超高压处理对毛栓菌多酚氧化酶的影响

 以茶多酚为底物,测定毛栓菌多酚氧化酶（Polyphenol Oxidase,PPO）活性的最适pH值为5.0、最适底物浓度为4 mg/mL,经100、300和500 MPa超高压处理10 min的PPO酶活分别是未处理的93%、87%和86%。圆二色谱和高效液相凝胶渗透色谱分析结果显示,100和300 MPa压力处理PPO的圆二色谱位于190~210 nm的正Cotton效应峰分别产生“褶皱”和“分蘖”;高效液相凝胶渗透色谱中的第一时间出峰组分（即最大分子量组分）的保留时间由6.678 min分别提前至1.514和2.296 min。500 MPa压力处理PPO的圆二色谱和高效液相凝胶渗透色谱与对照相比差异更加显著,其圆二色谱中只有一个正Cotton效应峰;其高效液相凝胶渗透色谱中主要组分的色谱峰面积减小（即280 nm的响应值降低）。不同压力处理的PPO的圆二色谱与高效液相凝胶渗透色谱图的差异程度具有对应关系。     

应用FTIR研究超声对牛血清白蛋白二级结构的影响

应用傅里叶变换红外光谱(FTIR)结合荧光光谱研究了牛血清白蛋白(BSA)在超声作用下的结构变化。荧光光谱表明,超声作用使BSA溶液荧光光谱最大发射峰发生了蓝移,表明超声改变了BSA中色氨酸(Trp)残基环境;荧光强度的降低表明超声改变了蛋白质分子的构象,具有荧光猝灭效应。采用对BSA红外光谱酰胺Ⅰ带进行曲线拟合的方法,定量分析了不同超声功率、时间对BSA二级结构的影响,发现超声作用对BSA中的α-螺旋、β-折叠、β-转角及无规卷曲的相对含量有不同程度的影响;超声作用具有使BSA的二级结构由α-螺旋向β-折叠、β-转角转化的趋势,而无规则卷曲含量则基本不受超声影响而保持相对稳定。     

FTIR分析共价偶联对抗体蛋白二级结构的影响

用共价偶联的方法制备了免疫胶乳,并利用傅里叶变换红外光谱技术,结合差谱、去卷积、二阶导和曲线拟合等计算机辅助分析方法,研究免疫胶乳中抗体蛋白的二级结构。结果表明,随着pH值的升高以及胶乳浓度的增大,抗体蛋白的有序结构含量增加。由此认为共价偶联会对抗体蛋白的二级结构产生较为显著的影响。     

FTIR光学信息研究PLD蛋白分子冻干胁迫下的二级结构变化

通过FTIR光学信息研究液态和固态磷脂酶D的二级结构,结果表明FTIR光学信息是一种有效的蛋白质分子二级结构研究方法。FTIR的光学信息分析表明冻干胁迫下,磷脂酶D的FTIR光学信息及二级结构发生较大变化。在磷脂酶D活性构象时的pH冻干使磷脂酶Dα螺旋大大下降,无规卷曲大大上升,β折叠几乎没有变化,说明PLD蛋白质分子空间有序构象解体严重。功能性试验表明,磷脂酶D二级结构的变化直接导致了其活性发生很大变化。     

傅里叶变换红外光谱与圆二色光谱表征磷脂酶D二级结构的比较

报道了傅里叶变换红外光谱与圆二色光谱光学信息表征蛋白质二级结构的比较,研究表明傅里叶变换红外光谱与圆二色光谱光学信息表征蛋白质二级结构时,要注意缓冲体系的选择,磷酸盐缓冲液比醋酸盐缓冲液更适合使用,利用FTIR与CD光谱信息表征蛋白质二级结构,两者的数据吻合较好。     

FTIR法研究核糖核酸酶（RNaseA）二级结构随温度的变化

本文利用ＦＴＩＲ法研究了牛胰核糖酸酶（ＲＮａｓｅＡ）二级结构随温度的变化。结果表明，α－螺旋结构随温度的变化较大，β－折叠的结构变化是一个比较复杂的过程。     

超高压处理对多酚氧化酶活性的影响

 研究了超高压中温协同处理对砀山梨汁中多酚氧化酶活性的影响,实验压力为0.1~500 MPa,温度为20~60 ℃。此外,考察了不同pH值（3~7）和保压时间（2~34 min）超高压处理对酶活性的影响。实验结果分析表明：在处理温度为50 ℃、保压时间为10 min和梨汁pH值为5的条件下,200~300 MPa处理梨汁时多酚氧化酶被激活,活性表现最高;500 MPa时酶的活性下降到75.3%。协同温度为30 ℃处理梨汁时,酶的活性反而增大;30 ℃以后,酶活性随温度升高而迅速降低;有效协同高压处理的温度为40 ℃。随着保压时间的延长,梨汁中过氧化物酶的活性减小;18 min以前下降速度较快些,之后下降速度变缓。pH在5~6之间,酶的残留活性最大;pH值为6时,梨汁中多酚氧化酶最为耐压。     

