柞蚕丝蛋白中酪氨酸分布的电子光谱研究

柞蚕丝和桑蚕丝均系由18种氨基酸组成的丝蛋白,但二者的性质差异较大。搞清酪氨酸在丝蛋白中的分布,涉及蛋白一级结构,才能有助于找出柞蚕丝与桑蚕丝性质差异的原因。为此我们研究了酪氨酸的分布。本文采用分级水解的方法,对柞蚕丝进行水解,测定各次水解液的紫外和荧光光谱,考察芳香氨基酸(指酪氨酸)在柞蚕丝中的分布。结果发现其分布呈“W”型。     

脉冲电场与牛血清白蛋白相互作用的同步荧光光谱和拉曼光谱比较研究

应用同步荧光光谱和拉曼光谱研究了脉冲电场与牛血清白蛋白的相互作用。同步荧光光谱研究表明,脉冲电场对牛血清白蛋白的发射荧光光谱强度产生影响,降低了处于其活性部位的色氨酸和酪氨酸残基的发射荧光强度。拉曼光谱进一步验证了这种实验结果。两种实验表明：脉冲电场改变了处于牛血清白蛋白活性中心的芳香族氨基酸所处的微环境,进而表明了蛋白质的构象发生了变化,从而影响它的生物学功能。     

核糖核酸酶溶液的拉曼光谱研究

报道了用硝基苯萃取处理前后的核糖核酸酶溶液的拉曼光谱。对其主链构象和侧链构象进行了分析,指出酰胺Ⅰ和酰胺Ⅱ中β一折迭和无规卷曲的构象较为明显,二硫桥键的构象是扭式—扭式—扭式,酪氨酸残基有部分是“埋藏”的。同时指出用硝基苯萃取处理蛋白质溶液是获得高质量拉曼谱图的一个简明有效的方法。     

茶刺蛾颗粒体病毒色涵体的拉曼光谱研究

得到了茶刺蛾颗粒体病毒(Darna trima Granulosis Virus 缩写为 DtGV)包涵体的拉曼光谱。分析表明,DtGV 包涵体蛋白的二级结构主要为β折叠和无规卷曲,后者含有大量β回折结构;蛋白的所有酪氨酸残基均“暴露”于溶剂中;其 C—C—S—S—C—C 构型属于扭曲—扭曲—扭曲式。     

[去-组氨酸~(35),去-甘氨酸~(40)]大鼠转化生长因子-α(34—43)的拉曼光谱

报导了粉末状结晶[des His~(35),des Gly~(40)]γ-TGF-α(34—43)的拉曼光谱。酰胺Ⅰ振动谱带在1669cm~(-1),酰胺Ⅲ振动谱带在1337、1262和1242 cm~(-1),硫—硫键的振动谱带在504cm~(-1),酪氨酸残基的特征谱带在847、830和642 cm~(-1)。它们定性地证明了肽链可能由β-回转和β-折叠织成,酪氨酸残基在肽链中呈全“暴露”式,硫—硫键部位的几何构型为扭式-扭式-扭式。据此提出了该肽链的结构模型。     

稻米陈化中谷蛋白变化光谱解析及其对功能性质的影响

稻米储藏陈化导致食用品质下降,蛋白质变化是重要诱因。谷蛋白是稻米中主要蛋白,采用拉曼和红外光谱表征陈化中谷蛋白的变化,并对其功能性质差异进行比较,利于阐明稻米的陈化机理。拉曼光谱表明,陈米谷蛋白1 665和1 218 cm-1处的拉曼归一化强度分别为1.01和0.25,明显低于新米谷蛋白,表明陈化后谷蛋白的α-螺旋减少;陈米谷蛋白中二硫键(516和527 cm-1处峰强度分别为0.45和0.42)、亚砜(1 035 cm-1处峰强度为0.48)和砜(1 124,1 152,1 159,1 316和1 334 cm-1处峰强度分别为0.47,0.22,0.26,0.50和0.63)的强度明显高于新米谷蛋白,表明含硫氨基酸残基发生明显氧化;陈米谷蛋白的酪氨酸Fermi共振857/830 cm-1的强度比值1.68明显高于新米谷蛋白,酪氨酸残基更加暴露;陈米谷蛋白751 cm-1附近色氨酸的拉曼强度为0.20,比新米谷蛋白的强度0.14显著提高,陈化后谷蛋白色氨酸残基更加埋藏;陈米谷蛋白3 423 cm-1处的O-H伸缩强度为0.05,比新米谷蛋白对应强度0.02显著增大,表明分子间结合程度升高,谷蛋白与淀粉分子结合更加紧密。除了酪氨酸的Fermi共振、1 333和1 152 cm-1处砜的吸收峰不高外,陈化谷蛋白的其余拉曼强度均高于陈米谷蛋白,说明陈化谷蛋白的氧化程度更高。红外光谱表明,陈米谷蛋白和陈化谷蛋白中1 153,1 078和1 026 cm-1处的硫氧化物吸收峰增大,进一步支持谷蛋白发生了氧化。与新米谷蛋白相比,陈米谷蛋白的溶解性、持水性、乳化性和乳化稳定性均显著降低,而持油性升高,支持陈米中谷蛋白发生了明显氧化。陈化谷蛋白的溶解性（除pH 9）、持水性和乳化性比陈米谷蛋白更低,持油性更高,表明新米谷蛋白被提取出来后单独陈化时氧化程度更深。陈化后谷蛋白功能性质的变化支持红外和拉曼光谱显示的氧化变化,这为阐明蛋白质在陈米品质劣变中的作用提供了光谱依据,为控制稻米陈化劣变以减少产后损失奠定基础。     

