Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21828
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dc.contributor鄭隨和zh_TW
dc.contributor王雯靜zh_TW
dc.contributor林榮流zh_TW
dc.contributor許清森zh_TW
dc.contributor詹迺立zh_TW
dc.contributor楊明德zh_TW
dc.contributor.advisor許文輝zh_TW
dc.contributor.advisorWen-Hwei Hsuen_US
dc.contributor.author許世光zh_TW
dc.contributor.authorHsu, Shih-Kuangen_US
dc.contributor.other中興大學zh_TW
dc.date2008zh_TW
dc.date.accessioned2014-06-06T07:16:39Z-
dc.date.available2014-06-06T07:16:39Z-
dc.identifierU0005-1207200710400900zh_TW
dc.identifier.citation1. Anderson, A. W., H. C. Nordan, R. F. Cain, G. Parrish, and D. Duggan. 1956. Studies on a radio-resistant Micrococcus. I. Isolation, morphology, culture characteristics, and resistance to r-radiation. Food Technol 10:575-577. 2. Babbitt, P. C., M. S. Hasson, J. E. Wedekind, D. R. Palmer, W. C. Barrett, G. H. Reed, I. Rayment, D. Ringe, G. L. Kenyon, and J. A. Gerlt. 1996. The enolase superfamily: a general strategy for enzyme-catalyzed abstraction of the alpha-protons of carboxylic acids. Biochemistry 35:16489-16501. 3. Babbitt, P. C., G. T. Mrachko, M. S. Hasson, G. W. Huisman, R. Kolter, D. Ringe, G. A. Petsko, G. L. Kenyon, and J. A. Gerlt. 1995. A functionally diverse enzyme superfamily that abstracts the alpha protons of carboxylic acids. Science 267:1159-61. 4. Barbara, S., F. Kurt, and K. Wolfgang. 2003. Enzymatic racemisation and its application to synthetic biotransformations. Adv Synth Catal 345:653-666. 5. Baumeister, W., M. Barth, R. Hegerl, R. Guckenberger, M. Hahn, and W. O. Saxton. 1986. Three-dimensional structure of the regular surface layer (HPI layer) of Deinococcus radiodurans carotenoids. Arch Biochem Biophys 275:244-251. 6. Bergmann, M., and L. Zervas. 1928. Catalytic racemization via aminosauren and peptide. Biochem 203:280-292. 7. Brooks, B. W., and R. G. E. Murray. 1981. Nomenclature for "Micrococcus radiodurans" and other radiation resistant cocci: Deinococcaceae fam. Nov. and Deinococcus gen. Nov., including five species. Int. J. Syst. Bacteriol 30:627-646. 8. Chibata, I., T. Kakimoto, and J. Kato. 1965. Enzymatic production of L-alanine by Pseudomonas dacunhae. Appl. Microbiol. 13:638-645. 9. Chibata, I., T. Kakimoto, J. Kato, T. Shibatani, and K. Nishimura. 1967. Crystalline aspartate-decarboxylase of Pseudomonas dacunhae. Biochem. Biophys. Res. Commun. 26:662-667. 10. Crosby, J. 1994. Chirality in industry: An overview. John Wiley and Sons, West Sussex, England. 11. Crueger, W., and A. Crueger. 1984. Biotechnology: A textbook of industrial microbiology. Science Tech, Inc. 12. Crump, S. P., and J. D. Rozzell. 1994. Biocatalytic production of amino acids by transamination. John Wiley and Sons, West Sussex, England. 13. Gerlt, J. A., P. C. Babbitt, and I. Rayment. 2005. Divergent evolution in the enolase superfamily: the interplay of mechanism and specificity. Arch Biochem Biophys 433:59-70. 14. Harwood, J. M., and R. E. Parales. 1996. The beta-ketoadipate pathway and the biology of self-identity. Annu Rev Microbiol 50:333-336. 