UPR信号通路遗传操作提高酵母异源表达
|
摘要
随着后基因组时代的来临,越来越多的蛋白质开发成药物或直接进行分析,这就需要获得合适的宿主来进行异源表达。而为便于后续的纯化过程,分泌性的表达宿主更为理想,虽然在毕赤酵母中获得了较低成本的大量蛋白的异源表达,但因其存在着诱导时间长、受专利保护等因素的限制,国际上针对非常规酵母及丝状真菌的研究逐渐成为热点。
UPR信号传导是一个进化上保守的通路,尤其是在酵母和丝状真菌中表现了相当的保守性,由内质网腔的未折叠蛋白累积造成胁迫来自然激活。通路上的转录因子Hac1p在激活、维持这一信号途径中起着关键作用,通过依赖于UPR信号响应元件(UPRE)调节蛋白折叠或降解等相关基因的转录过程,来缓解产生于内质网腔的这种胁迫。
为了研究UPR信号的激活对异源表达的影响,我们首先克隆了溶菌酶基因并与α信号肽融合后在酿酒酵母中表达。建立了一个基于平板计数的穿梭质粒拷贝数确定方法,通过质粒的稳定性及拷贝数的研究建立了一步培养的方法使溶菌酶快速表达、分析。
本文用逆转录方法克隆了HAC1icDNA,以不同拷贝数的载体或在基因组水平做HAC1的基因去内含子扩增的方式介导UPR信号的持续激活,结果表明UPR信号激活提高异源表达的同时存在着剂量效应。YEplac181介导的UPR激活提高异源表达程度最大,使溶菌酶的表达量跨越了微克级,提高到2.7 mg/L。
本文建立了一个新的高通量异源表达菌株筛选系统,本系统不依赖于流式细胞术及微孔板检测仪。依据色谱原理,展示在细胞表面的CBD或亮氨酸拉链与纤维素基质的相互作用成为抵抗来至流动相的“筛选压力”。基于这个系统,通过定向进化技术获得了一个Hac1p突变子(Gly48,Gly111),记为Hac1pm1,异源表达量达到了3.9 mg HEL/L。通过对比Hac1p m1与野生型结构分析推测,氨基酸突变,由于空间位阻减小,可能导致其折叠发生改变,影响其功能。更为重要的是Arg48Gly不但打破了一段α螺旋,使肽链趋向改变走向,更使得亮氨酸拉链基序与DNA的作用发生改变,导致突变子调节的靶基因发生变化。同时也导致实质“剂量”与野生型相比降低,进而调节靶基因的转录发生变化,导致异源表达量的提高。
最后,我们通过分析UPR信号途径在转录水平及翻译水平调节的靶基因性质,确定了UPR信号激活过程中菌体生长受抑制的主要原因是氧还平衡的破坏。为进一步提高异源表达量,通过UPRE介导的UPR信号扩增遗传操作,利用KAR2启动子替换MAE1启动子并扩增于ura3基因座,以及敲除基因GPD2来维持氧还平衡。最终,与未进行能荷及氧还平衡调节的对照菌株相比,前者异源表达量是原来的1.4倍(4.9 mg HEL/L),后者为1.2倍(4.4 mg HEL/L)。
本文针对UPR这一保守的信号途径在酿酒酵母中进行了系列遗传操作,期望在其他微生物中得到应用。
Many natural proteins were developed into drugs and produced for direct analysis, requiring improved hosts to achieve high-level heterologous proteins production. The unfolded protein response (UPR) is an evolutionarily conserved mechanism (especially the yeast and filamentous fungi) by which all eukaryotic cells adapt to the accumulation of unfolded proteins in endoplasmic reticulum (ER). Hac1p, the transcription factor can activate the UPR signal transduction, which regulates genes related the protein folding, degradation and etc.
To investigate the effects of engineering UPR pathway to heterologous expression, hen egg white lysozyme (HEL) cDNA cloned from tissue of mature hen oviduct fused toα-signal peptide was inserted into pYES2 and expressed as a modle protein. The shuttle plasmids copy number was assessed using a novel method based on the E.coli transformation and plate count technique, meanwhile, the plasmids stability was estimated. Results proposed one-step cultivation strategy was a feasible alternative.
HAC1icDNA was gained by RT-PCR and cloned into different plasmids or substituted the HAC1locus on genome. The results indicated the effect of UPR pathway activation on heterologous expression was dose-dependent, and yeast strain W303 harboring HAC1icDNA on a multi-copy plasmid YEplac181 achieved the highest yields, 2.7 mg HEL/L.
A novel high-throughput screening (HTS) system to obtain heterologous over-expression in Saccharomyces cerevisiae strains was developed. The protocol designed here was based on bio-macromolecular physical interaction between CBD or leucine zipper displayed on the surface of hosts and the cellulose substrate. Using the screening system, directed evolution of Hac1p was carried out in Saccharomyces cerevisiae, and an improved mutant (Hac1pm1, Arg48Gly, Ser111Gly, 3.9 mg HEL/L yields) were selected. Results proposed that through selective pressure, this assay may afford a more effective screening system compared with previous selection system. Moreover, it could be employed in general biochemical analysis without utilization of flow cytometry or well plate reader.
At last, through UPR pathway analysis, we found that the reduction-oxidation (Redox) balance disruption of strains by UPR activation was the main reason for growth deficit. UPRE (UPR element) mediating the signal augmentation of UPR pathway was performed to improve the production. Truncated form, MAE1s under the control of the promoter of KAR2, harboring UPRE sequence was amplified into the ura3 locus, and 1.4 fold yield was gained compared with the control (W303 +Hac1pm1) when UPR pathway was activated by Hac1pm1. Further, GPD2 was knocked out to balance the Redox and achieved more large heterologous expression.
Engineering UPR pathway is expected to improve the heterologous expression in other microorganisms (P.pastoris, K.lactis, filamentous fungi, etc.).
UPR信号传导是一个进化上保守的通路,尤其是在酵母和丝状真菌中表现了相当的保守性,由内质网腔的未折叠蛋白累积造成胁迫来自然激活。通路上的转录因子Hac1p在激活、维持这一信号途径中起着关键作用,通过依赖于UPR信号响应元件(UPRE)调节蛋白折叠或降解等相关基因的转录过程,来缓解产生于内质网腔的这种胁迫。
为了研究UPR信号的激活对异源表达的影响,我们首先克隆了溶菌酶基因并与α信号肽融合后在酿酒酵母中表达。建立了一个基于平板计数的穿梭质粒拷贝数确定方法,通过质粒的稳定性及拷贝数的研究建立了一步培养的方法使溶菌酶快速表达、分析。
本文用逆转录方法克隆了HAC1icDNA,以不同拷贝数的载体或在基因组水平做HAC1的基因去内含子扩增的方式介导UPR信号的持续激活,结果表明UPR信号激活提高异源表达的同时存在着剂量效应。YEplac181介导的UPR激活提高异源表达程度最大,使溶菌酶的表达量跨越了微克级,提高到2.7 mg/L。
本文建立了一个新的高通量异源表达菌株筛选系统,本系统不依赖于流式细胞术及微孔板检测仪。依据色谱原理,展示在细胞表面的CBD或亮氨酸拉链与纤维素基质的相互作用成为抵抗来至流动相的“筛选压力”。基于这个系统,通过定向进化技术获得了一个Hac1p突变子(Gly48,Gly111),记为Hac1pm1,异源表达量达到了3.9 mg HEL/L。通过对比Hac1p m1与野生型结构分析推测,氨基酸突变,由于空间位阻减小,可能导致其折叠发生改变,影响其功能。更为重要的是Arg48Gly不但打破了一段α螺旋,使肽链趋向改变走向,更使得亮氨酸拉链基序与DNA的作用发生改变,导致突变子调节的靶基因发生变化。同时也导致实质“剂量”与野生型相比降低,进而调节靶基因的转录发生变化,导致异源表达量的提高。
最后,我们通过分析UPR信号途径在转录水平及翻译水平调节的靶基因性质,确定了UPR信号激活过程中菌体生长受抑制的主要原因是氧还平衡的破坏。为进一步提高异源表达量,通过UPRE介导的UPR信号扩增遗传操作,利用KAR2启动子替换MAE1启动子并扩增于ura3基因座,以及敲除基因GPD2来维持氧还平衡。最终,与未进行能荷及氧还平衡调节的对照菌株相比,前者异源表达量是原来的1.4倍(4.9 mg HEL/L),后者为1.2倍(4.4 mg HEL/L)。
本文针对UPR这一保守的信号途径在酿酒酵母中进行了系列遗传操作,期望在其他微生物中得到应用。
Many natural proteins were developed into drugs and produced for direct analysis, requiring improved hosts to achieve high-level heterologous proteins production. The unfolded protein response (UPR) is an evolutionarily conserved mechanism (especially the yeast and filamentous fungi) by which all eukaryotic cells adapt to the accumulation of unfolded proteins in endoplasmic reticulum (ER). Hac1p, the transcription factor can activate the UPR signal transduction, which regulates genes related the protein folding, degradation and etc.