傅里叶红外结合拉曼分析卡拉胶寡糖对南美白对虾蛋白结构影响

为深入研究卡拉胶寡糖对反复冻融南美白对虾样品品质与蛋白变化影响,采用傅里叶变换中红外（Fourier transform infrared, FTIR）互补结合激光显微拉曼（micro Raman）光谱技术,以不同预处理方式（无菌蒸馏水、三聚磷酸盐溶液与卡拉胶寡糖溶液）下反复冻融南美白对虾肌原纤维蛋白为研究对象,深入研究了卡拉胶寡糖预处理对不同冻融阶段南美白对虾肌肉品质和蛋白的影响机制。分别对三种预处理方式下经0,2,4和6次冻融循环后的南美白对虾肌原纤维蛋白进行FTIR和拉曼光谱分析。FTIR和拉曼一阶谱图中各特征峰强度变化谱图可得出,随冻融次数增加,南美白对虾肌肉的蛋白损失和结构破坏明显加剧。样品肌肉蛋白二级结构的主链构象主要由酰胺Ⅰ带（1 600~1 700 cm-1）表征,FTIR分析显示新鲜虾肉蛋白二级结构以β-转角为主,其次为β-折叠,其弥补了拉曼光谱对β-折叠和β-转角的不敏感。FTIR与拉曼光谱酰胺Ⅰ带高斯拟合后谱图可定性定量显示冻融过程中样品的蛋白二级结构变化主要是α-螺旋结构的减少与无规卷曲结构的增加,而卡拉胶寡糖预处理能明显抑制冻融过程中α-螺旋结构的损失。FTIR对蛋白表面氨基酸变化不敏感,拉曼光谱则可互补显示样品蛋白侧链的构象变化。其表征酪氨酸残基的谱带出现在850和830 cm-1,峰强比表征了样品中酪氨酸的暴露程度在冻融期间呈增长趋势;脂肪族侧链氨基酸残基的C-H弯曲与伸缩振动分别在1 440~1 465和2 800~3 100 cm-1区间,1 448和2 935 cm-1处峰强变化表征了样品侧链氨基酸的疏水相互作用在冻融过程中逐渐增强。拉曼光谱中蛋白侧链的特征谱带变化表明冻融过程使虾肉蛋白中分子内、分子间氢键断裂和侧链酪氨酸、脂肪族氨基酸残基暴露,而卡拉胶寡糖处理后明显延缓了该变化。因此,卡拉胶寡糖可延缓反复冻融虾肉蛋白中氢键断裂、侧链疏水基暴露,进一步稳定蛋白二级结构,维持其蛋白功能特性,从而起到对反复冻融南美白对虾肌肉品质的保护作用。同时,本研究将FTIR与拉曼光谱技术结合应用在卡拉胶寡糖对冻融南美白对虾肌肉品质保护机制研究上,发现FTIR在南美白对虾蛋白二级结构的表征上更敏感,拉曼光谱则能为样品蛋白侧链构象变化提供参考,二者结合可通过提供互补信息,更好表征样品经处理后蛋白的结构变化。     

Cr(Ⅵ)与细胞色素c相互作用的研究

运用荧光光谱、圆二色(CD)谱法研究了K2Cr2O7与细胞色素c的相互作用。结果表明,Cr(Ⅵ)酸根离子与细胞色素c相互作用引起细胞色素c的α-螺旋含量降低,使酪氨酸残基所处微环境的极性增加,色氨酸残基的疏水性增加。Cr(Ⅵ)与细胞色素c的作用位点更接近于色氨酸残基,引起色氨酸残基的微环境变化的程度要大于酪氨酸残基。     

