Protection against Trp-P-2 DNA adduct formation in C57bl6 mice by purpurin is accompanied by induction of cytochrome P450

Purpurin, an anthraquinone constituent from madder root, has previously been reported as antimutagenic in the Ames Salmonella bacterial mutagenicity assay and as antigenotoxic in Drosophila melanogaster, against a range of environmental carcinogens. Short-term dietary supplementation with purpurin inhibits the formation of hepatic DNA adducts in male C57bl6 mice after a single dose of the heterocyclic amine dietary carcinogen Trp-P-2 (30 mg/kg). Inhibition of adduct formation was dose-dependent. No DNA adducts were observed in animals treated only with purpurin. The decrease in adduct formation was accompanied by significant, dose-dependent inductions of hepatic cytochrome P450-dependent dealkylations of methoxy- (CYP1A2), ethoxy- (CYP1A1), and pentoxy- (CYP2B) resorufins, total cytochrome P450, and NADPH cytochrome P450 reductase. It is hypothesized that purpurin exhibits chemopreventive potential by inhibiting the cytochrome P450-dependent metabolism of heterocyclic amines to their genotoxic N-hydroxylamines.     

Modulation of rat hepatic and pulmonary cytochromes P450 and phase II enzyme systems by erucin, an isothiocyanate structurally related to sulforaphane

Administration of dietary doses of the isothiocyanate erucin had no effect on rat hepatic cytochrome P450 activity or protein levels, but at higher doses a rise in CYP1A/B1 protein levels was evident. In lung, treatment with erucin, as well as sulforaphane, failed to modulate cytochrome P450 activities but elevated CYP1A/B1 protein levels. In liver, erucin stimulated quinone reductase activity accompanied by a rise in protein. Glutathione S-transferase activity was unaffected, but GSTalpha and GSTmu protein levels increased. In lung, both isothiocyanates increased quinone reductase paralleled by a rise in protein levels; at the higher dose both isothiocyanates elevated moderately GSTalpha levels. Hepatic microsomes converted both isothiocyanates to metabolites that impaired cytochrome P450 activity, which was antagonized by reduced glutathione. It may be concluded that erucin may protect against carcinogens by stimulating the detoxication of quinones but is unlikely to significantly influence reactive intermediate generation through modulation of cytochrome P450 activity.     

与甜瓜性别分化相关植物激素生物合成途径分析

甜瓜性别分化是由多因素构成的一个复杂的网络系统调控,受到多种环境因素及激素水平的影响。本研究利用甜瓜(Cucumis melo L.)WI 998×TopMark 构建重组自交系群体,通过第二代高通量测序技术对F2S6群体中雌雄异花同株及全雌系植株的转录组测序,比较差异表达基因,分析与性别差异表达相关的植物激素合成途径。结果显示,雌雄异花同株获得77376 条 Unigenes,平均长度为 435 bp;全雌系转录组测序获得 80 825 条 unigenes,平均长度为 509 bp。比较雌雄异花同株与全雌系转录组,发现共有 8966个基因差异表达,4 296 个基因下调,4 670 个基因上调。对差异基因进行基因本体论(GO)功能分类,发现2 352 条 Unigenes 归入到生物学过程,4 107 条 Unigenes 归入到细胞学过程,2 507 条 Unigenes 归入到分子功能。差异表达基因共参与 121 个 Pathway,发现赤霉素(gibberellin, GA)合成途径中 GA3 - 氧合成酶(GA3-ox)(MU3674)、GA7- 氧化酶(GA7-ox)(MU36987)、GA2- 氧化酶(GA2-ox)(MU13098/MU13099)、GA20-氧化酶(GA20-ox)和 GA2- 氧化酶(GA2-ox)(MU33020)等基因差异表达;共有 38 个基因在脱落酸(abscisicacid, ABA)合成途径中差异表达,其中 19 个上调,19 个下调;油菜素内酯(brassinolide, BR)合成途径中,MU22012 (cytochrome P450),MU26893 (cytochrome P450) 基因上调 ,MU56098 (cytochrome P450,CYP724B3),MU76596(CYP724A1)下调。本研究还发现参与甜瓜性别表达,分别与赤霉素、脱落酸、油菜素内酯及玉米素等多种激素合成、信号传导等代谢途径相关。研究结果为分析甜瓜决定性别分化的可能机理,为下一步研究甜瓜性别修饰基因的克隆,提供重要依据。     

Phytomonitoring and phytoremediation of agrochemicals and related compounds based on recombinant cytochrome P450s and aryl hydrocarbon receptors (AhRs)