pH诱导烟草多酚氧化酶二级结构变化的光谱学研究

为了获得更多的关于多酚氧化酶二级结构和活性中心结构的信息 ,应用FT -IR以及去卷积技术、荧光光谱、紫外差光谱研究了溶液 pH的变化对烟草多酚氧化酶的二级结构的影响并对其活性中心的光谱跃迁进行了归属 .实验结果表明 ,随着 pH值的降低或提高 ,多酚氧化酶的二级结构发生了变化 ,浕 螺旋和反平行 β 折叠含量减少 ,无规卷曲含量有所增加 ,同时色氨酸和酪氨酸残基的微环境发生了一定的变化 ;随pH的降低 ,活性中心的Cu2 + 与配位的组氨酸咪唑基的作用加强 ;此外 ,归属和讨论了酶活性中心金属离子与配体间全部电荷转移谱带     

FTIR和Raman用于盐酸曲马多的结构分析

采用AVATAR 360型傅里叶变换红外光谱仪和RM-1000型激光共聚焦拉曼光谱仪测定盐酸曲马多的红外光谱和拉曼光谱。拉曼光谱和红外光谱中均显示出了盐酸曲马多的特征峰,都能够用于其结构鉴别。采用红外、拉曼光谱这两种方法互相印证,互相补充,可增强鉴定的准确性、可靠性。     

硫酸铜诱导烟草多酚氧化酶抗变性的研究

为了进一步探讨外源铜与多酚氧化酶的相互作用 ,通过酶活性测定 ,荧光光谱和圆二色光谱 (CD)对外源Cu2 + 诱导烟草多酚氧化酶抗变性进行了研究 .荧光光谱结果表明 ,没有外源铜存在的情况下 ,盐酸胍诱导的烟草多酚氧化酶变性是一个二态过程 ,烟草多酚氧化酶在 6mol/L盐酸胍中变性 5min ,酶活性降至 18.6 % ,变性30min便彻底失活 ;10 .0mmol/L外源铜存在时 ,盐酸胍诱导的烟草多酚氧化酶变性是一个三态过程 ,上述相同盐酸胍浓度下变性相同时间 ,酶的剩余活性分别为 6 0 .6 %和 2 4 .9% .CD谱数据结果表明 ,6mol/L盐酸胍中变性 30min ,烟草PPO二级结构中α 螺旋、反平行 β 折叠、β 转角 /平行 β 折叠、芳香残基和二硫键、无规卷曲 /γ 转角的百分含量分别为 1.1、3.8、3.3、7.5和 84 .3;在 10 .0mmol/LCu2 + 存在时 ,上述相同变性条件下 ,各种二级结构百分含量分别为 34.2、13.7、2 1.0、9.5和 2 1.6 .适量外源铜可明显提高烟草多酚氧化酶结构的稳定性 .     

Effects of high hydrostatic pressure on secondary structure and emulsifying behavior of sweet potato protein

In this study, secondary structures of sweet potato protein (SPP) after high hydrostatic pressure (HHP) treatment (200–600?MPa) were evaluated and emulsifying properties of emulsions with HHP-treated SPP solutions in different pH values (3, 6, and 9) were investigated. Circular dichroism analysis confirmed the modification of the SPP secondary structure. Surface hydrophobicity increased at pH 3 and decreased at 6 and 9. Emulsifying activity index at pH 6 increased with an increase in pressure, whereas emulsifying stability index increased at pH 6 and 9. Oil droplet sizes decreased, while volume frequency distribution of the smaller droplets increased at pH 3 and 6 with the HHP treatment. Emulsion viscosity increased at pH 6 and 9 and pseudo-plastic flow behaviors were not altered for all emulsions produced with HHP-treated SPP. These results suggested that HHP could modify the SPP structure for better emulsifying properties, which could increase the use of SPP emulsion in the food industry.     

