苏芸金杆菌发酵生产工艺改进研究：Ⅰ芽孢接种和营养体接种比较

苏芸金杆菌液体深层发酵中用营养体接种二级发酵工艺和用二级发酵初期培养物作种子进行三级发酵工艺是可行的。试验结果表明：二级发酵种子罐营养体最佳移种茵龄为９ｈ左右,此时营养体数量多、整齐、染色均匀,显微镜下菌体有折光点存在,三级接种营养体菌龄约为４ｈ左右,菌体数量多,同步率高,少量染色不均匀。发酵液含菌数和发酵产品毒力均达到芽抱接种相同水平,但生产周期明显缩短４－５ｈ,因此相应提高发酵罐生产能力２０％。     

白细胞介素对大鼠海马培养神经元膜电特性的影响

用细胞内微电极记录方法研究了重组人白细胞介素－１β（ｒｈＩＬ－１β）和重组人白细胞介素－２（ｒｈｉＬ－２）对分散培养的新生大鼠海马神经元膜电性质的影响。采用压力脉冲微量给药技术将白介素施加于所记录的细胞表面。结果发现：１００Ｕ／ｍｌ浓度的ｒｈＩＬ－１β使受作用的海马神经元超极化４．２０±１．８６ｍＶ;１００Ｕ／ｍｌ浓度的ｒｈＩＬ－２使５０％受作用的海马神经元去极化１１．１２±３．７１ｍＶ,并伴有强烈的自发放电反应,而１０００Ｕ／ｍｌ浓度的ｒｈＩＬ－２使１００％受作用的神经元超极化３．２５±０．６３ｍＶ,这些神经元的膜阻抗均无明显变化。本实验结果提示ｒｈＩＬ－１β和ｒｈＩＬ－２可显著影响体外培养海马神经元的膜兴奋性。     

天花粉蛋白与FMP复合物的晶体结构

用浸泡法得到了天花粉蛋白（ＴＣＳ）与ＦＭＰ复合物的晶体,在ＳＩＭＥＮＮＳＸ－２００Ｂ面探测器系统上收集了一套２．０分辨率的Ｘ射线衍射数据。用同晶差值傅立叶法解析了复合物的结构,经Ｘ—ＰＬＯＲ程序修正得到了ＴＣＳ—ＦＭＰ复合物的分子结构并找出了１９７个水分子,最后的Ｒ因子为０．１７２,键长和键角的ＲＭＳ偏差分别为０．０１５和２．９２２度。ＴＣＳ—ＦＭＰ复合物中,ＦＭＰ与天花粉蛋白分子有较好的结合,其结合位置正处于根据三维结构和突变体信息推测的Ｎ一糖苷酶活性口袋之中。它的类嘌呤环夹在Ｙ７０和Ｙ１１１两个侧链环之间,与Ｙ７０环近乎平行,其Ｎ７和Ｎ６分别与ＴＣＳ分子的Ｇ１０９４羰基氧和Ｉ７１的Ｎ成氢键,Ｎ３靠近Ｒ１６３的侧链,其磷酸根则伸向活性口袋的底部,与Ｅ１８９、Ｅ１６０和Ｒ１６３等残基作用。     

湖羊瘤胃微生物GH9家族葡聚糖酶基因IDSGLUC9-25的表达与功能表征

【目的】葡聚糖酶是饲用添加剂的重要成分,本研究旨在从湖羊消化道微生物中挖掘性质优良的GH9家族葡聚糖酶基因,用于研发新型饲用酶制剂。【方法】从湖羊瘤胃微生物cDNA中扩增IDSGLUC9-25基因,在大肠杆菌中进行异源表达,对重组蛋白进行诱导表达和纯化,研究重组蛋白的酶学性质和底物水解模式。【结果】IDSGLUC9-25基因编码527个氨基酸,包含一个CelD_N结构和一个GH9家族催化结构域;重组蛋白rIDSGLUC9-25分子量约为62.7 kDa,最适反应温度和pH分别为40℃和6.0,在30-50℃下活性较高,在pH 4.0-8.0范围内能够保持较高的稳定性,经pH 4.0-8.0缓冲液处理1 h后残余活性均大于90%;底物谱分析表明,rIDSGLUC9-25能催化大麦β-葡聚糖、苔藓地衣多糖、魔芋胶和木葡聚糖,比活性分别为(443.55±24.48)、(65.56±5.98)、(122.37±2.85)和(159.16±7.73) U/mg;利用薄层色谱法(thin layer chromatography, TLC)和高效液相色谱法(high performance liquid chromatography, HPLC)分析水解产物发现,rIDSGLUC9-25降解大麦葡聚糖主要生成纤维三糖(占总还原糖64.19%±1.19%)和纤维四糖(占总还原糖26.24%±0.12%),催化地衣多糖主要生成纤维三糖(占总还原糖78.46%±0.89%)。【结论】本研究报道了一种来自密螺旋体属细菌的内切β-1,4-葡聚糖酶IDSGLUC9-25 (EC 3.2.1.4),能高效催化多糖底物生成纤维三糖和纤维四糖,为研发饲用酶制剂和制备低聚寡糖建立基础。     

