Evaluation of bacterial hosts for conversion of lignin-derived p-coumaric acid to 4-vinylphenol

AbstractHydroxycinnamic acids such as p-coumaric acid (CA) are chemically linked to lignin in grassy biomass with fairly labile ester bonds and therefore represent a straightforward opportunity to extract and valorize lignin components. In this work, we investigated the enzymatic conversion of CA extracted from lignocellulose to 4-vinylphenol (4VP) by expressing a microbial phenolic acid decarboxylase in Corynebacterium glutamicum, Escherichia coli, and Bacillus subtilis. The performance of the recombinant strains was evaluated in response to the substrate concentration in rich medium or a lignin liquor and the addition of an organic overlay to perform a continuous product extraction in batch cultures. We found that using undecanol as an overlay enhanced the 4VP titers under high substrate concentrations, while extracting > 97% of the product from the aqueous phase. C. glutamicum showed the highest tolerance to CA and resulted in the accumulation of up to 187 g/L of 4VP from pure CA in the overlay with a 90% yield when using rich media, or 17 g/L of 4VP with a 73% yield from CA extracted from lignin. These results indicate that C. glutamicum is a suitable host for the high-level production of 4VP and that further bioprocess engineering strategies should be explored to optimize the production, extraction, and purification of 4VP from lignin with this organism.

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Bioconversion of p-coumaric acid to p-hydroxystyrene using phenolic acid decarboxylase from B. amyloliquefaciens in biphasic reaction system

Applied Microbiology and Biotechnology ◽

10.1007/s00253-012-4358-8 ◽

2012 ◽

Vol 97 (4) ◽

pp. 1501-1511 ◽

Cited By ~ 41

Author(s):

Da-Hye Jung ◽

Wonji Choi ◽

Kwon-Young Choi ◽

Eunok Jung ◽

Hyungdon Yun ◽

...

Keyword(s):

Phenolic Acid ◽

Reaction System ◽

Acid Decarboxylase ◽

Coumaric Acid ◽

Biphasic Reaction ◽

Phenolic Acid Decarboxylase

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Improving the catalytic characteristics of phenolic acid decarboxylase from Bacillus amyloliquefaciens by the engineering of N-terminus and C-terminus

BMC Biotechnology ◽

10.1186/s12896-021-00705-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Qin Li ◽

Ying Xia ◽

Ting Zhao ◽

Yuanyuan Gong ◽

Shangling Fang ◽

...

Keyword(s):

Ferulic Acid ◽

Phenolic Acid ◽

Bacillus Amyloliquefaciens ◽

Sinapic Acid ◽

Alkali Resistance ◽

Acid Decarboxylase ◽

Coumaric Acid ◽

C Terminus ◽

N Terminus ◽

Phenolic Acid Decarboxylase

Abstract Background 4-vinylphenols produced by phenolic acid degradation catalyzed by phenolic acid decarboxylase can be used in food additives as well as flavor and fragrance industry. Improving the catalytic characters of phenolic acid decarboxylase is of great significance to enhance its practical application. Results A phenolic acid decarboxylase (P-WT) was created from Bacillus amyloliquefaciens ZJH-01. Mutants such as P-C, P-N, P-m1, P-m2, P-Nm1, and P-Nm2 were constructed by site-directed mutagenesis of P-WT. P-C showed better substrate affinities and higher turnover rates than P-WT for p-coumaric acid, ferulic acid, and sinapic acid; however, P-N had reduced affinity toward p-coumaric acid. The extension of the C-terminus increased its acid resistance, whereas the extension of the N-terminus contributed to the alkali resistance and heat resistance. The affinity of P-m1 to four substrates and that of P-m2 to p-coumaric acid and ferulic acid were greatly improved. However, the affinity of P-Nm2 to four phenolic acids was greatly reduced. The residual enzyme activities of P-Nm1 and P-Nm2 considerably improved compared with those of P-m1 and P-m2 after incubation at 50 °C for 60 min. Conclusions The extension of the N-terminus may be more conducive to the combination of the binding cavity with the substrate in an alkaline environment and may make its structure more stable.