Evaluation of high hydrostatic pressure effects on bovine red blood cells and platelets

The objective of this study was to investigate the effects of high hydrostatic pressure (HHP) on the stability of red blood cells (RBCs) and platelets. Bovine blood cells (n=5) were treated with the pressure of 55, 110, 154 and 220 MPa at 25 °C for 5 min. Light microscopy, atomic force microscopy and flow cytometry studies revealed that RBCs were morphologically stable up until the 220 MPa pressure treatments, at which surface modifications were observed. The platelets were found to be less stable than RBCs. HHP application did not cause any significant change in the signal intensity, band area and frequency values of the infrared bands with the exception that a significant variation was observed in the area of the cholesterol band. No statistically significant variations were observed in the secondary structure elements due to HHP treatment according to the artificial neural network study based on the FTIR data.     

癫痫大鼠海马神经元生化分子的同步辐射显微红外光谱成像研究

癫痫是影响所有年龄段的慢性脑功能障碍,主要特征为整个或局部脑区神经元异常同步化高频群集动作电位发放。利用神经毒素海人藻酸（KA）立体定位注射入大鼠海马,诱导大鼠产生癫痫持续状态,建立颞叶癫痫大鼠模型。应用同步辐射显微光谱和同步辐射显微光谱成像分析癫痫持续状态发作后24小时的颞叶癫痫大鼠海马角（CA）1区神经元生物化学分子的胞内浓度和分布是否改变。结果显示反映蛋白质二级结构的酰胺Ⅰ在1 655 cm-1的振动频率和属于脂类功能集团的2 800～3 000 cm-1振动频率,在正常对照大鼠海马CA1神经元胞体内呈高浓度分布,但在癫痫大鼠海马CA1神经元胞体,反映蛋白质二级内呈低浓度分布,并且以细胞核分布浓度最低,但在神经元胞体外围分布浓度相对较高。属于核酸集团的1 055～1 054 cm-1 PO₂反对称拉伸振动在正常和癫痫大鼠海马CA1神经元胞体内分布趋势没有差异,都在胞体内呈高浓度分布,尤其在细胞核分布浓度最高。对属于酰胺Ⅰ的吸收频率进行二级导数分析显示癫痫海马神经元的酰胺Ⅰ相对于正常对照多出1个1 653 cm-1附近的负峰。以上结果提示在发生癫痫持续发作后海马神经元生物化学分子的细胞分布会出现变化。     

五种杂交玉米籽粒品质的FTIR光谱研究

为了研究傅里叶变换红外光谱在杂交玉米品质评定上的可行性,使用傅里叶变换红外光谱仪获得五种杂交玉米籽粒的光谱。1 776～952 cm-1范围内的光谱主要由蛋白质、淀粉和油脂的红外吸收带组成,淀粉的吸收带较为强烈,蛋白质和油脂的吸收带稍弱。该范围光谱中特征吸收带相互重叠,导致一些重要信息被掩盖。通过对1 776～952 cm-1范围内的红外光谱进行曲线拟合分析,发现1 051和1 548 cm-1吸收子峰的面积在1 776～952 cm-1范围内吸收带总面积中占的百分比与玉米籽粒中淀粉和蛋白质的含量存在着线性关系。研究结果显示傅里叶变换红外光谱结合曲线拟合分析技术可用以分析杂交玉米籽粒中淀粉和蛋白质的含量,为玉米品质的快速评定提供一种方法。     

采用FTIR技术研究高压处理对冻干大豆分离蛋白构象的影响

采用傅里叶转换红外光谱技术(FTIR)研究了高压(HP)处理对冷冻干燥的大豆分离蛋白(SPI)构象的影响。在SPI的去卷积FTIR光谱的酰胺Ⅰ′区域(1 600～1 700 cm-1),观察到12个与蛋白构象相关的红外吸收峰,分别对应于CO键伸缩振动与肽键的C—N伸缩振动。通过对该区域的峰强度与波数分析显示,压力为200～400 MPa的HP处理导致SPI在该区域的峰发生明显的“红移”(约2 cm-1),强度也显著增加。更高的HP处理进一步增强了SPI的酰胺Ⅰ′区域的峰强度。对酰胺Ⅱ峰分析显示,HP处理导致酰胺Ⅱ峰(如1 560～1 500 cm-1)的强度、面积逐渐增加(与压力呈正相关)。以上分析显示,HP处理导致SPI的二级与三级结构逐渐打开,然而变性蛋白的结构在高压释放后经历一个“重构过程”。     