15. Helin, S., P. C. Kahn, B. L. Guha, D. G. Mallows, and A. Goldman. 1995. The refined X-ray structure of muconate lactonizing enzyme from Pseudomonas putida PRS2000 at 1.85 A Resolution. 254:918. 16. Henne, A., H. Bruggemann, C. Raasch, A. Wiezer, T. Hartsch, H. Liesegang, A. Johann, T. Lienard, O. Gohl, R. Martinez-Arias, C. Jacobi, V. Starkuviene, S. Schlenczeck, S. Dencker, R. Huber, H. P. Klenk, W. Kramer, R. Merkl, G. Gottschalk, and H. J. Fritz. 2004. The genome sequence of the extreme thermophile Thermus thermophilus. Nat Biotechnol 22:547-553. 17. Ho, S. N., H. D. Hunt, R. M. Horton, J. K. Pullen, and L. R. Pease. 1989. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51-59. 18. Hoch, J. A., and R. Losick. 1997. Panspermia, spores and the Bacillus subtilis genome. Nature 390:237-238. 19. Kamphuis, J., H. J. Boesten, K. B., H. F. M. Hermes, T. Sonke, Q. B. Broxterman, W. J. J. Van Den Tweel, and H. E. Schoemaker. 1994. The production and uses of optically pure nature and unnatural amino acids. John Wiley and Sons, West Sussex, England. 20. Kamphuis, J., H. J. Boesten, B. Kaptein, H. F. M. Hermes, T. Sonke, Q. B. Broxterman, W. J. J. Van Den Tweel, and H. E. Schoemaker. 1994. The production and uses of optically pure nature and unnatural amino acids. John Wiley and Sons, West Sussex, England. 21. Kleemann, W. L., and H. T. Hoppe. 1985. Ullmann''s Encyclopedia of Industrial Chemistry, 5th ed. VCH Verlagsgesellschaft, Weinheim. 22. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685. 23. Landro, J. A., J. A. Gerlt, J. W. Kozarich, C. W. Koo, V. J. Shah, G. L. Kenyon, D. J. Neidhart, S. Fujita, and G. A. Petsko. 1994. The role of lysine 166 in the mechanism of mendelate racemase from Pseudomonas putida: mechanistic and crystallographic evidence for stereospecific alkylation by (R)-alpha-phenylglycidate. Biochemistry 33:635-643. 24. Larsen, T. M., J. E. Wedekind, I. Rayment, and G. H. Reed. 1996. A carboxylate oxygen of the substrate bridges the magnesium ions at the active site of enolase: structure of the yeast enzyme complexed with the equilibrium mixture of 2-phosphoglycerare and phosphoenolpyruvate at 1.8A resolution. Biochemistry 35:4349-4358. 25. Lebioda, L., and B. Stec. 1988. Crystal structure of enolase indicates that enolase and pyruvate kinase evolved from a common ancestor. Nature 333:683-686. 26. May, O., P. T. Nguyen, and F. H. Arnold. 2000. Inverting enantioselectivity by directed evolution of hydantoinase for improved production of L-methionine. Nat Biotechnol 18:317-320. 27. Mazur, P., W. J. Henzel, S. Mattoo, and J. W. Kozarich. 1994. 3-Carboxy-cis,cis-muconate lactonizing enzyme from Neurospora crassa: an alternate cycloisomerase motif. J. Bacteriol. 176:1718-1728. 28. Neidhart, D. J., P. L. Howell, G. A. Petsko, V. M. Powers, R. S. Li, G. L. Kenyon, and J. A. Gerlt. 1991. Mechanism of the reaction catalyzed by mandelate racemase. 2. Crystal structure of mandelate racemase at 2.5-A resolution: identification of the active site and possible catalytic residues. Biochemistry 30:9264-73. 