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Realizing the Biotechnological Potential of Fungal Cellulosomes- [electronic resource]
Realizing the Biotechnological Potential of Fungal Cellulosomes- [electronic resource]
상세정보
- 자료유형
- 학위논문(국외)
- 자관 청구기호
- 기본표목-개인명
- 표제와 책임표시사항
- Realizing the Biotechnological Potential of Fungal Cellulosomes - [electronic resource] / Stephen Peter Lillington
- 발행, 배포, 간사 사항
- 발행, 배포, 간사 사항
- 형태사항
- 1 online resource(p.211 )
- 일반주기
- Source: Dissertations Abstracts International, Volume: 85-02, Section: B.
- 일반주기
- Advisor: O'Malley, Michelle A.;Shell, M. Scott.
- 학위논문주기
- Thesis (Ph.D.)--University of California, Santa Barbara, 2023.
- 이용제한주기
- This item must not be sold to any third party vendors.
- 요약 등 주기
- 요약Rising risks of climate change and supply chain insecurity highlight the need to develop alternative, greener synthesis routes to common materials currently sourced from petroleum. Biological systems excel at interconverting chemicals with exquisite specificity and speed, using networks of enzymes that perform catalysis at mild conditions. Protein complexes in nature colocalize complementary subunits to perform sophisticated biochemistry, and artificial, spatial organization of enzyme systems into synthetic complexes is an attractive strategy for improving biocatalytic process throughputs in industrial settings. While some sets of modular parts that enable designer protein complex construction exist, there is still a need to develop new components that are widely compatible with different enzymes and that are highly engineerable to impart desired self-assembly properties. Fungal cellulosomes, modular protein machines produced by anaerobic fungi in the guts of herbivores to rapidly free sugars from plant matter, represent an unexplored framework for synthetic protein complex construction. Cellulosomes synergistically incorporate enzymes involved in biomass degradation into discrete complexes via modular protein-protein interactions between enzyme fused dockerin domains and cohesin domains repeated on a central scaffoldin protein. Over 80% of the degradative power anaerobic fungi possess is attributed to cellulosomes, but the mechanistic nature of their activity and their assembly mechanism remain unknown. These knowledge gaps have precluded the development of fungal cellulosomes or their parts as biocatalytic technologies with real world applications.We apply a range of experimental techniques towards addressing how cellulosomes are produced in native anaerobic fungal cultures and characterizing the composition, nanostructure, and biochemical activity of purified, native cellulosomes. Immunofluorescence microscopy with cellulosome-labeling antibodies shows cellulosomes localize to the surfaces of cells, but that only cells at certain stages of the multi-staged life cycle produce cellulosomes under specific growth conditions. A robust cellulosome purification method we developed, in conjunction with mass spectrometry-based proteomics and biomass hydrolysis kinetic assays, provides high resolution details into the composition and lignocellulolytic activities of isolated cellulosomes produced by an anaerobic fungus, advancing our understanding of how cellulosomes can be engineered to enhance biomass hydrolysis rates.Towards leveraging the modular cellulosome assembly framework for synthetic biology applications, we develop a suite of modular interacting parts for constructing protein complexes with fungal cellulosome proteins. Through a combination of molecular modeling and high-throughput screening, we engineer interacting domains with a range of pH dependent binding behaviors for building protein complexes whose composition and therefore function are modulated with and environmental trigger, pH. Together, these tools and insights shed light on how cellulosomes make anaerobic fungi prolific biomass degraders and provide a framework for engineering protein complexes inspired by fungal cellulosomes designed for a wide range of applications.
- 주제명부출표목-일반주제명
- 주제명부출표목-일반주제명
- 주제명부출표목-일반주제명
- 비통제 색인어
- 비통제 색인어
- 비통제 색인어
- 비통제 색인어
- 비통제 색인어
- 부출표목-단체명
- 기본자료저록
- Dissertations Abstracts International. 85-02B.
- 기본자료저록
- Dissertation Abstract International
- 전자적 위치 및 접속
- 원문정보보기
- 소장사항
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202402 2024
MARC
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■090 ▼a전자도서(박사논문)
■1001 ▼aLillington, Stephen Peter.
