Safe preparation of beefy meaty peptide with Bacillus subtilis

Preparation of flavour peptides with microorganisms is an attractive choice for its large-scale production. However, beefy meaty peptide (BMP), as a research hotspot in the field of umami peptide, has been studied in few microorganisms, and furthermore, the safe preparation of BMP has not yet been reported. In this study, multi-copy BMP (8BMP) was successfully expressed and produced in the ‘generally recognized as safe’ Bacillus subtilis (B. subtilis). Firstly, 8BMP with the potentially intense umami was screened out based on molecular docking analysis. Then, it was successfully expressed in the B. subtilis 168 and verified through sensory evaluation. Finally, the scaling capacity of 8BMP was evaluated in a 5-L fermenter, with the highest yield (0.84 g L⁻¹) 6.4 times than that of shake flask, which is conductive for industrial production of BMP and other food peptides.     

Proteome analysis of tissues by mass spectrometry

Tissues and biofluids are important sources of information used for the detection of diseases and decisions on patient therapies. There are several accepted methods for preservation of tissues, among which the most popular are fresh‐frozen and formalin‐fixed paraffin embedded methods. Depending on the preservation method and the amount of sample available, various specific protocols are available for tissue processing for subsequent proteomic analysis. Protocols are tailored to answer various biological questions, and as such vary in lysis and digestion conditions, as well as duration. The existence of diverse tissue‐sample protocols has led to confusion in how to choose the best protocol for a given tissue and made it difficult to compare results across sample types. Here, we summarize procedures used for tissue processing for subsequent bottom‐up proteomic analysis. Furthermore, we compare protocols for their variations in the composition of lysis buffers, digestion procedures, and purification steps. For example, reports have shown that lysis buffer composition plays an important role in the profile of extracted proteins: the most common are tris(hydroxymethyl)aminomethane, radioimmunoprecipitation assay, and ammonium bicarbonate buffers. Although, trypsin is the most commonly used enzyme for proteolysis, in some protocols it is supplemented with Lys‐C and/or chymotrypsin, which will often lead to an increase in proteome coverage. Data show that the selection of the lysis procedure might need to be tissue‐specific to produce distinct protocols for individual tissue types. Finally, selection of the procedures is also influenced by the amount of sample available, which range from biopsies or the size of a few dozen of mm² obtained with laser capture microdissection to much larger amounts that weight several milligrams.     

The effects of washing a collagen sample prior to TEM examination

Transmission electron microscopy (TEM) is an important analysis technique to visualize (bio)macromolecules and their assemblies, including collagen fibers. Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2-propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1 min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.     

Laser-assisted preparation of monolithic acidic catalysts for biomass conversion

5-(Hydroxymethyl)furfural (5-HMF) is a vital platform molecule from which a variety of high-value-added fine chemicals and polymerizable monomers can be prepared. The use of solid acids to catalyze the conversion of biomass into 5-HMF is environmentally friendly and economical. However, exploiting the high yield of 5-HMF in a highly concentrated reactant system is challenging. Herein, we present a laser-assisted method for preparing highly acidic monolithic acidic catalysts. A monolithic acidic catalyst based on metal Zr sheets was synthesized and used to catalytically convert 30 wt% fructose into 5-HMF (conversion rate: 96%; yield: 95%). The catalyst was immediately separated from the reaction solution by direct removal at the end of the reaction. Catalytic efficiency was largely unaffected after 10 cycles of use, and the same catalytic efficiency was observed after laser-regeneration, highlighting the potential industrial applicability of the developed catalyst.     

一锅法制备纳米纤维素及其性能研究

采用一种操作简单、绿色环保的以对甲苯磺酸为催化剂对竹纤维进行水热处理制备纳米纤维素(NCC)的方法。考察了反应时间、反应温度和超声作用对纳米纤维素得率和性能的影响。结果表明,在反应温度110℃、反应时间45 min、超声时间120 min的条件下,纳米纤维素得率最高。     

海洋非成岩天然气水合物原位快速制备实验及评价

我国的海洋天然气水合物（以下简称水合物）资源主要分布在沿海大陆架水深介于300～3 000 m的深水区，具有弱胶结、非成岩的特征，为了测试和研究该类型水合物必须原位、大量、快速制备样品，而目前常用的搅拌法、喷淋法、鼓泡法等制备技术却存在着生成速度慢、储气密度低等问题。为此，自主设计、研制了1 062 L非成岩水合物快速制备釜，针对我国南海非成岩水合物的物性特征，开展了搅拌法、鼓泡法、喷淋法单一制备方法及“三合一”法（上述3种方法相结合）水合物制备实验，测试了实验过程中温度、压力、电阻率及反应时间等数据，分析比较了几种不同制备方法的水合物生成情况与制备效率。实验结果表明：①搅拌法、鼓泡法、喷淋法制备水合物过程中，生成的水合物均缓慢增加并逐渐铺满整个液面；②搅拌法制备过程中可以观测到明显的诱导期，而喷淋法、鼓泡法及“三合一”法却无明显的诱导期；③单一制备方法及“三合一”法制备水合物过程中，电阻率均随反应时间的增加而增加，其变化趋势亦与水合物制备速率基本一致；④“三合一”法的制备周期明显短于单一制备方法（搅拌法、喷淋法、鼓泡法单一方法的制备时间分别约为“三合一”法的5.14倍、3.59倍、3.16倍）。结论认为，所研制的1 062 L水合物快速制备釜能够实现海洋非成岩水合物样品的原位、快速制备；较之于单一制备方法，“三合一”法大大提高了水合物的制备效率。     

