{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/102824"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/102824","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Design and engineering of multi-step (bio)catalytic systems","abstract":"Nature has been a perpetual source of inspiration for biochemists. It is not only the vast diversity of compounds that living beings can create, but also the extraordinary strategies of synthesis deployed. Evidently, the catalysts used by living beings -enzymes- are key to nature’s synthesis strategies. Biocatalysis is undoubtfully one of the most invaluable gifts given by nature to flourish the development of green chemical and pharmaceutical industries. With the development of protein and metabolic engineering tools and strategies, more and more enzymes have been used in the industries to improve the chemical processing; and microbes such as Escherichia coli and Saccharomyces cerevisiae have been engineered to produce a wide variety of value-added and bulk chemicals to replace traditional chemical synthesis. However, researchers have just explored the tip of the iceberg in the biocatalysis area. Proteins with new catalytic functionality should be discovered or engineered to broaden current biotransformation boundaries. New in vitro enzymatic or chemoenzymatic cascade reactions need to be designed and optimized to realize stronger synthetic power and more stable systems. Metabolic networks of traditional or new microorganisms should be largely rewired to meet the manufacturing standards. In this work, I aimed at designing and engineering multi-step (bio) catalytic systems for selective synthesis of value-added chemicals. Microorganisms synthesize complex molecules from simple substrates by a series of enzymes working cooperatively. Inspired by how aromatic polyketides are synthesized by the teamwork between enzymes, I sought to couple biocatalysis with organometallic catalysis, two distinct catalytic disciplines, in one pot to realize synthetic power that cannot be achieved by either of them. I first developed a modular, one-pot, sequential chemoenzymatic system for the formal enantioselective construction of C-C bond in 2-aryl 1,4-dicarbonyl compounds. This sequence comprises a rhodium-catalyzed diazocoupling that provides >9:1 selectivity for heterocoupling of two diazoesters and a reduction mediated by an ene-reductase (ER), which occurs in up to 99% enantiomeric excess (ee). The high yield and enantioselectivity of this system were resulted from the preferential generation of an (E)-alkene from the diazo coupling reaction and selective reduction of the (E)-alkene in a mixture of (E) and (Z) isomers by the ER. This work demonstrates the benefit of combining organometallic and enzymatic catalysis to create unusual overall transformations that do not require the isolation and purification of intermediates. To make the system works better on a broader range of substrates, I later developed a new class of cooperative chemoenzymatic reactions that combine photocatalysts that isomerize alkenes with ene-reductases that reduce carbon-carbon double bonds to generate valuable enantioenriched products. I demonstrated that this method enables the stereoconvergent reduction of E/Z mixtures of alkenes or reduction of the unreactive stereoisomer of an alkene in yields and ee’s that match those obtained from the reduction of the pure, more reactive isomer. This new cooperative system overcomes the limitations of both individual catalysts and affords a range of synthetically valuable and biologically active enantioenriched compounds. More generally, these results illustrate the value of driving a chemical reaction with light to ensure compatibility between the chemical and enzymatic catalysts. In vitro biocatalytic reaction normally has poor tolerance to harsh conditions such as low pH or high substrate concentrations. Cells membrane is natural compartmentalization and protects the enzymes from extracellular inhibitors. In addition, cell factories-based production provides an attractive alternative to chemical synthesis of value-added chemicals. I also worked on engineering a S. cerevisiae strain as a whole-cell catalyst for L-lactic acid overproduction in industrially preferred low pH environment (pH 3) by metabolic engineering and genome-wide engineering methods. In addition, to establish an automated cellular engineering platform, I developed a growth-based L-lactic acid biosensor and an automated quantification