Molecular mechanisms of metabolism and modes of actions of agrochemicals and related compounds are important for understanding selective toxicity, biodegradability, and monitoring of biological effects on nontarget organisms. It is well-known that in mammals, cytochrome P450 (P450 or CYP) monooxygenases metabolize lipophilic foreign compounds. These P450 species are inducible, and both CYP1A1 and CYP1A2 are induced by aryl hydrocarbon receptor (AhR) combined with a ligand. Gene engineering of P450 and NADPH cytochrome P450 oxidoreductase (P450 reductase) was established for bioconversion. Also, gene modification of AhRs was developed for recombinant AhR-mediated β-glucronidase (GUS) reporter assay of AhR ligands. Recombinant P450 genes were transformed into plants for phytoremediation, and recombinant AhR-mediated GUS reporter gene expression systems were each transformed into plants for phytomonitoring. Transgenic rice plants carrying CYP2B6 metabolized the herbicide metolachlor and remarkably reduced the residues in the plants and soils under paddy field conditions. Transgenic Arabidopsis plants carrying recombinant guinea pig (g) AhR-mediated GUS reporter genes detected PCB126 at the level of 10 ng/g soils in the presence of biosurfactants MEL-B. Both phytomonitoring and phytoremediation plants were each evaluated from the standpoint of practical uses.     

Black tea polyphenols, theaflavins, prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P450 1A1 in cell cultures.

Tea polyphenols have been demonstrated as chemopreventive agents in a number of experimental models. However, less is known about the mechanism of chemoprevention by black tea compared with that of green tea. Some beneficial properties of theaflavins, the black tea polyphenols, were investigated in the present study. Theaflavins showed inhibitory effects on H(2)O(2)- and tert-butyl hydroperoxide (tBuOOH)-induced cytotoxicity (evaluated by tetrazolium bromide reduction), cellular oxidative stress (detected by oxidation of 2', 7'-dichlorofluorescin), and DNA damage (measured by amount of 8-OHdG and comet assay) in rat normal liver epithelium cell RL-34 cell lines. In addition, theaflavins also exhibited suppression of cytochrome P450 1A1 induced by omeprazole in the human hepatoma HepG2 cell line. Furthermore, when HepG2 cells were pretreated with omeprazole to induce CYP1A1, then exposed to benzo[a]pyrene (B[a]P), DNA damage was observed using the comet assay. However, theaflavins could inhibit this DNA damage. These results indicated that theaflavins could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P450 1A1 in cell cultures.     

Identification and quantification of astaxanthin esters in shrimp (Pandalus borealis) and in a microalga (Haematococcus pluvialis) by liquid chromatography-mass spectrometry using negative ion atmospheric pressure chemical ionization

Negative ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [negative ion LC-(APCI)MS] was used for the identification of astaxanthin esters in extracts of commercial shrimp (Pandalus borealis) and dried microalga (Haematococcus pluvialis) samples. A cleanup step using a normal phase solid phase extraction (SPE) cartridge was applied prior to analysis. Recovery experiments with astaxanthin oleate as model compound proved the applicability of this step (98.5 +/- 7.6%; n = 4). The assignment of astaxanthin esters in negative ion LC-(APCI)MS was based on the detection of the molecular ion (M*-) and the formation of characteristic fragment ions, resulting from the loss of one or two fatty acids. Quantification of individual astaxanthin esters was performed using an astaxanthin calibration curve, which was found to be linear over the required range (1-51 micromol/L; r2 = 0.9996). Detection limits, based on the intensity of M*-, a signal-to-noise ratio of 3:1, and an injection volume of 20 microL, were estimated to be 0.05 microg/mL (free astaxanthin), 0.28 microg/mL (astaxanthin-C16:0), and 0.78 microg/mL (astaxanthin-C16:0/C16:0), respectively. This LC-(APCI)MS method allows for the first time the characterization of native astaxanthin esters in P. borealis and H. pluvialis without using time-consuming isolation steps with subsequent gas chromatographic analyses of fatty acid methyl esters. The results suggest that the pattern of astaxanthin-bound polyunsaturated fatty acids of P. borealis does not reflect the respective fatty acid pattern found in triacylglycerides. Application of the presented LC-(APCI)MS technique in common astaxanthin ester analysis will forestall erroneous xanthophyll ester assignment in natural sources.     