肺癌患者胸水游离氨基酸分析

本文对11例肺癌患者胸水13种游离氨基酸作了分析,并与28例正常人血浆游离氨基酸水平作了对照,结果表明:肺癌患者胸水的必需及非必需氨基酸普遍高于正常人血浆游离氨基酸,但其胸水谷氨酰胺水平则明显低于正常人血浆水平。     

Complete ammonia oxidization in agricultural soils: High ammonia fertilizer loss but low N2O production

The contribution of agriculture to the sustainable development goals requires climate-smart and profitable farm innovations. Increasing the ammonia fertilizer applications to meet the global food demands results in high agricultural costs, environmental quality deterioration, and global warming, without a significant increase in crop yield. Here, we reported that a third microbial ammonia oxidation process, complete ammonia oxidation (comammox), is contributing to a significant ammonia fertilizer loss (41.9 ± 4.8%) at the rate of 3.53 ± 0.55 mg N kg⁻¹ day⁻¹ in agricultural soils around the world. The contribution of comammox to ammonia fertilizer loss, occurring mainly in surface agricultural soil profiles (0–0.2 m), was equivalent to that of bacterial ammonia oxidation (48.6 ± 4.5%); both processes were significantly more important than archaeal ammonia oxidation (9.5 ± 3.6%). In contrast, comammox produced less N₂O (0.98 ± 0.44 μg N kg⁻¹ day⁻¹, 11.7 ± 3.1%), comparable to that produced by archaeal ammonia oxidation (16.4 ± 4.4%) but significantly lower than that of bacterial ammonia oxidation (72.0 ± 5.1%). The efficiency of ammonia conversion to N₂O by comammox (0.02 ± 0.01%) was evidently lower than that of bacterial (0.24 ± 0.06%) and archaeal (0.16 ± 0.04%) ammonia oxidation. The comammox rate increased with increasing soil pH values, which is the only physicochemical characteristic that significantly influenced both comammox bacterial abundance and rates. Ammonia fertilizer loss, dominated by comammox and bacterial ammonia oxidation, was more intense in soils with pH >6.5 than in soils with pH <6.5. Our results revealed that comammox plays a vital role in ammonia fertilizer loss and sustainable development in agroecosystems that have been previously overlooked for a long term.     

Deciphering RNA m6A regulation in aging: Perspectives on current advances and future directions

N⁶-methyladenosine (m⁶A) is a dynamic and reversible RNA modification that has emerged as a crucial player in the life cycle of RNA, thus playing a pivotal role in various biological processes. In recent years, the potential involvement of RNA m⁶A modification in aging and age-related diseases has gained increasing attention, making it a promising target for understanding the molecular mechanisms underlying aging and developing new therapeutic strategies. This Perspective article will summarize the current advances in aging-related m⁶A regulation, highlighting the most significant findings and their implications for our understanding of cellular senescence and aging, and the potential for targeting RNA m⁶A regulation as a therapeutic strategy. We will also discuss the limitations and challenges in this field and provide insights into future research directions. By providing a comprehensive overview of the current state of the field, this Perspective article aims to facilitate further advances in our understanding of the molecular mechanisms underlying aging and to identify new therapeutic targets for aging-related diseases.     

Exercise enhancement by RGS14 disruption is mediated by brown adipose tissue

Enhanced exercise capacity is not only a feature of healthful aging, but also a therapy for aging patients and patients with cardiovascular disease. Disruption of the Regulator of G Protein Signaling 14 (RGS14) in mice extends healthful lifespan, mediated by increased brown adipose tissue (BAT). Accordingly, we determined whether RGS14 knockout (KO) mice exhibit enhanced exercise capacity and the role of BAT in mediating exercise capacity. Exercise was performed on a treadmill and exercise capacity was assessed by maximal running distance and work to exhaustion. Exercise capacity was measured in RGS14 KO mice and their wild types (WT), and also in WT mice with BAT transplantation from RGS14 KO mice or from other WT mice. RGS14 KO mice demonstrated 160 ± 9% increased maximal running distance and 154 ± 6% increased work to exhaustion, compared to WT mice. RGS14 KO BAT transplantation to WT mice, resulted in a reversal of phenotype, with the WT mice receiving the BAT transplant from RGS14 KO mice demonstrating 151 ± 5% increased maximal running distance and 158 ± 7% increased work to exhaustion, at three days after BAT transplantation, compared to RGS14 KO donors. BAT transplantation from WT to WT mice also resulted in increased exercise performance, but not at 3 days, but only at 8 weeks after transplantation. The BAT induced enhanced exercise capacity was mediated by (1) mitochondrial biogenesis and SIRT3; (2) antioxidant defense and the MEK/ERK pathway, and increased hindlimb perfusion. Thus, BAT mediates enhanced exercise capacity, a mechanism more powerful with RGS14 disruption.