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High level production of a cellulase-free xylanase in glucose-limited fed batch cultures of a thermophilic Bacillus strain

Journal of Biotechnology ◽

10.1016/s0168-1656(97)00140-5 ◽

1997 ◽

Vol 58 (2) ◽

pp. 71-78 ◽

Cited By ~ 14

Author(s):

E Samain ◽

Ph Debeire ◽

J.P Touzel

Keyword(s):

Fed Batch ◽

Batch Cultures ◽

Thermophilic Bacillus ◽

Bacillus Strain ◽

High Level ◽

Level Production

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Rewiring carbon metabolism in yeast for high level production of aromatic chemicals

Nature Communications ◽

10.1038/s41467-019-12961-5 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 29

Author(s):

Quanli Liu ◽

Tao Yu ◽

Xiaowei Li ◽

Yu Chen ◽

Kate Campbell ◽

...

Keyword(s):

Carbon Metabolism ◽

Maximum Yield ◽

Glycolytic Flux ◽

Biosynthesis Pathway ◽

Coumaric Acid ◽

Carbon Distribution ◽

High Level ◽

Level Production ◽

Plant Compounds ◽

Key Nodes

Abstract The production of bioactive plant compounds using microbial hosts is considered a safe, cost-competitive and scalable approach to their production. However, microbial production of some compounds like aromatic amino acid (AAA)-derived chemicals, remains an outstanding metabolic engineering challenge. Here we present the construction of a Saccharomyces cerevisiae platform strain able to produce high levels of p-coumaric acid, an AAA-derived precursor for many commercially valuable chemicals. This is achieved through engineering the AAA biosynthesis pathway, introducing a phosphoketalose-based pathway to divert glycolytic flux towards erythrose 4-phosphate formation, and optimizing carbon distribution between glycolysis and the AAA biosynthesis pathway by replacing the promoters of several important genes at key nodes between these two pathways. This results in a maximum p-coumaric acid titer of 12.5 g L−1 and a maximum yield on glucose of 154.9 mg g−1.

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Improving the catalytic characteristics of phenolic acid decarboxylase from Bacillus amyloliquefaciens by the engineering of N-terminus and C-terminus

10.21203/rs.3.rs-114079/v1 ◽

2020 ◽

Author(s):

Qin Li ◽

Ying Xia ◽

Ting Zhao ◽

Yuanyuan Gong ◽

Shangling Fang ◽

...

Keyword(s):

Ferulic Acid ◽

Phenolic Acid ◽

Food Additives ◽

Sinapic Acid ◽

Alkali Resistance ◽

Acid Decarboxylase ◽

Coumaric Acid ◽

C Terminus ◽

N Terminus ◽

Phenolic Acid Decarboxylase

Abstract Background: 4-vinylphenols produced by phenolic acid degradation catalyzed by phenolic acid decarboxylase can be used in food additives as well as flavor and fragrance industry. Improving the catalytic characters of phenolic acid decarboxylase is of great significance to enhance its practical application. Results: Mutants such as P-C, P-N, P-m1, P-m2, P-Nm1, and P-Nm2 were constructed by site-directed mutagenesis of P-WT. P-C showed better substrate affinities and higher turnover rates than P-WT for p-coumaric acid, ferulic acid, and sinapic acid; however, P-N had reduced affinity toward p-coumaric acid. The extension of the C-terminus increased its acid resistance, whereas the extension of the N-terminus contributed to the alkali resistance and heat resistance. The affinity of P-m1 to four substrates and that of P-m2 to p-coumaric acid and ferulic acid were greatly improved. However, the affinity of P-Nm2 to four phenolic acids was greatly reduced. The residual enzyme activities of P-Nm1 and P-Nm2 considerably improved compared with those of P-m1 and P-m2 after incubation at 50°C for 60 min.Conclusions: The extension of the N-terminus may be more conducive to the combination of the binding cavity with the substrate in an alkaline environment and may make its structure more stable.

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