29. Neidhart, D. J., G. L. Kenyon, J. A. Gerlt, and G. A. Petsko. 1990. Mandelate racemase and muconate lactonizing enzyme are mechanistically distinct and structurally homologous. Nature 347:692-694. 30. Palmer, D. R., J. B. Garrett, V. Sharma, R. Meganathan, P. C. Babbitt, and J. A. Gerlt. 1999. Unexpected divergence of enzyme function and sequence: N-acylamino acid racemase is o-succinylbenzoate synthase. Biochemistry 38:4252-4258. 31. Popp, J. L., C. Berliner, and R. Bentley. 1989. Vitamin K (menaquinone) biosynthesis in bacteria: high-performance liquid chromatographic assay of the overall synthesis of o-succinylbenzoic acid and of 2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylic acid synthase. Anal Biochem 178:306-310. 32. Quintela, J. C., F. Garcia-del Portillo, E. Pittenauer, G. Allmaier, and M. A. de Pedro. 1999. Peptidoglycan Fine Structure of the Radiotolerant Bacterium Deinococcus radiodurans Sark. J. Bacteriol. 181:334-337. 33. Saiki, R. K., P. S. Walsh, C. H. Levenson, and H. A. Erlich. 1989. Genetic Analysis of Amplified DNA with Immobilized Sequence-Specific Oligonucleotide Probes. Proc. Natl. Acad. Sci. USA 86:6230-6234. 34. Schmidt, D. M. Z., B. K. Hubbard, and J. A. Gerlt. 2001. Evolution of enzymatic activities in the enolase superfamily: functional assignment of unknown proteins in Bacillus subtilis and Escherichia coli as L-ALA-D/L-Glu epimerases. Biochemistry 40:15707-15715. 35. Schroeder, U., B. Henrich, J. Fink, and R. Plapp. 1994. Peptidase D of Escherichia coli K-12, a metallopeptidase of low substrate specificity. FEMS Microbiol Lett 123:153-159. 36. Senuma, M., O. Otsuki, N. Sakata, M. Furui, and T. Tosa. 1989. Industrial production of D-aspartic acid using a pressuried column reactor containing immobilized Pseudomonas dacunhae. J. Ferment. Bioeng. 67:233-237. 37. Stegink, L. D. 1987. The aspartame story: a model for the clinical testing of a food additive. Am J Clin Nutr 46:204-15. 38. Stinson, S. C. 1998. Counting on chiral drugs. Chem Engineer News 76:1-136. 39. Su, S. C., and C. Y. Lee. 2002. Cloning of the N-acylamino acid racemase gene from Amycolatopsis azurea and biochemical characterization of the gene product. Enzyme and Microbial Technology 30:647. 40. Syldatk, C., O. May, J. Altenbuchner, and R. Mattes. 1999. Microbial hydantoinase-industrial enzymes from the origin of life. Appl. Microbiol. Biotechnol 51:293-309. 41. Syldatk, C., R. Muller, M. Pietzsch, and F. Wagner. 1994. Microbial and enzymatic production of L-amino acids from DL-5-monosubstituted hydantoins. John Wiley and Sons., West Sussex, England. 42. Syldatk, C., R. Muller, M. Siemann-Herzberg, K. Krohn, and F. Wagner. 1994. Microbial and enzymatic production of D-amino acids from DL-5-monosubstituted hydantoins. John Wiley and Sons, West Sussex, England. 43. Takahashi, T., and K. Hatano. 1991. Acylamino acid racemase, production and use thereof. US. 44. Takuyama, S., and K. Hatano. 1995a. Purification and properties of thermostable N-acylamino acid racemase from Amycolatopsis sp. TS-1-60. Appl Microbiol Biotechnol 42:853-859. 45. Taylor, E. A., J. B. Garrett, J. B. Thoden, H. M. Holden, I. Rayment, and J. A. Gerlt. 2004. Evolution of enzymatic activity in the enolase superfamily: functional studies of the promiscuous o-succinylbenzoate synthase from Amycolatopsis. Biochemistry 43:224-229. 46. Taylor, P. P., D. P. Pantaleone, R. F. Senkpeil, and I. G. Fotheringham. 1998. Novel biosynthetic approaches to the production of unnatural amino acids using aminotransferase. Trends Biotechnol 16:412-418. 47. Taylor Ringia, E. A., J. B. Garrett, J. B. Thoden, H. M. Holden, I. Rayment, and J. A. Gerlt. 2004. Evolution of enzymatic activity in the enolase superfamily: functional studies of the promiscuous o-succinylbenzoate synthase from Amycolatopsis. Biochemistry 43:224-9. 48. Thoden, J. B., E. A. Taylor, J. B. Garrett, J. A. Gerlt, H. M. Holden, and I. Rayment. 2004. Evolution of enzymatic activity in the enolase superfamily: structural studies of the promiscuous o-succinylbenzoate synthase from Amycolatopsis. Biochemistry 43:5716-5727. 49. Thompson, T. B., J. B. Garrett, E. A. Taylor, R. Meganathan, J. A. Gerlt, and I. Rayment. 2000. Evolution of enzymatic activity in the enolase superfamily: structure of o-succinylbenzoate synthase from Escherichia coli in complex with Mg2+ and o-succinylbenzoate. Biochemistry 39:10662-10676. 50. Tokuyama, S., and K. Hatano. 1996. Overexpression of the gene for N-acylamino acid racemase from Amycolatopsis sp. TS-1-60 in Escherichia coli and continuous produciton of optically active methionine by a bioreactor. Appl Microbiol Biotechnol 44:774-7. 51. Tokuyama, S., and K. Hatano. 1995. Purification and properties of thermostable N-acylamino acid racemase from Amycolatopsis sp. TS-1-60. Appl Microbiol Biotechnol 42:853-9. 52. Tokuyama, S., H. Miya, K. Hatano, and T. Takahashi. 1994. Purification and properties of a novel enzyme, N-acylamino acid racemase, from Streptomyces atratus Y-53. Appl Microbiol Biotechnol 40:835. 53. Tuker, G. T. 2000. Chiral switch. Lancet 355:1085-1087. 54. Verseck, S., A. Bommarius, and M. R. Kula. 2001. Screening, overexpression and characterization of an N-acylamino acid racemase from Amycolatopsis orientalis subsp. lurida. Appl Microbiol Biotechnol 55:354-61. 55. Virtanin, A. I., and T. Laine. 1937. The decarboxylation of D-lysine and L-aspartic acid. Enzymology 3:266-270. 56. Wang, W. C., W. C. Chiu, S. K. Hsu, C. L. Wu, C. Y. Chen, J. S. Liu, and W. H. Hsu. 2004. Structural basis for catalytic racemization and substrate specificity of an N-acylamino acid racemase homologue from Deinococcus radiodurans. J Mol Biol 342:155-169. 57. Weetall, H. H., and A. M. Fibert. 1974. Porous glass for affinity chromatography applications. Methods. Enzymol. 34:59-72. 58. Work, E., and H. Griffiths. 1968. Morphology and chemistry of cell walls of Micrococcus radiodurans. J. Bacteriol 95:641-657.en_US
dc.identifier.urihttp://hdl.handle.net/11455/21828-
