Quantitative isotopic 13C nuclear magnetic resonance at natural abundance to probe enzyme reaction mechanisms via site-specific isotope fractionation: The case of the chain-shortening reaction for the bioconversion of ferulic acid to vanillin (Record no. 45250)

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fixed length control field 02576nam a2200325Ia 4500
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control field MX-MdCICY
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20250625140642.0
040 ## - CATALOGING SOURCE
Transcribing agency CICY
090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN)
Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) B-11023
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245 10 - TITLE STATEMENT
Title Quantitative isotopic 13C nuclear magnetic resonance at natural abundance to probe enzyme reaction mechanisms via site-specific isotope fractionation: The case of the chain-shortening reaction for the bioconversion of ferulic acid to vanillin
490 0# - SERIES STATEMENT
Volume/sequential designation Analytical BioChemistry, 393, p.182-188, 2009
520 3# - SUMMARY, ETC.
Summary, etc. Isotope fractionation is a powerful technique by which to probe the reaction mechanism of enzymes. The effect of a heavy isotope on the reaction energetics can be used to predict transition state architecture and reaction mechanism. In order to examine simultaneously the isotope fractionation in 13C at multiple sites within the substrate and product molecules without any need for site-selective isotope enrichment, a technique exploiting quantitative isotopic nuclear magnetic resonance (NMR)spectrometry at natural abundance (NAQ-NMR)has been developed. Here we report the first application of this technique to the study of an enzyme-catalyzed reaction, the bioconversion of ferulic acid to vanillin in cultures of Streptomyces setonii. We were able to show that the NAQ-NMR methodology is sufficiently precise and robust to measure the isotope shifts in the 13C/12C ratios in both substrate and product of this biotransformation, thereby permitting meaningful data to be obtained ven at carbon positions that take part only indirectly in the reaction and show only secondary isotope fractionation. The results obtained provide direct evidence in support of the current hypothesis for the reaction mechanism of the enzyme hydroxycinnamoyl-CoA hydratase/lyase, notably the proposed involvement of the quinone methide enolate of feruloyl-CoA as intermediate in the catalytic pathway.
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element BIOTRANSFORMATION
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element CHAIN-SHORTENING REACTION
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element ISOTOPIC 13C NMR SPECTROMETRY
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element FERULIC ACID
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element REACTION MECHANISMS
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element STREPTOMYCES
650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element VANILLIN
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Botosoa, E.P.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Blumenstein, C.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Mackenzie, D.A.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Silvestre, V.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Remaud, G.S.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Kwiecien, R.A.
700 12 - ADDED ENTRY--PERSONAL NAME
Personal name Robins, R.J.
856 40 - ELECTRONIC LOCATION AND ACCESS
Uniform Resource Identifier <a href="https://drive.google.com/file/d/1rhG-Gezitxt816h4iBJQnqmBqxDcNFBR/view?usp=drivesdk">https://drive.google.com/file/d/1rhG-Gezitxt816h4iBJQnqmBqxDcNFBR/view?usp=drivesdk</a>
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Source of classification or shelving scheme Clasificación local
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  Clasificación local     Ref1 CICY CICY Documento préstamo interbibliotecario 25.06.2025   B-11023 25.06.2025 25.06.2025 Documentos solicitados