不同发酵温度下虾油中细菌的16S rDNA分子生态分析以及蛋白酶系和产品品质的变化研究

本文采用16S r DNA高通量测序技术分析了不同温度发酵下虾油菌相的变化,采用酶谱电泳研究了虾油蛋白酶系的变化,并结合测定各质量指标以探索细菌在虾油发酵中的作用。研究发现虾油中细菌数量远高于真菌数量,显示了细菌对发酵的重要贡献。虾油菌相复杂,且不同温度下菌相差异很大;发酵中菌相及蛋白酶系均呈现动态变化,25℃和35℃发酵的虾油中蛋白酶失活较少,且存在大量Tetragenococcus,Virgibacillus等产生良好风味成分的细菌;而45℃的虾油中蛋白酶失活严重,且检出的细菌与产生良好风味无关,感官评定也最差,因此,45℃不适合发酵虾油。发酵30 d时,虾油中的氨基酸态氮、无盐可溶性固形物、挥发性盐基氮(TVB-N)和三甲胺(TMA)等均达到峰值,此后各指标基本恒定,但菌相仍变化较大且有1种新蛋白酶出现,而且虾油风味继续改善。     

基于rDNA ITS序列分析的桑黄真菌菌株分子鉴定

对6份桑黄真菌菌株的rDNA ITS区段进行克隆测序和序列特征比较分析,并与GenBank中获得的桑黄基源物种相关序列进行同源性比对,进一步将从GenBank检索获得的桑黄基源物种的ITS序列、复合外组ITS序列连同6份桑黄菌ITS序列一起作系统发育分析。结果表明:Sh-03、04、05、06与Phellinus linteus亲缘关系最近,Sh-01与Phellinus baumii亲缘关系最近,Sh-02与Phellinus baumii、Phellinus linteus、Phellinus igniarius的亲缘关系相差甚远,为一错误菌种。本实验的研究结果可以提供一种准确可靠且简单易行的桑黄真菌分子鉴定技术。     

Protein changes in shrimp (Metapenaeus ensis) frozen stored at different temperatures and the relation to water-holding capacity

To study the effects of freezing temperature on muscle proteins, shrimp (Metapenaeus ensis) were frozen stored at either −18 or −60 °C up to 90 and 210 days. Shrimp frozen at −18 °C had higher thawing and compression loss and poor myofibril water-holding capacity compared with those frozen at −60 °C. In terms of protein characteristics, shrimp frozen at −18 °C had higher levels of carbonyls and reduced sulphhydryls. Moreover, the shrimp frozen at −18 °C had higher surface hydrophobicity and reduced Ca²⁺-ATPase activity, indicating increased protein denaturation. Proteomics revealed that seventy-five proteins were classified as differentially abundant proteins (DAPs) following freezing. There were sixty-four DAPs in the F18-CON comparison group (shrimp frozen at −18 °C vs. control) and thirty-two DAPs in the F60-CON comparison group (shrimp frozen at −60 °C vs. control), suggesting that freezing at −18 °C results in more DAPs than freezing at −60 °C. A comparison between F18 and F60 revealed that ribosomal proteins (L44, L7a) and heat shock protein 21 were downregulated in F18. These results increase our understanding of the variable quality associated with shrimp frozen at different temperatures.     

浙东腌冬瓜产组胺微生物的分离及产组胺特性研究

为了解浙东地区传统腌冬瓜发酵体系中产组胺细菌及其组胺产生规律,本研究通过鉴别培养基结合HPLC筛选并确认了菌株的产组胺能力,并利用16S r DNA分子鉴定产组胺菌株;研究培养温度、p H和盐度对菌株生长和产组胺的影响。结果表明,从腌冬瓜发酵前、中期筛选出的5株菌属于2个菌种,命为M1和M3,均是肠杆菌属,且分别与Enterobacter aerogenes(产气肠杆菌)和Enterobacter xiangfangensis的亲缘关系最近。这两种菌的最适生长温度均为30℃,最适生长p H均为7,在小于5%盐度时生长均稳定;在组胺发酵培养基中,产组胺的最适p H均为4,在20~35℃的适宜生长温度内,最适产组胺温度均为35℃,随盐度的升高,产组胺量均降低。适宜的实验条件下,M1和M3的最大生长量分别为11.65 lg CFU/m L和14.30 lg CFU/m L,最大产组胺量分别为35.66mg/L和39.54 mg/L。研究结果可为浙东特色腌冬瓜的加工工艺及质量安全控制等提供参考依据。     

RNA-As-Graphs Motif Atlas—Dual Graph Library of RNA Modules and Viral Frameshifting-Element Applications

RNA motif classification is important for understanding structure/function connections and building phylogenetic relationships. Using our coarse-grained RNA-As-Graphs (RAG) representations, we identify recurrent dual graph motifs in experimentally solved RNA structures based on an improved search algorithm that finds and ranks independent RNA substructures. Our expanded list of 183 existing dual graph motifs reveals five common motifs found in transfer RNA, riboswitch, and ribosomal 5S RNA components. Moreover, we identify three motifs for available viral frameshifting RNA elements, suggesting a correlation between viral structural complexity and frameshifting efficiency. We further partition the RNA substructures into 1844 distinct submotifs, with pseudoknots and junctions retained intact. Common modules are internal loops and three-way junctions, and three submotifs are associated with riboswitches that bind nucleotides, ions, and signaling molecules. Together, our library of existing RNA motifs and submotifs adds to the growing universe of RNA modules, and provides a resource of structures and substructures for novel RNA design.     

Ribosomal Synthesis of Peptides with C‐Terminal Lactams,Thiolactones, and Alkylamides

All change at the C terminus : We have established a novel methodology for the ribosomal synthesis of peptides featuring C‐terminal cyclization and various modifications, including macrocyclization, by making use of genetic code reprogramming. The C‐terminal amide modification of linear and cyclic peptides should enhance their physiological stabilities, and open up the possibility of developing new drug‐like peptides.

     

Sparsomycin-linezolid conjugates can promote ribosomal translocation

Sparsomycin is an antibiotic that targets the peptidyl transferase center of the ribosome and has the ability to promote ribosomal translocation in the absence of EF-G and GTP. Here we show that changes in the configurations at the two chiral centers of sparsomycin, especially at the chiral carbon, can greatly affect its capability to promote ribosomal translocation. More importantly, the incorporation of the pseudo-uracil moiety of sparsomycin into linezolid through a covalent linkage conferred on linezolid derivatives the ability to promote translocation, thus indicating the importance of interactions between this pseudo-uracil moiety, rRNA, and tRNA for promoting translocation. In addition, these translocation promoters can also effectively inhibit spontaneous reverse translocation; this suggests that they might promote forward translocation by trapping the ribosome in the post-translocation state and shifting the equilibrium between the pre- and post-translocation ribosome in the forward direction.