assay by BioProfile Analyzer.","abstract_html":"Nature has been a perpetual source of inspiration for biochemists. It is not only the vast diversity of compounds that living beings can create, but also the extraordinary strategies of synthesis deployed. Evidently, the catalysts used by living beings -enzymes- are key to nature’s synthesis strategies. Biocatalysis is undoubtfully one of the most invaluable gifts given by nature to flourish the development of green chemical and pharmaceutical industries. With the development of protein and metabolic engineering tools and strategies, more and more enzymes have been used in the industries to improve the chemical processing; and microbes such as Escherichia coli and Saccharomyces cerevisiae have been engineered to produce a wide variety of value-added and bulk chemicals to replace traditional chemical synthesis. However, researchers have just explored the tip of the iceberg in the biocatalysis area. Proteins with new catalytic functionality should be discovered or engineered to broaden current biotransformation boundaries. New in vitro enzymatic or chemoenzymatic cascade reactions need to be designed and optimized to realize stronger synthetic power and more stable systems. Metabolic networks of traditional or new microorganisms should be largely rewired to meet the manufacturing standards. In this work, I aimed at designing and engineering multi-step (bio) catalytic systems for selective synthesis of value-added chemicals. Microorganisms synthesize complex molecules from simple substrates by a series of enzymes working cooperatively. Inspired by how aromatic polyketides are synthesized by the teamwork between enzymes, I sought to couple biocatalysis with organometallic catalysis, two distinct catalytic disciplines, in one pot to realize synthetic power that cannot be achieved by either of them. I first developed a modular, one-pot, sequential chemoenzymatic system for the formal enantioselective construction of C-C bond in 2-aryl 1,4-dicarbonyl compounds. This sequence comprises a rhodium-catalyzed diazocoupling that provides >9:1 selectivity for heterocoupling of two diazoesters and a reduction mediated by an ene-reductase (ER), which occurs in up to 99% enantiomeric excess (ee). The high yield and enantioselectivity of this system were resulted from the preferential generation of an (E)-alkene from the diazo coupling reaction and selective reduction of the (E)-alkene in a mixture of (E) and (Z) isomers by the ER. This work demonstrates the benefit of combining organometallic and enzymatic catalysis to create unusual overall transformations that do not require the isolation and purification of intermediates. To make the system works better on a broader range of substrates, I later developed a new class of cooperative chemoenzymatic reactions that combine photocatalysts that isomerize alkenes with ene-reductases that reduce carbon-carbon double bonds to generate valuable enantioenriched products. I demonstrated that this method enables the stereoconvergent reduction of E/Z mixtures of alkenes or reduction of the unreactive stereoisomer of an alkene in yields and ee’s that match those obtained from the reduction of the pure, more reactive isomer. This new cooperative system overcomes the limitations of both individual catalysts and affords a range of synthetically valuable and biologically active enantioenriched compounds. More generally, these results illustrate the value of driving a chemical reaction with light to ensure compatibility between the chemical and enzymatic catalysts. In vitro biocatalytic reaction normally has poor tolerance to harsh conditions such as low pH or high substrate concentrations. Cells membrane is natural compartmentalization and protects the enzymes from extracellular inhibitors. In addition, cell factories-based production provides an attractive alternative to chemical synthesis of value-added chemicals. I also worked on engineering a S. cerevisiae strain as a whole-cell catalyst for L-lactic acid overproduction in industrially preferred low pH environment (pH 3) by metabolic engineering and genome-wide engineering methods. In addition, to establish an automated cellular engineering platform, I developed a growth-based L-lactic acid biosensor and an automated quantification assay by BioProfile Analyzer.","abstract_has_math":false,"creators":["Wang, Yajie"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Chemical