scholarly journals The purification and properties of p-cresol-(acceptor) oxidoreductase (hydroxylating), a flavocytochrome from Pseudomonas putida

1977 ◽  
Vol 167 (1) ◽  
pp. 155-162 ◽  
Author(s):  
D J Hopper ◽  
D G Taylor

The enzyme that catalyses the hydroxylation of the methyl group of p-cresol was purified from Pseudomonas putida. It has mol.wt. 115000 and appears to contain two subunits of equal molecular weight. One subunit is a c-type cytochrome and the other is a flavoprotein. Reduction of the cytochrome occurred on addition of substrate. The same enzyme catalyses both p-cresol hydroxylation and the further oxidation of the product, 4-hydroxybenzyl alcohol. The stoicheiometry of acceptor reduced per molecule of substrate oxidized is that for two dehydrogenation reactions. The Km for p-cresol is 7.3 × 10(-6) M and that for 4-hydroxybenzyl alcohol is 47.6 × 10(-6) M. The enzyme, which is assayed with phenazine methosulphate as electron acceptor, was stimulated by particulate material, which probably contains the acceptor in vivo.

1978 ◽  
Vol 175 (2) ◽  
pp. 659-667 ◽  
Author(s):  
M J Keat ◽  
D J Hopper

Whole cells of Pseudomonas putida N.C.I.B 9869, when grown on either 3,5-xylenol or p-cresol, oxidized both m- and p-hydroxybenzyl alcohols. Two distinct NAD+-dependent m-hydroxybenzyl alcohol dehydrogenases were purified from cells grown on 3,5-xylenol. Each is active with a range of aromatic alcohols, including both m- and p-hydroxybenzyl alcohol, but differ in their relative rates with the various substrates. An NAD+-dependent alcohol dehydrogenase was also partially purified from p-cresol grown cells. This too was active with m- and p-hydroxybenzyl alcohol and other aromatic alcohols, but was not identical with either of the other two dehydrogenases. All three enzymes were unstable, but were stabilized by dithiothreitol and all were inhibited with p-chloromercuribenzoate. All were specific for NAD+ and each was shown to catalyse conversion of alcohol into aldehyde.


1978 ◽  
Vol 175 (1) ◽  
pp. 345-347 ◽  
Author(s):  
D J Hopper

In the hydroxylation of the methyl group of p-cresol by an enzyme from Pseudomonas putida the oxygen atom is derived from water. Although a second reaction by the same enzyme converts the product, p-hydroxybenzyl alcohol, into the aldehyde, the alcohol is an enzyme-free intermediate.


1973 ◽  
Vol 29 (02) ◽  
pp. 490-498 ◽  
Author(s):  
Hiroh Yamazaki ◽  
Itsuro Kobayashi ◽  
Tadahiro Sano ◽  
Takio Shimamoto

SummaryThe authors previously reported a transient decrease in adhesive platelet count and an enhancement of blood coagulability after administration of a small amount of adrenaline (0.1-1 µg per Kg, i. v.) in man and rabbit. In such circumstances, the sensitivity of platelets to aggregation induced by ADP was studied by an optical density method. Five minutes after i. v. injection of 1 µg per Kg of adrenaline in 10 rabbits, intensity of platelet aggregation increased to 115.1 ± 4.9% (mean ± S. E.) by 10∼5 molar, 121.8 ± 7.8% by 3 × 10-6 molar and 129.4 ± 12.8% of the value before the injection by 10”6 molar ADP. The difference was statistically significant (P<0.01-0.05). The above change was not observed in each group of rabbits injected with saline, 1 µg per Kg of 1-noradrenaline or 0.1 and 10 µg per Kg of adrenaline. Also, it was prevented by oral administration of 10 mg per Kg of phenoxybenzamine or propranolol or aspirin or pyridinolcarbamate 3 hours before the challenge. On the other hand, the enhancement of ADP-induced platelet aggregation was not observed in vitro, when 10-5 or 3 × 10-6 molar and 129.4 ± 12.8% of the value before 10∼6 molar ADP was added to citrated platelet rich plasma (CPRP) of rabbit after incubation at 37°C for 30 second with 0.01, 0.1, 1, 10 or 100 µg per ml of adrenaline or noradrenaline. These results suggest an important interaction between endothelial surface and platelets in connection with the enhancement of ADP-induced platelet aggregation by adrenaline in vivo.


