Antibacterial activity and action target of phenyllactic acid against Staphylococcus aureus and its application in skim milk and cheese

Phenyllactic acid (PLA) has been demonstrated to possess antibacterial activity and capacity to prolong food shelf life. However, studies on the performance of PLA in inhibiting Staphylococcus aureus and its effectiveness when applied to dairy products are largely lacking. Here, antibacterial activity (planktonic and biofilm states) of PLA against S. aureus CICC10145 (S. aureus_45) were investigated. The results showed that PLA inhibited growth of S. aureus_45 and formation of S. aureus_45 biofilm. Next, the antibacterial action target of PLA was uncovered from both physiological and phenotypic perspectives. The results showed that PLA decreased cell metabolic activity and cell viability, damaged cell membrane integrity, triggered leakage of intracellular contents (DNA, proteins, and ATP), and caused oxidative stress damage and morphological deformation of S. aureus_45. In practical application, the antibacterial activity of PLA against S. aureus_45 cells was further confirmed in skim milk and cheese as dairy food models, and the antibacterial effects can be adequately maintained during storage for 21 d, at least at 4°C. These findings suggested that PLA could be a potential candidate for controlling S. aureus outgrowth in dairy foods.     

Characteristic tryptic peptides and gelling properties of porcine skin gelatin affected by thermal action

Thermal action in extraction process had effects on characteristic tryptic peptides identification and gelling properties of porcine gelatin. SDS-PAGE, HPLC-LTQ/Orbitrap high-resolution mass spectrometry, texture analyser and rheometer were used to evaluate collagen depolymerisation degree, characteristic tryptic peptides and gelling properties of gelatins prepared in various thermal actions. Results showed that with increasing temperature and time, depolymerisation degree enlarged, while gel strength, gelling and melting temperature decreased. Mass spectra showed that 47 and 49 common characteristic tryptic peptides were identified in gelatins extracted at 50 °C and 100 °C with various times, respectively. Moreover, 34 common characteristic tryptic peptides were identified in all gelatin samples. Further comparison between this work and our previous investigations yielded 20 common characteristic tryptic peptides, which stably exist in various thermal actions. These common characteristic tryptic peptides may be very helpful for the accurate authentication of porcine gelatin.     

Structural Aspects and Prediction of Calmodulin-Binding Proteins

Calmodulin (CaM) is an important intracellular protein that binds Ca²⁺ and functions as a critical second messenger involved in numerous biological activities through extensive interactions with proteins and peptides. CaM’s ability to adapt to binding targets with different structures is related to the flexible central helix separating the N- and C-terminal lobes, which allows for conformational changes between extended and collapsed forms of the protein. CaM-binding targets are most often identified using prediction algorithms that utilize sequence and structural data to predict regions of peptides and proteins that can interact with CaM. In this review, we provide an overview of different CaM-binding proteins, the motifs through which they interact with CaM, and shared properties that make them good binding partners for CaM. Additionally, we discuss the historical and current methods for predicting CaM binding, and the similarities and differences between these methods and their relative success at prediction. As new CaM-binding proteins are identified and classified, we will gain a broader understanding of the biological processes regulated through changes in Ca²⁺ concentration through interactions with CaM.     

微波辅助孵育法富集麦胚多肽的工艺优化

目的 麦胚孵育过程中蛋白酶将蛋白质水解成肽和氨基酸，多肽具有明确的生理活性和营养调节作用。本研究以麦胚为试验原料，采用微波辅助预处理的方法，研究孵育的温度、时间、pH及料液比4种因素对孵育后的蛋白酶活力及多肽含量的影响。 方法 通过单因素和响应曲面试验进行工艺条件优化。 结果 与未进行微波辅助预处理的样品相比，微波辅助处理（600 W, 10 s）能显著提高孵育后样品中的蛋白酶酶活力（约9.4倍）和肽含量（约3.1倍）。经过微波辅助处理后，孵育温度为51.5℃、pH为4.0、时间为6.33 h、料液比为1:7时，蛋白酶活力达最高为3826.24 U/g；孵育温度为45.0 ℃、pH为4.8、时间为8 h、料液比为1:7时，肽含量达最高为262.63 mg/g。 结论 微波辅助处理能有效的激活麦胚孵育液中的内源性蛋白酶活性，促进蛋白水解反应，显著提高孵育后的肽含量。该研究结果为麦胚多肽的制备新工艺开发提供了研究基础。     

Synthesis of Long-Chain β-Lactones and Their Antibacterial Activities against Pathogenic Mycobacteria

In the quest for new antibacterial agents, a series of novel long- and medium-chain mono- and disubstituted β-lactones was developed. Their activity against three pathogenic mycobacteria—M. abscessus, M. marinum, and M. tuberculosis—was assessed by the resazurin microtiter assay (REMA). Among the 16 β-lactones synthesized, only 3-hexadecyloxetan-2-one (VM005) exhibited promising activity against M. abscessus, whereas most of the β-lactones showed interesting activities against M. marinum, similar to that of the classical antibiotic, isoniazid. Regarding M. tuberculosis, six compounds were found to be active against this mycobacterium, with β-lactone VM008 [trans-(Z)-3-(hexadec-7-en-1-yl)-4-propyloxetan-2-one] being the best growth inhibitor. The promising antibacterial activities of the best compounds in this series suggest that these molecules may serve as leads for the development of much more efficient antimycobacterial agents.     

