疏水性氨基酸的羟基化研究进展

羟基化氨基酸在生物技术和分子生物学中具有独特价值,具有抗真菌、抗菌、抗病毒和抗癌的特性。通过比较化学合成与生物催化合成羟基氨基酸的异同,选择具有高对映结构选择性的生物催化合成方法成为羟基氨基酸合成的首选。生物催化实现疏水性氨基酸的羟基化和羟化酶紧密相关,而羟化酶又是单核非血红素Fe(Ⅱ)和α-酮戊二酸依赖型双加氧酶(Fe/αKGDs)的一种,Fe/αKGDs存在共性催化机制。因此,疏水性氨基酸在被催化的过程中,会利用关键中间体高价铁-超氧复合体(Fe(Ⅳ)=O)引起多种氧化转化,从而完成羟基化过程。文中就疏水性氨基酸的羟基化合成及功能应用,尤其是(2S,3R,4S)-4-羟基-异亮氨酸(4-HIL)和羟脯氨酸,进行了详细的阐述,探讨了Fe/αKGDs的共性催化反应机制,并对羟基氨基酸在基础研究和工业中的应用进行了综述。     

An outcome model for human bladder cancer: A comprehensive study based on weighted gene co‐expression network analysis

The precision evaluation of prognosis is crucial for clinical treatment decision of bladder cancer (BCa). Therefore, establishing an effective prognostic model for BCa has significant clinical implications. We performed WGCNA and DEG screening to initially identify the candidate genes. The candidate genes were applied to construct a LASSO Cox regression analysis model. The effectiveness and accuracy of the prognostic model were tested by internal/external validation and pan‐cancer validation and time‐dependent ROC. Additionally, a nomogram based on the parameter selected from univariate and multivariate cox regression analysis was constructed. Eight genes were eventually screened out as progression‐related differentially expressed candidates in BCa. LASSO Cox regression analysis identified 3 genes to build up the outcome model in E‐MTAB‐4321 and the outcome model had good performance in predicting patient progress free survival of BCa patients in discovery and test set. Subsequently, another three datasets also have a good predictive value for BCa patients' OS and DFS. Time‐dependent ROC indicated an ideal predictive accuracy of the outcome model. Meanwhile, the nomogram showed a good performance and clinical utility. In addition, the prognostic model also exhibits good performance in pan‐cancer patients. Our outcome model was the first prognosis model for human bladder cancer progression prediction via integrative bioinformatics analysis, which may aid in clinical decision‐making.     

New class of IMP cyclohydrolases in Methanococcus jannaschii

The enzyme responsible for observed IMP cyclohydrolase activity in Methanococcus jannaschii was purified and sequenced: its genetic locus was found to correspond to gene MJ0626. The MJ0626 gene was cloned, and its protein product was expressed in Escherichia coli and shown to catalyze the cyclization of 5-formylamidoimidazole-4-carboxamide ribonucleotide to IMP. The enzyme has no sequence similarity to known enzymes, and its catalytic properties appear distinct from any characterized IMP cyclohydrolase. The purO gene for the enzyme is currently found only in the domain ARCHAEA:     

凋落物添加和移除对杉木人工林土壤水解酶活性及其化学计量比的影响

土壤生态酶化学计量比可用于评估微生物碳(C)、氮(N)和磷(P)养分的需求状况。以往从凋落物输入量变化对生态酶化学计量比的角度来探讨杉木人工林土壤养分状况的研究较少。以亚热带杉木人工林为研究对象,采用随机区组实验设计,对12块杉木人工林样地进行3种凋落物处理(凋落物添加(LA);凋落物移除(LR);对照(CK)),通过测定土壤C、N和P水解酶(β-葡糖苷酶(BG)、半纤维素酶(CB)、β-乙酰葡糖胺糖苷酶(NAG)、亮氨酸氨基肽酶(LAP)和酸性磷酸酶(AP))的活性及土壤理化性质,探讨凋落物添加和移除对杉木人工林土壤养分状况的影响。结果表明:LR显著抑制了AP、BG、CB、NAG和LAP活性,同时降低了土壤含水量(SMC)、有机碳(SOC)、总氮(TN)、总磷(TP)和有效氮含量;而LA对以上指标均存在显著的正效应,表明凋落物输入量变化主要是通过改变土壤水分和养分状况来影响水解酶的活性。LR和CK处理下土壤酶活性比ln(BG+CB):ln(AP)和ln(NAG+LAP):ln(AP)均低于全球尺度上土壤酶活性比ln(BG+CB):ln(AP)(0.62)和ln(NAG+LAP):ln...     

