.感受态还是芽孢?细胞命运决定的遗传调控网络[J].生物工程学报,2015,31(11):1543-1552
[3]张晓舟.枯草杆菌新型表达系统和遗传操作体系的建立及应用[D].南京:南京农业大学,2006.
[4]李信志,卢争辉,周玉玲,等.枯草芽孢杆菌SCK6超级感受态的制备和转化条件优化[J].生物工程学报,2017,33(4):692-698.
[5]You C,Zhang X Z,Zhang Y H P.Simple cloning via direct transformation of PCR product(DNA Multimer)to Escherichia coli and Bacillus subtilis[J].Appl Environ Microbiol,2012,78(5):1593.
[6]李瑞芳,薛雯雯,黄亮,等.枯草芽孢杆菌感受态细胞的制备及质粒转化方法研究[J].生物技术通报,2011,35(5):227-230.
[7]Zhang X Z,Zhang Y.Simple,fast and high-efficiency transformation system for directed evolution of cellulase in Bacillus subtilis.[J].Microb Biotechnol,2011,4(1):98.
[8]Piggot P J,Hilbert D W.Sporulation of Bacillus subtilis[J].Current Opin Microbiol,2004,7(6):579-586.
[9]王慧.芽孢杆菌遗传操作体系初建及工程菌株分子改良[D].北京:中国农业科学院,2016.
[10]何庆,王茜,朱清华,等.基因组工程在微生物细胞工厂构建中的应用进展[J].生物产业技术,2017(1):31-36.
[11]Toya Y,Hirasawa T,Morimoto T,et al.13 C-metabolic flux analysis in heterologous cellulase production by Bacillus subtilis,genome-reduced strain[J].J Biotech,2014,179(1):42-49.
[12]Li Y,Zhu X,Zhang X,et al.Characterization of genome-reduced Bacillus subtilis,strains and their application for the production of guanosine and thymidine[J].Microb Cell Factor,2016,15(1):94.
[13]夏雨,成玉梁,李达倩,吕源玲.枯草芽孢杆菌分泌载体构建及其对脂肪酶A的分泌表达[J].安徽农业科学,2011,39(30):18435-18437.
[14]赵君峰.淀粉液化芽孢杆菌ES-2-4基因组改组提高脂肽产量及其突变菌株差异蛋白组学分析[D].南京:南京农业大学,2012.
[15]Nguyen H D,Nguyen Q A,Ferreira R C,et al.Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability[J].Plasmid,2005,54(3):241-248.
[16]余小霞,田健,刘晓青,等.枯草芽孢杆菌表达系统及其启动子研究进展[J].生物技术通报,2015,31(2):35-44.
[17]张飞燕,何敏,王振华,等.铜绿假单胞菌脂肪酶基因(Lip A)在枯草芽孢杆菌pab02中的整合表达[J].湖南农业大学学报(自然科学版),2017,43(2):200-205.
[18]Yang M,Zhang W,Ji S,et al.Generation of an artificial double promoter for protein expression in Bacillus subtilisthrough a promoter trap system[J].Plos One,2013,8(2):e56321.
[19]杨韵霏,李由然,张梁,等.细菌麦芽糖淀粉酶在枯草芽孢杆菌中的诱导型异源表达[J].微生物学通报,2017,44(2):263-273.
[20]Phanaksri T,Luxananil P,Panyim S,et al.Synergism of regulatory elements inσ(B)-andσ(A)-dependent promoters enhances recombinant protein expression in Bacillus subtilis.[J].J Biosci Bioengin,2015,120(4):470-475.
[21]Lee S J,Pan J G,Park S H,et al.Development of a stationary phase-specific autoinducible expression system in Bacillus subtilis[J].J Biotech,2010,149(1):16-20.
[22]Zhang J,Kang Z,Ling Z,et al.High-level extracellular production of alkaline polygalacturonate lyase in Bacillus subtilis with optimized regulatory elements.[J].Bioresour Tech,2013,146(146C):543-548.
[23]Zanen G,Houben E N G,Rob M,et al.Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis[J].Feb J,2005,272(18):4617.
[24]Beena K,Jayant B,Udgaonkar R,et al.Effect of signal peptide on the stability and folding kinetics of maltose binding protein[J].Biochemistry,2004,43(12):3608.
[25]潘兴亮.枯草芽孢杆菌高效表达系统构建及饲用酶制剂的开发研究[D].北京:中国农业科学院,2016.
