RRC ID 53579
著者 Sonoki T, Morooka M, Sakamoto K, Otsuka Y, Nakamura M, Jellison J, Goodell B.
タイトル Enhancement of protocatechuate decarboxylase activity for the effective production of muconate from lignin-related aromatic compounds.
ジャーナル 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.
巻・号 192 Pt A
ページ 71-7
公開日 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
引用数 40
リソース情報
原核生物(大腸菌)