Sterols, a class of cyclopentane poly-hydrophenanthrene derivatives, are the predominant membrane constituent of eukaryotes. These substances have a variety of biological activities and have been widely used in food and pharmaceutical industries. The presence of endogenous ergosterol biosynthetic pathway in Saccharomyces cerevisiae cells make it an ideal chassis for the de novo synthesis of sterol and its derivatives. Most recently, the rational modification of organelles provides a novel strategy for the directed transportation and storage of target products and the ultimate enhanced product synthesis. This review summarizes the phenotypic responses of S. cerevisiae cells upon different physiological stimulations and the underlying molecular mechanisms. Reinforcement of sterol production through directed storage, transportation, and excretion of sterols offers a novel strategy for breaking the limitation of de novo biosynthesis of sterols in yeast.
Ergosterol ; Phytosterols ; Saccharomyces cerevisiae ; Sterols

In order to establish a method for the determination of the sterols of the oil in the freeze-dried acai (Euterpe oleracea Mart.) and to evaluate its antioxidant activities, a saponification/extraction procedure and high performance liquid chromatography (HPLC) analysis method were developed and validated for the analysis of phytosterols in PEE (Petroleum ether extract). Separation was achieved on a Purosper STAR LP C18 column with a binary, gradient solvent system of acetonitrile and isopropanol. Evaporative light scattering detection (ELSD) was used to quantify β-sitosterol and the total sterols. Peak identification was verified by retention times and spikes with external standards. Standard curves were constructed (r = 0.999 2) to allow for sample quantification. Recovery of the saponification and extraction was demonstrated via analysis of spiked samples. The highest content of total sterols is β-sitosterol. The antioxidant activities of the extracts were evaluated using the total oxyradical scavenging capacity assay (TOSC assay). The result showed that the PEE exhibited significant antioxidant properties, sample concentration and the antioxidant capacity had a certain relevance.
Antioxidants ; analysis ; Arecaceae ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Phytosterols ; analysis ; Sitosterols ; analysis

OBJECTIVETo investigate the dietary phytosterol intake of elderly women in three different cities of China, and to compare the main dietary sources, so that to discuss the relationship of dietary phytosterol intake and serum lipids.

METHODSBased on the dietary pattern, women more than 50 years old from Beijing, Hefei and Urumchi were chosen as testers, 80 - 100 people for each city respectively. The dietary survey was done by continues 24 hours review of two days, the plant food were collected and the phytosterol content (include beta-sitosterol, campesterol, stigmasterol, sitostanol) were analyzed by GC methods, the total phytosterols content were calculated. The dietary phytosterol intake were calculated and serum lipids were also analyzed in all the testers.

RESULTSTesters from Beijing, Hefei and Urumchi were 100, 101 and 84 respectively. The average dietary phytosterol intake of people in Beijing and Hefei were 340.3 mg/d and 313.5 mg/d, the main sources were plant oil and cereals, while the average dietary phytosterol intake of people in Urumchi were 550.4 mg/d, higher than the other two cities (t values were 9.369, 10.420, respectively, both P values < 0.01), the main source in Urumchi was cereal (provide 53.1% of the total phytosterol intake). The laboratory results showed, testers in Urumchi had significantly lower serum TC content ((4.04 +/- 0.78) mmol/L) than that in Beijing ((4.89 +/- 0.91) mmol/L) and Hefei ((4.71 +/- 0.83) mmol/L) (t value were 6.766 and 5.401 respectively, both P values < 0.01); serum TG content in Urumchi((1.01 +/- 0.48) mmol/L) was also lower than that in Beijing ((1.31 +/- 0.53) mmol/L) and Hefei ((1.66 +/- 0.75) mmol/L) (t values were 3.343 and 7.293 respectively, both P values < 0.01); the serum glucose is also lower in testers in Urumchi ((5.02 +/- 2.18) mmol/L) compared with testers in Beijing ((5.69 +/- 1.53) mmol/L, t = 2.561, P < 0.05) and Hefei ((5.78 +/- 1.53) mmol/L, t = 2.934, P < 0.01).

CONCLUSIONDifferent dietary pattern result in significantly different dietary phytosterol intake in elder women in three cities, higher, phytosterol intake seemed to contribute to lower serum lipids.

Aged ; Aged, 80 and over ; China ; Cholesterol ; analogs & derivatives ; blood ; Cholesterol, Dietary ; metabolism ; Female ; Humans ; Lipids ; blood ; Middle Aged ; Phytosterols ; blood ; metabolism ; Sitosterols ; blood ; Urban Population

OBJECTIVETo investigate the chemical constituents from aerial part of Curcuma wenyujin.

METHODCompounds were isolated by repeated column chromatography on silica gel. Their structures were elucidated on the basis of spectral analysis and comparison with literature data.

RESULTSix compounds were isolated and identified as codonolactone (1), voleneol (2), octacosanoic acid (3), beta-sitosterol (4), mangdesisterol (5), and daucosterol (6).

CONCLUSIONCompounds 1, 2, and 5 were isolated from the plant for the first time.