圆柱形超高静水压水听器设计与测试

(本文介绍了一种应用于20Hz~20kHz频段、最大工作深度5000米的超高静水压水听器,水听器敏感元件采用厚壁压电陶瓷圆管,高强度复合泡沫去耦,通过对敏感元件及水听器结构进行合理设计,保证水听器具有高的耐压强度。设计了一种圆柱形高静压耦合腔,根据耦合腔声学和力学条件,确定了腔体尺寸及厚度,在国内首次实现了最高50MPa压力下的水听器低频灵敏度测试。测试结果表明,水听器常压下接收灵敏度响应平坦,在常压到50MPa压力范围内,水听器低频接收灵敏度最大变化小于2dB,表明该型水听器具有良好的压力稳定性,可在深海资源勘测、深海通讯等方面获得广泛应用。)     

Inactivation of Staphylococcus aureus using high hydrostatic pressure

The aim of this work was to develop a high-pressure decontamination and sterilization process for pharmaceutical treatments as was developed in food processing in the late eighties. The lack of normalized biological indicators able to validate sterilizing treatments under high pressure led us to select representative pathogenic strains from flora and the European Pharmacopoeia. We selected the following four bacterial strains: Candida albicans (ATCC 10231), Psuedomonas aeruginosa (ATCC 9027), spores of Aspergillus niger (ATCC 16404) and Staphylococcus aureus (ATCC 6538).

This present study is focussed on S. aureus. Successive pressurization and depressurization cycles appeared to be more efficient than a continuous high-pressure treatment. Importantly, these pressure conditions, temperature and process duration are perfectly compatible with current industrial plants. These results show that HHP technology is a new alternative to inactivate pathogenic strains in accordance with pharmaceutical requirements.     

Effect of high hydrostatic pressure processing and squeezing pressure on some quality properties of pomegranate juice against thermal treatment

The aim of this study was to investigate the effect of high hydrostatic pressure (HHP) treatment (200, 300, 400?MPa; 5°C, 15°C and 25°C; 5 and 10 min) on some quality properties of pomegranate juice. Juice samples are obtained under industrial conditions at two different squeezing pressure levels (100 and 150?psi – 0.689 and 1.033?MPa, respectively). Results are compared against conventional thermal treatment (85°C/10 min) and raw sample. For all three processing temperature, HHP combinations at 400?MPa for 10 min were sufficient to decrease the microbial load around 4.0 log cycles for both squeeze levels. All HHP treatments showed no significant decrease at antioxidant activity, total phenolic content and monomeric anthocyanin pigment concentrations, while there was a significant decrease (p?≤?.05) in thermal-treated samples. Being the highest sugar alcohol in pomegranate juice, mannitol content must be considered for determining the authenticity, and mannitol content increased with squeezing pressure and thermal treatment.     

Effect of high hydrostatic pressure on enzyme activity in unpasteurized draft sake

ABSTRACT

Unpasteurized draft sake has a potentially high market value, due to its fresh flavor and fruity taste, compared with conventional thermal-pasteurized sake. However, the shelf life of draft sake is limited. To increase the shelf life of draft sake, it is necessary to suppress flavor and taste deterioration resulting from inactivation of enzymes produced by koji-mold. Draft sake was treated with high hydrostatic pressure (HHP) of 200 to 500?MPa at ?7 to 50°C to analyze the inactivation of α-amylases, glucose-forming enzymes, and acid carboxypeptidases. We found significant inactivation of enzymes produced by koji-mold in draft sake subjected to HHP treatment at both high and low temperature. However, HHP treatment at low temperature effectively inactivated enzymes while retaining the fresh flavor and fruity taste of draft sake.     

Effect of plating and incubation conditions on the detection of lactic acid bacteria after high hydrostatic pressure treatment

ABSTRACT

Lactic acid bacteria (Leuconostoc mesenteroides, Enterococcus faecalis, and Lactobacillus fermentum) were subjected to high hydrostatic pressures (HHPs) of 400 and 600?MPa at 25°C for 10?min in phosphate-buffered saline. Differential plating methods were applied to evaluate HHP-treated cell populations, assuming that healthy and injured cells during plate incubation survived maximum and minimal stress, respectively. The stress was altered by using several selective media in combination with aerobic or anaerobic incubation at 25°C or 30°C. E. faecalis was detectable after 600?MPa treatment while L. mesenteroides and L. fermentum were nondetectable. Specific combinations of incubation conditions were suggested to determine maximum and minimum viable counts of L. mesenteroides and E. faecalis. The difference between the maximum and minimum counts can be used to evaluate HHP-injured population with reduced risks to overestimate healthy and/or underestimate HHP-injured cells.