Engineering","degree_department":null,"school":null,"contributors":["Zhao, Huimin","Yang, Hong","Rao, Christopher V.","Lu, Yi"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2019,"date_issued":"2019-02-07T20:39:41Z","date_published":"2019-02-07T20:39:41Z","updated_at":"2026-07-22T22:24:42Z","subjects":["Protein engineering, Synthetic Biology, Chemical engineering, Tandem catalysis, Natural Product, Metabolic Engineering"],"languages":["en"],"rights":["Copyright 2018 Yajie Wang"],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/2142/102824","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Zhao, Huimin","Yang, Hong","Rao, Christopher V.","Lu, Yi"]},{"key":"dc:creator","label":"Author","values":["Wang, Yajie"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2019-02-07T20:39:41Z","2021-02-08T10:15:25Z","2018-12-06","2018-12"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Chemical Engineering"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Protein engineering, Synthetic Biology, Chemical engineering, Tandem catalysis, Natural Product, Metabolic Engineering"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 2018 Yajie Wang"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/102824"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Nature has been a perpetual source of inspiration for biochemists. It is not only the vast diversity of compounds that living beings can create, but also the extraordinary strategies of synthesis deployed. Evidently, the catalysts used by living beings -enzymes- are key to nature’s synthesis strategies. Biocatalysis is undoubtfully one of the most invaluable gifts given by nature to flourish the development of green chemical and pharmaceutical industries. With the development of protein and metabolic engineering tools and strategies, more and more enzymes have been used in the industries to improve the chemical processing; and microbes such as Escherichia coli and Saccharomyces cerevisiae have been engineered to produce a wide variety of value-added and bulk chemicals to replace traditional chemical synthesis. However, researchers have just explored the tip of the iceberg in the biocatalysis area. Proteins with new catalytic functionality should be discovered or engineered to broaden current biotransformation boundaries. New in vitro enzymatic or chemoenzymatic cascade reactions need to be designed and optimized to realize stronger synthetic power and more stable systems. Metabolic networks of traditional or new microorganisms should be largely rewired to meet the manufacturing standards. In this work, I aimed at designing and engineering multi-step (bio) catalytic systems for selective synthesis of value-added chemicals. Microorganisms synthesize complex molecules from simple substrates by a series of enzymes working cooperatively. Inspired by how aromatic polyketides are synthesized by the teamwork between enzymes, I sought to couple biocatalysis with organometallic catalysis, two distinct catalytic disciplines, in one pot to realize synthetic power that cannot be achieved by either of them. I first developed a modular, one-pot, sequential chemoenzymatic system for the formal enantioselective construction of C-C bond in 2-aryl 1,4-dicarbonyl compounds. This sequence comprises a rhodium-catalyzed diazocoupling that provides >9:1 selectivity for heterocoupling of two diazoesters and a reduction mediated by an ene-reductase (ER), which occurs in up to 99% enantiomeric excess (ee). The high yield and enantioselectivity of this system were resulted from the preferential generation of an (E)-alkene from the diazo coupling reaction and selective reduction of the (E)-alkene in a mixture of (E) and (Z) isomers by the ER. This work demonstrates the benefit of combining organometallic and enzymatic catalysis to create unusual overall transformations that do not require the isolation and purification of intermediates. To make the system works better on a broader range of substrates, I later developed a new class of cooperative chemoenzymatic reactions that combine photocatalysts that isomerize alkenes with ene-reductases that reduce carbon-carbon double bonds to generate valuable enantioenriched products. I demonstrated that this method enables the stereoconvergent reduction of E/Z mixtures of alkenes or reduction of the unreactive stereoisomer of an alkene in yields and ee’s that match those obtained from the reduction of the pure, more reactive isomer. This new cooperative system overcomes the limitations of both individual catalysts and affords a