Author(s):  
Nguyễn Thỵ Đan Huyền ◽  
Lê Thanh Long ◽  
Trần Thị Thu Hà ◽  
Nguyễn Cao Cường ◽  
Nguyễn Hiền Trang
Keyword(s):  

Chủng T1 phân lập từ các mẫu ngô nếp NK66 nhiễm nấm mốc tự nhiên được sử dụng để nghiên cứu khả năng kháng nấm của dịch chiết vi khuẩn Pseudomonas putida 199B. Đặc điểm hình thái của chủng T1 đã được quan sát đại thể (màu sắc, hình dáng, kích thước khuẩn lạc) trên môi trường PDA và vi thể (hình dáng bào tử) trên kính hiển vi kết hợp so sánh với loài Aspergilus flavus đối chứng. Kết quả phân tích trình tự gen mã hóa 28S rRNA của chủng T1 cho thấy sự tương đồng trình tự cao với các trình tự tương ứng của loài Aspergilus flavus trên ngân hàng gen. Kết quả khảo sát ảnh hưởng của dịch chiết vi khuẩn P. putida lên sự phát triển của nấm A.  flavus gây bệnh trên hạt ngô sau thu hoạch và bảo quản ở điều kiện in vitro cho thấy, ở nồng độ P. putida 24% đã ức chế 74,50% sự phát triển đường kính tản nấm sau 10 ngày nuôi cấy, ức chế 79,63% sự hình thành sinh khối sợi nấm sau 7 ngày nuôi cấy. Ở điều kiện in vivo, sự nảy mầm của hạt giống ngô sau 30 ngày được tạo màng bao sinh học bằng dịch chiết vi khuẩn P. putida nồng độ 18% đạt 97,91%, tỉ lệ hạt nhiễm nấm mốc giảm còn 20% so với 72% ở mẫu đối chứng.


1996 ◽  
Vol 34 (10) ◽  
pp. 67-72 ◽  
Author(s):  
Lu Chih-Jen ◽  
Lee Chi-Mei ◽  
Huang Chiou-Zong

The biodegradation of phenol and chlorophenols by immobilized pure-culture cells was conducted by a series of batch reactors. The microorganisms used in this study were Pseudomonas putida, Psuedomonas testosteroni, Pseudomonas aeruginosa, and Agrobacterium radiobacter. All four species showed the ortho-cleavage pathway to metabolize chlorophenols. Among the four species, P. testosteroni, P. putida, and P. aeruginosa could effectively remove phenol at 200 mg/l. P. testosteroni could effectively remove 2-chlorophenol at 10mg/l. However, the other three species, P. putida, P. aeruginosa, and A. radiobacter, could not effectively remove 2-chlorophenol. Although 3-chlorophenol is a recalcitrant compound, P. testosteroni also could rapidly metabolize 3-chlorophenol at 10 mg/l. The removal of 4-chlorophenol at 10 mg/l by P. testosteroni reached 98% within one day. P. aeruginosa and A. radiobacter also could metabolize 4-chlorophenol after 2 and 7 days of lag period, respectively.


1987 ◽  
Vol 52 (9) ◽  
pp. 2317-2325 ◽  
Author(s):  
Jan Hlaváček ◽  
Jan Pospíšek ◽  
Jiřina Slaninová ◽  
Walter Y. Chan ◽  
Victor J. Hruby

[8-Neopentylglycine]oxytocin (II) and [8-cycloleucine]oxytocin (III) were prepared by a combination of solid-phase synthesis and fragment condensation. Both analogues exhibited decreased uterotonic potency in vitro, each being about 15-30% that of oxytocin. Analogue II also displayed similarly decreased uterotonic potency in vivo and galactogogic potency. On the other hand, analogue III exhibited almost the same potency as oxytocin in the uterotonic assay in vivo and in the galactogogic assay.


2006 ◽  
Vol 50 (6) ◽  
pp. 2261-2264 ◽  
Author(s):  
Hee-Soo Park ◽  
Hyun-Joo Kim ◽  
Min-Jung Seol ◽  
Dong-Rack Choi ◽  
Eung-Chil Choi ◽  
...  

ABSTRACT DW-224a showed the most potent in vitro activity among the quinolone compounds tested against clinical isolates of gram-positive bacteria. Against gram-negative bacteria, DW-224a was slightly less active than the other fluoroquinolones. The in vivo activities of DW-224a against gram-positive bacteria were more potent than those of other quinolones.


2013 ◽  
Vol 2013 ◽  
pp. 1-21 ◽  
Author(s):  
Giuseppe Sautto ◽  
Nicasio Mancini ◽  
Giacomo Gorini ◽  
Massimo Clementi ◽  
Roberto Burioni

More than 150 arboviruses belonging to different families are known to infect humans, causing endemic infections as well as epidemic outbreaks. Effective vaccines to limit the occurrence of some of these infections have been licensed, while for the others several new immunogens are under development mostly for their improvements concerning safety and effectiveness profiles. On the other hand, specific and effective antiviral drugs are not yet available, posing an urgent medical need in particular for emergency cases. Neutralizing monoclonal antibodies (mAbs) have been demonstrated to be effective in the treatment of several infectious diseases as well as in preliminaryin vitroandin vivomodels of arbovirus-related infections. Given their specific antiviral activity as well-tolerated molecules with limited side effects, mAbs could represent a new therapeutic approach for the development of an effective treatment, as well as useful tools in the study of the host-virus interplay and in the development of more effective immunogens. However, before their use as candidate therapeutics, possible hurdles (e.g., Ab-dependent enhancement of infection, occurrence of viral escape variants) must be carefully evaluated. In this review are described the main arboviruses infecting humans and candidate mAbs to be possibly used in a future passive immunotherapy.


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