To investigate the effects of engineering UPR pathway to heterologous expression, hen egg white lysozyme (HEL) cDNA cloned from tissue of mature hen oviduct fused toα-signal peptide was inserted into pYES2 and expressed as a modle protein. The shuttle plasmids copy number was assessed using a novel method based on the E.coli transformation and plate count technique, meanwhile, the plasmids stability was estimated. Results proposed one-step cultivation strategy was a feasible alternative.
HAC1icDNA was gained by RT-PCR and cloned into different plasmids or substituted the HAC1locus on genome. The results indicated the effect of UPR pathway activation on heterologous expression was dose-dependent, and yeast strain W303 harboring HAC1icDNA on a multi-copy plasmid YEplac181 achieved the highest yields, 2.7 mg HEL/L.
A novel high-throughput screening (HTS) system to obtain heterologous over-expression in Saccharomyces cerevisiae strains was developed. The protocol designed here was based on bio-macromolecular physical interaction between CBD or leucine zipper displayed on the surface of hosts and the cellulose substrate. Using the screening system, directed evolution of Hac1p was carried out in Saccharomyces cerevisiae, and an improved mutant (Hac1pm1, Arg48Gly, Ser111Gly, 3.9 mg HEL/L yields) were selected. Results proposed that through selective pressure, this assay may afford a more effective screening system compared with previous selection system. Moreover, it could be employed in general biochemical analysis without utilization of flow cytometry or well plate reader.
At last, through UPR pathway analysis, we found that the reduction-oxidation (Redox) balance disruption of strains by UPR activation was the main reason for growth deficit. UPRE (UPR element) mediating the signal augmentation of UPR pathway was performed to improve the production. Truncated form, MAE1s under the control of the promoter of KAR2, harboring UPRE sequence was amplified into the ura3 locus, and 1.4 fold yield was gained compared with the control (W303 +Hac1pm1) when UPR pathway was activated by Hac1pm1. Further, GPD2 was knocked out to balance the Redox and achieved more large heterologous expression.
Engineering UPR pathway is expected to improve the heterologous expression in other microorganisms (P.pastoris, K.lactis, filamentous fungi, etc.).
引文
[1] Itakura K, Hirose T, Crea R, et al. Expression in Escherichia coli of a chemically synthesized gene for the hormone somatostatin. Science, 1977, 198:1056~1063.
[2] Klein S, Geiger T, Linchevski I, et al. Expression and purification of active PKB kinase from Escherichia coli. Protein Express. Purif., 2005, 41:162~169.
[3] Julia Scerbo M, Bibolini MJ, Barra JL, et al. Expression of a bioactive fusion protein of Escherichia coli heat-labile toxin B subunit to a synapsin peptide. Protein Expression and Purification, 2008, 59:320~326.
[4] Magliery TJ, Wilson CG, Pan W, et al. Detecting protein-protein interactions with a green fluorescent protein fragment reassembly trap: scope and mechanism. J. Am. Chem. Soc., 2005, 127:146~157.
[5] Clark EB. Refolding of recombinant proteins. Curr. Opin. Biotechnol., 1998, 9: 157~163.
[6] Yamabhai M, Emrat S, Sukasem S, et al. Secretion of recombinant Bacillus hydrolytic enzymes using Escherichia coli expression systems. Journal of Biotechnology, 2008, 133:50~57.
[7] Ferreira LC, Ferreira RC, Schumann W. Bacillus subtilis as a tool for vaccine development: from antigen factories to delivery vectors. Ann. Braz. Acad. Sci., 2005, 77:113~124.
[8] Barnard GC, Henderson GE, Srinivasan S, Gerngross TU. High level recombinant protein expression in Ralstonia eutropha using T7 RNA polymerase based amplication. Protein Expression and Purifcation, 2004, 38:264 ~ 271.
[9] Carvalho H, Lima L, Lescure N, et al. Differential expression of the two cytosolic glutamine synthetase genes in various organs of Medicago truncatula. Plant Science, 2000, 159, 301~312.
[10] Yang LJ, Tada Y, Yamamoto MP et al. A transgenic rice seed accumulating an anti-hypertensive peptide reduces the blood pressure of spontaneously hypertensive rats. FEBS Letters, 2006, 580:3315~3320.
[11] Yang L, Kajiura H, Suzuki K et al. Generation of a transgenic rice seed-based edible vaccine against house dust mite allergy. Biochemical and Biophysical Research Communications, 2008, 365:334~339.
[12] Youma JW, Won YS, Jeon JH et al. Oral immunogenicity of potato-derived HbsAg middle protein in BALB/c mice. Vaccine, 2007, 25:577~584.
[13] Moravec T, Schmidt MA, Herman EM et al. Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine. Vaccine, 2007, 25:1647~1657.
[14] Rigano MM, Dreitz S, Kipnis AP et al. Oral immunogenicity of a plant-made, subunit, tuberculosis vaccine. Vaccine, 2006, 24:691~695.
[15] Ritala A, Wahlstr?m EH, Holkeri H. et al. Production of a recombinant industrial protein using barley cell cultures. Protein Expression and Purification, 2008, 59:274~281.
[16] Smith GE, Summers MD, Fraser MJ. Production of human beta interferon in insect cells in fected with a baculovirus expression vector. Mol. Cell. Biol., 1983, 3: 2156~2165.
[17] Honjo E, Shoyama Y, Tamada T, et al. Expression of the extracellular region of the human interleukin-4 receptor a chain and interleukin-13 receptor a1 chain by a silkworm–baculovirus system. Protein Expression and Purification, 2008, 60:25~30.
[18] Donald L, Jarvis DL. Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology, 2003, 310:1~7.
[19] Jarvis DL. Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology, 2003, 310:1~7.
[20] Taylor AL, Haze-Filderman A, Blumenfel A et al. High yield of biologically active recombinant human amelogenin using the baculovirus expression system. Protein Expression and Purification, 2006, 45:43~53.
[21] Peng S, Lva N, Zhang Y, et al. HeImmune response induced by spike protein from transmissible gastroenteritis coronavirus expressed in mouse mammary cells. Virus Research, 2007, 128:52~57.
[22] Kevin E, Cott V, Stephen P. et al. Transgenic pigs as bioreactors: a comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland. Genetic Analysis: Biomolecular Engineering, 1999, 15: 155~160.
[23] Takami H, Watanabe H, Ohmori Y, et al. Human alkaline phosphatase expression and secretion into chicken eggs after in vivo gene electroporation in the oviduct of laying hens. Biochemical and Biophysical Research Communications, 2002, 292:88~93.
[24] Ziomek CA. Commercialization of proteins produced in the mammary gland.Theriogenology, 1996, 49:139~144.
[25] European Medicines Agency London, Question and answers on atryn. International Non-proprietary Name (INN): antithrombin alfa. 2006, Doc.Ref. EMEA/191862/2006.
[26] Makrides SC, Makrides SC. Virus-based vectors for gene expression in mammalian cells: SV40. Gene Transfer and Expression in Mammalian Cells: New Comprehensive Biochemistry, 2003, 38:71~91.
[27] Gillard M, Chatelain P, Fuks B. Binding characteristics of levetiracetam to synaptic vesicle protein 2A (SV2A) in human brain and in CHO cells expressing the human recombinant protein. European Journal of Pharmacology, 2006, 536:102~108.
[28] Bork K, Reutter W, Weidemann W, Horstkorte R. Enhanced sialylation of EPO by overexpression of UDP-GlcNAc 2-epimerase/ManAc kinase containing a sialuria mutation in CHO cells. FEBS Letters, 2007, 581:4195~4198.
