马里亚纳海沟可培养水生细菌的多样性

【目的】马里亚纳海沟是地球表面最深的海沟,环境极端多样,如高压、低温及无光,拥有独特的微生物资源。本研究旨在探究马里亚纳海沟不同深度水生细菌形态特征并挖掘可培养细菌资源。【方法】采集马里亚纳海沟7个层位海水(2–8727 m),利用原子力显微镜与扫描电镜观察水生微生物的形态特征;采用2种常规培养基(1/5×2216E和1/30×2216E)及6种选择性培养基(有机碳氮组合),结合切向流与高压富集培养进行水生细菌分离与鉴定。【结果】从不同深度水样中发现多种大小不一的细菌类群(130 nm–1.5μm),以球菌和杆菌为主。在表层水体中常见颗粒附着的细菌,在深层水体中常见自由游动的细菌。共鉴定365株可培养水生细菌,隶属于3个门、31个属与56个种。γ-变形菌纲(Gammaproteobacteria)是绝对优势类群(占据可培养细菌总数的62.7%),相对丰度在深层水体中高于浅层。交替单胞菌属(Alteromonas,21.8%)和亚硫酸杆菌属(Sulfitobacter,19.1%)是主要优势属,在浅层水体中占绝对优势。稀释的2216E与氨基酸培养基对海杆菌属的选择性更好,葡萄糖-甘露糖培养基与牛磺酸-乙醇酸培养基对稀有细菌的选择性更好。7株菌(5种)是潜在的新型细菌。此外,通过切向流富集培养与压力筛选培养分别分离得到70株(22属)可通过0.22-μm细菌(0.22-μm-passable bacteria)与33株(8属)耐压细菌。【结论】马里亚纳海沟不同深度水样中不同营养利用型细菌、可通过0.22-μm细菌与耐压细菌及其形态均具有丰富的多样性。本研究所获得的不同类型的细菌菌株为研究细菌在马里亚纳海沟中生物地球化学功能及其营养类型差异和高压适应机制奠定了菌株基础。     

导电材料强化微生物直接种间电子传递产甲烷的研究进展

厌氧条件下,微生物可以通过厌氧代谢产生甲烷(CH_4),由此衍生的厌氧消化技术可实现能源的回收利用。产CH_4的关键步骤是刺激发酵细菌和产甲烷古菌之间的有效电子转移,电活性微生物可以取代传统的氢/甲酸盐实现直接种间电子传递,其电子传递效率更高。添加导电材料可以促进直接种间电子传递并提高CH_4产率,是一种更有效的强化电子传递方式。本文在梳理直接种间电子传递发展和机理的基础上,综述了常见的促进直接种间电子传递的碳基和铁基导电材料,对其结构特征、电子传递机理、强化产CH_4和中间产物消耗等方面进行了系统总结。旨在为导电材料促进直接种间电子传递的研究提供参考,并探讨了未来可能的研究方向。     

产大米蛋白水解酶的菌株筛选、酶学性质及制备大米寡肽

【目的】旨在分离、筛选并鉴定具有大米蛋白降解作用的菌株及其关键蛋白酶,为高效制备大米寡肽提供制备酶及最优制备条件。【方法】以"水解圈"为评价指标,从粮食仓库附近土壤筛选获得具有降解大米蛋白能力的菌株;通过16S rRNA序列分析确定菌株归属;利用单因素实验获得最佳氮源并初步分析酶学性质;利用HPLC检测寡肽得率,并对制备条件进一步优化。【结果】经鉴定具有大米蛋白降解作用的菌株为沙雷氏菌(Serratia sp. JWG-D15),以大米蛋白为氮源培养菌株JWG-D15,蛋白酶So PRO产量最高,其最适温度40℃,最适pH 8.0;在加酶量20 U/mg,大米蛋白浓度40 mg/mL,40℃,4 h条件下寡肽得率最高72.38%。【结论】以大米蛋白为氮源培养菌株JWG-D15,蛋白酶So PRO产量最高;蛋白酶So PRO制得的大米寡肽,其得率是目前行业最高。本研究既丰富了大米寡肽的制备用酶的种类,又为深入大米寡肽产业化提供一定理论基础。     

Are cultured human myotubes far from home?

Satellite cells can be isolated from skeletal muscle biopsies, activated to proliferating myoblasts and differentiated into multinuclear myotubes in culture. These cell cultures represent a model system for intact human skeletal muscle and can be modulated ex vivo. The advantages of this system are that the most relevant genetic background is available for the investigation of human disease (as opposed to rodent cell cultures), the extracellular environment can be precisely controlled and the cells are not immortalized, thereby offering the possibility of studying innate characteristics of the donor. Limitations in differentiation status (fiber type) of the cells and energy metabolism can be improved by proper treatment, such as electrical pulse stimulation to mimic exercise. This review focuses on the way that human myotubes can be employed as a tool for studying metabolism in skeletal muscles, with special attention to changes in muscle energy metabolism in obesity and type 2 diabetes.     