range of synthetically valuable and biologically active enantioenriched compounds. More generally, these results illustrate the value of driving a chemical reaction with light to ensure compatibility between the chemical and enzymatic catalysts. In vitro biocatalytic reaction normally has poor tolerance to harsh conditions such as low pH or high substrate concentrations. Cells membrane is natural compartmentalization and protects the enzymes from extracellular inhibitors. In addition, cell factories-based production provides an attractive alternative to chemical synthesis of value-added chemicals. I also worked on engineering a S. cerevisiae strain as a whole-cell catalyst for L-lactic acid overproduction in industrially preferred low pH environment (pH 3) by metabolic engineering and genome-wide engineering methods. In addition, to establish an automated cellular engineering platform, I developed a growth-based L-lactic acid biosensor and an automated quantification assay by BioProfile Analyzer.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2020-12-01","The student, Yajie Wang, accepted the attached license on 2018-11-30 at 15:19.","The student, Yajie Wang, submitted this Dissertation for approval on 2018-11-30 at 15:36.","This Dissertation was approved for publication on 2018-12-06 at 08:56.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13164 on 2019-02-07 at 14:18:42","Made available in DSpace on 2019-02-07T20:39:41Z (GMT). No. of bitstreams: 6 WANG-DISSERTATION-2018.pdf: 11708977 bytes, checksum: 11bef31153c5271ad34195e62df49852 (MD5) Chapter 1 Permission from Taylor_francis.pdf: 134494 bytes, checksum: bc48dbc76d15b46b9d1dc49abf83d625 (MD5) Chapter 2 Permission from Springer Nature.pdf: 316399 bytes, checksum: 6c8b04dd1f33bf27f977758b8a7fcbdc (MD5) Chapter 3 Permission from ACS Catalysis.pdf: 263759 bytes, checksum: 04ef550745b68ee98304f9ac8129135c (MD5) Chapter 4 Permission from Nature.pdf: 309938 bytes, checksum: 5f7f17688c2d15fe15ef41ec6aa8e11d (MD5) LICENSE.txt: 4207 bytes, checksum: a626d9962cb3a101a5ffd9ff60b0677d (MD5) Previous issue date: 2018-12-06","Embargo set by: Seth Robbins for item 109849 Lift date: 2021-02-07T20:39:46Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 109849 Lift date: 2021-02-07T20:44:35Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 109849 on 2021-02-08T10:15:25Z."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Design and engineering of multi-step (bio)catalytic systems"]}]}],"canonical_facts":{"dc:contributor":["Zhao, Huimin","Yang, Hong","Rao, Christopher V.","Lu, Yi"],"dc:creator":["Wang, Yajie"],"dc:date":["2019-02-07T20:39:41Z","2021-02-08T10:15:25Z","2018-12-06","2018-12"],"dc:description":["Nature has been a perpetual source of inspiration for biochemists. It is not only the vast diversity of compounds that living beings can create, but also the extraordinary strategies of synthesis deployed. Evidently, the catalysts used by living beings -enzymes- are key to nature’s synthesis strategies. Biocatalysis is undoubtfully one of the most invaluable gifts given by nature to flourish the development of green chemical and pharmaceutical industries. With the development of protein and metabolic engineering tools and strategies, more and more enzymes have been used in the industries to improve the chemical processing; and microbes such as Escherichia coli and Saccharomyces cerevisiae have been engineered to produce a wide variety of value-added and bulk chemicals to replace traditional chemical synthesis. However, researchers have just explored the tip of the iceberg in the biocatalysis area. Proteins with new catalytic functionality should be discovered or engineered to broaden current biotransformation boundaries. New in vitro enzymatic or chemoenzymatic cascade reactions need to be designed and optimized to realize stronger synthetic power and more stable systems. Metabolic networks of traditional or new microorganisms should be largely rewired to meet the manufacturing standards. In this work, I aimed at designing and engineering multi-step (bio) catalytic systems for selective synthesis of value-added chemicals. Microorganisms synthesize complex molecules from simple substrates by a series of enzymes working cooperatively. Inspired by how aromatic polyketides are synthesized by the teamwork between enzymes, I sought to couple biocatalysis with organometallic catalysis, two distinct catalytic disciplines, in one pot to realize synthetic power that cannot be achieved by either of