[29] Teixeira A, Cunha AE, Clemente JJ, et al. Modelling and optimization of a recombinant BHK-21 cultivation process using hybrid grey-box systems. Journal of Biotechnology, 2005, 118:290~303.
[30] Jardin BA, Zhao Y, Selvaraj M, et al. Expression of SEAP (secreted alkaline phosphatase) by baculovirus mediated transduction of HEK 293 cells in a hollow fiber bioreactor system. Journal of Biotechnology, 2008, doi:10.1016/j.jbiotec.2008.04.006.
[31] Müller D, Katinger H, Grillari J. MicroRNAs as targets for engineering of CHO cell factories. Trends in Biotechnology, 2008, doi:10.1016/j.tibtech.2008.03.010.
[32] Hitzeman RA, Hagie FE, Levine HL, et al. Expression of a human gene for interferon in yeast. Nature, 1981, 293:717~722.
[33] Thor D, Xiong S, Orazem CC, et al. Cloning and characterization of the Pichia pastoris MET2 gene as a selectable marker. FEMS Yeast Research, 2005, 5:935~942.
[34] Steinborna G, Gellissenb G, Kunze G. Assessment of Hansenula polymorpha and Arxula adeninivorans-derived rDNA-targeting elements for the design of Arxula adeninivorans expression vectors. FEMS Yeast Research, 2005, 5:1047~1054.
[35] Broach JR, Li Y, Feldman J, et al. Localization and sequence analysis of yeast origins of DNA replication. Cold Spring Harbor Symposium on Quantitative Biology, 1982, 47:1165~1173.
[36] Falco SC, Li Y, Broach JR and Botstein D. Genetic properties of chromosomally integrated 2μplasmid DNA in yeast. Cell, 1982, 29:573~584.
[37] Jayaram M, Li Y, Broach JR. The yeast plasmid 2μencodes componentsrequired for its high copy propagation, Cell, 1983, 34:95~104.
[38] Jayaram, M, Li, YY., McLeod M, Broach JR. Analysis of site-specific recombination associated with the yeast plasmid 2 micron circle. in mechanisms of DNA replication and recombination UCLA Symposia on Molecular and Cellular Biology (Cozzelli and Fox, eds.) New York, Alan R. Liss, Inc., 1983. 685~694.
[39] Huo KK, Yu LL, Chen XJ, Li YY. A stable vector for high level expression and secretion of human interferonαA gene in yeast. Science in China, 1993, 36:557~567.
[40] Suzuki K, Ichikawa K, Jigami Y. Yeast mutants with enhanced ability to secrete human lysozyme:Isolation and identification of a protease-deficient mutant. Mol Gen Genet, 1989, 219:58~64.
[41] Sampaio PN, Fortes AM, Cabral JM, et al. Production and Characterization of Recombinant Cyprosin B in Saccharomyces cerevisiae (W303-1A) Strain. Journal of Bioscience and Bioengineering, 2008, 105:305~312.
[42] Griffith DA, Delipala C, Leadsham J et al. A novel yeast expression system for the overproduction of quality-controlled membrane proteins. FEBS Letters, 2003, 553:45~50.
[43] Juozapaitis M, Zvirbliene A, Kucinskaite I. Synthesis of recombinant human parainfluenza virus 1 and 3 nucleocapsid proteins in yeast Saccharomyces cerevisiae. Virus Research, 2008, 133:178~186.
[44] Juozapaitis M, Serva A, Kucinskaite I. Generation of menangle virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae. Journal of Biotechnology, 2007, 130:441~447.
[45] Juozapaitis M, Serva A, Zvirbliene A. Generation of henipavirus nucleocapsid proteins in yeast Saccharomyces cerevisiae. Virus Research, 2007, 124:95~102.
[46] Gasser B, Maurer M, Gach J et al. Engineering of Pichia pastoris for improved production of antibody fragments. Biotechnology and Bioengineering, 2006, 94: 353~361.
[47] Wetterholm A, Molina DM, Nordlund P, et al. High-level expression, purification, and crystallization of recombinant rat leukotriene C4 synthase from the yeast Pichia pastoris. Protein Expression and Purification, 2008, 60:1~6.
[48] Zámocky M, Schümann C, Sygmund C, O'Callaghan J, et al. Cloning, sequence analysis and heterologous expression in Pichia pastoris of a gene encoding a thermostable cellobiose dehydrogenase from Myriococcum thermophilum. Protein Expression and Purification, 2008, 59:258~265.
[49] Madzak C, Gaillardin C, Beckerich JM. Heterologous protein expression andsecretion in the non-conventional yeast Yarrowia lipolytica: a review. Journal of Biotechnology, 2004, 109:63~81.
[50] Fickers P, Fudalej F, Nicaud JM, Destain J, Thonart P. Selection of new over-producing derivatives for the improvement of extracellular lipase production by the non-conventional yeast Yarrowia lipolytica. Journal of Biotechnology, 2005, 115: 379~386.
[51] Platko JD, Deeg M, Thompson V, et al. Heterologous expression of Mytilus californianus foot protein three (Mcfp-3) in Kluyveromyces lactis. Protein Expression and Purification, 2008, 57:57~62.
[52] Nevalainen KM, Te'o VS, Bergquist PL. Heterologous protein expression in filamentous fungi. Trends in Biotechnology, 2005, 23:468~474.
[53] Wang Y, Xue W, Sims AH, et al. Isolation of four pepsin-like protease genes from Aspergillus niger and analysis of the effect of disruptions on heterologous laccase expression. Fungal Genetics and Biology, 2008, 45:17~27.
[54] Koda A, Bogaki T, Minetoki T, et al. High expression of a synthetic gene encoding potatoα-glucan phosphorylase in Aspergillus niger. Journal of bioscience and bioengineering, 2005, 100:531~537.
[55] Ahamed A, Vermette P. Culture-based strategies to enhance cellulase enzyme production from Trichoderma reesei RUT-C30 in bioreactor culture conditions. Biochemical Engineering Journal, 2008, 40:399~407.
[56] Wiebe MG. Stable production of recombinant proteins in filamentous fungi- problems and improvements. Mycologist, 2003, 17:140~144.
[57] Meyer V. Genetic engineering of filamentous fungi-progress, obstacles and future trends. Biotechnology Advances, 2008, 26:177~185.
[58] Tamalampudi S, Talukder MR, Hama S, et al. Enzymatic production of biodiesel from Jatropha oil: a comparative study of immobilized-whole cell and commercial lipases as a biocatalyst. Biochemical Engineering Journal, 2008, 39:185~189.
[59] Novick P, Ferro S, Schekman R. Order of events in the yeast secretory pathway. Cell, 1981, 25:461~419.
[60] Novick P, Garrett MD, Brennwald P, et al. Control of exocytosis in yeast. Cold Spring Harbor Symposia On Quantitative Biology, 1995, 60:171~177.
[61] Rapoport TA, Rolls MM and Jungnickel B. Approaching the mechanism of protein transport across the ER membrane. Curr Opin Cell Biol, 1996, 8:499~504.
[62] Wiertz EJ, Tortorella D, Bogyo M, et al. Sec61-mediated transfer proteasome for destruction. Nature, 1996, 384: 432~438.
[63] Hartmann E, Sommer T, Prehn S, et al. Evolutionary conservation ofcomponents of the protein translocation complex. Nature, 1994, 367: 654~657.
[64] Glick BS. Can Hsp70 proteins act as force-generating motors? Cell, 1995, 80:11~4.
[65] Ng DT, Brown JD and Walter P. Signal sequences specify the targeting route to the endoplasmic reticulum membrane. J Cell Biol, 1996, 134:269~78.
[66] B?hni PC, Deshaies RJ and Schekman RW. SEC11 is required for signal peptide processing and yeast cell growth. J Cell Biol, 1988, 106:1035~1042.
[67] Evans EA, Gilmore R, and Blobel G. Purification of microsomal signal peptidase as a complex. Proc Natl Acad Sci USA, 1986, 83:581~585.
[68] Gavel Y, von Heijne G. Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering. Prot Eng, 1990, 3:433~442.
[69] Nilsson IM and von Heijne G. Determination of the distance between the oligosaccharyl transferase active site and the endoplasmic reticulum membrane. J Biol Chem, 1993, 268:5798~5801.
[70] Shelness GS, Lin L and Nicchitta CV. Membrane topology and biogenesis of eukaryotic signal peptidase. J Biol Chem, 1993, 268:5201~5208.
