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Structure and Function of a Type III Polyketide Synthase Involved in Cannabinoid Biosynthesis
Structure and Function of a Type III Polyketide Synthase Involved in Cannabinoid Biosynthesis
상세정보
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211152016
- ISBN
- 9798382837314
- DDC
- 574
- 서명/저자
- Structure and Function of a Type III Polyketide Synthase Involved in Cannabinoid Biosynthesis
- 발행사항
- [Sl] : University of California, Los Angeles, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 297 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-12, Section: B.
- 주기사항
- Advisor: Abramson, Jeff S.
- 학위논문주기
- Thesis (Ph.D.)--University of California, Los Angeles, 2024.
- 초록/해제
- 요약Cannabinoids are produced in Cannabis sativa L. via a highly modular process involving chemical precursors originating from the polyketide synthase and monoterpene biosynthetic pathways. In the polyketide synthase pathway, tetraketide synthase (TKS) catalyzes an imprecise reaction to produce a diffusible, linear tetra-β-ketide that is subsequently cyclized by an accessory protein to olivetolic acid. Alkylation, oxidative cyclization, and non-enzymatic decarboxylation of molecules derived from olivetolic acid produce the pharmaceutically important cannabinoids (-)-trans-Δ9 - tetrahydrocannabinol (Δ9 -THC) and cannabidiol (CBD). Besides producing a required pathway precursor, TKS generates multiple aberrant by-products that derail cannabinoid biosynthesis. Structure determination and site-directed mutagenesis experiments were conducted to clarify the TKS reaction mechanism. Crystal structures of TKS in complex with coenzyme A and a pentyl mono-β-ketide, pentyl di-β-ketide, or pentyl tri-β-ketide revealed that polyketide synthesis is facilitated by horizontal expansion of the cyclization pocket. Horizontal expansion is mediated by a single residue rotating outwards from the cyclization pocket. Substitutions truncating this residue's sidechain improved olivetolic acid formation by 176%. Orthogonal structure-function experiments were conducted on hexaketide synthase (Drosophyllum lusitanicum) (DluHKS) and implicated a structural basis for aberrant by-products catalyzed by TKS. A crystal structure was determined for DluHKS and revealed a pentyl tri-β-ketide covalently attached in monomer A and a covalent pentyl di-β-ketide in monomer B. The pentyl tri-β-ketide is stabilized within the cyclization pocket via a hydrogen bond that additionally predisposes polyketide intermediates to lactonization. These collective efforts to determine the structural basis of TKS and DluHKS catalytic activity provide an updated understanding of the TKS reaction mechanism as well as suggestions for rationally mutating TKS for improved cannabinoid bioproduction.
- 일반주제명
- Biochemistry
- 일반주제명
- Biology
- 일반주제명
- Molecular biology
- 일반주제명
- Bioengineering
- 키워드
- Cannabinoids
- 키워드
- Crystallography
- 키워드
- Olivetolic acid
- 기타저자
- University of California, Los Angeles Molecular Biology 0573
- 기본자료저록
- Dissertations Abstracts International. 85-12B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520250211152016
■006m o d
■007cr#unu||||||||
■020 ▼a9798382837314
■035 ▼a(MiAaPQ)AAI31331723
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a574
■1001 ▼aGonzalez-DeWhitt, Kristofer R.
■24510▼aStructure and Function of a Type III Polyketide Synthase Involved in Cannabinoid Biosynthesis
■260 ▼a[Sl]▼bUniversity of California, Los Angeles▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a297 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-12, Section: B.
■500 ▼aAdvisor: Abramson, Jeff S.
■5021 ▼aThesis (Ph.D.)--University of California, Los Angeles, 2024.
■520 ▼aCannabinoids are produced in Cannabis sativa L. via a highly modular process involving chemical precursors originating from the polyketide synthase and monoterpene biosynthetic pathways. In the polyketide synthase pathway, tetraketide synthase (TKS) catalyzes an imprecise reaction to produce a diffusible, linear tetra-β-ketide that is subsequently cyclized by an accessory protein to olivetolic acid. Alkylation, oxidative cyclization, and non-enzymatic decarboxylation of molecules derived from olivetolic acid produce the pharmaceutically important cannabinoids (-)-trans-Δ9 - tetrahydrocannabinol (Δ9 -THC) and cannabidiol (CBD). Besides producing a required pathway precursor, TKS generates multiple aberrant by-products that derail cannabinoid biosynthesis. Structure determination and site-directed mutagenesis experiments were conducted to clarify the TKS reaction mechanism. Crystal structures of TKS in complex with coenzyme A and a pentyl mono-β-ketide, pentyl di-β-ketide, or pentyl tri-β-ketide revealed that polyketide synthesis is facilitated by horizontal expansion of the cyclization pocket. Horizontal expansion is mediated by a single residue rotating outwards from the cyclization pocket. Substitutions truncating this residue's sidechain improved olivetolic acid formation by 176%. Orthogonal structure-function experiments were conducted on hexaketide synthase (Drosophyllum lusitanicum) (DluHKS) and implicated a structural basis for aberrant by-products catalyzed by TKS. A crystal structure was determined for DluHKS and revealed a pentyl tri-β-ketide covalently attached in monomer A and a covalent pentyl di-β-ketide in monomer B. The pentyl tri-β-ketide is stabilized within the cyclization pocket via a hydrogen bond that additionally predisposes polyketide intermediates to lactonization. These collective efforts to determine the structural basis of TKS and DluHKS catalytic activity provide an updated understanding of the TKS reaction mechanism as well as suggestions for rationally mutating TKS for improved cannabinoid bioproduction.
■590 ▼aSchool code: 0031.
■650 4▼aBiochemistry
■650 4▼aBiology
■650 4▼aMolecular biology
■650 4▼aBioengineering
■653 ▼aCannabinoids
■653 ▼aCrystallography
■653 ▼aOlivetolic acid
■653 ▼aStructural biology
■653 ▼aTetraketide synthase
■690 ▼a0487
■690 ▼a0306
■690 ▼a0202
■690 ▼a0307
■71020▼aUniversity of California, Los Angeles▼bMolecular Biology 0573.
■7730 ▼tDissertations Abstracts International▼g85-12B.
■790 ▼a0031
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17162468▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