Dissecting the in vivo assembly of the 30S ribosomal subunit reveals the role of RimM and general features of the assembly process

Ribosome biogenesis is a tightly regulated, multi-stepped process. The assembly of ribosomal subunits is a central step of the complex biogenesis process, involving nearly 30 protein factors in vivo in bacteria. Although the assembly process has been extensively studied in vitro for over 40 years, very limited information is known for the in vivo process and specific roles of assembly factors. Such an example is ribosome maturation factor M (RimM), a factor involved in the late-stage assembly of the 30S subunit. Here, we combined quantitative mass spectrometry and cryo-electron microscopy to characterize the in vivo 30S assembly intermediates isolated from mutant Escherichia coli strains with genes for assembly factors deleted. Our compositional and structural data show that the assembly of the 3′-domain of the 30S subunit is severely delayed in these intermediates, featured with highly underrepresented 3′-domain proteins and large conformational difference compared with the mature 30S subunit. Further analysis indicates that RimM functions not only to promote the assembly of a few 3′-domain proteins but also to stabilize the rRNA tertiary structure. More importantly, this study reveals intriguing similarities and dissimilarities between the in vitro and the in vivo assembly pathways, suggesting that they are in general similar but with subtle differences.     

Stimulation of σ1-receptor restores abnormal mitochondrial Ca mobilization and ATP production following cardiac hypertrophy

Background

We previously reported that the σ₁-receptor (σ₁R) is down-regulated following cardiac hypertrophy and dysfunction in transverse aortic constriction (TAC) mice. Here we address how σ₁R stimulation with the selective σ₁R agonist SA4503 restores hypertrophy-induced cardiac dysfunction through σ₁R localized in the sarcoplasmic reticulum (SR).

Methods

We first confirmed anti-hypertrophic effects of SA4503 (0.1–1 μM) in cultured cardiomyocytes exposed to angiotensin II (Ang II). Then, to confirm the ameliorative effects of σ₁R stimulation in vivo, we administered SA4503 (1.0 mg/kg) and the σ₁R antagonist NE-100 (1.0 mg/kg) orally to TAC mice for 4 weeks (once daily).

Results

σ₁R stimulation with SA4503 significantly inhibited Ang II-induced cardiomyocyte hypertrophy. Ang II exposure for 72 h impaired phenylephrine (PE)-induced Ca^2 + mobilization from the SR into both the cytosol and mitochondria. Treatment of cardiomyocytes with SA4503 largely restored PE-induced Ca^2 + mobilization into mitochondria. Exposure of cardiomyocytes to Ang II for 72 h decreased basal ATP content and PE-induced ATP production concomitant with reduced mitochondrial size, while SA4503 treatment completely restored ATP production and mitochondrial size. Pretreatment with NE-100 or siRNA abolished these effects. Chronic SA4503 administration also significantly attenuated myocardial hypertrophy and restored ATP production in TAC mice. SA4503 administration also decreased hypertrophy-induced impairments in LV contractile function.

Conclusions

σ₁R stimulation with the specific agonist SA4503 ameliorates cardiac hypertrophy and dysfunction by restoring both mitochondrial Ca^2 + mobilization and ATP production via σ₁R stimulation.

General significance

Our observations suggest that σ₁R stimulation represents a new therapeutic strategy to rescue the heart from hypertrophic dysfunction.     

Honokiol: A non-adipogenic PPARγ agonist from nature

Background

Peroxisome proliferator-activated receptor gamma (PPARγ) agonists are clinically used to counteract hyperglycemia. However, so far experienced unwanted side effects, such as weight gain, promote the search for new PPARγ activators.

Methods

We used a combination of in silico, in vitro, cell-based and in vivo models to identify and validate natural products as promising leads for partial novel PPARγ agonists.

Results

The natural product honokiol from the traditional Chinese herbal drug Magnolia bark was in silico predicted to bind into the PPARγ ligand binding pocket as dimer. Honokiol indeed directly bound to purified PPARγ ligand-binding domain (LBD) and acted as partial agonist in a PPARγ-mediated luciferase reporter assay. Honokiol was then directly compared to the clinically used full agonist pioglitazone with regard to stimulation of glucose uptake in adipocytes as well as adipogenic differentiation in 3T3-L1 pre-adipocytes and mouse embryonic fibroblasts. While honokiol stimulated basal glucose uptake to a similar extent as pioglitazone, it did not induce adipogenesis in contrast to pioglitazone. In diabetic KKAy mice oral application of honokiol prevented hyperglycemia and suppressed weight gain.

Conclusion

We identified honokiol as a partial non-adipogenic PPARγ agonist in vitro which prevented hyperglycemia and weight gain in vivo.

General significance

This observed activity profile suggests honokiol as promising new pharmaceutical lead or dietary supplement to combat metabolic disease, and provides a molecular explanation for the use of Magnolia in traditional medicine.