[71] Meldolesi J and Pozzan T. The endoplasmic reticulum Ca2+ store: a view from the lumen. Trends Biochem Sci, 1998, 23:10~14.
[72] Ellgaard L, Molinari M, Helenius A. Setting the standards: quality control in the secretory pathway. Science, 1999, 286:1882~1888.
[73] Orci L, Stamnes M, Ravazzola M, et al. Bidirectional transport by distinct populations of COPI-coated vesicles. Cell, 1997, 90:335~349.
[74] Scales SJ, Pepperkok R, Kreis TE. Visualization of ER-to-Golgi transport in living cells reveals a sequential mode of action for COPII and COPI. Cell, 1997, 90: 1137~1148.
[75] Barlowe C, Schekman R. SEC12 encodes a guanine-nucleotide exchange factor essential for transport vesicle budding from the ER. Nature, 1993, 365:347~349.
[76] Antonny B and Schekman R. ER export: public transportation by the COPII coach. Current Opinion in Cell Biology, 2001, 13:438~443.
[77] Harter C, Wieland F. The secretory pathway: mechanisms of protein sorting and transport. Biochim Biophys Acta, 1996, 1286:75~93.
[78] Lledo PM. Exocytosis in excitable cells: a conserved molecular machinery from yeast to neuron. European Journal of Endocrinology, 1997, 137:1~9.
[79] Gordon CL, Archer DB, Jeenes DJ, et al. A glucoamylase::GFP gene fusion to study protein secretion by individual hyphae of Aspergillus niger. J Microbiol Methods, 2000, 42:39~48.
[80] Nyk?nen M, Saarelainen R, Raudaskoski M, et al. Expression and secretion ofbarley Cysteine Endopeptidase B and Cellobiohydrolase I in Trichoderma reesei. Appl Environ Microbiol, 1997, 63:4929~4937.
[81] Nyk?nen M. Protein secretion in Trichoderma reesei. Ph.D. thesis, Dept Biol Environ Sci. Univ. of Jyv?skyl?, 2002.
[82] Gierz G and Bartnicki-Garcia S. A three-dimensional model of fungal morphogenesis based on the vesicle supply center concept. J Theor Biol, 2001, 208: 151~164.
[83] Rupes I, Mao WZ, ?str?m H and Raudaskoski M. Effects of nocodazole and brefeldin A on microrubule cytoskeleton and membrane organization in the homobasidiomycete Schizophyllum. Commune. Protoplasma, 1995, 185:212~221.
[84] Thompson SA, Golightly EJ and Yaver DS. Nucleotide sequence of the Aspergillus niger srpA gene. Gene, 1995, 167:337~338.
[85] Zakrzewska A, Migdalski A, Saloheimo M, et al. cDNA encoding protein Omannosyltransferase from the filamentous fungus Trichoderma reesei; functional equivalence to Saccharomyces cerevisiae PMT2. Curr Genet, 2003, 43:11~16.
[86] Veldhuisen G, Saloheimo M, Fiers MA, et al. Isolation and analysis of functional homologues of the secretion- related SAR1 gene of Saccharomyces cerevisiae from Aspergillus niger and Trichoderma reesei. Mol Gen Genet, 1997, 256:446~455.
[87] Vasara T, Ker?nen S, Penttil? M, Saloheimo M. Characterisation of two 14-3-3 genes from Trichoderma reesei: interactions with yeast secretory pathway components. Biochim Biophys Acta., 2002, 1590:27~40.
[88] Gething MJ, Sambrook J. Protein folding in the cell. Nature, 1992, 355:33~45.
[89] Mori K, Sant A, Kohno K, et al. A 22bp cis-acting element is necessary and sufficient for the induction of the yeast KAR2 (BiP) gene by unfolded proteins. EMBO J, 1992, 11:2583~2593.
[90] Patil C, Walter P. Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr Opin Cell Biol, 2001, 13:349~355.
[91] Cox JS and Walter P. A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response. Cell, 1996, 87:391~404.
[92] Kaufman RJ. Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev., 1999, 13:1211~1233.
[93] Sidrauski C, Walter P. The transmembrane kinase Ire1p is a sitespecific endonuclease that initiates mRNA splicing in the unfolded protein response. Cell, 1997, 90:1031~1039.
[94] Sidrauski C, Cox JS, Walter P. tRNA ligase is required for regulated mRNA splicing in the unfolded protein response. Cell, 1996, 87:405~413.
[95] Chapman RE, Walter P. Translational attenuation mediated by an mRNA intron. Curr Biol., 1997, 7:850-859.
[96] Kawahara T, Yanagi H, Yura T et al. Unconventional splicing of HAC1/ERN4 mRNA required for the unfolded protein response. The Journal of Biological Chemistry, 1998, 273:1802~1807.
[97] Ruegsegger U, Leber JH, Walter P. Block of HAC1 mRNA translation by long-range base pairing is released by cytoplasmic splicing upon induction of the unfolded protein response. Cell, 2001, 107:103~114.
[98] Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol, 2007, 8:519~529.
[99] Mori K, Ogawa N, Kawahara T et al. Palindrome with spacer of one nucleotide is characteristic of the cis-acting unfolded protein response element in Saccharomyces cerevisiae. The Journal of Biological Chemistry, 1998, 273:9912~9920.
[100] Mori K, Sant A, Kohno K et al. A 22 bp cis-acting element is necessary and sufficient for the induction of the yeast KAR2 (BiP) gene by unfolded proteins. The EMBO Journal, 1993, 11:2583~2593.
[101] Casagrande R, Stern P, Brown DM et al. Degradation of proteins from the ER of S. cerevisiae requires an intact unfolded protein response pathway. Mol Cell, 2000, 5:729~735.
[102] Xua P, Radena D, Doyle FJ et al. Analysis of unfolded protein response during single-chain antibody expression in Saccaromyces cerevisiae reveals different roles for BiP and PDI in folding. Metabolic Engineering, 2005, 7: 269~279.
[103] Wang JD, Herman C, Tipton KA. Directed evolution of substrate-optimized GroEL/S chaperonins. Cell, 2002, 111:1027~1039.
[104] Hibbert EG, Baganz F, Hailes HC. Directed evolution of biocatalytic processes. Biomolecular Engineering, 2005, 22:11~19.
[105] Alper H, Moxley J, Nevoigt E et al. Engineering yeast transcription machinery for improved ethanol tolerance and production. Science, 2006, 314:1565~1568.
[106] Jin S, Ye K, Shimizu K. Metabolic flux distributions in recombinant Saccharomyces cerevisiae during foreign protein production. Journal of Biotechnology, 1997, 54:161~174.
[107] Jensen R, Sprague GF, Herskowitz JI. Control of cell type in yeast by the mating type locus. J Mol Biol, 1981, 147:357~372.
[108] Song Y, Sata J, Saito A, Usui M, et al. Effects of calnexin deletion in Saccharomyces cerevisiae on the secretion of glycosylated lysozymes. J. Biochem., 2001, 130:757~764.
[109] Wormald MR, Dwek RA. Glycoproteins: glycan presentation and protein-fold stability. Structure, 1999, 15:150~160.
[110] Shibasaki S, Tanaka A, Ueda M. Development of combinatorial bioengineering using yeast cell surface display-order-made design of cell and protein for bio-monitoring. Biosensors and Bioelectronics, 2003, 19:123~130.
[111] Tanino T, Noguchi E, Kimura S. Effect of cultivation conditions on cell-surface display of Flo1 fusion protein using sake yeast. Biochemical Engineering Journal, 2007, 33:232~237.
[112] Wu CH, Mulchandani A, Chen W. Versatile microbial surface-display for environmental remediation andbiofuels production. Trends in Microbiology, 2008, 16, doi:10.1016/j.tim.2008.01.003.
[113] Khaw TS, Katakura Y, Ninomiya K, et al. Enhancement of ethanol production by promoting surface contact between starch granules and arming yeast in direct ethanol fermentation. Journal of bioscience and bioengineering, 2007, 103:95~97.
[114] Dürauer A, Berger E, Zandian M, et al. Yeast cell surface display system for determination of humoral response to active immunization with a monoclonal antibody against EpCAM. J. Biochem. Biophys Methods, 2008, 70:1109~1115.
[115] Bordes F, Fudalej F, Dossat V, et al. A new recombinant protein expression system for high-throughput screening in the yeast Yarrowia lipolytica. Journal of Microbiological Methods, 2007, 70:493~502.
