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Title: Biocatalytic system for comparatively assessing the functional association of monolignol cytochrome P450 monooxygenases with their redox partners

Book ·

Lignin is a complex heterogenous polymer derived from oxidative radical polymerization of three monolignols, i.e., p-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol. These lignin monomeric precursors structurally differ in their methoxy groups of the benzene rings. In phenylpropanoid-monolignol biosynthetic pathway, the endoplasmic reticulum (ER)-resident cytochrome P450 monooxygenases, cinnamate 4-hydroxylase, coumaroyl ester 3´-hydroxylase and ferulate 5-hydroxylase, establish the key structural characteristics of monolignols. The catalysis of cytochrome P450 monooxygenase requires reducing power, which is supplied by the ER electron transfer chains, composed of cytochrome P450 oxidoreductase (CPR), cytochrome b5 reductase (CBR) and/or cytochrome b5 protein (CB5), from cofactor NADPH or NADH. While NADPH-dependent CPR serves as the typical electron donor for most P450 enzymes, in some cases, the CBR-CB5 or CPR-CB5 electron transfer system also transfers electrons to the terminal P450 enzymes. There are tremendous studies focusing on the discovery and characterization of cytochrome P450 monooxygenases. However, very limited attention has been paid to the versatility and the roles of electron transfer components in the P450 catalytic system. Due to the membrane-residence property of both P450 enzymes and electron transfer components, it is challenging to establish an effective experimental system to evaluate the functional association of P450s with their redox partners. This chapter describes a yeast cell biocatalytic system and the related experimental procedures for comparatively assessing the functional relationship of monolignol biosynthetic P450 enzymes and different redox partners in their catalysis.

Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
DOE Contract Number:
SC0012704
OSTI ID:
1898599
Report Number(s):
BNL-223713-2022-BOOK
Country of Publication:
United States
Language:
English

References (39)

Cytochromes P450 journal January 2011
Cytochromes P450 as versatile biocatalysts journal June 2006
Role of cytochrome b5 in the modulation of the enzymatic activities of cytochrome P450 17α-hydroxylase/17,20-lyase (P450 17A1) journal June 2017
Lignin Biosynthesis journal June 2003
Toward Engineering the Stability and Hemin-Binding Properties of Microsomal Cytochromes b5 into Rat Outer Mitochondrial Membrane Cytochrome b5:  Examining the Influence of Residues 25 and 71 journal August 2002
A cytochrome b5 is required for full activity of flavonoid 3',5'-hydroxylase, a cytochrome P450 involved in the formation of blue flower colors journal January 1999
Molecular evolution of P450 superfamily and P450-containing monooxygenase systems journal October 1993
Evidence that cytochrome b5 acts as a redox donor in CYP17A1 mediated androgen synthesis journal August 2016
Cytochrome b5 reduction by NADPH-cytochrome P-450 reductase. journal September 1979
Cytochrome P450 17A1 Interactions with the FMN Domain of Its Reductase as Characterized by NMR journal December 2015
Microsomal Electron Transfer in Higher Plants: Cloning and Heterologous Expression of NADH-Cytochromeb5Reductase from Arabidopsis journal January 1999
Enzymatic assembly of DNA molecules up to several hundred kilobases journal April 2009
Cytochrome b 5 Is an Obligate Electron Shuttle Protein for Syringyl Lignin Biosynthesis in Arabidopsis journal April 2019
The scaffold proteins of lignin biosynthetic cytochrome P450 enzymes journal April 2018
Cytochrome P450 systems—biological variations of electron transport chains journal March 2007
Evidence That Cytochromeb5and Cytochromeb5Reductase Can Act as Sole Electron Donors to the Hepatic Cytochrome P450 System journal March 2013
Evidence for the participation of cytochrome b5 in hepatic microsomal mixed-function oxidation reactions journal March 1971
Novel Targeting Signals Mediate the Sorting of Different Isoforms of the Tail-Anchored Membrane Protein Cytochrome b 5 to Either Endoplasmic Reticulum or Mitochondria journal October 2004
Heterologous expression and mechanistic investigation of a fungal cytochrome P450 (CYP5150A2): Involvement of alternative redox partners journal February 2012
Molecular mechanism of metabolic NAD(P)H-dependent electron-transfer systems: The role of redox cofactors journal March 2019
Plant NADPH-cytochrome P450 oxidoreductases journal February 2010
Reactive Intermediates in Cytochrome P450 Catalysis journal April 2013
Higher Plant Cytochrome b5 Polypeptides Modulate Fatty Acid Desaturation journal February 2012
Intracellular sorting of the tail-anchored protein cytochrome b5 in plants: a comparative study using different isoforms from rabbit and Arabidopsis journal February 2007
The Cytochrome P450 Homepage journal October 2009
Discovery and engineering of colchicine alkaloid biosynthesis journal July 2020
Structure of the Arabidopsis thaliana NADPH ‐cytochrome P450 reductase 2 (ATR2) provides insight into its function journal February 2017
[6] Yeast expression of animal and plant P450s in optimized redox environments book January 1996
The roles of cytochromeb5 in cytochrome P450 reactions journal December 2002
Cytochrome P-450. Multiplicity of isoforms, substrates, and catalytic and regulatory mechanisms journal July 1991
The many roles of cytochrome b5 journal February 2003
Cloning, functional analysis, and subcellular localization of two isoforms of NADH:cytochrome b5 reductase from developing seeds of tung (Vernicia fordii) journal August 2005
NADH:Cytochrome b5 Reductase and Cytochrome b5 Can Act as Sole Electron Donors to Human Cytochrome P450 1A1-Mediated Oxidation and DNA Adduct Formation by Benzo[a]pyrene journal July 2016
Mutation of the Inducible ARABIDOPSIS THALIANA CYTOCHROME P450 REDUCTASE2 Alters Lignin Composition and Improves Saccharification journal October 2014
Characterization of recombinant plant cinnamate 4-hydroxylase produced in yeast. Kinetic and spectral properties of the major plant P450 of the phenylpropanoid pathway journal June 1994
Cytochrome b5, its functions, structure and membrane topology journal January 1995
Cytochrome b5 Reductase Encoded by CBR1 Is Essential for a Functional Male Gametophyte in Arabidopsis journal August 2013
Lack of Electron Transfer from Cytochrome b5 in Stimulation of Catalytic Activities of Cytochrome P450 3A4 journal November 1996
Roles of NADPH-P450 Reductase and Apo- and Holo-Cytochrome b5 on Xenobiotic Oxidations Catalyzed by 12 Recombinant Human Cytochrome P450s Expressed in Membranes of Escherichia coli journal April 2002