Curcuma ; chemistry ; Fatty Acids ; chemistry ; isolation & purification ; Phytosterols ; chemistry ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Sesquiterpenes ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification

OBJECTIVETo study the chemical constituents of Ervatamia hainanensis.

METHODThe compounds were separated and purified by column chromatography with silica gel, and identified by IR, MS, NMR and 2D-NMR.

RESULTFive compounds were identified as I (isolariciresinol 9-O-beta-D-glucopyranoside), II (cycloartenol), III (beta-amyrin acetate), IV (beta-sitosterol), V (daucosterol), respectively.

CONCLUSIONAll the compounds were isolated from this plant for the first time.

Apocynaceae ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Phytosterols ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Triterpenes

OBJECTIVETo study the chemical constituents in rhizome of Pinellia ternata.

METHODThe constituents were isolated by silica-gel and Sephadex LH-20 chromatography. The structures were identified by spectroscopic analysis including 2D NMR techniques.

RESULTSix compounds were obtained and identified as stigmast-4-en-3-one(I), cycloartenol(II), 5alpha,8alpha-epidioxyergosta-6,22-dien-3-ol(III), beta-sitosterol-3-O-beta-D-glucoside-6'-eicosanate(IV), alpha-monpalmitin(V), beta-sitosterol(VI). The bioactive assay indicated that: compound III was active against the human tumor cell lines HCT-8, Bel-7402, BGC-823, A549, A2780.

CONCLUSIONCompounds I-IV were isolated from Pinellia ternata for the first time, compound II was the first triterpene isolated from this genus. Compound III may be one of the antitumor constituents of P. ternata.

Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ergosterol ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Humans ; Phytosterols ; chemistry ; isolation & purification ; Pinellia ; chemistry ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Stigmasterol ; analogs & derivatives ; chemistry ; isolation & purification ; Triterpenes

Steroids are a class of medicines with important physiological and pharmacological effects. In pharmaceutical industry, steroidal intermediates are mainly prepared through Mycobacteria transformation, and then modified chemically or enzymatically into advanced steroidal compounds. Compared with the "diosgenin-dienolone" route, Mycobacteria transformation has the advantages of abundant raw materials, cost-effective, short reaction route, high yield and environmental friendliness. Based on genomics and metabolomics, the key enzymes in the phytosterol degradation pathway of Mycobacteria and their catalytic mechanisms are further revealed, which makes it possible for Mycobacteria to be used as chassis cells. This review summarizes the progress in the discovery of steroid-converting enzymes from different species, the modification of Mycobacteria genes and the overexpression of heterologous genes, and the optimization and modification of Mycobacteria as chassis cells.
Mycobacterium/metabolism* ; Steroids/metabolism* ; Phytosterols/metabolism* ; Genomics

OBJECTIVEA new method for simultaneous determination of solasonine (1), solamargine (2) and khasianine (3) in Solanum Nigrum by reversed-phase HPLC was developed.

METHODThe samples were separated at 30 degrees C on Agilent Zorbax SB C18 (4.6 mm x 150 mm, 5 microm) column with acetonitrile-water-phosphoric as mobile phase. Flow rate was 1.0 mL x min(-1) and the detection wavelength was 205 nm.

RESULTThere was good linearity between the peak area and concentration at the ranges of 0.860-10.320 microg (r = 0.999 7), 0.726-8.710 microg (r = 0.999 7), 0.856-10.270 microg (r = 0.999 7) for 1, 2 and 3 respectively. The average recoveries of 1, 2 and 3 were 101.04%, 99.65%, 100.17%.

CONCLUSIONThe method is rapid, simple and accurate, and it can be used for the evaluation of Solanum Nigrum L.

Alkaloids ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Phytosterols ; chemistry ; Solanaceous Alkaloids ; chemistry ; Solanum nigrum ; chemistry

We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

Sterols are a class of cyclopentano-perhydrophenanthrene derivatives widely present in living organisms. Sterols are important components of cell membranes. In addition, they also have important physiological and pharmacological activities. With the development of synthetic biology and metabolic engineering technology, yeast cells are increasingly used for the heterologous synthesis of sterols in recent years. Nevertheless, since sterols are hydrophobic macromolecules, they tend to accumulate in the membrane fraction of yeast cells and consequently trigger cytotoxicity, which hampers the further improvement of sterols yield. Therefore, revealing the mechanism of sterol transport in yeast, especially understanding the working principle of sterol transporters, is vital for designing strategies to relieve the toxicity of sterol accumulation and increasing sterol yield in yeast cell factories. In yeast, sterols are mainly transported through protein-mediated non-vesicular transport mechanisms. This review summarizes five types of sterol transport-related proteins that have been reported in yeast, namely OSBP/ORPs family proteins, LAM family proteins, ABC transport family proteins, CAP superfamily proteins, and NPC-like sterol transport proteins. These transporters play important roles in intracellular sterol gradient distribution and homeostasis maintenance. In addition, we also review the current status of practical applications of sterol transport proteins in yeast cell factories.
Saccharomyces cerevisiae/genetics* ; Sterols ; Phytosterols ; Biological Transport ; ATP-Binding Cassette Transporters/genetics*