Biosynthesis of eriodictyol from tyrosine by Corynebacterium glutamicum

Abstract

Background Eriodictyol is a bioactive flavonoid compound that shows potential applications in medicine development and food processing. Microbial synthesis of eriodictyol has been attracting increasing attention due to several benefits. In this study, we employed a GRAS strain Corynebacterium glutamicum as the host to produce eriodictyol directly from tyrosine. Results We firstly optimized the biosynthetic module of naringenin, the upstream intermediate for eriodictyol production, through screening of different gene orthologues. Next, to improve the level of the precursor malonyl-CoA necessary for naringenin production, we introduced matB and matC from Rhizobium trifolii into C. glutamicum to convert extracellular malonate to intracellular malonyl-CoA. This combinatorial engineering resulted in around 35-fold increase in naringenin production from tyrosine compared to the initial recombinant C. glutamicum. Subsequently, the hpaBC genes from E. coli encoding 4-hydroxyphenylacetate 3-hydroxylase were expressed in C. glutamicum to synthesize eriodictyol from naringenin. Further optimization of the biotransformation process parameters led to the production of 14.10 mg/L eriodictyol. Conclusions The biosynthesis of the ortho-hydroxylated flavonoid eriodictyol in C. glutamicum was achieved for the first time via functional expression of E. coli hpaBC, providing a baseline strain for biosynthesis of other complex flavonoids. Our study demonstrates the potential application of C. glutamicum as a host microbe for the biosynthesis of value-added natural compounds from tyrosine.

Promoting FADH2 Regeneration of Hydroxylation for High-Level Production of Hydroxytyrosol from Glycerol in Escherichia coli

“…HpaBC is widely used for the hydroxylation of some aromatic compounds. − HpaB utilizes FADH 2 , which is reduced by HpaC, adding a hydroxyl group to the structure of the substrate to give FAD, and continues to be used in the next cycle. HpaBC has often been selected to catalyze the hydroxylation step in the biosynthesis of hydroxytyrosol. ,, Therefore, HpaBC from E.…”

Flavonoid Production: Current Trends in Plant Metabolic Engineering and De Novo Microbial Production

“…Gao et al [ 144 ] reported the highest eriodictyol titer (3.3 g/L) to date by identifying and optimizing efficient Sm CPR and Sm F3′H from Silybum marianum in S. cerevisiae , demonstrating that S. cerevisiae is an attractive platform for P450 enzyme expression involved in flavonoid biosynthesis. In another study, Corynebacterium glutamicum (a GRAS strain) was used as a host for the first time to directly produce eriodictyol (14.10 mg/L) from tyrosine by introducing matC and matB genes from Rhizobium trifolii , as well as hpaBC genes expressing 4-hydroxyphenylacetate 3-hydroxylase from Escherichia coli [ 145 ].…”

High-Level De Novo Production of (2S)-Eriodictyol in Yarrowia Lipolytica by Metabolic Pathway and NADPH Regeneration Engineering

“…Therefore, the production of (2 S )-eriodictyol in prokaryotes is restricted, resulting in only low-level titers. To overcome this limitation, Wu et al applied the non-P450 enzyme HpaBC to catalyze the conversion between (2 S )-naringenin and (2 S )-eriodictyol in Corynebacterium glutamicum, obtaining 14.10 mg/L (2 S )-eriodictyol . Furthermore, our group previously truncated the transmembrane regions of F3′H and CPR and fused them to generate a soluble expression enzyme in E.…”

“…To overcome this limitation, Wu et al applied the non-P450 enzyme HpaBC to catalyze the conversion between (2S)-naringenin and (2S)eriodictyol in Corynebacterium glutamicum, obtaining 14.10 mg/L (2S)-eriodictyol. 14 Furthermore, our group previously truncated the transmembrane regions of F3′H and CPR and fused them to generate a soluble expression enzyme in E. coli, resulting in a 107 mg/L (2S)-eriodictyol titer. 15 Despite the efforts made, the high activity of F3′H/CPR in prokaryotes still faces challenges.…”