糖性白内障晶状体付立叶变换拉曼光谱的研究

本文应用付立叶变换拉曼光谱技术测定了正常鼠晶状体和半乳糖诱发糖性白内障鼠晶状体激光拉曼光谱。并将两种谱结果相比较，这些谱表明，随着半乳糖诱发糖性白内障的形成，主要表现在８４０ｃｍ＾－１附近的酷氨酸残基双峰强度比明显不同，说明由于晶状体浑浊时，某些酷氨酸残基氨键强度随着形成大分子量蛋白质聚集体而增加。     

甲状腺癌组织的傅里叶变换拉曼光谱研究

利用傅里叶变换拉曼光谱仪得到了甲状腺癌和甲状腺正常组织样品的拉曼光谱,这2种组织样本的拉曼特征峰主要出现在400～3 500 cm-1的波长范围内。在甲状腺正常组织的拉曼光谱中位于503和758 cm-1处有两个特征峰,758 cm-1处峰指认为苯环的环振动,503 cm-1处的拉曼峰经分析初步指认为C—Ⅰ伸缩振动,这2个特征峰在甲状腺癌的拉曼光谱中均消失。在甲状腺正常组织的拉曼光谱中位于3 062和1 003 cm-1处酪氨酸的特征峰,在甲状腺癌组织的拉曼光谱中,这2处峰的相对强度明显降低。因此,拉曼光谱法有望成为甲状腺癌诊断的有效方法。     

红外光谱在纤维质文物材料鉴别中的应用研究

纺织纤维和纸张纤维是常见纤维质文物材料,是构成博物馆精美文物如服饰手稿书画的基本材料,近年来寻求通过无损或微损方法对这一类材料的鉴别以及劣化状况评价备受文物鉴赏家和文物保护工作者的关注。借助傅里叶变换红外光谱,研究博物馆常见纺织纤维材料棉、麻、桑蚕丝、柞蚕丝、羊毛的红外光谱特征和它们的分子结构组成异同,研究传统纸纤维稻草、麦草、龙须草、龙旗松、桑皮红外光谱特征。结果表明：衰减全反射傅里叶变换红外光谱无损分析技术可通过比较3 300～2 800 cm-1 CH,NH,OH振动区间光谱形状以及指纹区峰位以区别不同种类纺织品纤维;碳氧振动纸张纤维最明显光谱差异位置出现在与纤维素OH伸缩振动相关波数3 300 cm-1和与C—O—C相关波数1 332,1 203,1 050 cm-1。文章探索研究红外光谱技术结合主成分分析法在快速鉴别纤维材料中的应用。通过对全光谱数据多元散射校正(MSC)预处理后进行主成分分析,可以把红外光谱十分相似的纺织纤维棉和亚麻、桑蚕丝和柞蚕丝明显分类;对光谱相似的纸纤维,可采用选择不同光谱波数段进行主成分分析,比较发现能够把五种纸纤维明显区分的光谱区间为3 800～2 800 cm-1。本研究为分子光谱无损分析技术应用于文物材料鉴别、科学评估纤维材料保存状况提供基础研究。     

Effect of degumming time on silkworm silk fibre for biodegradable polymer composites

Recently, many studies have been conducted on exploitation of natural materials for modern product development and bioengineering applications. Apart from plant-based materials (such as sisal, hemp, jute, bamboo and palm fibre), animal-based fibre is a kind of sustainable natural materials for making novel composites. Silkworm silk fibre extracted from cocoon has been well recognized as a promising material for bio-medical engineering applications because of its superior mechanical and bioresorbable properties. However, when producing silk fibre reinforced biodegradable/bioresorbable polymer composites, hydrophilic sericin has been found to cause poor interfacial bonding with most polymers and thus, it results in affecting the resultant properties of the composites. Besides, sericin layers on fibroin surface may also cause an adverse effect towards biocompatibility and hypersensitivity to silk for implant applications. Therefore, a proper pre-treatment should be done for sericin removal. Degumming is a surface modification process which allows a wide control of the silk fibre's properties, making the silk fibre possible to be used for the development and production of novel bio-composites with unique/specific mechanical and biodegradable properties. In this paper, a cleaner and environmentally friendly surface modification technique for tussah silk in polymer based composites is proposed. The effectiveness of different degumming parameters including degumming time and temperature on tussah silk is discussed through the analyses of their mechanical and morphological properties. Based on results obtained, it was found that the mechanical properties of tussah silk are affected by the degumming time due to the change of the fibre structure and fibroin alignment.     

