Probing and Engineering the Fatty Acyl Substrate Selectivity of Starter Condensation Domains of Nonribosomal Peptide Synthetases in Lipopeptide Biosynthesis

Lipopeptides are produced by nonribosomal peptide synthetases (NRPSs) and contain diverse fatty acyl moieties that are major determinants of antibiotic potency. The lipid chains are incorporated into peptidyl backbones via lipoinitiation, a process comprising free fatty acid activation and the subsequent starter condensation domain (C1)‐catalyzed conjugation of fatty acyl moieties onto the aminoacyl substrates. Thus, a thorough understanding of lipoinitiation biocatalysts would significantly expand their potential to produce novel antibiotics. Here, biochemical assays, in silico analysis, and mutagenesis studies are used to ultimately identify the specific amino acid residues that control the fatty acyl substrate selectivity of C1 in lipopeptide A54145. In silico docking study has identified four candidate amino acids, and subsequent in vitro assays confirmed their functional contribution to the channel that controls substrate selectivity. Two engineered variants with single point mutations in C1 are found to alter the substrate selectivity toward nonnatural fatty acyl substrates. The detailed mechanistic insights into the catalytic contribution of C1 obtained from the present study will facilitate future NPRS biocatalyst efforts     

西藏大团囊菌属及其生态分布

首次报道西藏分布的大团囊菌3个种与1个变种：Elaphoopcesgranulatusvar．granulatus,E.granulatusvar．asperulusE.tuberculatus和E.muricatus,讨论了种间和种内的一些特征分化或变异。其中,E．tuberculatus被确认为新种。这些种类均生于林下土中,与冷杉（Abies）、云杉（Picea）、栎属（Quercus）树种共生形成外生苗根,其垂直分布海拔高度可达3600m,然而西藏分布较广的松林下目前尚未发现该属种类的存在。大团囊菌种类在生态适应性上存在很宽的分布幅度,一般认为该类菌物是属于全球广布的热带、亚热带、温带直至亚高山寒温带均有分布的、典型的地下生生态类群。研究的标本全部保存于西藏高原生态研究所标本室（HXZE）。     

蛋白质糖基化修饰的研究方法及其应用

蛋白质糖基化是一种重要的翻译后修饰,它参与和调控生物体的许多生命活动。随着蛋白质组技术的不断发展,蛋白质糖基化研究越来越受到广泛的重视。本文介绍了蛋白质糖基化修饰的研究内容与方法,并综述了最近的研究进展。     

广东大宝山矿区生态环境退化现状及治理途径探讨

根据对广东大宝山矿区的实地调查、测试和所采集土、水样品的化学分析结果,对该区生态环境退化和向下游排放污染物现状进行报道.矿坑似乎是大宝山矿最重要的污染源地,据有限土壤样品分析结果推算,当其所含无机还原态硫彻底氧化后,平均1t矿坑表土可产出207kg浓硫酸.大部分矿坑表土和排土场废土已经历不同程度的酸化.实验室模拟试验显示:5g强烈酸化的矿坑表土在接力水提条件下可产出超过750mL的酸性“矿水”(pH<3.9).实测数据表明:排入翁江支流的矿水,其酸度大大超过国家规定的排放标准,大多数重金属元素的浓度也超过了国家规定的污水综合排放标准的最高允许值.从技术的角度来看,对大宝山矿区生态环境退化和向下游排放污染物状况的治理,宜采用环境地球化学工程与生物修复技术相结合的综合治理方法.     

Three-dimensional Biomimetic Technology: Novel Biorubber Creates Defined Micro- and Macro-scale Architectures in Collagen Hydrogels

Tissue scaffolds play a crucial role in the tissue regeneration process. The ideal scaffold must fulfill several requirements such as having proper composition, targeted modulus, and well-defined architectural features. Biomaterials that recapitulate the intrinsic architecture of in vivo tissue are vital for studying diseases as well as to facilitate the regeneration of lost and malformed soft tissue. A novel biofabrication technique was developed which combines state of the art imaging, three-dimensional (3D) printing, and selective enzymatic activity to create a new generation of biomaterials for research and clinical application. The developed material, Bovine Serum Albumin rubber, is reaction injected into a mold that upholds specific geometrical features. This sacrificial material allows the adequate transfer of architectural features to a natural scaffold material. The prototype consists of a 3D collagen scaffold with 4 and 3 mm channels that represent a branched architecture. This paper emphasizes the use of this biofabrication technique for the generation of natural constructs. This protocol utilizes a computer-aided software (CAD) to manufacture a solid mold which will be reaction injected with BSA rubber followed by the enzymatic digestion of the rubber, leaving its architectural features within the scaffold material.     

