Concerning the role of 24,25-dihydrolanosterol and lanostanol in sterol biosynthesis by cultured cells

Wd, Nes; Ra, Norton; Ej, Parish; Meenan, A; Popják, G.

doi:10.1016/0039-128x(89)90025-1

Cited by 5 publications

(2 citation statements)

References 15 publications

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“…F1, F2, and F3 were further confirmed for their structures by GC‐MS (Supporting Information Figure S2) and 1 H‐NMR (Supporting Information Figure S3). The identity of F1, F2, and F3 were assigned as dihydrolanosterol, lanosterol, and cholesterol based on agreement of MS data (Supporting Information Figure S2) with literature . NMR data were in agreement with that of target compounds lanosterol, dihydrolanosterol, and cholesterol reported in the literatures .…”

Section: Resultssupporting

confidence: 77%

Separation and purification of lanosterol, dihydrolanosterol, and cholesterol from lanolin by high‐performance counter‐current chromatography dual‐mode elution method

Pei

Pan

et al. 2019

J of Separation Science

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Lanosterol is a potential drug for cataracts treatment, which can reverse the aggregation of intracrystalline proteins. The low concentration in lanolin calls for high‐performance separation methods. In this study, a counter‐current chromatography dual‐mode elution method was developed for the first time to separate and purify lanosterol from hexane extract of lanolin after saponification, in which the column was first eluted with the lower phase as mobile phase in head‐to‐tail mode, followed by the upper phase in the tail‐to‐head mode. High purity of lanosterol, dihydrolanosterol, and cholesterol can be obtained simultaneously. A solvent system composed of n‐heptane/acetonitrile/ethyl acetate (5:5:1, v/v/v) was selected and optimized via partition coefficient determination. Compounds such as 111 mg lanosterol, 84 mg dihydrolanosterol, and 183 mg cholesterol with high purity of 99.77, 95.71, and 91.43%, respectively, analyzed by high‐performance liquid chromatography were obtained within 80 min from 700 mg crude extract from 1.78 g lanolin. The method was also used to improve the purity of commercial lanosterol product from 66.97 to above 99%. Counter‐current chromatography could serve as a potential and powerful technique for commercial production of highly pure lanosterol.

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Section: Resultssupporting

confidence: 77%

Separation and purification of lanosterol, dihydrolanosterol, and cholesterol from lanolin by high‐performance counter‐current chromatography dual‐mode elution method

Pei

Pan

et al. 2019

J of Separation Science

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“…While doubtful the new metabolite converts to Δ 5,7 -sterol under physiological conditions, it does shows mechanistically that the Δ 8 -bond is not essential for CYP51 activity. In contrast, a Δ 8 -bond in the CYP51 substrate is required in plant and animal sterol biosynthesis [41–43]. Except for cycloeucalenol as substrate, there was general agreement between affinity and catalytic competence determined by the sterol difference spectra and binding constant ( K d ) and efficiency for substrate to be converted to 14-desmethyl product ( k cat ).…”

Section: Resultsmentioning

confidence: 98%

Functional importance for developmental regulation of sterol biosynthesis in Acanthamoeba castellanii

Zhou

Warrilow

Thomas

et al. 2018

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

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The sterol metabolome of Acanthamoeba castellanii (Ac) yielded 25 sterols. Substrate screening of cloned AcCYP51 revealed obtusifoliol as the natural substrate which converts to ∆-sterol (<95%). The combination of [H-methyl]methionine incubation to intact cultures showing C-ergosterol incorporates 2-H atoms and C-7-dehydroporiferasterol incorporates 5 H-atoms, the natural distribution of sterols, CYP51 and previously published sterol methyltransferase (SMT) data indicate separate ∆- and ∆-olefin pathways to C- and C-sterol products from the protosterol cycloartenol. In cell-based culture, we observed a marked change in sterol compositions during the growth and encystment phases monitored microscopically and by trypan blue staining; trophozoites possess C/C-∆-sterols, viable encysted cells (mature cyst) possess mostly C-∆-sterol and non-viable encysted cells possess C/C-∆-sterols that turnover variably from stress to 6-methyl aromatic sterols associated with changed membrane fluidity affording lysis. An incompatible fit of steroidal aromatics in membranes was confirmed using the yeast sterol auxotroph GL7. Only viable cysts, including those treated with inhibitor, can excyst into trophozoites. 25-Azacycloartanol or voriconazole that target SMT and CYP51, respectively, are potent enzyme inhibitors in the nanomolar range against the cloned enzymes and amoeba cells. At minimum amoebicidal concentration of inhibitor amoeboid cells rapidly convert to encysted cells unable to excyst. The correlation between stage-specific sterol compositions and the physiological effects of ergosterol biosynthesis inhibitors suggests that amoeba fitness is controlled mainly by developmentally-regulated changes in the phytosterol B-ring; paired interference in the ∆-sterol biosynthesis (to ∆) - metabolism (to ∆ or 6-methyl aromatic) congruence during cell proliferation and encystment could be a source of therapeutic intervention for Acanthamoeba infections.

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Control of Sterol Biosynthesis and its Importance to Developmental Regulation and Evolution

Nes¹

1990

Biochemistry of the Mevalonic Acid Pathway to Terpenoids

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Concerning the role of 24,25-dihydrolanosterol and lanostanol in sterol biosynthesis by cultured cells

Cited by 5 publications

References 15 publications

Separation and purification of lanosterol, dihydrolanosterol, and cholesterol from lanolin by high‐performance counter‐current chromatography dual‐mode elution method

Separation and purification of lanosterol, dihydrolanosterol, and cholesterol from lanolin by high‐performance counter‐current chromatography dual‐mode elution method

Functional importance for developmental regulation of sterol biosynthesis in Acanthamoeba castellanii

Control of Sterol Biosynthesis and its Importance to Developmental Regulation and Evolution

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