Cleavage products of lycopene produced by in vitro oxidations: characterization and mechanisms of formation

The aim of this study was to produce in vitro oxidation products of lycopene, which could be possible in vivo metabolites. An oxidation of lycopene with potassium permanganate gave a range of lycopene degradation compounds resulting from the oxidative cleavage of one or two carbon-carbon double bonds. Eleven apo-lycopenals/ones and six apo-carotendials were obtained and tentatively characterized by HPLC-DAD-MS. Apo-11-lycopenal and apo-8,6'-carotendial were isolated and characterized by (1)H NMR for the first time. Lycopene was submitted to an oxidation by atmospheric oxygen catalyzed by a metalloporphyrin, a model system of the active center of cytochrome P450 enzymes. (Z)-Isomers, monoxides, and cleavage compounds of (E)-lycopene were formed. We propose a mechanism of oxidation of lycopene by this system.     

Metabolism of biochanin A and formononetin by human liver microsomes in vitro

Biochanin A and formononetin are abundant in legumes. These proestrogenic isoflavones can be converted by 4'-O-demethylation to the more potent phytoestrogens genistein and daidzein. Incubation of biochanin A or formononetin with human liver microsomes resulted in 4'-O-demethylation and the production of additional metabolites. Three new hydroxylated formononetin derivatives, 6,7-dihydroxy-4'-methoxyisoflavone, 7,8-dihydroxy-4'-methoxyisoflavone, and 7,3'-dihydroxy-4'-methoxyisoflavone, were isolated and characterized. We surveyed the O-demethylase competence of cytochrome P450 isoforms found in human liver. Human cytochrome P450 isoforms 1A2, 2E1, 2C9*1, 2C19, and 2D6*1 catalyzed biochanin A consumption and genistein production. Human cytochrome P450 isoforms 1A2, 2C9*1, 2A6, 2D6*1, and 2C19 catalyzed formononetin consumption and daidzein production. These isoforms also generated other hydroxylated metabolites. Although O-demethylation of isoflavones has been attributed to metabolism by gut microflora, our study demonstrates that human hepatic microsomal enzymes can perform the same transformation and may play a key role in the conversion of 4'-O-methylated isoflavones to more potent phytoestrogens.     

Effect of organic-phase solvents on physicochemical properties and cellular uptake of astaxanthin nanodispersions

A simplex centroid mixture design was used to study the interactions between two chosen solvents, dichloromethane (DCM) and acetone (ACT), as organic-phase components in the formation and physicochemical characterization and cellular uptake of astaxanthin nanodispersions produced using precipitation and condensation processes. Full cubic or quadratic regression models with acceptable determination coefficients were obtained for all of the studied responses. Multiple-response optimization predicted that the organic phase with 38% (w/w) DCM and 62% (w/w) ACT yielded astaxanthin nanodispersions with the minimum particle size (106 nm), polydispersity index (0.191), and total astaxanthin loss (12.7%, w/w) and the maximum cellular uptake (2981 fmol/cell). Astaxanthin cellular uptake from the produced nanodispersions also showed a good correlation with their particle size distributions and astaxanthin trans/cis isomerization ratios. The absence of significant (p > 0.05) differences between the experimental and predicted values of the response variables confirmed the adequacy of the fitted models.     

Phytoremediation of the herbicides atrazine and metolachlor by transgenic rice plants expressing human CYP1A1, CYP2B6, and CYP2C19

This study evaluated the expression of human cytochrome P450 genes CYP1A1, CYP2B6, and CYP2C19 in rice plants (Oryza sativa cv. Nipponbare) introduced using the plasmid pIKBACH. The transgenic rice plants (pIKBACH rice plants) became more tolerant toward various herbicides than nontransgenic Nipponbare rice plants. Rice plants expressing pIKBACH grown in soil showed tolerance to the herbicides atrazine, metolachlor, and norflurazon and to a mixture of the three herbicides. The degradation of atrazine and metolachlor by pIKBACH rice plants was evaluated to confirm the metabolic activity of the introduced P450s. Although both pIKBACH and nontransgenic Nipponbare rice plants could decrease the amounts of the herbicides in plant tissue and culture medium, pIKBACH rice plants removed greater amounts in greenhouse experiments. The ability of pIKBACH rice plants to remove atrazine and metolachlor from soil was confirmed in large-scale experiments. The metabolism of herbicides by pIKBACH rice plants was enhanced by the introduced P450 species. Assuming that public and commercial acceptance is forthcoming, pIKBACH rice plants may become useful tools for the breeding of herbicide-tolerant crops and for phytoremediation of environmental pollution by organic chemicals.     

