TAILIEUCHUNG - Báo cáo Y học: Kinetic analysis of hydroxylation of saturated fatty acids by recombinant P450foxy produced by an Escherichia coli expression system

Cytochrome P450foxy (P450foxy, CYP505) is a fused protein of cytochrome P450 (P450) and its reductase isolated from the fungus Fusarium oxysporum, which catalyzes the subterminal (x-1 x-3) hydroxylation of fatty acids. Here, we produced, purified and characterized a fused recombinant protein (rP450foxy) using the Escherichia coli expression system. Purified rP450foxy was catalytically and spectrally indistinguishable from the native protein, but most of the rP450foxy was recovered in the soluble fraction of E. coli cells unlike the membrane-bound native protein | Eur. J. Biochem. 269 2075-2082 2002 FEBS 2002 doi Kinetic analysis of hydroxylation of saturated fatty acids by recombinant P450foxy produced by an Escherichia coli expression system Tatsuya Kitazume1 Akinori Tanaka1 Naoki Takaya1 Akira Nakamura1 Shigeru Matsuyama1 Takahisa Suzuki1 and Hirofumi Shoun2 1 Institute of Applied Biochemistry University of Tsukuba Tsukuba Ibaraki Japan department of Biotechnology Graduate School of Agricultural and Life Sciences The University of Tokyo Japan Cytochrome P450foxy P450foxy CYP505 is a fused protein of cytochrome P450 P450 and its reductase isolated from the fungus Fusarium oxysporum which catalyzes the subterminal -1 -3 hydroxylation of fatty acids. Here we produced purified and characterized a fused recombinant protein rP450foxy using the Escherichia coli expression system. Purified rP450foxy was catalytically and spectrally indistinguishable from the native protein but most of the rP450foxy was recovered in the soluble fraction of E. coli cells unlike the membrane-bound native protein. The results are consistent with our notion that the native protein is targeted to the membrane by a post-translational modification mechanism. We also discovered that P450foxy could use shorter saturated fatty acid chains C9 and C10 as a substrate. The regiospecificity -1 -3 of hydroxylation due to the enzymatic reaction for the short substrates decanoate C10 undecanoate C11 was the same as that for longer substrates. Steady state kinetic studies showed that the kcat values for all substrates tested C9-C16 were of the same magnitude 1200-1800 min-1 whereas the catalytic efficiency kcat Km was higher for longer fatty acids. Substrate inhibition was observed with fatty acid substrates longer than C13 and the degree of inhibition increased with increasing chain length. This substrate inhibition was not apparent with P450BM3 a bacterial counterpart of P450foxy which was the first obvious difference in their .

• Báo cáo khoa học
• Traditional medicine
• Report medicine
• scientific reports
• Structural biochemistry
• oxidation
• bacteria
• Commercial use
• Tadehagi triquetrum (L ) H Ohashi
• Indigenous medicine in Myanmar
• Medicinal plants
• Calotropis gigantea R
• Br
• Traditional medicinal plants
• Local development in Myanmar
• Important role in health care system
• BMC Chemistry
• Trace analysis
• Dispersive solvent
• Traditional Chinese medicine
• Chinese medicine oral liquid
• Veterinary Medicine
• Herbal Medicine
• Varughese George
• Andrographis paniculata
• Chemical Constituents
• alternative medicine
• diagnostic techniques
• Diagnostic therapies
• Chinese medicine
• Medical herbalism
• Massage therapy review
• Lecture Massage therapy review
• Bodywork assessment
• Massage therapy
• Traditional chinese medicine modality
• Acanthospermum hispidum
• Burkina faso
• Pharmacognosic characterization
• Acanthospermum hispidum (Asteraceae)
• Medicinal plant widely
• Active components
• Plasma protein binding
• Research methods
• Doctoral thesis abstract
• Improving accounting work organization
• Accounting work organization
• Traditional medicine hospitals
• Hanoi city
• Vietnamese traditional medicine
• Medicinal herbs
• Ethnic groups
• Community healthcare
• Mountainous area
• Cultural Perspective
• Modern Medicine
• Funding Perspective
• Traditional Oriental
• medicine
• cell proliferation
• breast cancer
• chemical reaction
• gastric cancer
• hormone medicine
• tumor cells
• biochemical studies
• environmental medicine
• Crystal structure