dc.description.abstract由D. radiodurans菌體中選殖出一條長約1.2 kb之NAAAR基因,可轉譯出375個胺基酸,其分子量約為40 kDa。此酵素在60℃、pH8.0反應條件下具有最佳之酵素活性。NAAAR活性可明顯被二價金屬離子Co2+與Mn2+所提升。基質選擇性分析顯示,NAAAR具有廣泛之作用基質範圍。NAAAR屬於enolase superfamily中MLE (muconate lactonizing enzyme) subgroup類蛋白質,此類蛋白質的酵素催化反應包含有cycloisomerization (MLE)、dehydration [o-succinylbenzoate synthase (OSBS)]與1,1-proton transfer [L-Ala-D/L-Glu epimerase (AEE)]。本研究探討NAAAR在D. radiodurans菌體中扮演之生理功能。利用Dixon plot分析結果得知,NAAAR之活性易被OSBS之類似物 (salicyl hydroxamate) 所抑制,測得其Ki值為0.68 mM,此結果顯示,salicyl hydroxamate與NAAAR之反應基質作用在酵素相同的位置上,當少量之salicyl hydroxamate存在下,即可與NAAAR的反應基質競爭酵素活性部位。以LC/MS/MS與結合酵素反應分析顯示,NAAAR無法催化L-Ala-D/L-Glu dipeptide進行消旋化反應,也就是不具有AEE酵素活性。NAAAR也無法催化cis,cis-muconate進行cycloisomerization反應,顯示無MLE酵素活性。進一步分析D. radiodurans基因體序列,也無法由資料庫中獲取可提供研判生理功能的相關資訊。因此NAAAR在D. radiodurans中真正的生理功能值得再深入探討。依據NAAAR蛋白質結構分析顯示在酵素活性區域中包含有Lys170、Asp195、Glu220、Asp245與Lys269,其中Asp195、Glu220與Asp245推測與二價金屬離子結合有關,而Lys170與Lys269剛好位於活性部位兩相對位置上,可能負責催化消旋反應的進行。以定點突變方式分析Lys170與Lys269兩個殘基,並測試所有變異酵素之生化活性,結果顯示經突變後所有變異酵素之活性明顯降低約200至1400倍左右,因此,推測Lys170與Lys269在酵素催化消旋反應中扮演非常重要的功能。zh_TW
dc.description.abstractThe N-acylamino acid racemase (NAAAR) gene from Deinococcus radiodurans BCRC12827 consists of an 1.2 kb open reading frame, encoding a protein of 375-amino acid residues with a calculated molecular mass of about 40 kDa. NAAAR had maximal activity at 60℃and pH 8.0. The high enzyme activity could be observed by the addition of 2mM Co2+ and Mn2+ ion. Substrate specificity analysis revealed that the NAAAR has a broad substrate range. The NAAAR is a member of the MLE (muconate lactonizing enzyme) subgroup of the enolase superfamily, catalyzing the reactions including cycloisomerization (MLE), dehydration [o-succinylbenzoate synthase (OSBS)], and 1,1-proton transfer [L-Ala-D/L-Glu epimerase (AEE)]. Dixon plot analysis showed that NAAAR activity was competitively blocked by the OSBS inhibitor, salicyl hydroxamate, with a Ki of 0.68 mM, indicating that NAc-Met and salicyl hydroxamate bind to the same substrate site of NAAAR. Based on the amino acid sequences identity and protein structure, we proposed that the NAAAR might has AEE function. LC/MS/MS analysis and coupling enzyme assay revealved that NAAAR cannot catalyze the racemization of L-Ala-D/L-Glu. The NAAAR also showed no cycloisomerization activity to cis,cis-muconate. Our data indicated that the physiological function of NAAAR in D. radiodurans is still unclear. Analysis of NAAAR 3-D structure and site-directed mutagenesis implied Lys170 and Lys269 located at opposite side of the active site might be involved in the reversible racemization reaction.en_US