[116] Shusta EV, Kieke MC, Parke E et al. Yeast polypeptide fusion surface display levels predict thermal stability and soluble secretion efficiency. J Mol Biol, 1999, 292:949~956.
[117] Daugherty PS, Iverson BL. and Georgiou G. Flow cytometric screening of cell-based libraries. Journal of Immunological Methods, 2000, 243:211~227.
[118] Hamilton SR, Davidson RC, Sethuraman N. Humanization of yeast to produce complex terminally sialylated glycoproteins. Science, 2006, 313:1441~1443.
[119] Masuda T, Ueno Y, Kitabatake N. High yield secretion of the sweet-tasting protein lysozyme from the yeast Pichia pastoris. Protein Expression and Purification, 2005, 39:35~42.
[120] Gupta JC, Mukherjee KJ. Stable maintenance of plasmid in continuous culture of yeast under non-selective conditions. Journal of Bioscience and Bioengineering,2001, 92:317~323.
[121] Skulj M, Okr?lar V, Jalen ?, et al. Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes. Microbial Cell Factories, 2008, 7:6 doi:10.1186/1475-2859-7-6.
[122] Bicknell AA, Babour A, Federovitch CM. A novel role in cytokinesis reveals a housekeeping function for the unfolded protein response. The Journal of Cell Biology, 2007, 177:1017~1027.
[123] Ogawa N. and Mori K. Autoregulation of the HAC1 gene is required for sustained activation of the yeast unfolded protein response. Genes to Cells, 2004, 9:95~104.
[124] Ruegsegger U, Leber JH, Walter P. Block of HAC1 mRNA translation by long-range base pairing is released by cytoplasmic splicing upon induction of the unfolded protein response. Cell, 2001, 107:103~114.
[125] Travers KJ, Patil CK, Wodicka L et al. Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell, 2000, 101:249~258.
[126] Andreaus J, Azevedo H, Cavaco-Paulo A. Effects of temperature on the cellulose binding ability of cellulase enzymes. Journal of Molecular Catalysis B: Enzymatic, 1999, 7:233~239.
[127] Santiago-Hernandez Ja, Vasquez-Bahena Jm, Calixto-Romo Ma. Direct immobilization of a recombinant invertase to Avicel by E. coli overexpression of a fusion protein containing the extracellular invertase from Zymomonas mobilis and the carbohydrate-binding domain CBDcex from Cellulomonas fimi. Enzyme and Microbial Technology, 2006, 40:172~176.
[128] Shang J and Geva E. Computational study of a single surface-immobilized two-stranded coiled-coil polypeptide. J. Phys. Chem. B, 2007, 111:4178~4188.
[129] Wang J and Somasundaran P. Mechanisms of ethyl (hydroxyethyl) cellulose-solid interaction: Influence of hydrophobic modification. Journal of Colloid and Interface Science, 2006, 293:322~332.
[130] Levy I, Shoseyov O. Cellulose-binding domains: Biotechnological applications. Biotechnology Advances, 2002, 20:191~213.
[131] Tomme P, Boraston A, McLean B. Characterization and affinity applications of cellulose-binding domains. Journal of Chromatography B, 1998, 715:283~296.
[132] Linder M, Salovuori I, Ruohonen L, Teeri TT. Characterization of a double cellulose-binding domain. Synergistic high affinity binding to crystalline cellulose. J Biol Chem., 1996, 271:21268~72.
[133] Stocker-Majd G, Hilbrig F, Freitag R. Extraction of haemoglobin from human blood by affinity precipitation using a haptoglobin-based stimuli–responsive affinitymacroligand. Journal of Chromatography A, 2008, 1194:57~65.
[134] Dengis, PB, Nélissen LR, and Rouxhet PG. Mechanisms of Yeast Flocculation: comparison of top and bottom-fermenting strains. Applied and Environmental Microbiology, 1995, 61:718~728.
[135] Leber JH, Bernales S, Walter P. IRE1-independent gain control of the unfolded protein response. PloS Biol, 2004, 2:1197~1207.
[136] Mulder HJ, Nikolaev I, Madrid SM. HACA, the transcriptional activator of the unfolded protein response (UPR) in Aspergillus niger, binds to partly palindromic UPR elements of the consensus sequence 50-CAN(G/A) NTGT/GCCT-30. Fungal Genetics and Biology, 2006, 43:560~572.
[137] Payne T, Hanfrey C, Bishop AL, Michael AJ, et al. Transcript-specific translational regulation in the unfolded protein response of Saccharomyces cerevisiae. FEBS Letters, 2008, 582:503~509.
[138] Heux S, Cachon R, Dequin S. Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism. Metabolic Engineering, 2006, 8:303~314.
[139] Ricky A, Granath K, Hohmann S. The two isoenzymes for yeast NAD+-dependent glycerol 3-phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaptation and redox regulation. The EMBO Journal, 1997, 16:2179~2187.
[140] Santosa MM, Raghevendrana V, Kotter P et al. Manipulation of malic enzyme in Saccharomyces cerevisiae for increasing NADPH production capacity aerobically in different cellular compartments. Metabolic Engineering, 2004, 6:352~363.
[141] Boles E, De Jong-Gubbels P, Pronk JT. Identification and characterization of MAE1, the Saccharomyces cerevisiae structural gene encoding mitochondrial malic enzyme. J Bacteriol, 1998, 180:2875~2882.
[2] Klein S, Geiger T, Linchevski I, et al. Expression and purification of active PKB kinase from Escherichia coli. Protein Express. Purif., 2005, 41:162~169.
[3] Julia Scerbo M, Bibolini MJ, Barra JL, et al. Expression of a bioactive fusion protein of Escherichia coli heat-labile toxin B subunit to a synapsin peptide. Protein Expression and Purification, 2008, 59:320~326.
[4] Magliery TJ, Wilson CG, Pan W, et al. Detecting protein-protein interactions with a green fluorescent protein fragment reassembly trap: scope and mechanism. J. Am. Chem. Soc., 2005, 127:146~157.
[5] Clark EB. Refolding of recombinant proteins. Curr. Opin. Biotechnol., 1998, 9: 157~163.
[6] Yamabhai M, Emrat S, Sukasem S, et al. Secretion of recombinant Bacillus hydrolytic enzymes using Escherichia coli expression systems. Journal of Biotechnology, 2008, 133:50~57.
[7] Ferreira LC, Ferreira RC, Schumann W. Bacillus subtilis as a tool for vaccine development: from antigen factories to delivery vectors. Ann. Braz. Acad. Sci., 2005, 77:113~124.
[8] Barnard GC, Henderson GE, Srinivasan S, Gerngross TU. High level recombinant protein expression in Ralstonia eutropha using T7 RNA polymerase based amplication. Protein Expression and Purifcation, 2004, 38:264 ~ 271.
[9] Carvalho H, Lima L, Lescure N, et al. Differential expression of the two cytosolic glutamine synthetase genes in various organs of Medicago truncatula. Plant Science, 2000, 159, 301~312.
[10] Yang LJ, Tada Y, Yamamoto MP et al. A transgenic rice seed accumulating an anti-hypertensive peptide reduces the blood pressure of spontaneously hypertensive rats. FEBS Letters, 2006, 580:3315~3320.
[11] Yang L, Kajiura H, Suzuki K et al. Generation of a transgenic rice seed-based edible vaccine against house dust mite allergy. Biochemical and Biophysical Research Communications, 2008, 365:334~339.
[12] Youma JW, Won YS, Jeon JH et al. Oral immunogenicity of potato-derived HbsAg middle protein in BALB/c mice. Vaccine, 2007, 25:577~584.
[13] Moravec T, Schmidt MA, Herman EM et al. Production of Escherichia coli heat labile toxin (LT) B subunit in soybean seed and analysis of its immunogenicity as an oral vaccine. Vaccine, 2007, 25:1647~1657.
[14] Rigano MM, Dreitz S, Kipnis AP et al. Oral immunogenicity of a plant-made, subunit, tuberculosis vaccine. Vaccine, 2006, 24:691~695.
[15] Ritala A, Wahlstr?m EH, Holkeri H. et al. Production of a recombinant industrial protein using barley cell cultures. Protein Expression and Purification, 2008, 59:274~281.
[16] Smith GE, Summers MD, Fraser MJ. Production of human beta interferon in insect cells in fected with a baculovirus expression vector. Mol. Cell. Biol., 1983, 3: 2156~2165.
