Borcalein: a Carborane-Based Analogue of Baicalein with 12-Lipoxygenase-Independent Toxicity

12-Lipoxygenase is crucial for tumour angiogenesis. 5,6,7-Trihydroxy-2-phenyl-4H-1-benzopyran-4-one (baicalein) is a suitable inhibitor for this enzyme but is rapidly metabolised in vivo. Thus, an improvement of the metabolic stability is necessary to enhance the therapeutic efficiency. An emerging approach to enhance metabolic stability of carbon-based pharmaceuticals is the use of metabolically stable, non-toxic boron clusters, such as dicarba-closo-dodecaborane(12)s (carboranes) as phenyl mimetics. Therefore, the unsubstituted phenyl ring of baicalein was replaced by meta-carborane, resulting in borcalein, the carborane analogue of baicalein. This substitution resulted in a decreased inhibitory activity toward 12-lipoxygenase, but led to increased toxicity in melanoma (A375, B16, B16F10) and colon cancer cell lines (SW480, HCT116, CT26CL25) with decreased tumour selectivity in comparison to baicalein. Surprisingly, borcalein displays a different mechanism of cytotoxicity with increased intracellular production of reactive oxygen species (ROS), reactive nitrogen species (RNS) and nitric oxide (NO).     

猪肉脂肪氧化酶在高压结合热处理过程中失活动力学模型

为了解猪肉高压加工过程中脂肪氧化酶(lipoxygenase,LOX)活性的变化规律,以猪背最长肌为原料,经不同压力(200~700 MPa)、温度(20~60℃)结合处理后,研究LOX失活常数的变化规律,并分别用Arrhenius和Eyring方程为基础来建立LOX失活的动力学模型。结果表明,当处理温度不变时,失活常数随压力的增加而增加;同样,当处理压力不变时,失活常数随温度升高而增加。Arrhenius和Eyring方程分别适用于整个研究的温度区间和压力区间,分别以两者为基础可以建立LOX失活的动力学方程,两个方程的拟合程度均较高,决定系数分别达到0.989和0.965。因此在一定的处理温度(20~60℃)和压力(200~700 MPa)范围内,猪肉高压结合热处理过程中LOX活性的变化可以用以Arrhenius和Eyring方程为基础建立的数学模型进行预测。     

Co-Oxidative Transformation of Piperine to Piperonal and 3,4-Methylenedioxycinnamaldehyde by a Lipoxygenase from Pleurotus sapidus

The valuable aroma compound piperonal with its vanilla-like olfactory properties is of high interest for the fragrance and flavor industry. A lipoxygenase (LOX_Psa1) of the basidiomycete Pleurotus sapidus was identified to convert piperine, the abundant pungent principle of black pepper (Piper nigrum), to piperonal and a second volatile product, 3,4-methylenedioxycinnamaldehyde, with a vanilla-like odor through an alkene cleavage. The reaction principle was co-oxidation, as proven by its dependence on the presence of linoleic or α-linolenic acid, common substrates of lipoxygenases. Optimization of the reaction conditions (substrate concentrations, reaction temperature and time) led to a 24-fold and 15-fold increase of the piperonal and 3,4-methylenedioxycinnamaldehyde concentration using the recombinant enzyme. Monokaryotic strains showed different concentrations of and ratios between the two reaction products.     

Bread and durum wheat compared for antioxidants contents,and lipoxygenase and peroxidase activities

Potential beneficial components, including proteins, total phenolics, total flavonoids, carotenoids, tocopherols, and DPPH radical scavenging activity, were investigated in wholemeal of ten bread (T. aestivum L.) and ten durum (T. durum Desf.) novel wheat genotypes. In addition, the activity rate of lipoxygenase (LOX) and peroxidase (POD) enzymes implicated in the antioxidant metabolism was determined. The protein contnet and the antioxidant properties varied according to the two different wheat species, as well as, between the different bread and durum wheat genotypes themselves. The results indicated significant differences in proteins and antioxidant compounds between bread and durum wheat. Higher total proteins, wet gluten and antioxidants contents, combined with lower LOX and POD activities, point to a higher nutritive value of durum wheat than bread wheat.     

黑豆脂氧合酶的制备及其酶学活性的鉴定

从黑豆种子中分离纯化1 种黑豆脂氧合酶--bsLOX,对其酶学活性及降解花色苷等性质进行初步研究。采用缓冲液抽提、硫酸铵沉淀、透析、阳离子交换层析及凝胶层析等方法纯化出具有β-葡萄糖苷酶活性的bsLOX;分别以亚油酸钠和对硝基苯基-β-D-吡喃葡萄糖苷（p-nitrophenyl β-D-glucopyranoside,pNPG）为反应底物鉴定bsLOX的脂氧合酶活性和β-葡萄糖苷酶活性。从黑豆种子中纯化获得的bsLOX在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electropheresis,SDS-PAGE）图谱显示单一条带,分子质量约为95 kD。酶活性分析表明,纯化的bsLOX以亚油酸钠为底物时,酶活力为13 U/mL;当以pNPG为底物在405 nm波长处检测到了对硝基苯酚的生成,从而推测bsLOX具有β-葡萄糖苷酶活性。高效液相色谱实验结果表明bsLOX可以降解花色苷（矢车菊素-3-O-葡萄糖苷、矮牵牛素-3-O-葡萄糖苷）为花色素（矢车菊素、矮牵牛素）。由于降解作用不完全,继而发现花色素能抑制bsLOX的酶活性。bsLOX是黑豆中的一种兼具β-葡萄糖苷酶功能的蛋白质,而花色素可能是天然的bsLOX抑制剂。     