Raman and surface enhanced Raman scattering of a black dyed silk

The Raman and surface enhanced Raman scattering (SERS) spectra of a black dyed silk sample (BDS) were registered. The spectral analysis was performed on the basis of Raman and SERS spectral data of isolated samples of Bombyx mori silk fibroin, its motif peptide component (GAGAGS) and the synthetic reactive black 5 dye (RB5). The macro FT‐Raman spectrum of the silk sample is consistent with a silk II‐Cp crystalline fraction of Bombyx mori silk fibroin; the SERS spectrum is highly consistent with conformational modifications of the fibroin due to the interactions with the Ag nanoparticles. The GAGAGS peptide sequence dominates the Raman spectrum of the silk. The SERS spectrum of the peptide suggests a random coil conformation imposed by the surface interaction; the serine residue in the new conformation is exposed to the surface. Quantum chemical calculations for a model of the GAGAGS–Ag surface predict a nearly extended conformation at the Ag surface. The Raman spectrum of the dye was analysed, and a complete band assignment was proposed; it was not possible to propose a preferential orientation or organization of the molecule on the metal surface. Quantum chemical calculations for a model of the dye interacting with a silver surface predict a rather coplanar orientation of the RB5 on the Ag metal surface. The Raman spectrum of the BDS sample is dominated by signals from the dye; the general spectral behaviour indicates that the dye mainly interacts with the silk through the sulphone (–SO₂–) and sulphonate (–SO₂–O–) groups. Besides the presence of dye signals, mainly ascribed to the sulphone and sulphonate bands, the SERS spectrum of the BDS sample also displays bands belonging to the amino acids alanine, glycine, serine and particularly tyrosine. Copyright © 2013 John Wiley & Sons, Ltd.     

ESR法考察龙胆酸的氧化——柞蚕茧丝中龙胆酸的鉴定

本文用电子自旋共振波谱法研究了龙胆酸在碱性水溶液中的氧化,发现了一种新的自由基谱,通过自旋密度和超精细偶合常数的计算,给出了该自由基物种的结构模式,推测出龙胆酸在碱性水溶液中的氧化过程。作为柞蚕茧中龙胆酸的ESR检测方法,对测定的条件和一些影响因素进行了考察、     

用拉曼光谱研究蚕丝蛋白的结构与功能的关系

本文用拉曼光谱研究了蚕丝蛋白在不同状态时的构象，发现丝纤维的构象是以ｐ一折迭为主，而蚕丝腺体中未拉伸过的丝蛋白是以无规卷曲和α－螺旋为主。同时也用拉曼光谱方法验证了蚕吐丝过程中丝蛋白产生变住的机制。     

Comparative Raman spectroscopic analysis of orientation in fibers and regenerated films of Bombyx mori silk fibroin

This study is focused on the Raman spectroscopic analysis of degummed silk fibroin (SF) fibers and regenerated Bombyx mori silk fibroin films: a correlation was found between some spectral features related to the methylene deformation modes and the molecular orientation of the samples. Polarized Raman spectra on SF fibers were used to obtain the orientation distribution function of carbonyl groups along the protein backbone. The variation of the intensity ratio of 1400/1450 cm⁻¹ for the peaks attributed to the wagging and bending deformation modes of CH₂ groups with respect to the angular orientation of the fiber was measured and quantitatively correlated with the orientation distribution function of the carbonyl groups. Unpolarized Raman spectra were measured for regenerated silk fibroin films and lyophilized solutions. The variation of the intensity ratio of 1415/1455 cm⁻¹, which is related to the deformation modes of CH₂ groups in SF regenerated materials, was qualitatively related to the microstructural orientation of the samples observed by scanning electron microscopy (SEM), and to the presence of Silk I phase as suggested by the analysis of samples obtained in different casting conditions and also by the measurements on mechanically deformed films. The results obtained showed the utility of the spectroscopic intensity ratio of 1400/1450 cm⁻¹ for the rapid assessment of molecular orientation in silk fibers, which could be useful for quality and process control of regenerated silk‐based textiles. Moreover, the qualitative dependence of the intensity ratio of 1415/1455 cm⁻¹ was found to be sensitive to both the microstructural orientation and Silk I content of regenerated silk fibroin films, suggesting a possible correlation of this Raman marker of the Silk I phase with the degree of molecular order brought about by this polymorph. Copyright © 2006 John Wiley & Sons, Ltd.     