Differential effects of garlic oil and its three major organosulfur components on the hepatic detoxification system in rats.

The objective of this study was to compare the modulatory effect of garlic oil and its three organosulfur compounds, diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS), on rat hepatic detoxification enzyme activity, and protein and mRNA expression. Rats were orally administered garlic oil (80 or 200 mg/kg bw), DAS (20 or 80 mg/kg bw), DADS (80 mg/kg bw), or DATS (70 mg/kg bw) three times a week for 6 weeks. Control rats received corn oil. According to the results, garlic oil and DAS in dosages of 200 and 80 mg/kg bw, respectively, significantly increased pentoxyresorufin O-dealkylase (PROD) activity as compared with the that of the control rats (P < 0.05). In contrast, N-nitrosodimethylamine demethylase activity in rats that received DADS and DATS was significantly lower than that in the control rats (P < 0.05). Ethoxyresorufin O-deethylase and erythromycin demethylase activities were not influenced by garlic oil, DAS, DADS, or DATS. To the phase II enzyme, garlic oil, DADS, and DATS significantly increased the glutathione S-transferase (GST) activity toward ethacrynic aicd (P < 0.05). Immunoblot assay showed that the protein contents of cytochrome P450 1A1, 2B1, and 3A1 were increased by garlic oil and each of three allyl sulfides, and the change among the allyl sulfides was in the order of DAS > DADS > DATS. The placental form of GST (PGST) level was also increased by garlic oil and the three allyl sulfides, but the increase among the allyl sulfides was DATS congruent with DADS > DAS. P450 2E1, however, was suppressed by each garlic component. Northern blot results indicated that the changes in P450 1A1, 2B1, 3A1, and PGST mRNA levels by garlic components were similar to those noted in the protein levels. These results indicate that the modulatory effect of garlic oil on hepatic drug-metabolizing enzymes can be attributed to its three major allyl sulfide components DAS, DADS, and DATS. These three allyl sulfides vary in modulatory activity, and this variation is related to the number of sulfur atoms in the molecule.     

利用农业废弃物碳源的红法夫酵母生产虾青素研究进展

利用红法夫酵母产虾青素工艺具有绿色、安全、低成本等优点,但培养红法夫酵母的培养基(尤其是碳源)成本是制约该工艺实现大规模生产的主要因素之一。传统的利用纯糖如葡萄糖、蔗糖、木糖等作为红法夫酵母生产虾青素的碳源,效果虽好,但因其价格较高,仅适用于实验室研究和小型发酵生产,不适于大规模工业化生产。如何寻找来源广、价廉且发酵效果好的碳源,降低虾青素的生产成本是发展虾青素产业亟待解决的课题。利用廉价且来源广泛的原料如水果类原料、糖质原料、甘油及废弃木质纤维素类原料等作为碳源能有效降低成本。但这些原料都存在一些问题,比如水果类原料存在季节性及地域性限制,糖蜜类原料供应稳定性方面存在问题,甘油类原料杂质多且存在食品安全隐患,木质纤维素类原料来源广泛、价格低廉,不存在占用耕地等问题,是最有希望用于规模化低成本发酵生产虾青素的原料,但其利用存在一些技术瓶颈。该文介绍了目前利用红法夫酵母生产虾青素的基本情况,并系统地对国内外不同碳源条件下利用红法夫酵母生产虾青素的研究状况进行了综述、比较及分析。最后,对利用红法夫酵母生产虾青素在碳源方面的未来发展趋势及重点研究方向进行了展望。     

Metabolism of polychlorinated dibenzo-p-dioxins (PCDDs) by human cytochrome P450-dependent monooxygenase systems