熔融制样-X射线荧光光谱法测定硅铁中硅磷锰铝钙铬

熔融制样-X射线荧光光谱法(XRF)测定硅铁合金样品,需重点解决样品前处理中合金样品侵蚀铂-黄坩埚的难题。硅铁样品以四硼酸锂-碳酸锂预氧化剂在石墨垫底瓷坩埚中高温预氧化熔融后,再将熔融物转移至铂-黄坩埚中,用四硼酸锂熔融制成玻璃熔片,实现了熔融制样-X射线荧光光谱法对硅铁合金中硅、磷、锰、铝、钙、铬的测定。实验讨论了预氧化熔融的熔剂体系及氧化方法、试样与熔剂的稀释比,结果表明,试样与熔剂以1∶35的稀释比,以10滴300g/L碘化钾溶液为脱模剂,在1100℃熔融30min,熔融制得的玻璃片均匀、透明、无气泡,符合测定要求。用具有浓度梯度的系列硅铁有证标准样品制作校准曲线,各待测元素校准曲线的线性相关系数均大于0.9995。方法应用于硅铁合金实际样品中硅、磷、锰、铝、钙、铬的测定, 结果的相对标准偏差(RSD,n=11)在0.1%~5.8%之间;正确度试验表明,硅铁标准样品的测定结果与认定值相符,硅铁实际样品的测定结果与国家标准方法测定值一致,能满足常规分析要求。     

熔融制样-能量色散X射线荧光光谱仪分析硅酸盐中32 种组分

为充分发挥小型实验室设备的功能,实验采用熔融制样,使用台式能量色散X射线荧光光谱仪(10W, Rh靶)定量分析了硅酸盐样品中的Na₂O、MgO、Al₂O₃、SiO₂、P₂O₅、SO₃、Cl、K₂O、CaO、TiO₂、V、Cr、MnO、Fe₂O₃、Co、Ni、Cu、Zn、Ga、Rb、Sr、Y、Zr、Nb、As、Ba、La、Ce、Th、U、Hf、Pb等32种主、次、痕量组分。对3种不同的解谱方法进行比较,确定Na、Mg、Al、Si、P、As、Ce、Cr、Cu、Ga、La、Nb、Ni、Pb、Th、U、V、Y选择感兴趣区解谱,其余元素用高斯或经改进的高斯函数的最小二乘法拟合解谱。选用有代表性的多个硅酸盐类样品,比较了理论检出限及10次重复测定计算3倍标准偏差得出的检出限,发现以3倍标准偏差作为检出限再取它们的平均值则更具代表性和使用意义。各组分的检出限为0.2~1740μg/g。精密度试验表明,各组分测定结果的相对标准偏差(RSD)为0.12%~10.5%(Na为轻元素,由于含量低,小能谱测定的精密度稍差);对两个土壤标准样品进行正确度验证,测定值与标准值一致。     

Ion substitution induced formation of spherical ceramic particles

How to precipitate ceramic nano- and microspheres in water based solutions only using inorganic ions is a challenge. In this study, spherical particles of alkaline earth phosphates and fluorides were synthesized using a precipitation reaction. Substituting ions, through inhibition of crystal growth, was used to induce sphere formation and to alter the morphology, size and composition of the spheres. The difference in ionic radius between the substituting ion (Mg, Ca and Sr) and the main cation (Sr and Ba) influenced the critical concentration to allow for sphere formation as well as the crystallinity. The larger difference, the lower was the concentration needed to form spheres. Low concentrations of Mg was enough to generate amorphous spheres of Sr- and Ba-phosphates whereas higher concentrations were needed if the radius difference were smaller. An increasing degree of substitution leads to a decrease in crystallinity of precipitated particles. The degree of substitution was determined to 16–55% where a low degree of ion substitution in the phosphates resulted in the formation of spheres (500–800?nm) with rough surfaces composed of apatite like phases. A higher degree of substitution resulted in amorphous spheres (500?nm- 1?μm) with smooth surfaces.     

高熵合金制备及热处理工艺研究进展

与传统合金相比,高熵合金表现出了高强韧、良好的软磁性、高温稳定性、耐腐蚀性等突出特点,在许多的领域应用潜力巨大。本文主要介绍真空熔炼、粉末冶金、激光熔覆和磁控溅射等制备高熵合金方法的研究进展及优缺点,分析了不同热处理工艺对组织结构和力学性能的影响,并对高熵合金制备和模拟计算等方面提出了展望。