旅游生态学研究进展

作为生态学和旅游学相交叉的一门新兴分支学科,旅游生态学关注旅游发展中的生态环境问题,以生态学原理指导旅游生态系统管理,是促进旅游业可持续发展的重要理论支撑。论文采用引文可视化分析软件CiteSpace5.3和文献阅读相结合的方法分析旅游生态学研究的国际进展。分析发现,文献数量在进入21世纪后快速增长,文献来源地集中在北美和欧洲国家,研究方法呈现多学科综合性和最新科学技术成果应用及时等特点,研究内容主要集中在旅游活动的生态环境影响、旅游生态系统管理和旅游可持续发展的测度及实现途径3个方面。基于国际进展的分析,论文最后提出我国旅游生态学研究应在学科的基础理论、旅游影响的系统性和持续性、旅游生态修复、旅游开发活动的生态环境影响、生态环境变化对旅游业发展的影响和本土性旅游生态系统管理方法等方面加强。     

EFFECTS ON FITNESS COMPONENTS OF P-ELEMENT INSERTS IN DROSOPHILA MELANOGASTER: ANALYSIS OF TRADE-OFFS

We analyzed the trade-offs between fitness components detected in four experiments in which traits were manipulated by inserting small (control) and large (treatment) P-elements into the Drosophila melanogaster genome. Treatment effects and the interactions of treatment with temperature, experiment, and line were caused by the greater length and different positions of the treatment insert. In inbred flies, the treatment decreased early and total fecundity. Whether it increased the lifespan of mated females depended upon adult density. Analysis of line-by-treatment-by-temperature interactions revealed hidden trade-offs that would have been missed by other methods. They included a significant trade-off between lifespan and early fecundity. At 25°C high early fecundity was associated with decreased reproductive rates and increased mortality rates 10–15 days later and persisting throughout life, but not at 29.5°C. Correlations with Gompertz coefficients suggested that flies that were heavier at eclosion also aged more slowly and that flies that aged more slowly had higher fecundity late in life at 25°C. The results support the view that lifespan trades off with fecundity and that late fecundity trades off with rate of aging in fruitflies. Genetic engineering is an independent method for the analysis of trade-offs that complements selection experiments.     

耐受偏二甲肼的芦苇变异株系的筛选

四氧化二氮/偏二甲肼(Unsymmetrical dimethyl-hybrazine, 简称UDMH)为常用的航天器双组元液体推进剂。偏二甲肼沸点低,具有“三致”毒性,可在使用过程中释放到环境中,对污染地的生物多样性和人类健康构成威胁,因此迫切需要清除环境中的偏二甲肼。该文采用细胞工程的技术手段,以芦苇幼苗的下胚轴为材料诱导愈伤组织,继而通过逐步提高筛选压力选育出可耐受偏二甲肼的变异细胞系,再诱导变异细胞系分化,为可治理含偏二甲肼的废水的人工湿地处理系统的构建提供理想的工程植物。结果表明:芦苇的愈伤组织在含有0.5 mg·L^-1 2,4-D的MS培养基上生长良好;偏二甲肼对该愈伤组织生长的半致死剂量为16.3 mmol·L^-1;在分别含有1.63、3.26和8.15 mmol·L^-1偏二甲肼的筛选培养基上进行3~6次继代培养后,可得到较稳定的抗性细胞系,培养23 d后的相对生长量分别为对照的90.4%、84.3%和43.4%,培养43 d后的相对生长量分别为对照的95.6%、91.7%和46.8%;但是只有前两类抗性细胞系可在含0.1 g·L^-1 KT、0.01 g·L^-1 NAA和相应浓度的偏二甲肼的MS培养基中诱导分化;将绿色再生苗转移到不含激素但含偏二甲肼的培养基上强化根的生长,再经过35 d左右的适应性驯化,70%以上的再生苗可成功地转移至温室中培养,为日后人工湿地系统的构建奠定了基础。     

新工科背景下生物反应工程课程的多元化教学模式改革与探索

生物反应工程作为一门理论性与应用性都很强的专业课程,在生物工程等相关专业的课程设置中处于桥梁和纽带地位,对新型应用型工科人才的培养发挥着重要的作用.但由于该课程中公式等抽象理论知识过多,导致学生学习效率十分低下.因此,为了适应新工科教育背景下对创新型人才培养的需求,提高学生的学习兴趣和积极性,并培养学生的自主学习等创新...