[17] Honjo E, Shoyama Y, Tamada T, et al. Expression of the extracellular region of the human interleukin-4 receptor a chain and interleukin-13 receptor a1 chain by a silkworm–baculovirus system. Protein Expression and Purification, 2008, 60:25~30.
[18] Donald L, Jarvis DL. Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology, 2003, 310:1~7.
[19] Jarvis DL. Developing baculovirus-insect cell expression systems for humanized recombinant glycoprotein production. Virology, 2003, 310:1~7.
[20] Taylor AL, Haze-Filderman A, Blumenfel A et al. High yield of biologically active recombinant human amelogenin using the baculovirus expression system. Protein Expression and Purification, 2006, 45:43~53.
[21] Peng S, Lva N, Zhang Y, et al. HeImmune response induced by spike protein from transmissible gastroenteritis coronavirus expressed in mouse mammary cells. Virus Research, 2007, 128:52~57.
[22] Kevin E, Cott V, Stephen P. et al. Transgenic pigs as bioreactors: a comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland. Genetic Analysis: Biomolecular Engineering, 1999, 15: 155~160.
[23] Takami H, Watanabe H, Ohmori Y, et al. Human alkaline phosphatase expression and secretion into chicken eggs after in vivo gene electroporation in the oviduct of laying hens. Biochemical and Biophysical Research Communications, 2002, 292:88~93.
[24] Ziomek CA. Commercialization of proteins produced in the mammary gland.Theriogenology, 1996, 49:139~144.
[25] European Medicines Agency London, Question and answers on atryn. International Non-proprietary Name (INN): antithrombin alfa. 2006, Doc.Ref. EMEA/191862/2006.
[26] Makrides SC, Makrides SC. Virus-based vectors for gene expression in mammalian cells: SV40. Gene Transfer and Expression in Mammalian Cells: New Comprehensive Biochemistry, 2003, 38:71~91.
[27] Gillard M, Chatelain P, Fuks B. Binding characteristics of levetiracetam to synaptic vesicle protein 2A (SV2A) in human brain and in CHO cells expressing the human recombinant protein. European Journal of Pharmacology, 2006, 536:102~108.
[28] Bork K, Reutter W, Weidemann W, Horstkorte R. Enhanced sialylation of EPO by overexpression of UDP-GlcNAc 2-epimerase/ManAc kinase containing a sialuria mutation in CHO cells. FEBS Letters, 2007, 581:4195~4198.
[29] Teixeira A, Cunha AE, Clemente JJ, et al. Modelling and optimization of a recombinant BHK-21 cultivation process using hybrid grey-box systems. Journal of Biotechnology, 2005, 118:290~303.
[30] Jardin BA, Zhao Y, Selvaraj M, et al. Expression of SEAP (secreted alkaline phosphatase) by baculovirus mediated transduction of HEK 293 cells in a hollow fiber bioreactor system. Journal of Biotechnology, 2008, doi:10.1016/j.jbiotec.2008.04.006.
[31] Müller D, Katinger H, Grillari J. MicroRNAs as targets for engineering of CHO cell factories. Trends in Biotechnology, 2008, doi:10.1016/j.tibtech.2008.03.010.
[32] Hitzeman RA, Hagie FE, Levine HL, et al. Expression of a human gene for interferon in yeast. Nature, 1981, 293:717~722.
[33] Thor D, Xiong S, Orazem CC, et al. Cloning and characterization of the Pichia pastoris MET2 gene as a selectable marker. FEMS Yeast Research, 2005, 5:935~942.
[34] Steinborna G, Gellissenb G, Kunze G. Assessment of Hansenula polymorpha and Arxula adeninivorans-derived rDNA-targeting elements for the design of Arxula adeninivorans expression vectors. FEMS Yeast Research, 2005, 5:1047~1054.
[35] Broach JR, Li Y, Feldman J, et al. Localization and sequence analysis of yeast origins of DNA replication. Cold Spring Harbor Symposium on Quantitative Biology, 1982, 47:1165~1173.
[36] Falco SC, Li Y, Broach JR and Botstein D. Genetic properties of chromosomally integrated 2μplasmid DNA in yeast. Cell, 1982, 29:573~584.
[37] Jayaram M, Li Y, Broach JR. The yeast plasmid 2μencodes componentsrequired for its high copy propagation, Cell, 1983, 34:95~104.
[38] Jayaram, M, Li, YY., McLeod M, Broach JR. Analysis of site-specific recombination associated with the yeast plasmid 2 micron circle. in mechanisms of DNA replication and recombination UCLA Symposia on Molecular and Cellular Biology (Cozzelli and Fox, eds.) New York, Alan R. Liss, Inc., 1983. 685~694.
[39] Huo KK, Yu LL, Chen XJ, Li YY. A stable vector for high level expression and secretion of human interferonαA gene in yeast. Science in China, 1993, 36:557~567.
[40] Suzuki K, Ichikawa K, Jigami Y. Yeast mutants with enhanced ability to secrete human lysozyme:Isolation and identification of a protease-deficient mutant. Mol Gen Genet, 1989, 219:58~64.
[41] Sampaio PN, Fortes AM, Cabral JM, et al. Production and Characterization of Recombinant Cyprosin B in Saccharomyces cerevisiae (W303-1A) Strain. Journal of Bioscience and Bioengineering, 2008, 105:305~312.
[42] Griffith DA, Delipala C, Leadsham J et al. A novel yeast expression system for the overproduction of quality-controlled membrane proteins. FEBS Letters, 2003, 553:45~50.
[43] Juozapaitis M, Zvirbliene A, Kucinskaite I. Synthesis of recombinant human parainfluenza virus 1 and 3 nucleocapsid proteins in yeast Saccharomyces cerevisiae. Virus Research, 2008, 133:178~186.
[44] Juozapaitis M, Serva A, Kucinskaite I. Generation of menangle virus nucleocapsid-like particles in yeast Saccharomyces cerevisiae. Journal of Biotechnology, 2007, 130:441~447.
[45] Juozapaitis M, Serva A, Zvirbliene A. Generation of henipavirus nucleocapsid proteins in yeast Saccharomyces cerevisiae. Virus Research, 2007, 124:95~102.
[46] Gasser B, Maurer M, Gach J et al. Engineering of Pichia pastoris for improved production of antibody fragments. Biotechnology and Bioengineering, 2006, 94: 353~361.
[47] Wetterholm A, Molina DM, Nordlund P, et al. High-level expression, purification, and crystallization of recombinant rat leukotriene C4 synthase from the yeast Pichia pastoris. Protein Expression and Purification, 2008, 60:1~6.
[48] Zámocky M, Schümann C, Sygmund C, O'Callaghan J, et al. Cloning, sequence analysis and heterologous expression in Pichia pastoris of a gene encoding a thermostable cellobiose dehydrogenase from Myriococcum thermophilum. Protein Expression and Purification, 2008, 59:258~265.
[49] Madzak C, Gaillardin C, Beckerich JM. Heterologous protein expression andsecretion in the non-conventional yeast Yarrowia lipolytica: a review. Journal of Biotechnology, 2004, 109:63~81.
[50] Fickers P, Fudalej F, Nicaud JM, Destain J, Thonart P. Selection of new over-producing derivatives for the improvement of extracellular lipase production by the non-conventional yeast Yarrowia lipolytica. Journal of Biotechnology, 2005, 115: 379~386.
[51] Platko JD, Deeg M, Thompson V, et al. Heterologous expression of Mytilus californianus foot protein three (Mcfp-3) in Kluyveromyces lactis. Protein Expression and Purification, 2008, 57:57~62.
[52] Nevalainen KM, Te'o VS, Bergquist PL. Heterologous protein expression in filamentous fungi. Trends in Biotechnology, 2005, 23:468~474.
[53] Wang Y, Xue W, Sims AH, et al. Isolation of four pepsin-like protease genes from Aspergillus niger and analysis of the effect of disruptions on heterologous laccase expression. Fungal Genetics and Biology, 2008, 45:17~27.
[54] Koda A, Bogaki T, Minetoki T, et al. High expression of a synthetic gene encoding potatoα-glucan phosphorylase in Aspergillus niger. Journal of bioscience and bioengineering, 2005, 100:531~537.
[55] Ahamed A, Vermette P. Culture-based strategies to enhance cellulase enzyme production from Trichoderma reesei RUT-C30 in bioreactor culture conditions. Biochemical Engineering Journal, 2008, 40:399~407.