[26]Barnes A G,Cerovic V,Hobson P S,et al.Bacillus subtilis,spores:A novel microparticle adjuvant which can instruct a balanced Th1 and Th2 immune response to specific antigen[J].Europ J Immunol,2007,37(6):1538.
[27]李志会,岳盈盈,李鹏,等.EV71VP1黏膜疫苗制备及其诱导黏膜免疫的实验研究[J].山东医药,2011,51(43):4-5.
[28]苗雨,张珍珍,郭彦,等.一种展示SARS冠状病毒受体结合区的重组枯草杆菌芽孢的制备及免疫原性分析[J].生物技术通讯,2013(3):342-346.
[29]Amuguni H,Lee S,Kerstein K,et al.Sublingual immunization with an engineered Bacillus subtilis strain expressing tetanus toxin fragment C induces systemic and mucosal immune responses in piglets.[J].Microb Infect,2012,14(5):447.
[30]Cuburu N,Kweon M N,Song J H,et al.Sublingual immunization induces broad-based systemic and mucosal immune responses in mice[J].Vaccine,2007,25(51):8598.
[31]任晓冰,窦小龙,魏婕,等.口蹄疫病毒表位嵌合蛋白在枯草芽孢杆菌中的分泌表达[J].中国兽医科学,2014(3):287-292.
[32]戴茜茜,王振华,谷笑笑,等.枯草芽孢杆菌芽孢表面展示猪流行性腹泻病毒S蛋白保护性抗原片段的研究[J].中国兽医科学,2016,46(1):38-45
[33]Mou C,Zhu L,Xing X,et al.Immune responses induced by recombinant Bacillus subtilis expressing the spike protein of transmissible gastroenteritis virus in pigs[J].Antivir Res,2016,131:74.
[34]刘瑞瑞.口服禽流感重组枯草芽孢杆菌对鸡免疫和生长的影响[D].南京:南京农业大学,2015.
[35]刘明刚,刘红露,戴茜茜,等.以CotB作为融合基序在枯草芽孢杆菌芽孢表面展示雏沙门菌Omp C的研究[J].中国兽医科学,2015,45(11):1159-1165.
[2]卢争辉,周玉玲,张晓舟,等.感受态还是芽孢?细胞命运决定的遗传调控网络[J].生物工程学报,2015,31(11):1543-1552
[3]张晓舟.枯草杆菌新型表达系统和遗传操作体系的建立及应用[D].南京:南京农业大学,2006.
[4]李信志,卢争辉,周玉玲,等.枯草芽孢杆菌SCK6超级感受态的制备和转化条件优化[J].生物工程学报,2017,33(4):692-698.
[5]You C,Zhang X Z,Zhang Y H P.Simple cloning via direct transformation of PCR product(DNA Multimer)to Escherichia coli and Bacillus subtilis[J].Appl Environ Microbiol,2012,78(5):1593.
[6]李瑞芳,薛雯雯,黄亮,等.枯草芽孢杆菌感受态细胞的制备及质粒转化方法研究[J].生物技术通报,2011,35(5):227-230.
[7]Zhang X Z,Zhang Y.Simple,fast and high-efficiency transformation system for directed evolution of cellulase in Bacillus subtilis.[J].Microb Biotechnol,2011,4(1):98.
[8]Piggot P J,Hilbert D W.Sporulation of Bacillus subtilis[J].Current Opin Microbiol,2004,7(6):579-586.
[9]王慧.芽孢杆菌遗传操作体系初建及工程菌株分子改良[D].北京:中国农业科学院,2016.
[10]何庆,王茜,朱清华,等.基因组工程在微生物细胞工厂构建中的应用进展[J].生物产业技术,2017(1):31-36.
[11]Toya Y,Hirasawa T,Morimoto T,et al.13 C-metabolic flux analysis in heterologous cellulase production by Bacillus subtilis,genome-reduced strain[J].J Biotech,2014,179(1):42-49.
[12]Li Y,Zhu X,Zhang X,et al.Characterization of genome-reduced Bacillus subtilis,strains and their application for the production of guanosine and thymidine[J].Microb Cell Factor,2016,15(1):94.
[13]夏雨,成玉梁,李达倩,吕源玲.枯草芽孢杆菌分泌载体构建及其对脂肪酶A的分泌表达[J].安徽农业科学,2011,39(30):18435-18437.
[14]赵君峰.淀粉液化芽孢杆菌ES-2-4基因组改组提高脂肽产量及其突变菌株差异蛋白组学分析[D].南京:南京农业大学,2012.