dc.description.tableofcontents表 次 VII 圖 次 VIII 縮寫對照表 X 第一章:Deinococcus radiodurans BCRC12827的N-acylamino acid racemase酵素之生化性質與生理功能 1 中文摘要 2 英文摘要 3 一、緒 論 4 (一) 單一旋光性胺基酸及其應用 4 (二) 光學活性胺基酸的合成 4 (三) Aminoacylase結合N-acylamino acid racemase生產光學活性胺基酸 7 (四) N-acylamino acid racemase的生化特性與功能 7 (五) Enolase superfamily的相關研究 8 (六) N-acylamino acid racemase的生理角色 10 (七) Deinococcus radiodurans的特性 13 (八) 研究目的 14 二、實驗材料與方法 15 (一) 實驗材料 15 (二) 菌株、質體及培養條件 15 (三) D.radioduransi 染色體 DNA 之快速抽取法 15 (四) 聚合酶連鎖反應 (Polymerase chain reaction, PCR) 17 (五) naaar 基因選殖、表現與純化 17 (六) 蛋白質之定量 19 (七) 蛋白質的電泳分析 20 (八) 酵素活性分析 21 (九) o-succinylbenzoate synthase (OSBS) 酵素活性分析 21 (十) L-Ala-D/L-Glu epimerase (AEE) 酵素活性分析 22 (十一) Muconate lactonizing enzyme活性分析 23 (十二) NAAAR之定點突變 23 三、結 果 25 (一) N-acylamino acid racemase基因的選殖與表現 25 (二) 相關N-acylamino acid racemase蛋白質序列分析及比對 25 (三) N-acylamino acid racemase酵素之生化性質 25 (四) N-acylamino acid racemase生理功能之探討 29 (五) 以定向演化法改造NAAAR酵素之活性 37 四、討 論 42 (一) N-acylamino acid racemase基因之表現與生化性質 42 (二) N-acylamino acid racemase之生理功能 42 (三) N-acylamino acid racemase變異酵素之催化機制與基質光學選擇性 47 五、結 論 50 六、參考文獻 51 第二章:利用表現L-aminoacylase和N-acylamino acid racemase的重組E. coli細胞進行光學選擇性合成L-homophenylalanine 56 中文摘要 57 英文摘要 58 一、緒 論 59 (一) 前 言 59 (二) 血管收縮素轉換酶抑制劑的發展 59 (三) L-Homophenylalanine的生合成 64 (四) Aminoacylase 65 (五) 研究目的 67 二、實驗材料與方法 69 (一) 實驗材料 69 (二) laa 基因在 E. coli 菌體中的表現 69 (三) 酵素活性分析 69 (四) 構築naaar和laa雙基因在單一宿主細胞E. coli中的共同表現 73 (五) 利用全細胞生物催化系統轉換生產 L-HPA 74 三、結 果 75 (一) L-aminoacylase與N-acylamino acid racemase的生化性質 75 (二) naaar和laa基因在單一宿主細胞E. coli中的共同表現 75 (三) 利用雙基因在同一宿主細胞中共同表現的全細胞生物催化反應生產L-HPA 79 (四) 利用各別基因表現的宿主細胞生產L-HPA 80 (五) 重複使用全細胞生物催化反應生產L-HPA 80 四、討 論 87 (一) LAA之基質光學選擇性 87 (二) naaar和laa基因在單一宿主細胞E. coli中的共同表現 87 (三) 利用雙基因在同一宿主細胞中共同表現的全細胞生物催化反應生產L-HPA 87 (四) 利用各別基因表現的宿主細胞生產L-HPA 88 五、結 論 90 六、參考文獻 91 第三章:利用carbamoylase酵素法結合N-acylamino acid racemase的消旋化作用進行立體選擇性生合成L-homophenylalanine 96 中文摘要 97 英文摘要 98 一、緒 論 99 (一) 前 言 99 (二) N-Carbamoylamino acid amidohydrolase (N-carbamoylase) 99 (三) 全細胞生物催化反應之應用 100 (四) 增加微生物對物質通透性之方法 101 (五) 研究目的 103 二、實驗材料與方法 104 (一) 實驗材料 104 (二) 菌株、質體與分離純化DNA 104 (三) 基因之構築與表現 104 (四) 酵素活性分析 107 (五) 細胞通透性之處理 107 (六) 全細胞生物催化法生產L-HPA 109 (七) 利用高濃度基質生產L-HPA 109 三、結 果 110 (一) NAAAR酵素性質分析 110 (二) 利用雙基因(NAAAR與LNCA)在同一宿主細胞中共同表現的全細胞生物催化系統生產L-HPA 113 (三) 通透劑的選擇 113 (四) 以具通透性之E. coli全細胞生物催化反應進行生物轉換作用 118 (五) 重複使用具通透性之E. coli全細胞生物催化反應生產L-HPA 121 四、討 論 123 (一) NAAAR與LNCA酵素對基質光學選擇性之分析 123 (二) 利用雙基因在同一宿主細胞中共同表現的全細胞生物催化反應生產L-HPA 123 (三) 通透劑的選擇及最適反應條件 125 (四) 以具通透性之E. coli全細胞生物催化反應生產L-HPA 126 (五) 重複使用具通透性之E. coli全細胞生物催化反應生產L-HPA 127 五、結 論 129 六、參考文獻 130zh_TW
dc.language.isoen_USzh_TW
dc.publisher分子生物學研究所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1207200710400900en_US
dc.subjectN-acylamino acid racemaseen_US
dc.subjectMuconate lactonizing enzymeen_US
dc.subjecto-succinylbenzoate synthaseen_US
dc.subjectL-homophenylalanineen_US
dc.titleBiochemical characterization of N-acylamino acid racemase from Deinococcus radiodurans BCRC12827 and its application in the enantioselective synthesis of L-homophenylalanineen_US
dc.titleDeinococcus radiodurans BCRC12827的N-acylamino acid racemase酵素生化性質之分析及應用此酵素參與光學選擇性合成L-homophenylalaninezh_TW
dc.typeThesis and Dissertationzh_TW
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item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.languageiso639-1en_US-
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item.openairetypeThesis and Dissertation-
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