Metabolism of polychlorinated dibenzo-p-dioxins (PCDDs) by monooxygenase systems dependent on 12 forms of human cytochrome P450 (CYP) was examined with the recombinant yeast microsomes containing each of the human CYP. The metabolites of PCDDs were analyzed by HPLC and GC-MS. Remarkable metabolism by the multiple CYP forms was observed toward dibenzo-p-dioxin (DD) and mono-, di-, and trichloroDDs. The metabolism contained multiple reactions such as hydroxylation at an unsubstituted position, hydroxylation with migration of a chloride substituent, and hydroxylation with elimination of a chloride substituent. Although major CYPs in human liver such as CYP2C8, CYP2C9, and CYP3A4 showed no significant metabolism toward the PCDDs, CYP1A1 and CYP1A2 showed high catalytic activity toward DD and mono-, di-, and trichloroDDs. The kinetic parameters K(m)(app) and V(max) of the CYP1A1-dependent 8-hydroxylation activity of 2,3,7-trichloro-DD (2,3,7-triCDD) were estimated to be 0.30 microM and 51 (mol/min/mol of P450), respectively, suggesting that 2,3,7-triCDD was a good substrate for CYP1A1. However, none of the CYPs showed any detectable activity [<0.01 mol/min/mol of P450)] toward 2,3,7,8-tetraCDD. Substrate-induced absorption spectrum and inhibition studies indicated that CYP1A1 could bind 2,3,7,8-tetraCDD with considerably high affinity. It was strongly suggested that the long half-life (7.1 years) of 2,3,7,8-tetraCDD in humans was due to an extremely low activity of CYPs toward 2,3,7,8-tetraCDD in addition to its chemical stability.     

Substrates Influence Stimulatory Effect of Mevalonic Acid on Carotenoid Production in Red Yeasts

Stimulation of carotenogenesis in carotenoid producing red yeasts, algae, or bacteria for enhanced carotenoid production has been achieved by mevalonic acid addition. Recently, carotenoid‐enriched feed was produced by Phaffia rhodozyma fermentation of inexpensive animal feeds. Because mevalonate improves carotenoid yield in P. rhodozyma in synthetic medium, this study tested whether a similar enhancement was possible in biobased substrates. Four concentrations, 0, 0.02, 0.04, and 0.1% of mevalonate as a precursor of P. rhodozyma production of astaxanthin and β‐carotene were evaluated in five substrates: defatted rice bran, full fat rice bran, wheat bran, corn whole stillage, and synthetic media. Additionally, four concentrations, 0, 0.05, 0.1, and 0.5% of apple pomace and tomato pomace were also evaluated as precursors of carotenogenesis in P. rhodozyma fermentation of corn whole stillage and rice bran. Mevalonic acid, tomato pomace, and apple pomace enhanced carotenoid yields in all substrates in that order. However, the optimal concentration of precursor and the percent increase of carotenoid yield in each substrate were variable, potentially indicating substrate influence on carotenoid stimulation. Among animal feed substrates, mevalonic acid in corn whole stillage resulted in the best astaxanthin yield of 220 μg/g and β‐carotene of 904 μg/g. Tomato pomace resulted in 29% astaxanthin and β‐carotene enhancement in corn whole stillage, and apple pomace increased β‐carotene production by 26% in whole stillage. The use of expensive mevalonate is offset by the inexpensive process of producing carotenoid‐enriched DDGS. Optimization of tomato or apple pomace addition may further enhance the carotenoid yields.     

诸葛菜OvCYP86MF基因的克隆及其特性分析

为进一步阐明细胞色素P450基因CYP86MF在诸葛菜发育中的分子机理,本文根据已知同源基因保守序列设计特异引物,利用RT-PCR和RACE技术从诸葛菜中获得一个细胞色素P450基因(OvCYP86MF)全长cDNA序列,该序列全长为1876bp,含有1605bp的完整开放阅读框,可编码534个氨基酸,分子量和等电点分别为61.4kDa和6.90,具有细胞色素P450蛋白的典型特征,即保守结构域FNAGPRLCIG;原核表达显示该基因的融合蛋白在体外可以诱导表达;DANSTAR和Clustal W软件分析表明该基因的全长cDNA序列及其编码氨基酸序列与十字花科物种拟南芥相似性很高,达到80%以上,亲缘关系最近;与CYP86C亚家族成员在氨基酸水平上的相似性均高于50%,因此推断该基因属于CYP86C这个亚家族。Northern杂交分析表明该基因在花蕾中特异表达。     

Functional expression of geraniol 10-hydroxylase reveals its dual function in the biosynthesis of terpenoid and phenylpropanoid

Geraniol 10-hydroxylase (G10H), a cytochrome P450 monooxygenase, has been reported to be involved in the biosynthesis of terpenoid indole alkaloids. The gene for Catharanthus roseus G10H (CrG10H) was cloned and heterologously expressed in baculovirus-infected insect cells. A number of substrates were subjected to assay the enzyme activity of CrG10H. As reported in a previous study, CrG10H hydroxylated the monoterpenoid geraniol at the C-10 position to generate 10-hydroxygeraniol. Interestingly, CrG10H also catalyzed 3'-hydroxylation of naringenin to produce eriodictyol. Coexpression of an Arabidopsis NADPH P450 reductase substantially increased the ability of CrG10H to hydroxylate naringenin. The catalytic activity of CrG10H was approximately 10 times more efficient with geraniol than with naringenin, judged by the k(cat)/K(m) values. Thus, G10H also plays an important role in the biosynthetic pathway of flavonoids, in addition to its previously described role in the metabolism of terpenoids.     

