Mixotrophy in a Local Strain of Nannochloropsis granulata for Renewable High-Value Biomass Production on the West Coast of Sweden

A local strain of Nannochloropsis granulata (Ng) has been reported as the most productive microalgal strain in terms of both biomass yield and lipid content when cultivated in photobioreactors that simulate the light and temperature conditions during the summer on the west coast of Sweden. To further increase the biomass and the biotechnological potential of this strain in these conditions, mixotrophic growth (i.e., the simultaneous use of photosynthesis and respiration) with glycerol as an external carbon source was investigated in this study and compared with phototrophic growth that made use of air enriched with 1–2% CO₂. The addition of either glycerol or CO₂-enriched air stimulated the growth of Ng and theproduction of high-value long-chain polyunsaturated fatty acids (EPA) as well as the carotenoid canthaxanthin. Bioassays in human prostate cell lines indicated the highest antitumoral activity for Ng extracts and fractions from mixotrophic conditions. Metabolomics detected betaine lipids specifically in the bioactive fractions, suggesting their involvement in the observed antitumoral effect. Genes related to autophagy were found to be upregulated by the most bioactive fraction, suggesting a possible therapeutic target against prostate cancer progression. Taken together, our results suggest that the local Ng strain can be cultivated mixotrophically in summer conditions on the west coast of Sweden for the production of high-value biomass containing antiproliferative compounds, carotenoids, and EPA.     

代谢组学及其在动物营养中的应用

代谢组学是对内源性代谢物的综合评估,可以实现来自生物样品的代谢物及其相关代谢途径的分离、检测、表征和定量,以此来反映生物机体的营养代谢变化、营养状况甚至一些疾病的发展程度等。然而,代谢组学技术在动物营养中的研究起步较晚,目前尚处于起始阶段,不过随着代谢组学研究平台的不断完善,该技术在动物营养中的应用价值必将日渐突出。本文就代谢组学的概念特点、分析技术及其在动物营养中的应用作一综述。     

Investigation of Interspecies Interactions within Marine Micromonosporaceae Using an Improved Co-Culture Approach

With respect to bacterial natural products, a significant outcome of the genomic era was that the biosynthetic potential in many microorganisms surpassed the number of compounds isolated under standard laboratory growth conditions, particularly among certain members in the phylum Actinobacteria. Our group, as well as others, investigated interspecies interactions, via co-culture, as a technique to coax bacteria to produce novel natural products. While co-culture provides new opportunities, challenges exist and questions surrounding these methods remain unanswered. In marine bacteria, for example, how prevalent are interspecies interactions and how commonly do interactions result in novel natural products? In an attempt to begin to answer basic questions surrounding co-culture of marine microorganisms, we have tested both antibiotic activity-based and LC/MS-based methods to evaluate Micromonosporaceae secondary metabolite production in co-culture. Overall, our investigation of 65 Micromonosporaceae led to the identification of 12 Micromonosporaceae across three genera that produced unique metabolites in co-culture. Our results suggest that interspecies interactions were prevalent between marine Micromonosporaceae and marine mycolic acid-containing bacteria. Furthermore, our approach highlights a sensitive and rapid method for investigating interspecies interactions in search of novel antibiotics, secondary metabolites, and genes.     

Taurine alone or in combination with fish protein hydrolysate affects growth performance,taurine transport and metabolism in juvenile turbot (Scophthalmus maximus L.)

The study was conducted to investigate the effects of taurine (Tau) alone or in combination with fish protein hydrolysate (FPH) on growth performance, the expression of Tau transporter (TauT) and metabolic profile in juvenile turbot. FM, FPH0, FPH0+T, FPH10 and FPH10+T diets, respectively, contained 300, 150, 150, 80, and 80 g/kg fishmeal. FPH10 and FPH10+T diets contained 62 g/kg FPH. FPH0+T and FPH10+T diets were, respectively, prepared by supplementing the FPH0 and FPH10 diet formulations with 8 g/kg Tau. Specific growth rate was the highest in FM group and the lowest in FPH10 group. TauT mRNA levels in fish fed Tau supplemented diets were significantly lower than that in Tau unsupplemented diets. NMR‐based metabolomics analysis showed that Tau contents in liver of FPH0+T and FPH10+T were significantly higher than that of FM, FPH0 and FPH10. In muscle, Tau contents were significantly decreased in the FPH10+T versus FPH0 and the FPH10+T versus FPH10 comparisons. In conclusion, 62 g/kg FPH to replace fishmeal may not affect Tau synthesis, transport and metabolism. However, Tau supplemented alone or in combination with a certain level of FPH could reduce the requirement for Tau synthesis and transport and increased Tau levels in muscle and liver.     