摇青机械力对乌龙茶脂肪族类香气形成的影响

做青是乌龙茶香气品质形成的核心工艺,为明确做青过程中摇青机械力对茶叶脂肪族类香气形成的影响,以乌龙茶品种毛蟹（Camellia sinensis cv. Maoxie）的鲜叶、摇青叶及未摇青等时间摊放叶（简称摊放叶）为供试材料,采用超高效液相色谱-串联质谱法对脂肪族香气前体物质亚油酸、亚麻酸及愈伤酸进行检测,结果表明,摇青机械力能显著降低鲜叶中的亚麻酸及亚油酸含量（P＜0.05）,摊放处理仅能显著降低鲜叶中亚油酸的含量（P＜0.05）,摊放叶中愈伤酸的含量虽然较鲜叶有所降低,但仍旧显著高于摇青叶（P＜0.05）。通过顶空气相色谱-质谱联用法检测脂肪族类香气物质,结果表明,摇青叶的C6醛类相对含量明显降低,C6醇类和叶醇酯类衍生物相对含量增多;而摊放叶中C6醛类、C6醇类和叶醇酯类的衍生物相对含量均远低于摇青叶。实时荧光定量聚合酶链式反应结果表明,LOX1、ADH基因在摇青叶中表达量最高,LOX2、LOX3、HPL基因在摊放叶中表达量最高。相关性分析表明,不同叶态中LOX1基因的表达水平与C6醛类含量间存在极显著的负相关性（P＜0.01）,与C6醇、叶醇酯类衍生物含量间均存在显著正相关性（P＜0.05）,ADH基因表达水平与叶醇酯类衍生物含量间均存在极显著的正相关性（P＜0.01）,而LOX2、HPL基因表达水平与脂肪族类香气含量间相关性均未达显著水平（P＞0.05）。LOX1、ADH基因在做青过程中的上调表达可能与乌龙茶花果香的形成密切相关。     

调味剂对核桃仁脂氧合酶活性和过氧化值的影响

研究了海盐、蔗糖、柠檬酸3种调味剂及其处理条件对核桃仁脂氧合酶活性和过氧化值的影响。结果显示,调味剂处理可降低核桃仁的脂氧合酶活性和过氧化值。在55℃真空浸泡处理3. 0 h条件下,1. 9%海盐、19%蔗糖和2. 4%柠檬酸使核桃仁脂氧合酶相对酶活较原料分别降低了37. 37%、48. 76%和36. 59%,过氧化值较空白对照分别降低了8. 99%、11. 61%和8. 61%,调味后核桃仁均具有良好的感官特性。表明3种调味剂对核桃仁氧化均具有一定的抑制效果。     

超声波、超高压对白鲢鱼肌肉脂肪氧合酶构象及酶活力的影响

本研究采用超声波、超高压对白鲢鱼肌肉中的脂肪氧合酶（lipoxygenase,LOX）进行处理,通过测定 LOX活力、圆二色谱、荧光光谱变化,考察超声波、超高压处理对白鲢鱼肌肉LOX构象及活力的影响。实验结 果表明：随着超声波功率的增加、超声时间的延长,LOX分子内α-螺旋和β-折叠含量降低,荧光强度不断增强, 说明其二级结构和三级结构明显发生变化,使LOX活力不断降低,最适超声条件为300 W、3 h,此时酶活力降低 66.07%;随着超高压压力的增加、超高压时间的延长,LOX分子内α-螺旋和β-折叠含量降低,荧光强度不断减弱, 说明LOX分子二级结构和三级结构发生显著变化,LOX活力迅速降低,最适超高压条件为300 MPa、20 min,此时 酶活力降低93.10%。     

脂肪氧合酶催化亚油酸氧化与大豆蛋白相互作用过程中自由基迁移的电子顺磁共振研究(I)自由基的检测

应用电子顺磁共振(EPR)波谱研究了模拟体系中导致大豆蛋白聚集体形成的自由基由氧化亚油酸到大豆蛋白的迁移.经脂肪氧合酶催化亚油酸的诱导,在大豆蛋白EPR波谱中检测到一个很强的源于肽链骨架α-碳原子或其侧链其它碳原子的自由基信号,其g值范围在2.0041～2.0054之间.此外,在碳自由基中场信号的低场区尚有一个额外的肩峰,它是硫自由基信号,其g值范围在2.019～2.028之间.在大豆蛋白与脂肪氧合酶和亚油酸混合反应前,添加天然抗氧化剂如VC和α-生育酚或人工合成的抗氧化剂2,6-二叔丁基对甲酚(BHT)均能抑制自由基信号和荧光的发展,可使中场碳自由基信号降低35%～65%.