基于生态供给-消耗平衡关系的中尼廊道地区生态承载力研究

中尼廊道地区作为南亚通道建设的关键环节,具有生态环境脆弱且对生态资源依赖性强的基本特征,中尼廊道建设占用生态空间,将增加生态系统对农牧业生产活动的承载压力;因此,开展中尼廊道生态承载力研究对青藏高原生态保护和南亚通道建设具有重要的实践价值和指导意义。本文基于生态供给与消耗平衡关系,通过计算农牧业生产对生态系统净初级生产力的消耗,评估中尼廊道地区的生态承载力,结果表明：（1）中尼廊道地区生态承载力处于富富有余状态,2015年有7个县域生态承载力处于临界超载或平衡有余状态;整体上生态系统尚有较大的生态空间来支撑中尼廊道建设,但建设可能給局部县域带来超载风险。（2）中尼廊道地区生态系统供给量处于稳定波动状态,畜牧业生态消耗在生态消耗中占主导地位（占比超80%）;得益于2005年起实施的草畜平衡政策,中尼廊道地区及其24个县域生态消耗量呈下降趋势,农牧业生产活动对生态系统造成的压力降低,生态承载状态向优发展;但地处粮食主产区、口岸区、城市化辐射区的7个县域,在本地及周边地区日益增长的对农牧业产品的生活需求驱动下生态消耗量处于增加态势。（3）若以实现生物多样性公约所倡导的50%生态资源得到保护为生态保护目标,目前中尼廊道农牧业生产模式产生的生态消耗将超出其可利用供给,区域可持续发展需要关注如何通过中尼廊道建设带动当地产业结构调整以缓解农牧业生产对生态系统的压力。     

滇南喀斯特地区灌木群落和人工林土壤元素化学计量特征

在云南喀斯特地区,为提升退化灌木群落的生态系统服务功能,营造了不同树种的人工林分。这些人工林分如何影响土壤化学性质还未得到充分认识。以云南泸西县灌木群落及三种常见人工林(云南松(Pinus yunnanensis)、赤杨(Alnus japonica)和侧柏(Platycladus orientalis))土壤为研究对象探讨喀斯特地区在人工林建造后土壤的13种元素全量、可利用性含量和化学计量学特征变异格局,为喀斯特石漠化治理提供理论依据。结果表明,1)基于判别分析,四种群落土壤化学计量特征可以显著区分。土壤Fe、P、K、Mn全量及交换性Ca、交换性Mg和NH_4~+-N对区分四种群落土壤贡献最大。2)四种群落之间相比,侧柏林土壤C、N、S、Na全量和NO_3~--N含量均低于其他三种群落,土壤肥力较低;赤杨林铵态氮含量最高;云南松林有效Fe、有效Cu含量/N、C素具有显著相关性,占所有元素对数的38.5%,说明该地土壤元素积累的相互依赖性。与灌木群落相比,人工林土壤元素全量和可利用性含量相关性比例均更高。这些研究结果对今后基于适地适树人工林营造、生态系统服务功能提升和经营利用,均具有重要指导意义。     

营养物质对桑黄菌丝生物量及胞外多糖产量的影响

研究了不同碳源、氮源和无机盐对桑黄深层培养菌丝生物量和胞外多糖产量的影响,结果表明：在培养温度为26℃、摇床转速为160r／min、发酵时间为10d的条件下,以桑黄菌丝生物量为指标,最适碳源、氮源和无机盐分别是果糖或葡萄糖、酵母粉或酵母膏、KH2P04或MgSO4·7H2O;菌丝生物量分别达到1．51、1．31、1．69、1．52、1．52、1．36g／100mL;以胞外多糖为指标,最适碳源、氮源和无机盐分别是葡萄糖、酵母膏、ZnSO4·7H2O,胞外多糖产量分别达到0．49、0．45、0．22g／100mL。     

Cloning, characterization, and engineering of fungal L-arabinitol dehydrogenases

L-Arabinitol 4-dehydrogenase (LAD) catalyzes the conversion of L-arabinitol to L-xylulose with concomitant NAD⁺ reduction in fungal L-arabinose catabolism. It is an important enzyme in the development of recombinant organisms that convert L-arabinose to fuels and chemicals. Here, we report the cloning, characterization, and engineering of four fungal LADs from Penicillium chrysogenum, Pichia guilliermondii, Aspergillus niger, and Trichoderma longibrachiatum, respectively. The LAD from P. guilliermondii was inactive, while the other three LADs were NAD⁺-dependent and showed high catalytic activities, with P. chrysogenum LAD being the most active. T. longibrachiatum LAD was the most thermally stable and showed the maximum activity in the temperature range of 55–65°C with the other LADs showed the maximum activity in the temperature range of 40–50°C. These LADs were active from pH 7 to 11 with an optimal pH of 9.4. Site-directed mutagenesis was used to alter the cofactor specificity of these LADs. In a T. longibrachiatum LAD mutant, the cofactor preference toward NADP⁺ was increased by 2.5 × 10⁴-fold, whereas the cofactor preference toward NADP⁺ of the P. chrysogenum and A. niger LAD mutants was also drastically improved, albeit at the expense of significantly reduced catalytic efficiencies. The wild-type LADs and their mutants with altered cofactor specificity could be used to investigate the functionality of the fungal L-arabinose pathways in the development of recombinant organisms for efficient microbial L-arabinose utilization.     

Selective reduction of xylose to xylitol from a mixture of hemicellulosic sugars

The biocatalytic reduction of d-xylose to xylitol requires separation of the substrate from l-arabinose, another major component of hemicellulosic hydrolysate. This step is necessitated by the innate promiscuity of xylose reductases, which can efficiently reduce l-arabinose to l-arabinitol, an unwanted byproduct. Unfortunately, due to the epimeric nature of d-xylose and l-arabinose, separation can be difficult, leading to high production costs. To overcome this issue, we engineered an E. coli strain to efficiently produce xylitol from d-xylose with minimal production of l-arabinitol byproduct. By combining this strain with a previously engineered xylose reductase mutant, we were able to eliminate l-arabinitol formation and produce xylitol to near 100% purity from an equiweight mixture of d-xylose, l-arabinose, and d-glucose.