基于血浆非靶代谢组学探究急性笼养应激对蛋鸭代谢的影响

随着环保政策改革和养殖业的集约化、规模化发展,蛋鸭叠层式笼养模式的应用越来越广泛,笼养应激对蛋鸭的影响逐渐受到重视。本研究旨在利用非靶向代谢组学分析蛋鸭平养与笼养模式下血浆代谢物的差异。随机挑选平养组(floor-water rearing system, FWR组)和叠层式笼养组(cage-rearing system, CR组)蛋鸭各10只,跖骨静脉采血,通过高效液相色谱-质谱(HPLC-HRMS)非靶向代谢组学技术获得20个在不同饲养模式下的血浆代谢物图谱,经T检验等分析,取差异倍数(Fold-change)ratio≥2或ratio≤1/2、变量重要性(variable importance in projection, VIP)>1、q值<0.05为筛选条件得到显著差异代谢物并作通路富集分析。结果共筛选到差异代谢物30种,其中上调表达的差异代谢物14种(P<0.05),下调表达的差异代谢物16种(P<0.05)。差异代谢物种类主要有甘油磷脂(glycerophospholipids, 16种)、脂肪酰基(fatty acyls, 4种)、羧酸及其衍生物...     

γ-氨基丁酸和2-乙酰-1-吡咯啉代谢通路在水稻籽粒发育过程中的变化分析

【目的】结合代谢组和转录组技术,分析香稻和非香稻不同发育时期籽粒中γ-氨基丁酸（GABA）和2-乙酰-1-吡咯啉（2AP）代谢通路主要化合物和基因的动态变化,为高含量GABA和2AP的水稻育种提供理论依据。【方法】本研究收集优质香籼稻美香占2号和优质籼稻黄华占花后8 d（乳熟期）、花后15 d（蜡熟期）、30 d（完熟期）的脱壳籽粒和花后40 d收获的籽粒的精米,对各样本采用LC-MS/MS和RNA-seq技术进行代谢产物检测和转录表达分析,分析两个水稻品种GABA和2AP代谢通路相关化合物的空间分布以及含量变化特征。【结果】代谢组数据PCA分析表明两个水稻品种乳熟期籽粒和花后40 d收获的籽粒的精米明显分离,蜡熟期和完熟期籽粒分离不明显。代谢组共鉴定出623种代谢物,四个时期差异代谢物合计161个。GABA和2AP通路中9个代谢物被检出,两个水稻品种的相关代谢物变化趋势相似,其中腐胺主要定位于籽粒的糊粉层,而亚精胺在内胚乳特异累积。转录组共检出3.3万个左右基因,其中检出GABA通路18个酶中的14个酶对应的基因有较高的表达水平,除Badh2外其他基因均不是差异表达基因,同一基因在两份水稻材料中的表达水平和变化趋势相似,GAD1、GABA-T1、DAO4和PAO4在相应同源基因中表达水平较高。黄华占籽粒中L-精氨酸和腐胺的代谢在蜡熟期已基本结束,美香占2号在籽粒整个发育期具有持续较强的多胺降解水平。美香占2号籽粒具有合成2AP的物质基础,多胺降解途径和谷氨酸-脯氨酸转化通路同时有助于2AP的积累。【结论】水稻籽粒GABA代谢以降解为主,GABA-T1和GABA-T2共同调控GABA的降解。GABA相关代谢物主要集中在糊粉层,黄华占籽粒中代谢物分布和基因表达特征与美香占2号相似,美香占2号籽粒成熟过程中具有持续较高的线粒体活性。     

Metabolomic Tools for Secondary Metabolite Discovery from Marine Microbial Symbionts

