Characterization of Thiamine Diphosphate-Dependent 4-Hydroxybenzoylformate Decarboxylase Enzymes from Rhodococcus jostii RHA1 and Pseudomonas fluorescens Pf-5 Involved in Degradation of Aryl C2 Lignin Degradation Fragments.
Characterization of Thiamine Diphosphate-Dependent 4-Hydroxybenzoylformate Decarboxylase Enzymes from Rhodococcus jostii RHA1 and Pseudomonas fluorescens Pf-5 Involved in Degradation of Aryl C2 Lignin Degradation Fragments.
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红球菌 jostii RHA1 和荧光假单胞菌 Pf-5 参与芳基 C2 木质素降解片段降解的硫胺素二磷酸依赖性 4-羟基苯甲酰甲酸脱羧酶的表征。
DOI：
10.1021/acs.biochem.9b00177
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发表时间：
2019
期刊：
影响因子：
2.9
通讯作者：
Wei Z
中科院分区：
文献类型：
--
作者：
Wei Z
一种被注释为苯甲酰甲酸脱羧酶的二磷酸硫胺素依赖性酶由Rhodococcus jostii RHA 1中的基因簇ro 02984-ro 02986编码，该基因簇先前显示从木质素氧化产生香草醛和4-羟基苯甲醛，并且在荧光假单胞菌Pf-5的基因组中也发现了一个密切相关的基因簇。两种假说的可能途径，涉及硫胺素二磷酸依赖的裂解，无论是C-C裂解的酮醇或二酮芳基C3底物或脱羧的芳基C2底物，进行了研究的表达和纯化的重组酶和表达的脱氢酶和氧化酶也发现在基因簇。ThDP依赖性酶没有表现出裂解芳基C3酮醇或二酮底物的活性，但表现出苯甲酰甲酸和4-羟基苯甲酰甲酸脱羧的活性。由基因ro 02984编码的黄素依赖氧化酶被发现氧化扁桃酸或苯乙二醛。对P.荧光脱羧酶的测定在1.69 μ m分辨率，显示与已知的苯甲酰甲酸脱羧酶的结构相似。P。荧光脱羧酶表现出增强的香草醛和乙醛之间的carboligase活性，合理的丙氨酸相对于丝氨酸的存在下，在残基73在酶活性位点，这是进一步研究通过定点诱变该残基。提出了木质素中苯香豆素和二芳基丙烷结构氧化裂解产生的芳基C2片段降解途径的假说。
A thiamine diphosphate-dependent enzyme annotated as a benzoylformate decarboxylase is encoded by gene cluster ro02984–ro02986 inRhodococcus jostiiRHA1 previously shown to generate vanillin and 4-hydroxybenzaldehyde from lignin oxidation, and a closely related gene cluster is also found in the genome ofPseudomonas fluorescensPf-5. Two hypotheses for possible pathways involving a thiamine diphosphate-dependent cleavage, either C–C cleavage of a ketol or diketone aryl C3substrate or decarboxylation of an aryl C2substrate, were investigated by expression and purification of the recombinant enzymes and expression of dehydrogenase and oxidase enzymes also found in the gene clusters. The ThDP-dependent enzymes showed no activity for cleavage of aryl C3ketol or diketone substrates but showed activity for decarboxylation of benzoylformate and 4-hydroxybenzoylformate. A flavin-dependent oxidase encoded by gene ro02984 was found to oxidize either mandelic acid or phenylglyoxal. The crystal structure of theP. fluorescensdecarboxylase enzyme was determined at 1.69 Å resolution, showing similarity to structures of known benzoylformate decarboxylase enzymes. TheP. fluorescensdecarboxylase enzyme showed enhanced carboligase activity between vanillin and acetaldehyde, rationalized by the presence of alanine versus serine at residue 73 in the enzyme active site, which was investigated further by site-directed mutagenesis of this residue. A hypothesis for a pathway for degradation of aryl C2fragments arising from oxidative cleavage of phenylcoumaran and diarylpropane structures in lignin is proposed.
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影响因子：
4
作者：
Sainsbury, Paul D.;Hardiman, Elizabeth M.;Bugg, Timothy D. H.
通讯作者：
Bugg, Timothy D. H.
DOI：
10.1107/s0907444907050433
发表时间：
2007-12-01
影响因子：
2.2
作者：
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通讯作者：
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影响因子：
4
作者：
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DOI：
10.1016/j.bbapap.2015.04.027
发表时间：
2015-08-01
影响因子：
3.2
作者：
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通讯作者：
McLeish, Michael J.
影响因子：
2.9
作者：
Ahmad, Mark;Roberts, Joseph N.;Bugg, Timothy D. H.
通讯作者：
Bugg, Timothy D. H.