Subtracted shifted Raman spectroscopy of organic dyes and lakes

Resonance Raman spectroscopy has been utilized to identify vegetal and animal dyes and lakes. These compounds have been used by artists since antiquity, and their identification has important implications in art history and conservation. The resonance condition is useful when working with real ancient objects so as to enhance the Raman scattering of chromophores with respect to the strong signal of the matrix, such as the textile or parchment over which the colorant is supported. The strong fluorescence that generally characterizes the resonance Raman spectra was eliminated by using subtracted shifted Raman spectroscopy (SSRS). A systematic study on reference organic dyes was primarily carried out to evaluate the suitability of the method and to estimate the strengths and limitations of the spectrum reconstruction process. The method was then applied to the noninvasive identification of madder on ancient dyed silk textiles such as an altar table cloth and a manutergium attributed to the Egyptian artistic textile production of the 10th–11th Century A.D. Copyright © 2009 John Wiley & Sons, Ltd.     

A vibrational analysis of trichloroiodomethane

The infrared and Raman spectra of CCl₃I have been recorded. Band assignments have been made on the basis of the infrared results, and the Raman spectrum has been interpreted in terms of the fundamentals of CCl₃I, with additional bands being assigned to decomposition products of CCl₃I. A Urey-Bradley force constant calculation has been made, transferring results from the earlier work of Ngai and Mann.     

Identification of berberine in ancient and historical textiles by surface‐enhanced Raman scattering

The highly fluorescent natural dye berberine can be easily identified in microscopic textile samples by surface‐enhanced Raman spectroscopy employing citrate‐reduced Ag colloid. The ordinary Raman (OR) and SERS spectra of berberine are presented and discussed in the light of a DFT calculation. Using FT‐Raman and FT‐SERS we could reliably compare relative intensity shifts and investigate the adsorption geometry of berberine on Ag nanoparticles. The significant enhancement in the FT‐SERS spectrum of the out‐of‐plane ring system bending deformation mode at 729 cm⁻¹ relative to a group of in‐plane vibrations at around 1500 cm⁻¹ was interpreted as evidence of a ‘flat‐on’ adsorption geometry. SERS was successfully used to identify berberine in silk fiber samples coated with colloidal Ag following a pretreatment with HCl vapor. The SERS method allowed us to detect berberine in a microscopic sample of a single silk fiber from a severely degraded and soiled 17th Century Chinese textile fragment. Copyright © 2007 John Wiley & Sons, Ltd.     

拉曼光谱和色差法在丝质文物色彩损伤评估中的比较

丝质文物是博物馆内的重要藏品之一,具有极高的文化、艺术、历史价值。作为一种性质不稳定的蛋白质有机构成材料,丝质文物极易受到光学辐射而发生泛黄等色彩损伤,尤其是在Light Emitting Diode(LED)得到广泛应用的博物馆光环境中。如何针对馆藏丝质文物色彩损伤进行科学评估是本研究要解决的主要问题。色差评估法是分析博物馆照明对文物色彩损伤的有效手段,但是存在相应的局限性,无法对色彩损伤的诱导期进行评估。由于文物发生色彩损伤的根本原因在于材料内部分子结构发生光化学反应,理论上说,从微观分子层面研究的拉曼光谱法能够更加科学地评估丝质文物色彩损伤。本研究将拉曼光谱引入博物馆照明领域,对比色差评估结果验证拉曼光谱色彩损伤评估的可行性和科学性。通过构成四基色LED的450, 510, 583和650 nm四种窄带光对丝质样品开展长周期照射实验,分别以CIE L*a*b*色差和拉曼光谱作为评估指标,计算得到基于两种评估方法的不同窄带光对丝质文物的色彩相对损伤系数,分别为450 nm∶510 nm∶583 nm∶650 nm=1.00∶0.63∶0.48∶0.32和450 nm∶510 nm∶583 nm∶650 nm=1.00∶0.69∶0.47∶0.27。结果一方面表明,两种方法得到的四种窄带光对丝质文物的色彩损伤趋势是一致的,即450 nm>510 nm>583 nm>650 nm,波长越短对丝质文物的色彩损伤程度越大,说明拉曼光谱是一种能够量化评估丝质文物色彩损伤的可行方法;另一方面表明,基于拉曼参数计算的色彩相对损伤系数的比例差异更大。丝质样品的老化过程中存在诱导期。色差法难以分析丝质样品诱导期的色彩变化,而拉曼光谱分析则可灵敏地检测出相应的分子结构变化,包括诱导期。因此,拉曼光谱法能够对丝质文物的色彩损伤进行更加科学地评估。同时,研究得到的构成LED光谱的四种窄带光对丝质文物色彩相对损伤系数,可以为这类光源在丝质文物照明中的损伤度评估和博物馆准入评估提供依据。