Marine invertebrate-associated symbiotic bacteria produce a plethora of novel secondary metabolites which may be structurally unique with interesting pharmacological properties. Selection of strains usually relies on literature searching, genetic screening and bioactivity results, often without considering the chemical novelty and abundance of secondary metabolites being produced by the microorganism until the time-consuming bioassay-guided isolation stages. To fast track the selection process, metabolomic tools were used to aid strain selection by investigating differences in the chemical profiles of 77 bacterial extracts isolated from cold water marine invertebrates from Orkney, Scotland using liquid chromatography-high resolution mass spectrometry (LC-HRMS) and nuclear magnetic resonance (NMR) spectroscopy. Following mass spectrometric analysis and dereplication using an Excel macro developed in-house, principal component analysis (PCA) was employed to differentiate the bacterial strains based on their chemical profiles. NMR ¹H and correlation spectroscopy (COSY) were also employed to obtain a chemical fingerprint of each bacterial strain and to confirm the presence of functional groups and spin systems. These results were then combined with taxonomic identification and bioassay screening data to identify three bacterial strains, namely Bacillus sp. 4117, Rhodococcus sp. ZS402 and Vibrio splendidus strain LGP32, to prioritize for scale-up based on their chemically interesting secondary metabolomes, established through dereplication and interesting bioactivities, determined from bioassay screening.     

不同温度下褪黑素处理对‘阳光玫瑰'葡萄采后品质的影响

【目的】冷藏是提高葡萄耐储性、延长货架期的有效方法,但低温会降低糖酸和香气含量等果实品质。本研究旨在测定不同温度条件下褪黑素处理对‘阳光玫瑰'葡萄储藏期品质的影响,并探讨其调控储藏品质的代谢基础。【方法】设置室温和1℃两个储藏温度,葡萄采后利用5和50 μmol∙L^-1褪黑素浸果,于不同储藏期取样测定果实品质指标。利用HPLC-MS测定褪黑素含量,利用GC-MS测定香气组分及含量,利用毛细管电泳测定可溶性糖和有机酸含量,利用质构仪测定果实质地,利用广靶代谢组学分析褪黑素处理导致的差异代谢物。【结果】外源褪黑素处理能显著提高葡萄果实褪黑素含量;50 µmol∙L ^-1处理效果显著高于5 µmol∙L ^-1,并且低温能大幅提高褪黑素处理效果,低温、50 mol∙L^-1褪黑素处理的果实中褪黑素含量为室温同浓度处理的2.6倍。低温下,果实失水率降低;在室温和低温储藏下,褪黑素对果实失水率、果皮硬度和果肉硬度未产生显著影响。室温条件下,5 µmol∙L ^-1褪黑素处理显著提高了葡萄糖和果糖含量,而50 µmol∙L ^-1褪黑素产生了相反的结果。与室温储藏相比,低温储藏显著降低了果实糖含量;低温条件下,与对照相比,5 µmol∙L ^-1和50 µmol∙L ^-1褪黑素处理均显著提高了可溶性糖含量,储藏40 d时增幅在19.2%以上。与室温相比,低温提高了可滴定酸尤其是苹果酸含量。低温条件下,两种浓度褪黑素处理较对照显著降低了可滴定酸尤其是苹果酸含量,苹果酸含量降幅超过53.5%,对酒石酸含量影响较小。与室温储藏相比,低温储藏大幅度降低了未经褪黑素处理的果实香气含量。但是,褪黑素处理能有效增加低温条件下果实香气总量及香气组分含量,且以5 µmol∙L ^-1褪黑素处理效果较好,在处理后30和40 d,较低温对照香气总量增幅分别达2.12和1.6倍;同时显著增加了反式-2-己烯醛、里那醇、2,4-二叔丁基苯酚等特征香气含量。低温下对照和5 µmol∙L ^-1褪黑素处理的果实代谢组学分析表明,232种代谢物含量存在差异,涉及的主要代谢途径包括氨基酸生化合成、氨酰基-tRNA生化合成、精氨酸生化合成、丙氨酸、天冬氨酸和谷氨酸代谢及苯丙胺代谢。【结论】与室温储藏相比,低温冷藏降低了部分糖和绝大多数香气物质的含量。褪黑素处理能够显著提高冷藏果实的可溶性糖含量,降低有机酸水平,大幅提高香气水平,能有效提高果实品质,其中5 µmol∙L ^-1褪黑素处理效果更明显。褪黑素主要通过影响氨基酸代谢提高香气含量。