[56] Wiebe MG. Stable production of recombinant proteins in filamentous fungi- problems and improvements. Mycologist, 2003, 17:140~144.
[57] Meyer V. Genetic engineering of filamentous fungi-progress, obstacles and future trends. Biotechnology Advances, 2008, 26:177~185.
[58] Tamalampudi S, Talukder MR, Hama S, et al. Enzymatic production of biodiesel from Jatropha oil: a comparative study of immobilized-whole cell and commercial lipases as a biocatalyst. Biochemical Engineering Journal, 2008, 39:185~189.
[59] Novick P, Ferro S, Schekman R. Order of events in the yeast secretory pathway. Cell, 1981, 25:461~419.
[60] Novick P, Garrett MD, Brennwald P, et al. Control of exocytosis in yeast. Cold Spring Harbor Symposia On Quantitative Biology, 1995, 60:171~177.
[61] Rapoport TA, Rolls MM and Jungnickel B. Approaching the mechanism of protein transport across the ER membrane. Curr Opin Cell Biol, 1996, 8:499~504.
[62] Wiertz EJ, Tortorella D, Bogyo M, et al. Sec61-mediated transfer proteasome for destruction. Nature, 1996, 384: 432~438.
[63] Hartmann E, Sommer T, Prehn S, et al. Evolutionary conservation ofcomponents of the protein translocation complex. Nature, 1994, 367: 654~657.
[64] Glick BS. Can Hsp70 proteins act as force-generating motors? Cell, 1995, 80:11~4.
[65] Ng DT, Brown JD and Walter P. Signal sequences specify the targeting route to the endoplasmic reticulum membrane. J Cell Biol, 1996, 134:269~78.
[66] B?hni PC, Deshaies RJ and Schekman RW. SEC11 is required for signal peptide processing and yeast cell growth. J Cell Biol, 1988, 106:1035~1042.
[67] Evans EA, Gilmore R, and Blobel G. Purification of microsomal signal peptidase as a complex. Proc Natl Acad Sci USA, 1986, 83:581~585.
[68] Gavel Y, von Heijne G. Sequence differences between glycosylated and non-glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering. Prot Eng, 1990, 3:433~442.
[69] Nilsson IM and von Heijne G. Determination of the distance between the oligosaccharyl transferase active site and the endoplasmic reticulum membrane. J Biol Chem, 1993, 268:5798~5801.
[70] Shelness GS, Lin L and Nicchitta CV. Membrane topology and biogenesis of eukaryotic signal peptidase. J Biol Chem, 1993, 268:5201~5208.
[71] Meldolesi J and Pozzan T. The endoplasmic reticulum Ca2+ store: a view from the lumen. Trends Biochem Sci, 1998, 23:10~14.
[72] Ellgaard L, Molinari M, Helenius A. Setting the standards: quality control in the secretory pathway. Science, 1999, 286:1882~1888.
[73] Orci L, Stamnes M, Ravazzola M, et al. Bidirectional transport by distinct populations of COPI-coated vesicles. Cell, 1997, 90:335~349.
[74] Scales SJ, Pepperkok R, Kreis TE. Visualization of ER-to-Golgi transport in living cells reveals a sequential mode of action for COPII and COPI. Cell, 1997, 90: 1137~1148.
[75] Barlowe C, Schekman R. SEC12 encodes a guanine-nucleotide exchange factor essential for transport vesicle budding from the ER. Nature, 1993, 365:347~349.
[76] Antonny B and Schekman R. ER export: public transportation by the COPII coach. Current Opinion in Cell Biology, 2001, 13:438~443.
[77] Harter C, Wieland F. The secretory pathway: mechanisms of protein sorting and transport. Biochim Biophys Acta, 1996, 1286:75~93.
[78] Lledo PM. Exocytosis in excitable cells: a conserved molecular machinery from yeast to neuron. European Journal of Endocrinology, 1997, 137:1~9.
[79] Gordon CL, Archer DB, Jeenes DJ, et al. A glucoamylase::GFP gene fusion to study protein secretion by individual hyphae of Aspergillus niger. J Microbiol Methods, 2000, 42:39~48.
[80] Nyk?nen M, Saarelainen R, Raudaskoski M, et al. Expression and secretion ofbarley Cysteine Endopeptidase B and Cellobiohydrolase I in Trichoderma reesei. Appl Environ Microbiol, 1997, 63:4929~4937.
[81] Nyk?nen M. Protein secretion in Trichoderma reesei. Ph.D. thesis, Dept Biol Environ Sci. Univ. of Jyv?skyl?, 2002.
[82] Gierz G and Bartnicki-Garcia S. A three-dimensional model of fungal morphogenesis based on the vesicle supply center concept. J Theor Biol, 2001, 208: 151~164.
[83] Rupes I, Mao WZ, ?str?m H and Raudaskoski M. Effects of nocodazole and brefeldin A on microrubule cytoskeleton and membrane organization in the homobasidiomycete Schizophyllum. Commune. Protoplasma, 1995, 185:212~221.
[84] Thompson SA, Golightly EJ and Yaver DS. Nucleotide sequence of the Aspergillus niger srpA gene. Gene, 1995, 167:337~338.
[85] Zakrzewska A, Migdalski A, Saloheimo M, et al. cDNA encoding protein Omannosyltransferase from the filamentous fungus Trichoderma reesei; functional equivalence to Saccharomyces cerevisiae PMT2. Curr Genet, 2003, 43:11~16.
[86] Veldhuisen G, Saloheimo M, Fiers MA, et al. Isolation and analysis of functional homologues of the secretion- related SAR1 gene of Saccharomyces cerevisiae from Aspergillus niger and Trichoderma reesei. Mol Gen Genet, 1997, 256:446~455.
[87] Vasara T, Ker?nen S, Penttil? M, Saloheimo M. Characterisation of two 14-3-3 genes from Trichoderma reesei: interactions with yeast secretory pathway components. Biochim Biophys Acta., 2002, 1590:27~40.
[88] Gething MJ, Sambrook J. Protein folding in the cell. Nature, 1992, 355:33~45.
[89] Mori K, Sant A, Kohno K, et al. A 22bp cis-acting element is necessary and sufficient for the induction of the yeast KAR2 (BiP) gene by unfolded proteins. EMBO J, 1992, 11:2583~2593.
[90] Patil C, Walter P. Intracellular signaling from the endoplasmic reticulum to the nucleus: the unfolded protein response in yeast and mammals. Curr Opin Cell Biol, 2001, 13:349~355.
[91] Cox JS and Walter P. A novel mechanism for regulating activity of a transcription factor that controls the unfolded protein response. Cell, 1996, 87:391~404.
[92] Kaufman RJ. Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls. Genes Dev., 1999, 13:1211~1233.
[93] Sidrauski C, Walter P. The transmembrane kinase Ire1p is a sitespecific endonuclease that initiates mRNA splicing in the unfolded protein response. Cell, 1997, 90:1031~1039.
[94] Sidrauski C, Cox JS, Walter P. tRNA ligase is required for regulated mRNA splicing in the unfolded protein response. Cell, 1996, 87:405~413.
[95] Chapman RE, Walter P. Translational attenuation mediated by an mRNA intron. Curr Biol., 1997, 7:850-859.
[96] Kawahara T, Yanagi H, Yura T et al. Unconventional splicing of HAC1/ERN4 mRNA required for the unfolded protein response. The Journal of Biological Chemistry, 1998, 273:1802~1807.
[97] Ruegsegger U, Leber JH, Walter P. Block of HAC1 mRNA translation by long-range base pairing is released by cytoplasmic splicing upon induction of the unfolded protein response. Cell, 2001, 107:103~114.
[98] Ron D, Walter P. Signal integration in the endoplasmic reticulum unfolded protein response. Nat Rev Mol Cell Biol, 2007, 8:519~529.
[99] Mori K, Ogawa N, Kawahara T et al. Palindrome with spacer of one nucleotide is characteristic of the cis-acting unfolded protein response element in Saccharomyces cerevisiae. The Journal of Biological Chemistry, 1998, 273:9912~9920.
[100] Mori K, Sant A, Kohno K et al. A 22 bp cis-acting element is necessary and sufficient for the induction of the yeast KAR2 (BiP) gene by unfolded proteins. The EMBO Journal, 1993, 11:2583~2593.
