Equol,an Isoflavone Metabolite,Regulates Cancer Cell Viability and Protein Synthesis Initiation via c-Myc and eIF4G

Epidemiological studies implicate dietary soy isoflavones as breast cancer preventives, especially due to their anti-estrogenic properties. However, soy isoflavones may also have a role in promoting breast cancer, which has yet to be clarified. We previously reported that equol, a metabolite of the soy isoflavone daidzein, may advance breast cancer potential via up-regulation of the eukaryotic initiation factor 4GI (eIF4GI). In estrogen receptor negative (ER−) metastatic breast cancer cells, equol induced elevated levels of eIF4G, which were associated with increased cell viability and the selective translation of mRNAs that use non-canonical means of initiation, including internal ribosome entry site (IRES), ribosome shunting, and eIF4G enhancers. These mRNAs typically code for oncogenic, survival, and cell stress molecules. Among those mRNAs translationally increased by equol was the oncogene and eIF4G enhancer, c-Myc. Here we report that siRNA-mediated knockdown of c-Myc abrogates the increase in cancer cell viability and mammosphere formation by equol, and results in a significant down-regulation of eIF4GI (the major eIF4G isoform), as well as reduces levels of some, but not all, proteins encoded by mRNAs that are translationally stimulated by equol treatment. Knockdown of eIF4GI also markedly reduces an equol-mediated increase in IRES-dependent mRNA translation and the expression of specific oncogenic proteins. However, eIF4GI knockdown did not reciprocally affect c-Myc levels or cell viability. This study therefore implicates c-Myc as a potential regulator of the cancer-promoting effects of equol via up-regulation of eIF4GI and selective initiation of translation on mRNAs that utilize non-canonical initiation, including certain oncogenes.     

Proteins of the penicillin biosynthesis pathway

Two sequential steps are common to the biosynthesis of all penicillin-derived antibiotics: the reaction of three l-amino acids to give lδ-(α-aminoadipoyl)-l-cysteinyl-d-valine, and the oxidation of this tripeptide to give isopenicillin N. Recent studies on the peptide synthetase and oxidase enzymes responsible for these steps have implications for the mechanisms and structures of related enzymes involved in a range of metabolic processes.     

酶解大豆蛋白替代鱼粉对星斑川鲽幼鱼生长、血液生化和体组成的影响

正近年来,水产品消费需求量的增长促进了水产养殖业的快速发展,相应的也推动了水产饲料工业的发展,使我国水产饲料产量由2001年的627万吨增长到2011年的1855万吨。然而,鱼粉作为海水鱼类配合饲料的主要组成(占干物质的7%—70%)[1],水产饲料工业的壮大也加大了对其需求     

Solubility of soy lipophilic proteins: comparison with other soy protein fractions

Solubility of soy lipophilic proteins (LP) was studied as compared with that of other soy protein fractions. LP, β-conglycinin, glycinin, and soy protein isolate (N-SPI) were prepared under the condition to avoid heat denaturation. Solubility of LP was lower than that of other soy protein fractions under all the tested conditions varying in pH values and ionic strength. The solubility of LP was increased constantly by elevating temperature until 90 °C, whereas that of β-conglycinin and glycinin dropped at high temperature. Temperature-dependent change in solubility of N-SPI might reflect the balance among that of glycinin, β-conglycinin and LP. Based on the results of SDS-PAGE, determination of phospholipid content and Fourier Transform Infrared spectroscopy, we discussed the solubilization behavior of LP relating to its origin and composition.     

Contribution of 2-methyl-3-furanthiol to the cooked meat-like aroma of fermented soy sauce

The cooked meat-like aroma compound, 2-methyl-3-furanthiol (2M3F), was detected in fermented soy sauce (FSS) by GC-olfactometry and GC-MS. 2M3F was present in FSS at a concentration considerably greater than the perception threshold, and the 2M3F concentration increased with heating temperature. Sensory analysis indicated that with the addition of only 0.2 μg/L of 2M3F to the soy sauce sample, the cooked meat-like aroma is significantly stronger than that of sample without the addition of 2M3F. Hence, 2M3F contributes to the cooked meat-like aroma of FSS, which constitutes the key aroma component of FSS. In addition, 2M3F was generated from the addition of ribose and cysteine in FSS by heating at 120 °C, but it was not detected in a phosphate buffer under the same condition. Furthermore, 2M3F was not detected in acid-hydrolyzed vegetable-protein-mixed soy sauce (ASS) and heated ASS. These results indicated that fermentation by micro-organisms facilitates the generation of 2M3F in FSS.     