■24510▼aRealizing the Biotechnological Potential of Fungal Cellulosomes▼h[electronic resource]▼cStephen Peter Lillington
■260 ▼a[S.l.]▼bUniversity of California, Santa Barbara. ▼c2023
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2023
■300 ▼a1 online resource(p.211 )
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-02, Section: B.
■500 ▼aAdvisor: O'Malley, Michelle A.;Shell, M. Scott.
■5021 ▼aThesis (Ph.D.)--University of California, Santa Barbara, 2023.
■506 ▼aThis item must not be sold to any third party vendors.
■520 ▼aRising risks of climate change and supply chain insecurity highlight the need to develop alternative, greener synthesis routes to common materials currently sourced from petroleum. Biological systems excel at interconverting chemicals with exquisite specificity and speed, using networks of enzymes that perform catalysis at mild conditions. Protein complexes in nature colocalize complementary subunits to perform sophisticated biochemistry, and artificial, spatial organization of enzyme systems into synthetic complexes is an attractive strategy for improving biocatalytic process throughputs in industrial settings. While some sets of modular parts that enable designer protein complex construction exist, there is still a need to develop new components that are widely compatible with different enzymes and that are highly engineerable to impart desired self-assembly properties. Fungal cellulosomes, modular protein machines produced by anaerobic fungi in the guts of herbivores to rapidly free sugars from plant matter, represent an unexplored framework for synthetic protein complex construction. Cellulosomes synergistically incorporate enzymes involved in biomass degradation into discrete complexes via modular protein-protein interactions between enzyme fused dockerin domains and cohesin domains repeated on a central scaffoldin protein. Over 80% of the degradative power anaerobic fungi possess is attributed to cellulosomes, but the mechanistic nature of their activity and their assembly mechanism remain unknown. These knowledge gaps have precluded the development of fungal cellulosomes or their parts as biocatalytic technologies with real world applications.We apply a range of experimental techniques towards addressing how cellulosomes are produced in native anaerobic fungal cultures and characterizing the composition, nanostructure, and biochemical activity of purified, native cellulosomes. Immunofluorescence microscopy with cellulosome-labeling antibodies shows cellulosomes localize to the surfaces of cells, but that only cells at certain stages of the multi-staged life cycle produce cellulosomes under specific growth conditions. A robust cellulosome purification method we developed, in conjunction with mass spectrometry-based proteomics and biomass hydrolysis kinetic assays, provides high resolution details into the composition and lignocellulolytic activities of isolated cellulosomes produced by an anaerobic fungus, advancing our understanding of how cellulosomes can be engineered to enhance biomass hydrolysis rates.Towards leveraging the modular cellulosome assembly framework for synthetic biology applications, we develop a suite of modular interacting parts for constructing protein complexes with fungal cellulosome proteins. Through a combination of molecular modeling and high-throughput screening, we engineer interacting domains with a range of pH dependent binding behaviors for building protein complexes whose composition and therefore function are modulated with and environmental trigger, pH. Together, these tools and insights shed light on how cellulosomes make anaerobic fungi prolific biomass degraders and provide a framework for engineering protein complexes inspired by fungal cellulosomes designed for a wide range of applications.
■590 ▼aSchool code: 0035.
■650 4▼aChemical engineering.
■650 4▼aBioengineering.
■650 4▼aMicrobiology.
■653 ▼aAnaerobic fungi
■653 ▼aCellulosome
■653 ▼aNanobody
■653 ▼aProtein engineering
■653 ▼aYeast surface display
■690 ▼a0542
■690 ▼a0202
■690 ▼a0410
■71020▼aUniversity of California, Santa Barbara▼bChemical Engineering.
■7730 ▼tDissertations Abstracts International▼g85-02B.
■773 ▼tDissertation Abstract International
■790 ▼a0035
■791 ▼aPh.D.
■792 ▼a2023
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T16933252▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
■980 ▼a202402▼f2024
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