[15]Nguyen H D,Nguyen Q A,Ferreira R C,et al.Construction of plasmid-based expression vectors for Bacillus subtilis exhibiting full structural stability[J].Plasmid,2005,54(3):241-248.
[16]余小霞,田健,刘晓青,等.枯草芽孢杆菌表达系统及其启动子研究进展[J].生物技术通报,2015,31(2):35-44.
[17]张飞燕,何敏,王振华,等.铜绿假单胞菌脂肪酶基因(Lip A)在枯草芽孢杆菌pab02中的整合表达[J].湖南农业大学学报(自然科学版),2017,43(2):200-205.
[18]Yang M,Zhang W,Ji S,et al.Generation of an artificial double promoter for protein expression in Bacillus subtilisthrough a promoter trap system[J].Plos One,2013,8(2):e56321.
[19]杨韵霏,李由然,张梁,等.细菌麦芽糖淀粉酶在枯草芽孢杆菌中的诱导型异源表达[J].微生物学通报,2017,44(2):263-273.
[20]Phanaksri T,Luxananil P,Panyim S,et al.Synergism of regulatory elements inσ(B)-andσ(A)-dependent promoters enhances recombinant protein expression in Bacillus subtilis.[J].J Biosci Bioengin,2015,120(4):470-475.
[21]Lee S J,Pan J G,Park S H,et al.Development of a stationary phase-specific autoinducible expression system in Bacillus subtilis[J].J Biotech,2010,149(1):16-20.
[22]Zhang J,Kang Z,Ling Z,et al.High-level extracellular production of alkaline polygalacturonate lyase in Bacillus subtilis with optimized regulatory elements.[J].Bioresour Tech,2013,146(146C):543-548.
[23]Zanen G,Houben E N G,Rob M,et al.Signal peptide hydrophobicity is critical for early stages in protein export by Bacillus subtilis[J].Feb J,2005,272(18):4617.
[24]Beena K,Jayant B,Udgaonkar R,et al.Effect of signal peptide on the stability and folding kinetics of maltose binding protein[J].Biochemistry,2004,43(12):3608.
[25]潘兴亮.枯草芽孢杆菌高效表达系统构建及饲用酶制剂的开发研究[D].北京:中国农业科学院,2016.
[26]Barnes A G,Cerovic V,Hobson P S,et al.Bacillus subtilis,spores:A novel microparticle adjuvant which can instruct a balanced Th1 and Th2 immune response to specific antigen[J].Europ J Immunol,2007,37(6):1538.
[27]李志会,岳盈盈,李鹏,等.EV71VP1黏膜疫苗制备及其诱导黏膜免疫的实验研究[J].山东医药,2011,51(43):4-5.
[28]苗雨,张珍珍,郭彦,等.一种展示SARS冠状病毒受体结合区的重组枯草杆菌芽孢的制备及免疫原性分析[J].生物技术通讯,2013(3):342-346.
[29]Amuguni H,Lee S,Kerstein K,et al.Sublingual immunization with an engineered Bacillus subtilis strain expressing tetanus toxin fragment C induces systemic and mucosal immune responses in piglets.[J].Microb Infect,2012,14(5):447.
[30]Cuburu N,Kweon M N,Song J H,et al.Sublingual immunization induces broad-based systemic and mucosal immune responses in mice[J].Vaccine,2007,25(51):8598.
[31]任晓冰,窦小龙,魏婕,等.口蹄疫病毒表位嵌合蛋白在枯草芽孢杆菌中的分泌表达[J].中国兽医科学,2014(3):287-292.
[32]戴茜茜,王振华,谷笑笑,等.枯草芽孢杆菌芽孢表面展示猪流行性腹泻病毒S蛋白保护性抗原片段的研究[J].中国兽医科学,2016,46(1):38-45
[33]Mou C,Zhu L,Xing X,et al.Immune responses induced by recombinant Bacillus subtilis expressing the spike protein of transmissible gastroenteritis virus in pigs[J].Antivir Res,2016,131:74.
[34]刘瑞瑞.口服禽流感重组枯草芽孢杆菌对鸡免疫和生长的影响[D].南京:南京农业大学,2015.
[35]刘明刚,刘红露,戴茜茜,等.以CotB作为融合基序在枯草芽孢杆菌芽孢表面展示雏沙门菌Omp C的研究[J].中国兽医科学,2015,45(11):1159-1165.