Reference - Detail
| RRC ID | 53579 |
|---|---|
| Author | Sonoki T, Morooka M, Sakamoto K, Otsuka Y, Nakamura M, Jellison J, Goodell B. |
| Title | Enhancement of protocatechuate decarboxylase activity for the effective production of muconate from lignin-related aromatic compounds. |
| Journal | J Biotechnol |
| Abstract |
The decarboxylation reaction of protocatechuate has been described as a bottleneck and a rate-limiting step in cis,cis-muconate (ccMA) bioproduction from renewable feedstocks such as sugar. Because sugars are already in high demand in the development of many bio-based products, our work focuses on improving protocatechuate decarboxylase (Pdc) activity and ccMA production in particular, from lignin-related aromatic compounds. We previously had transformed an Escherichia coli strain using aroY, which had been used as a protocatechuate decarboxylase encoding gene from Klebsiella pneumoniae subsp. pneumoniae A170-40, and inserted other required genes from Pseudomonas putida KT2440, to allow the production of ccMA from vanillin. This recombinant strain produced ccMA from vanillin, however the Pdc reaction step remained a bottleneck during incubation. In the current study, we identify a way to increase protocatechuate decarboxylase activity in E. coli through enzyme production involving both aroY and kpdB; the latter which encodes for the B subunit of 4-hydroxybenzoate decarboxylase. This permits expression of Pdc activity at a level approximately 14-fold greater than the strain with aroY only. The expression level of AroY increased, apparently as a function of the co-expression of AroY and KpdB. Our results also imply that ccMA may inhibit vanillate demethylation, a reaction step that is rate limiting for efficient ccMA production from lignin-related aromatic compounds, so even though ccMA production may be enhanced, other challenges to overcome vanilate demethylation inhibition still remain. |
| Volume | 192 Pt A |
| Pages | 71-7 |
| Published | 2014-12-20 |
| DOI | 10.1016/j.jbiotec.2014.10.027 |
| PII | S0168-1656(14)00935-3 |
| PMID | 25449108 |
| MeSH | Bacterial Proteins / genetics Bacterial Proteins / metabolism* Benzaldehydes / metabolism* Carboxy-Lyases / genetics Carboxy-Lyases / metabolism* Escherichia coli / enzymology* Escherichia coli / genetics Genes, Bacterial / genetics Hydroxybenzoates / metabolism* Klebsiella pneumoniae / enzymology Klebsiella pneumoniae / genetics Lignin Plasmids Pseudomonas putida / genetics Recombinant Proteins / metabolism Sorbic Acid / analogs & derivatives* Sorbic Acid / metabolism |
| IF | 3.503 |
| Times Cited | 40 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | Patent(IFI CLAIMS) |
| Total number of mentions | 14 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Prokaryotes E. coli | |