[101] Casagrande R, Stern P, Brown DM et al. Degradation of proteins from the ER of S. cerevisiae requires an intact unfolded protein response pathway. Mol Cell, 2000, 5:729~735.
[102] Xua P, Radena D, Doyle FJ et al. Analysis of unfolded protein response during single-chain antibody expression in Saccaromyces cerevisiae reveals different roles for BiP and PDI in folding. Metabolic Engineering, 2005, 7: 269~279.
[103] Wang JD, Herman C, Tipton KA. Directed evolution of substrate-optimized GroEL/S chaperonins. Cell, 2002, 111:1027~1039.
[104] Hibbert EG, Baganz F, Hailes HC. Directed evolution of biocatalytic processes. Biomolecular Engineering, 2005, 22:11~19.
[105] Alper H, Moxley J, Nevoigt E et al. Engineering yeast transcription machinery for improved ethanol tolerance and production. Science, 2006, 314:1565~1568.
[106] Jin S, Ye K, Shimizu K. Metabolic flux distributions in recombinant Saccharomyces cerevisiae during foreign protein production. Journal of Biotechnology, 1997, 54:161~174.
[107] Jensen R, Sprague GF, Herskowitz JI. Control of cell type in yeast by the mating type locus. J Mol Biol, 1981, 147:357~372.
[108] Song Y, Sata J, Saito A, Usui M, et al. Effects of calnexin deletion in Saccharomyces cerevisiae on the secretion of glycosylated lysozymes. J. Biochem., 2001, 130:757~764.
[109] Wormald MR, Dwek RA. Glycoproteins: glycan presentation and protein-fold stability. Structure, 1999, 15:150~160.
[110] Shibasaki S, Tanaka A, Ueda M. Development of combinatorial bioengineering using yeast cell surface display-order-made design of cell and protein for bio-monitoring. Biosensors and Bioelectronics, 2003, 19:123~130.
[111] Tanino T, Noguchi E, Kimura S. Effect of cultivation conditions on cell-surface display of Flo1 fusion protein using sake yeast. Biochemical Engineering Journal, 2007, 33:232~237.
[112] Wu CH, Mulchandani A, Chen W. Versatile microbial surface-display for environmental remediation andbiofuels production. Trends in Microbiology, 2008, 16, doi:10.1016/j.tim.2008.01.003.
[113] Khaw TS, Katakura Y, Ninomiya K, et al. Enhancement of ethanol production by promoting surface contact between starch granules and arming yeast in direct ethanol fermentation. Journal of bioscience and bioengineering, 2007, 103:95~97.
[114] Dürauer A, Berger E, Zandian M, et al. Yeast cell surface display system for determination of humoral response to active immunization with a monoclonal antibody against EpCAM. J. Biochem. Biophys Methods, 2008, 70:1109~1115.
[115] Bordes F, Fudalej F, Dossat V, et al. A new recombinant protein expression system for high-throughput screening in the yeast Yarrowia lipolytica. Journal of Microbiological Methods, 2007, 70:493~502.
[116] Shusta EV, Kieke MC, Parke E et al. Yeast polypeptide fusion surface display levels predict thermal stability and soluble secretion efficiency. J Mol Biol, 1999, 292:949~956.
[117] Daugherty PS, Iverson BL. and Georgiou G. Flow cytometric screening of cell-based libraries. Journal of Immunological Methods, 2000, 243:211~227.
[118] Hamilton SR, Davidson RC, Sethuraman N. Humanization of yeast to produce complex terminally sialylated glycoproteins. Science, 2006, 313:1441~1443.
[119] Masuda T, Ueno Y, Kitabatake N. High yield secretion of the sweet-tasting protein lysozyme from the yeast Pichia pastoris. Protein Expression and Purification, 2005, 39:35~42.
[120] Gupta JC, Mukherjee KJ. Stable maintenance of plasmid in continuous culture of yeast under non-selective conditions. Journal of Bioscience and Bioengineering,2001, 92:317~323.
[121] Skulj M, Okr?lar V, Jalen ?, et al. Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes. Microbial Cell Factories, 2008, 7:6 doi:10.1186/1475-2859-7-6.
[122] Bicknell AA, Babour A, Federovitch CM. A novel role in cytokinesis reveals a housekeeping function for the unfolded protein response. The Journal of Cell Biology, 2007, 177:1017~1027.
[123] Ogawa N. and Mori K. Autoregulation of the HAC1 gene is required for sustained activation of the yeast unfolded protein response. Genes to Cells, 2004, 9:95~104.
[124] Ruegsegger U, Leber JH, Walter P. Block of HAC1 mRNA translation by long-range base pairing is released by cytoplasmic splicing upon induction of the unfolded protein response. Cell, 2001, 107:103~114.
[125] Travers KJ, Patil CK, Wodicka L et al. Functional and genomic analyses reveal an essential coordination between the unfolded protein response and ER-associated degradation. Cell, 2000, 101:249~258.
[126] Andreaus J, Azevedo H, Cavaco-Paulo A. Effects of temperature on the cellulose binding ability of cellulase enzymes. Journal of Molecular Catalysis B: Enzymatic, 1999, 7:233~239.
[127] Santiago-Hernandez Ja, Vasquez-Bahena Jm, Calixto-Romo Ma. Direct immobilization of a recombinant invertase to Avicel by E. coli overexpression of a fusion protein containing the extracellular invertase from Zymomonas mobilis and the carbohydrate-binding domain CBDcex from Cellulomonas fimi. Enzyme and Microbial Technology, 2006, 40:172~176.
[128] Shang J and Geva E. Computational study of a single surface-immobilized two-stranded coiled-coil polypeptide. J. Phys. Chem. B, 2007, 111:4178~4188.
[129] Wang J and Somasundaran P. Mechanisms of ethyl (hydroxyethyl) cellulose-solid interaction: Influence of hydrophobic modification. Journal of Colloid and Interface Science, 2006, 293:322~332.
[130] Levy I, Shoseyov O. Cellulose-binding domains: Biotechnological applications. Biotechnology Advances, 2002, 20:191~213.
[131] Tomme P, Boraston A, McLean B. Characterization and affinity applications of cellulose-binding domains. Journal of Chromatography B, 1998, 715:283~296.
[132] Linder M, Salovuori I, Ruohonen L, Teeri TT. Characterization of a double cellulose-binding domain. Synergistic high affinity binding to crystalline cellulose. J Biol Chem., 1996, 271:21268~72.
[133] Stocker-Majd G, Hilbrig F, Freitag R. Extraction of haemoglobin from human blood by affinity precipitation using a haptoglobin-based stimuli–responsive affinitymacroligand. Journal of Chromatography A, 2008, 1194:57~65.
[134] Dengis, PB, Nélissen LR, and Rouxhet PG. Mechanisms of Yeast Flocculation: comparison of top and bottom-fermenting strains. Applied and Environmental Microbiology, 1995, 61:718~728.
[135] Leber JH, Bernales S, Walter P. IRE1-independent gain control of the unfolded protein response. PloS Biol, 2004, 2:1197~1207.
[136] Mulder HJ, Nikolaev I, Madrid SM. HACA, the transcriptional activator of the unfolded protein response (UPR) in Aspergillus niger, binds to partly palindromic UPR elements of the consensus sequence 50-CAN(G/A) NTGT/GCCT-30. Fungal Genetics and Biology, 2006, 43:560~572.
[137] Payne T, Hanfrey C, Bishop AL, Michael AJ, et al. Transcript-specific translational regulation in the unfolded protein response of Saccharomyces cerevisiae. FEBS Letters, 2008, 582:503~509.
[138] Heux S, Cachon R, Dequin S. Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism. Metabolic Engineering, 2006, 8:303~314.
[139] Ricky A, Granath K, Hohmann S. The two isoenzymes for yeast NAD+-dependent glycerol 3-phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaptation and redox regulation. The EMBO Journal, 1997, 16:2179~2187.
[140] Santosa MM, Raghevendrana V, Kotter P et al. Manipulation of malic enzyme in Saccharomyces cerevisiae for increasing NADPH production capacity aerobically in different cellular compartments. Metabolic Engineering, 2004, 6:352~363.
[141] Boles E, De Jong-Gubbels P, Pronk JT. Identification and characterization of MAE1, the Saccharomyces cerevisiae structural gene encoding mitochondrial malic enzyme. J Bacteriol, 1998, 180:2875~2882.