低温水等离子体活化和表面接枝DMAE聚氯乙烯中空纤维膜研究

采用低温水等离子体技术，在三通道聚氯乙烯(PVC)膜表面接枝了甲基丙烯氧基苄基二甲基氯化铵(DMAE)单体，增强了膜亲水和抗菌性能。通过红外分析，表明DMAE成功接枝到了PVC膜上，水通量提高两倍，PVC-ir-H₂O膜(通过水等离子体处理的膜)对牛血清蛋白(BSA)的吸附能力下降67%，对BSA溶液的通量从7.7提高至40 kg?m^-2?h^-1，并且对BSA的截留能力不变。通过静态及动态抗菌实验，接枝后的PVC膜（PVC-g-PMAE膜）抗菌率达到100%，膜组件运行中的抗菌率也达到82%以上。在保证细菌截留率100%的同时，其渗透通量提高三倍。该膜表面修饰工程技术能实现膜表面的均一化改性，且绿色环保、操作简便、成本低，改性膜在饮用水处理领域，尤其是家用净水器中展现了很好的应用前景。     

Purification,structural data and biological properties of polysaccharide from Prunus amygdalus gum

This work demonstrates the efficiency of almond gum polysaccharides (AGPs) as bioactive compounds. AGPs were first extracted using H₂O₂, in the presence of NaOH, at different times and temperatures. The optimal extraction conditions were 4% H₂O₂ and 2 N NaOH, for 7 h at 50 °C, leading to an extraction yield of 58.2% (w/w). After a purification step, the retained AGPs were characterised using high‐performance liquid chromatography showing a molecular weight of 99.3 kDa. The monosaccharide composition of AGPs were assessed using gas chromatography–mass spectrometry. AGPs were found to be a complex heteropolysaccharide with a repeating unit mainly composed of galactose, arabinose, xylose, mannose, rhamnose, and glucuronic acid with the respective ratios: 45:26:7:10:1:11. The acidic nature of the polysaccharide is due to the presence of glucuronic acid. Total antioxidant activity, free radical‐scavenging activity and reducing power assay of AGPs were investigated. The obtained results showed high antioxidant activities of AGPs. Furthermore, beyond 60 mg mL^?1, AGPs exhibited bacterial growth inhibition for five pathogenic strains: Escherichia coli, Staphylococcus aureus, Enterococcus feacalis, Pseudomonas aeruginosa and Salmonella typhimurium.     

Sequence-Dependent Nanofiber Structures of Phenylalanine and Isoleucine Tripeptides

The molecular design of short peptides to achieve a tailor-made functional architecture has attracted attention during the past decade but remains challenging as a result of insufficient understanding of the relationship between peptide sequence and assembled supramolecular structures. We report a hybrid-resolution model to computationally explore the sequence–structure relationship of self-assembly for tripeptides containing only phenylalanine and isoleucine. We found that all these tripeptides have a tendency to assemble into nanofibers composed of laterally associated filaments. Molecular arrangements within the assemblies are diverse and vary depending on the sequences. This structural diversity originates from (1) distinct conformations of peptide building blocks that lead to different surface geometries of the filaments and (2) unique sidechain arrangements at the filament interfaces for each sequence. Many conformations are available for tripeptides in solution, but only an extended β-strand and another resembling a right-handed turn are observed in assemblies. It was found that the sequence dependence of these conformations and the packing of resulting filaments are determined by multiple competing noncovalent forces, with hydrophobic interactions involving Phe being particularly important. The sequence pattern for each type of assembly conformation and packing has been identified. These results highlight the importance of the interplay between conformation, molecular packing, and sequences for determining detailed nanostructures of peptides and provide a detailed insight to support a more precise design of peptide-based nanomaterials.     

含膦酸酯结构单元的磺胺衍生物合成与抗菌活性

为了寻找抗菌候选化合物,采用基于片段的药物发现方法,以氨基膦酸酯和磺酰氯为原料,设计合成了15个含膦酸酯结构单元的磺胺衍生物,经IR、1HNMR和13CNMR确认结构。采用两倍稀释法测定目标化合物的MIC(最小抑菌浓度)。结果表明:部分目标化合物呈潜在的抗菌活性,对所测试标准菌和耐药菌均有抑制活性。其中,化合物Ⅱf〔N-[(二乙氧基膦酰基)-4-氟苯甲基]-4-甲氧基苯磺酰胺〕对金黄色葡萄球菌(S. aureus)、大肠埃希菌(E. coli)、耐甲氧西林金黄色葡萄球菌(MRSA)及耐氟喹诺酮类大肠杆菌(MREC)的MIC分别为32、64、128和128μg/mL,化合物Ⅱl〔N-[(二乙氧基膦酰基)-4-氟苯甲基]-4-氟苯磺酰胺〕对S. aureus、E. coli、MRSA及MREC的MIC分别为32、32、64和64μg/mL,抗菌活性优于对照药磺胺嘧啶。