Dataset Information

Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position.

ABSTRACT: Cytochrome P450(SP?) (CYP152B1) isolated from Sphingomonas paucimobilis is the first P450 to be classified as a H(2)O(2)-dependent P450. P450(SP?) hydroxylates fatty acids with high ?-regioselectivity. Herein we report the crystal structure of P450(SP?) with palmitic acid as a substrate at a resolution of 1.65 ?. The structure revealed that the C(?) of the bound palmitic acid in one of the alternative conformations is 4.5 ? from the heme iron. This conformation explains the highly selective ?-hydroxylation of fatty acid observed in P450(SP?). Mutations at the active site and the F-G loop of P450(SP?) did not impair its regioselectivity. The crystal structures of mutants (L78F and F288G) revealed that the location of the bound palmitic acid was essentially the same as that in the WT, although amino acids at the active site were replaced with the corresponding amino acids of cytochrome P450(BS?) (CYP152A1), which shows ?-regioselectivity. This implies that the high regioselectivity of P450(SP?) is caused by the orientation of the hydrophobic channel, which is more perpendicular to the heme plane than that of P450(BS?).

SUBMITTER: Fujishiro T

PROVIDER: S-EPMC3191035 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position.

Fujishiro Takashi T Shoji Osami O Nagano Shingo S Sugimoto Hiroshi H Shiro Yoshitsugu Y Watanabe Yoshihito Y

The Journal of biological chemistry 20110630 34

Cytochrome P450(SPα) (CYP152B1) isolated from Sphingomonas paucimobilis is the first P450 to be classified as a H(2)O(2)-dependent P450. P450(SPα) hydroxylates fatty acids with high α-regioselectivity. Herein we report the crystal structure of P450(SPα) with palmitic acid as a substrate at a resolution of 1.65 Å. The structure revealed that the C(α) of the bound palmitic acid in one of the alternative conformations is 4.5 Å from the heme iron. This conformation explains the highly selective α-hy ...[more]

PMID: 21719702

Similar Datasets

Project description:BackgroundCo-fermentation is an attractive technology for improving volatile fatty acids (VFAs) production by treatment of solid organic wastes. However, it remains unclear how the composition of different organic matters in solid waste influences the VFAs distribution, microbial community structure, and metabolic pathway during acidogenic co-fermentation. In this study, different organic wastes were added into waste activated sludge (WAS) as co-fermentation substrates to explore the impact of organic matter composition on VFAs pattern and the microbiological mechanism .ResultsAcetate was the most dominant VFA produced in all fermentation groups, making up 41.3-57.6% of the total VFAs produced during acidogenic co-fermentation under alkaline condition. With the increased addition of potato peel waste, the concentrations of propionate and valerate decreased dramatically, while ethanol and butyrate concentrations increased. The addition of food waste caused gradual decreases of valerate and propionate, but ethanol increased and butyrate was relatively stable. Some inconsistency was observed between hydrolysis efficiency and acidification efficiency. Our results revealed that starch was mainly responsible for butyrate and ethanol formation, while lipids and protein favored the synthesis of valerate and propionate. Microbial community analysis by high-throughput sequencing showed that Firmicutes had the highest relative abundance at phylum level in all fermentation groups. With 75% potato peel waste or 75% food waste addition to WAS, Bacilli (72.2%) and Clostridia (56.2%) were the dominant respective classes. In fermentation using only potato peel waste, the Bacilli content was 64.1%, while the Clostridia content was 53.6% in the food-only waste fermentation.ConclusionsAcetate was always the dominant product in acidogenic co-fermentation, regardless of the substrate composition. The addition of carbon-rich substrates significantly enhanced butyrate and ethanol accumulation, while protein-rich substrate substantially benefited propionate and valerate generation. Potato peel waste substantially favored the enrichment of Bacilli, while food waste dramatically increased Clostridia content in the sludge.

Dataset Information

Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position.

Publications

Crystal structure of H2O2-dependent cytochrome P450SPalpha with its bound fatty acid substrate: insight into the regioselective hydroxylation of fatty acids at the alpha position.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets