Increasing the Carbon Flux toward Synthesis of Short-Chain-Length—Medium-Chain-Length Polyhydroxyalkanoate in the Peroxisome of Saccharomyces cerevisiae through Modification of the β-Oxidation Cycle

Abstract

Short-chain-length-medium-chain-length polyhydroxyalkanoates were synthesized in Saccharomyces cerevisiae from intermediates of the ␤-oxidation cycle by expressing the polyhydroxyalkanoate synthases from Aeromonas caviae and Ralstonia eutropha in the peroxisomes. The quantity of polymer produced was increased by using a mutant of the ␤-oxidation-associated multifunctional enzyme with low dehydrogenase activity toward R-3-hydroxybutyryl coenzyme A.

Harnessing Yeast Peroxisomes for Biosynthesis of Fatty-Acid-Derived Biofuels and Chemicals with Relieved Side-Pathway Competition

“…We are the first to show that organelle compartmentalization of multiple-step biosynthetic pathways can significantly decrease the accumulation of byproducts by secluding the biosynthetic pathways from efficient competing enzymes (Figure ). Peroxisomes have been used for production of polyhydroxyalkanoate (PHA) − and fatty alcohols by expressing a single enzyme to transform the intermediates of fatty acid β-oxidation. Peroxisomes have also been harnessed for production of penicillin in Aspergillus nidulans by targeting a cytosolic step of penicillin biosynthesis into peroxisomes for enhanced substrate channeling .…”

Control of D-lactic acid content in P(LA-3HB) copolymer in the yeast Saccharomyces cerevisiae using a synthetic gene expression system

“…In previous studies polymers with D-lactic acid contents from 3 to 30 mol% ( Kadoya et al, 2015 ), 20–50 mol% ( Yang et al, 2010 ), 8–73 mol% ( Nduko et al, 2014 ), and from 55 to 86 mol% ( Jung et al, 2010 ), were obtained by controlling physiological parameters such as aeration and monomer feeding, and by expressing different PHA synthases. In the yeasts S. cerevisiae and Yarrowia lipolytica physiological parameters, substrate feeding, and changes in β-oxidation were used for controlling monomer content in medium chain length (mcl) and short chain length (scl) PHA copolymers ( De Oliveira et al, 2004 ; Gao et al, 2015 ; Haddouche et al, 2010 , 2011 ; Poirier et al, 2001 ; Rigouin et al, 2019 ; Zhang et al, 2006 ). However, none of the studied methods is readily applicable for ubiquitous control of production of other monomers in vivo.…”

Production of Polyhydroxyalkanoates with High 3-Hydroxydodecanoate Monomer Content by fadB and fadA Knockout Mutant of Pseudomonas putida KT2442

“…Many species of Pseudomonas have been reported to have the abilities of accumulating MCL PHA from fatty acids or glucose. ,− When fatty acids were used as the sole carbon sources, the β-oxidation pathway played an important role in providing intermediates for PHA synthesis . Since the β-oxidation pathway has a close relationship with MCL PHA synthesis, many researchers have studied the metabolic engineering of the β-oxidation pathway to increase PHA accumulation. − 3-Ketoacyl-CoA thiolase (FadA) and 3-hydroxyacyl-CoA dehydrogenase (FadB) are two important enzymes catalyzing the last two steps in the β-oxidation pathway. Snell et al and Park et al.…”

“…14 Since the β-oxidation pathway has a close relationship with MCL PHA synthesis, many researchers have studied the metabolic engineering of the β-oxidation pathway to increase PHA accumulation. [14][15][16][17][18][19][20] 3-Ketoacyl-CoA thiolase (FadA) and 3-hydroxyacyl-CoA dehydrogenase (FadB) are two important enzymes catalyzing the last two steps in the β-oxidation pathway. Snell et al and Park et al reported that MCL PHA production could be enhanced in fadB mutant E. coli harboring only the phaC gene.…”