基于包裹染料木黄酮的甘露糖靶向长循环纳米脂质体抗肿瘤效应研究

采用超声薄膜水化法制备包裹大豆异黄酮主要功能成分染料木黄酮(Ge)的甘露糖修饰聚乙二醇化纳米脂质体(man-PEG@LP/Ge)。伴刀豆球蛋白A(Con A)凝集实验证实甘露糖能够成功修饰在纳米脂质体表面。man-PEG@LP/Ge呈规则的球形,大小分布均一,颗粒分散性好。其粒径为(270±15)nm,Zeta电位为+7.5m V。由于聚乙二醇(PEG)长链的空间位阻效应和颗粒所带正电荷的排斥作用实现了较强的结构刚性和较好的稳定性。流式细胞术的结果表明:man-PEG@LP/Ge能够特异性的识别甘露糖受体高表达的人肝癌Huh-7细胞。细胞毒理实验证明:药物载体man-PEG@LP本身没有毒副作用,生物相容性较好。细胞水平和动物水平的肿瘤杀伤实验结果表明:运载药物之后的man-PEG@LP/Ge在甘露糖介导的"主动靶向"作用下高效而特异性的进入肿瘤细胞,对肿瘤组织进行了有效地杀伤。     

Cryopreservation of three Saprolegnia species (Oomycota): Preliminary evidence for the long-term archiving of water mould species

Saprolegnia spp. water moulds are opportunistic pathogens that can cause economic losses to aquaculture. The diseases caused by them are difficult to control since use of the effective drug, malachite green oxalate, is no longer permitted in several regions (including the European Union and USA). To develop an effective control strategy, Saprolegnia isolates must be maintained in the laboratory. Cryopreservation is a useful solution for long-term maintenance; however, at present, there is no developed protocol for the cryopreservation of Saprolegnia spp. Here, we isolated and identified three Saprolegnia species, S. parasitica, S. australis and S. ferax, and developed a deep-freezing protocol that enables the long-term archiving of these species. The survival and growth rates of isolates kept at −80 °C for 3, 6, 9 and 12 months, were tested and compared among the species examined. Although the growth rates of frozen isolates were significantly lower than those of the control (i.e. non-frozen) isolates, the overall survival rate (>90%) indicated the effectiveness of the technique developed. Thus, the protocol developed appears to be a promising method for the long-term preservation of Saprolegnia isolates and may facilitate the creation of stock collections.     

Soybean β-conglycinin improves carbohydrate and lipid metabolism in Wistar rats

The effects of dietary soybean β-conglycinin on lipid metabolism and energy consumption were studied in Wistar adult rats. Rats were fed, a diet containing casein (control group) or β-conglycinin (β-conglycinin group), for 4 weeks. Carbohydrate consumption was higher and fat consumption was lower in the β-conglycinin group than in the control group, whereas the total energy consumption was the same between the two groups. Serum adiponectin was higher in the β-conglycinin group than in the control group. Serum triacylglycerol levels in the β-conglycinin group were significantly lower than those in the control group. The secretion rate of triacylglycerols from the liver after the administration of tyloxapol, an inhibitor of lipolysis, was significantly lower in the β-conglycinin group than in the control group. These results suggest the possibility that β-conglycinin exerts hypolipidemic effects through an acceleration in carbohydrate consumption associated with an increase in adiponectin in rats.     

Development toward rapid and efficient screening for high performance hydrolysate lots in a recombinant monoclonal antibody manufacturing process

Plant‐derived hydrolysates are widely used in mammalian cell culture media to increase yields of recombinant proteins and monoclonal antibodies (mAbs). However, these chemically varied and undefined raw materials can have negative impact on yield and/or product quality in large‐scale cell culture processes. Traditional methods that rely on fractionation of hydrolysates yielded little success in improving hydrolysate quality. We took a holistic approach to develop an efficient and reliable method to screen intact soy hydrolysate lots for commercial recombinant mAb manufacturing. Combined high‐resolution ¹H nuclear magnetic resonance (NMR) spectroscopy and partial least squares (PLS) analysis led to a prediction model between product titer and NMR fingerprinting of soy hydrolysate with cross‐validated correlation coefficient R² of 0.87 and root‐mean‐squared‐error of cross‐validation RMSECV% of 11.2%. This approach screens for high performance hydrolysate lots, therefore ensuring process consistency and product quality in the mAb manufacturing process. Furthermore, PLS analysis was successful in discerning multiple markers (DL‐lactate, soy saccharides, citrate and succinate) among hydrolysate components that positively and negatively correlate with titer. Interestingly, these markers correlate to the metabolic characteristics of some strains of taxonomically diverse lactic acid bacteria (LAB). Thus our findings indicate that LAB strains may exist during hydrolysate manufacturing steps and their biochemical activities may attribute to the titer enhancement effect of soy hydrolysates. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 1061–1068, 2012     

Soy protein recovery in a solvent-free process using continuous liquid-solid circulating fluidized bed ion exchanger

Soy protein concentrates and soy protein isolates act as ingredients in bakery, meat and dairy products, baby formulas, starting materials for spun textured vegetable products, and other nutritional supplements. In this study, the effectiveness of a liquid-solid circulating fluidized bed (LSCFB) ion exchanger is demonstrated for the recovery of soluble soy proteins from full fat and defatted soy flour. Under steady-state operating conditions, about 50% of the proteins could be recovered from the feed streams entering the ion exchanger. The LSCFB was shown to be a promising system for the recovery of soy protein from both defatted and full fat soy flour solutions. As the ion exchange process captures dissolved proteins, the system may offer a less damaging form of processing compared with the acid precipitation process where soy protein aggregates form and functionality is affected. In addition, the LSCFB allows simultaneous adsorption and desorption of the proteins allowing for a continuous operation. No prefiltration of feed containing suspended particles is required as well, because fluidization is used in place of packed bed technology to improve on current ion exchange processes.