scholarly journals Primary alkylsulphatase activities of the detergent-degrading bacterium Pseudomonas C12B. Purification and properties of the P1 enzyme

1986 ◽  
Vol 236 (2) ◽  
pp. 401-408 ◽  
Author(s):  
T J Bateman ◽  
K S Dodgson ◽  
G F White
Keyword(s):  
Chain Length ◽  
Hydrophobic Interactions ◽  
Deae Cellulose ◽  
Carbon Chain ◽  
Maximum Activity ◽  
Carbon Chain Length ◽  
Streptomycin Sulphate ◽  
Degrading Bacterium ◽  
Purification And Properties ◽  
Hydrophobic Binding

The P1 primary alkylsulphatase of Pseudomonas C12B was purified 1500-fold to homogeneity by a combination of streptomycin sulphate precipitation of nucleic acids, (NH4)2SO4 fractionation and chromatography on columns of DEAE-cellulose, Sephacryl S-300 and butyl-agarose. The protein was tetrameric with an Mr of 181000-193000, and exhibited maximum activity at pH 6.1. Primary alkyl sulphates of carbon-chain length C1-C5 or above C14 were not substrates, but the intermediate homologues were shown to be substrates, either by direct assay (C6-C9 and C12) or by gel zymography (C10, C11, C13 and C14). Increasing the chain length from C6 to C12 led to diminishing Km. Values of delta G0′ for binding substrates to enzyme were dependent linearly on chain length, indicating high dependence on hydrophobic interactions. Vmax./Km values increased with increasing chain length. Inhibition by alk-2-yl sulphates and alkane-sulphonates was competitive and showed a similar dependence on hydrophobic binding. The P1 enzyme was active towards several aryl sulphates, including o-, m- and p-chlorophenyl sulphates, 2,4-dichlorophenyl sulphate, o-, m- and p-methoxyphenyl sulphates, m- and p-hydroxyphenyl sulphates and p-nitrophenyl sulphate, but excluding bis-(p-nitrophenyl) sulphate and the O-sulphate esters of tyrosine, nitrocatechol and phenol. The arylsulphatase activity was weak compared with alkylsulphatase activity, and it was distinguishable from the de-repressible arylsulphatase activity of Pseudomonas C12B reported previously. Comparison of the P1 enzyme with the inducible P2 alkylsulphatase of this organism, and with the Crag herbicide sulphatase of Pseudomonas putida, showed that, although there are certain similarities between any two of the three enzymes, very few properties are common to all three.

scholarly journals Purification and properties of the P2 primary alkylsulphohydrolase of the detergent-degrading bacterium pseudomonas C12B

1980 ◽  
Vol 185 (1) ◽  
pp. 23-31 ◽  
Author(s):  
J M Cloves ◽  
K S Dodgson ◽  
G F White ◽  
J W Fitzgerald
Keyword(s):  
Free Energy ◽  
Chain Length ◽  
Alkyl Chain ◽  
Competitive Inhibition ◽  
Deae Cellulose ◽  
Intact Protein ◽  
Alkyl Sulphate ◽  
Degrading Bacterium ◽  
Purification And Properties ◽  
Considerable Length

The P2 primary alkylsulphohydrolase of the soil bacterium Pseudomonas C12B was purified to homogeneity (200-250-fold) by column chromatography on DEAE-cellulose, Sephadex G-100 and butyl-agarose. The intact protein is a dimer with a mol. wt. of 160 000. Activity towards primary alkyl sulphate esters was maximal at pH 8.3, varied little in the range pH 7.8-8.7, but decreased sharply at higher pH. For a homologous series of primary alkyl sulphate substrates (C6-C12), logKm decreased linearly with increasing chain length, corresponding to a contribution to the free energy of association between enzyme and substrate of ‒2.5kJ/mol for each additional CH2 group in the alkyl chain. logKi for the competitive inhibition by secondary alkyl 2-sulphate esters followed a similar pattern (-2.4kJ/mol for each additional CH2 group) except that only n-1 carbon atoms effectively participate in hydrophobic bonding, implying that the C-1 methyl group is not involved. logKi values for inhibition primary alkanesulphonates also depended linearly on chain length but with a diminished gradient, indicating a free-energy increment of ‒1.2kJ/mol per additional CH2 group. The collective results showed the presence of a hydrophobic site on the enzyme capable of accomodating an alkyl chain of considerable length. Cationic structures (in the form of arginine, lysine or histidine), whose presence might be expected for binding the anionic sulphate group, were not detectable at the active site.

Candida cylindracea Lipase-Catalyzed Synthesis of Retinyl and Oleyl Palmitates; Carbon Chain Length Dependence of Esterase Activity

1992 ◽  
Vol 45 (4) ◽  
pp. 641 ◽  
Author(s):  
J Oconnor ◽  
SF Petricevic ◽  
RA Stanley
Keyword(s):  
Chain Length ◽  
Esterase Activity ◽  
Aqueous Ethanol ◽  
Carbon Chain ◽  
Bulk Water ◽  
Maximum Activity ◽  
Ethanol Solution ◽  
Carbon Chain Length ◽  
Candida Cylindracea ◽  
Candida Cylindracea Lipase

The ester activity of lipase from Candida cylindracea has been investigated in paraffin/water biphasic mixtures and in an aqueous ethanol solution. In the biphasic mixture with paraffin as an inert solvent, the equilibrium was found to be at approximately 75% esterification of palmitic acid and oleyl alcohol. Analysis was by a titrimetric assay. In a monophasic solvent of aqueous ethanol, more than 50% of retinol was converted into retinyl palmitate in 1 h. Analysis was by spectrofluorophotometry. Thus the Candida lipase can synthesize esters in the presence of a bulk water phase. The formation of the esters over the hydrolysis products is thought to be due to the partitioning of the hydrophobic reactants between the surface and interior of the organic phase. The esterase activity of the lipase has been determined against a range of 4 nitrophenyl alkanoates (carbon chain length 2-18) at pH 7 and 37�C in Tris buffer. Maximum activity occurred with the butanoate ester. The Michaelis-Menten parameters were determined for this substrate in the presence and absence of butan-1-ol which inhibited the reaction. The range of these data illustrates the potential for using this enzyme for biocatalytic purposes.