Carotenoid Value Addition of Cereal Products by Monoculture and Mixed‐Culture Fermentation of Phaffia rhodozyma and Sporobolomyces roseus

Carotenoid value addition of corn whole stillage by red yeast fermentation has yielded astaxanthin‐ and β‐carotene‐enriched distillers dried grains with solubles (DDGS) for animal feed. In this study, commonly used animal feeds (rice bran, wheat bran, milo whole stillage, and soybean products) were subjected to carotenoid value addition. Phaffia rhodozyma and Sporobolomyces roseus monoculture and mixed‐culture submerged fermentation of these substrates supplemented with 5% glycerol were analyzed for astaxanthin, β‐carotene, and residual glycerol. Among all the substrates, full‐fat rice bran and full‐fat soy flour resulted in the highest astaxanthin (80 μg/g by P. rhodozyma) and β‐carotene yields (836 μg/g by S. roseus). P. rhodozyma produced the highest astaxanthin yield on each substrate, whereas depending on the substrate, either the mixed culture or S. roseus monoculture produced the highest β‐carotene yield. Soy hull was a poor substrate for carotenoid value addition. Both yeasts used glycerol as a carbon source for carotenoid production. This study shows that substrates influence the carotenoid yield. However, it is impossible to dissect the effect of specific nutrients on carotenoid production in complex biological substrates. Carotenoid value addition of these substrates provides as much as or more than the required daily dosage of carotenoids in animal feed.     

Metalloporphyrins as biomimetic models for cytochrome p-450 in the oxidation of atrazine

The aim of this work was to evaluate whether metalloporphyrin models could mimic the action of cytochrome P-450 in the oxidation of atrazine, a herbicide. The commercially available second-generation metalloporphyrins 5,10,15,20-tetrakis(2,6-dichlorophenyl)porphyrin metal(III) chloride [M(TDCPP)Cl] and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin metal(III) chloride [M(TFPP)Cl] (metal = Fe or Mn) and the oxidants iodosylbenzene and metachloroperbenzoic acid were employed in this study. Results showed that the metalloporphyrins used here can oxidize atrazine. Yields as high as 32% were obtained for the Mn(TFPP)Cl/PhIO system, which shows that these catalysts can mimic both the in vivo and the in vitro action of cytochrome P-450, with formation of the metabolites DEA and DIA. The formation of five other unknown products was also detected, but only one of them could be identified, since the other four were present in very low concentrations. The compound COA, identified by mass spectrometry, was the main product in most of the oxidation reactions.     

Soybean meal fermented by Aspergillus awamori increases the cytochrome P-450 content of the liver microsomes of mice

The effect of soybean meal fermented by Aspergillus awamori on the acute lethality of acetaldehyde, pentobarbital sleeping time, and cytochrome P-450 content of the hepatic microsomes was studied in mice. Most of the daidzin and genistin in soybean meal (SBM) were converted into the respective aglycones, daidzein and genistein, by fermentation. In experiment 1, mice were fed isonitrogenic test diets with one of the following five protein sources for 28 d: casein, SBM, fermented and hot-air-dried SBM (FSBM-HD), fermented and freeze-dried SBM (FSBM-FD), or methanol-extracted FSBM-FD (FSMB-FD-R). The acute lethality of acetaldehyde in mice fed the FSBM-FD diet was significantly lower than that in mice fed the SBM, FSBM-HD, or FSBM-FD-R diet. In experiments 2 and 3, mice were fed isonitrogenic test diets with one of the following four protein sources for 28 d: casein, SBM, FSBM-FD, and FSBM-FD-R. The pentobarbital sleeping time was significantly shorter and the cytochrome P-450 content was significantly higher in the mice fed the FSBM-FD diet than the respective value in mice fed the other test diets. In experiment 4, mice were fed one of eight diets which contained different levels of aglycone obtained by varying the proportion of FSBM-FD and FSBM-FD-R, for 28 d. The cytochrome P-450 content in hepatic microsomes increased as the dietary level of isoflavonoid aglycones increased, but there was a saturation phenomenon. These results suggest that soy isoflavonoid aglycones are more potent inducers of cytochrome P-450 than isoflavonoid glycosides.