them. I first developed a modular, one-pot, sequential chemoenzymatic system for the formal enantioselective construction of C-C bond in 2-aryl 1,4-dicarbonyl compounds. This sequence comprises a rhodium-catalyzed diazocoupling that provides >9:1 selectivity for heterocoupling of two diazoesters and a reduction mediated by an ene-reductase (ER), which occurs in up to 99% enantiomeric excess (ee). The high yield and enantioselectivity of this system were resulted from the preferential generation of an (E)-alkene from the diazo coupling reaction and selective reduction of the (E)-alkene in a mixture of (E) and (Z) isomers by the ER. This work demonstrates the benefit of combining organometallic and enzymatic catalysis to create unusual overall transformations that do not require the isolation and purification of intermediates. To make the system works better on a broader range of substrates, I later developed a new class of cooperative chemoenzymatic reactions that combine photocatalysts that isomerize alkenes with ene-reductases that reduce carbon-carbon double bonds to generate valuable enantioenriched products. I demonstrated that this method enables the stereoconvergent reduction of E/Z mixtures of alkenes or reduction of the unreactive stereoisomer of an alkene in yields and ee’s that match those obtained from the reduction of the pure, more reactive isomer. This new cooperative system overcomes the limitations of both individual catalysts and affords a range of synthetically valuable and biologically active enantioenriched compounds. More generally, these results illustrate the value of driving a chemical reaction with light to ensure compatibility between the chemical and enzymatic catalysts. In vitro biocatalytic reaction normally has poor tolerance to harsh conditions such as low pH or high substrate concentrations. Cells membrane is natural compartmentalization and protects the enzymes from extracellular inhibitors. In addition, cell factories-based production provides an attractive alternative to chemical synthesis of value-added chemicals. I also worked on engineering a S. cerevisiae strain as a whole-cell catalyst for L-lactic acid overproduction in industrially preferred low pH environment (pH 3) by metabolic engineering and genome-wide engineering methods. In addition, to establish an automated cellular engineering platform, I developed a growth-based L-lactic acid biosensor and an automated quantification assay by BioProfile Analyzer.","Submission published under a 24 month embargo labeled 'U of I Access', the embargo will last until 2020-12-01","The student, Yajie Wang, accepted the attached license on 2018-11-30 at 15:19.","The student, Yajie Wang, submitted this Dissertation for approval on 2018-11-30 at 15:36.","This Dissertation was approved for publication on 2018-12-06 at 08:56.","DSpace SAF Submission Ingestion Package generated from Vireo submission #13164 on 2019-02-07 at 14:18:42","Made available in DSpace on 2019-02-07T20:39:41Z (GMT). No. of bitstreams: 6 WANG-DISSERTATION-2018.pdf: 11708977 bytes, checksum: 11bef31153c5271ad34195e62df49852 (MD5) Chapter 1 Permission from Taylor_francis.pdf: 134494 bytes, checksum: bc48dbc76d15b46b9d1dc49abf83d625 (MD5) Chapter 2 Permission from Springer Nature.pdf: 316399 bytes, checksum: 6c8b04dd1f33bf27f977758b8a7fcbdc (MD5) Chapter 3 Permission from ACS Catalysis.pdf: 263759 bytes, checksum: 04ef550745b68ee98304f9ac8129135c (MD5) Chapter 4 Permission from Nature.pdf: 309938 bytes, checksum: 5f7f17688c2d15fe15ef41ec6aa8e11d (MD5) LICENSE.txt: 4207 bytes, checksum: a626d9962cb3a101a5ffd9ff60b0677d (MD5) Previous issue date: 2018-12-06","Embargo set by: Seth Robbins for item 109849 Lift date: 2021-02-07T20:39:46Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","Embargo set by: Seth Robbins for item 109849 Lift date: 2021-02-07T20:44:35Z Reason: Author requested U of Illinois access only (OA after 2yrs) in Vireo ETD system","U of I Only Restriction Lifted for Item 109849 on 2021-02-08T10:15:25Z."],"dc:format":["application/pdf"],"dc:identifier":["http://hdl.handle.net/2142/102824"],"dc:language":["en"],"dc:rights":["Copyright 2018 Yajie Wang"],"dc:subject":["Protein engineering, Synthetic Biology, Chemical engineering, Tandem catalysis, Natural Product, Metabolic Engineering"],"dc:title":["Design and engineering of multi-step (bio)catalytic systems"],"dc:type":["text"],"thesis:degree_discipline":["Chemical Engineering"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:24:42Z"}