Cyanoacrylate Inhibitors of the Hill Reaction IV. Binding Characteristics of the Hydrophobic Domain

1987 ◽  
Vol 42 (6) ◽  
pp. 679-683 ◽  
Author(s):  
John L. Huppatz ◽  
John N. Phillips
Keyword(s):  
Chain Length ◽  
Carbon Chain ◽  
Hill Reaction ◽  
Carbon Chain Length ◽  
Hydrophobic Domain ◽  
Binding Characteristics ◽  
Hydrophobic Binding ◽  
Amino Derivatives ◽  
The Hill ◽  
Derivatives Of

Aryl- and aralkyl-amino derivatives of 2-cyanoacrylic esters were synthesized and assayed as inhibitors of the Hill reaction in isolated pea chloroplast fragments. Aryl- and aralkyl-amino 2-cyanoacrylates unsubstituted in the β-position were weak inhibitors but activity could be greatly enhanced by inclusion of a β-alkyl group with the increase in potency being dependent on the carbon chain length of the β-substituent. The magnitude of this effect appeared independent of the nature of the aryl- or aralkyl-amino function. Inclusion of a carbon chain between the phenyl and amino functions of phenylamino-2-cyano- acrylates produced a stepwise increase in activity with increasing chain length, indicating a prefer- red orientation for the phenyl ring within the hydrophobic binding domain.

Ionic liquid-biosurfactant blends as effective dispersants for oil spills: Effect of carbon chain length and degree of saturation

2021 ◽  
pp. 117119
Author(s):  
Mansoor Ul Hassan Shah ◽  
Ambavaram Vijaya Bhaskar Reddy ◽  
Suzana Yusup ◽  
Masahiro Goto ◽  
Muhammad Moniruzzaman
Keyword(s):  
Ionic Liquid ◽  
Chain Length ◽  
Oil Spills ◽  
Carbon Chain ◽  
Degree Of Saturation ◽  
Carbon Chain Length

Controlling the Secondary Assembly of Porous Anionic Uranyl-organic Polyhedra through Organic Cationic Templates

2021 ◽  
Author(s):  
Liwen Zeng ◽  
Kong-qiu Hu ◽  
Zhi-wei Huang ◽  
Lei Mei ◽  
Xianghe Kong ◽  
...  
Keyword(s):  
Chain Length ◽  
Carbon Chain ◽  
Carbon Chain Length ◽  
Organic Templates ◽  
Secondary Assembly

Herein, we report a new uranyl-organic polyhedron U4L4 (L=BTPCA) assembled from uranyl and a semirigid tritopic ligand. By adjusting the carbon chain length of the organic templates, two complexes can...

Effect of carbon chain length on biological activity of novel palladium (II) complexes

2014 ◽  
Vol 82 ◽  
pp. 172-180 ◽  
Author(s):  
En-Jun Gao ◽  
Hong Fu ◽  
Ming-Chang Zhu ◽  
Chi Ma ◽  
Shi-Kai Liang ◽  
...  
Keyword(s):  
Biological Activity ◽  
Chain Length ◽  
Carbon Chain ◽  
Carbon Chain Length

scholarly journals Studies on Surfactants, Cosurfactants, and Oils for Prospective Use in Formulation of Ketorolac Tromethamine Ophthalmic Nanoemulsions

Pharmaceutics ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 467
Author(s):  
Shahla S. Smail ◽  
Mowafaq M. Ghareeb ◽  
Huner K. Omer ◽  
Ali A. Al-Kinani ◽  
Raid G. Alany
Keyword(s):  
Chain Length ◽  
Molecular Descriptors ◽  
Chorioallantoic Membrane ◽  
Carbon Chain ◽  
Log P ◽  
Carbon Chain Length ◽  
Ketorolac Tromethamine ◽  
Hen’S Egg ◽  
Oil Water ◽  
Tween 60

Nanoemulsions (NE) are isotropic, dispersions of oil, water, surfactant(s) and cosurfactant(s). A range of components (11 surfactants, nine cosurfactants, and five oils) were investigated as potential excipients for preparation of ketorolac tromethamine (KT) ocular nanoemulsion. Diol cosurfactants were investigated for the effect of their carbon chain length and dielectric constant (DEC), Log P, and HLB on saturation solubility of KT. Hen’s Egg Test—ChorioAllantoic Membrane (HET-CAM) assay was used to evaluate conjunctival irritation of selected excipients. Of the investigated surfactants, Tween 60 achieved the highest KT solubility (9.89 ± 0.17 mg/mL), followed by Cremophor RH 40 (9.00 ± 0.21 mg/mL); amongst cosurfactants of interest ethylene glycol yielded the highest KT solubility (36.84 ± 0.40 mg/mL), followed by propylene glycol (26.23 ± 0.82 mg/mL). The solubility of KT in cosurfactants was affected by four molecular descriptors: carbon chain length, DEC, log P and HLB. KT solubility was directly proportional to DEC and the HLB yet, inversely proportional to carbon chain length and log P. All surfactants, except Labrasol ALF, were non-irritant. The majority of cosurfactants were slightly irritant, butylene glycol was a moderate irritant, pentylene and hexylene glycols were strong irritants. These findings will inform experiments aimed at developing NE formulations for ocular administration of KT.

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