木质纤维素预处理抑制物产生及脱除方法的研究进展

利用纤维素酶将木质纤维素降解成可发酵性糖,然后发酵生产氢气、乙醇、丁醇等生物燃料及高附加值产品,是当今全球研究的热点。预处理是生物质转化过程中至关重要的步骤,而预处理过程中产生的抑制物对木质纤维素后续的酶解和发酵微生物有负面影响。因此了解预处理方法及其过程中产生的抑制物及脱除方法是能否高效转化生物质的基础。文中首先介绍了木质纤维素常用的两类预处理方法即化学法和物理化学法。随后阐述了不同抑制物的产生及其抑制机制,并重点介绍了多种脱毒方法。最后展望了脱除木质纤维素预处理抑制物的研究趋势:应用交联聚乙烯亚胺和金属有机骨架化合物等新型材料脱除抑制物或通过基因工程、代谢工程技术等构建抑制物耐受性菌株等。     

大肠杆菌对木质纤维素水解液抑制物的胁迫耐受性

木质纤维素类生物质是前景广阔的化石原料替代品，其生物炼制可生产生物能源、生物基化学品和生物材料等多种产品，可降低碳排放，有助于实现“双碳”目标，因此受到越来越多的关注。然而，木质纤维素生物炼制需要经过预处理、微生物发酵和产物纯化等多个步骤，其中，预处理过程产生的多种化合物抑制微生物的细胞生长和发酵性能，是制约生物转化效率的瓶颈之一。大肠杆菌是木质纤维素生物炼制常用的宿主，被广泛应用于多种化合物的生产，研究其对木质纤维素水解液中抑制物的耐受性，对于提高木质纤维素生物炼制效率具有重要意义。本文首先介绍了木质纤维素的主要成分和基本结构，对木质纤维素的预处理方法以及预处理后水解液中的主要抑制物种类进行了简单阐述；随后，总结了木质纤维素水解液中几类主要抑制物呋喃类、羧酸类和酚类对大肠杆菌细胞的毒性，以及大肠杆菌对上述抑制物的胁迫响应机制和基于机制的菌株改造靶点；最后，综述了提高大肠杆菌对上述抑制物的胁迫耐受性的菌株改造策略，包括随机突变、实验室适应性进化和组学辅助的理性设计等，为利用代谢工程构建用于木质纤维素生物炼制的高效大肠杆菌菌株提供参考。     

微生物利用木质纤维素的研究进展

木质纤维素原料是世界上最为丰富的资源之一,可用作微生物发酵生产高附加值生物化学品的原料。与传统用于微生物发酵的可食用生物质原料相比,目前微生物利用木质纤维素还存在以下几个关键问题:开发经济有效的木质纤维素预处理工艺、提高微生物对木质纤维素水解液中第二大单糖木糖的有效利用水平、增强微生物对木质纤维素水解液中混糖的综合利用能力以及提高微生物对木质纤维素水解液中糠醛、乙酸等发酵抑制物的耐受能力。综述了近年来国内外针对这几个关键问题的最新研究成果。为今后微生物大规模利用木质纤维素进行商业生产提出了展望和建议。     

木质纤维素预处理液膜分离脱毒技术研究进展

木质纤维素预处理会产生各种抑制物,严重影响后续的酶水解和微生物发酵。因此,对木质纤维素预处理液进行脱毒处理成为木质纤维素高效经济生物转化的前提条件之一。重点介绍了膜分离技术应用于木质纤维素预处理液的脱毒过程所取得的研究进展。这些膜分离技术包括膜萃取、膜吸附、纳滤、反渗透、电渗析、电去离子、膜电容脱盐、渗透汽化和膜蒸馏等。膜分离技术在木质纤维素预处理液脱毒领域具有十分广阔的应用前景。     

纤维素乙醇生物加工过程中的抑制物对酿酒酵母的影响及应对措施

以木质纤维素为原料生产乙醇,预处理是必需的环节,这一过程中不可避免产生了多种对微生物有抑制作用的化合物,这些抑制物主要有3大类:弱酸、呋喃醛类和酚类化合物。这些化合物影响后续乙醇发酵微生物酿酒酵母(Saccharomyces cerevisiae)的生长及发酵性能,降低了乙醇的得率和产量,是木质纤维素原料大规模生产乙醇的一个主要障碍。以下介绍了3类抑制物的形成及作用机制,并介绍了应对抑制物作用、提高酵母发酵能力的措施及研究进展,包括发酵前预处理原料脱毒、通过进化工程驯化菌种或通过对抑制物耐受性相关基因的代谢工程操作提高酿酒酵母耐受性,及通过发酵过程控制减少抑制物影响等。     

玉米芯半纤维素水解液的制备及成分测定

木质纤维素原料是自然界中含量丰富的可再生资源,可在水解糖化后被微生物发酵生产乙醇等有用产品。将玉米芯等木质纤维素类农业废料用稀硫酸水解糖化过程中,受反应条件的影响,水解液中除生成可发酵的单糖外还会产生一些可能对后续乙醇发酵微生物有抑制作用的副产物。本课题采用正交试验优化了制备玉米芯半纤维素水解液的工艺参数,并确立了水解液中的五种主要副产物。     

利用木质纤维素生产丁醇的研究进展

随着化石燃料的逐年减少,以生物质为原料的生物能源研究近年来成为能源领域的研究热点,充分利用可再生生物质为发展经济的生物燃料生产工艺提供了一个极好的机会。与燃料乙醇和生物柴油相比,生物丁醇更具有优越性,以可再生木质纤维素生物质为原料进行发酵生产丁醇在近年来被广泛的研究。对于利用可再生生物质为原料生产丁醇,需要解决原料的选择、产品收率低、抑制物对生产菌株毒性等问题。本文对以木质纤维素生物质为原料进行生物丁醇发酵过程中的原料预处理、抑制物对丁醇生产菌的影响,以及水解液的脱毒和耐抑制物菌株的选育等方面进行综述,并对以木质纤维素生产燃料丁醇所面临的机遇与问题进行了简要评述。     

工业酵母抗逆机理研究进展

工业酵母利用木质纤维素等生物质资源发酵生产醇、酮、醛、酸等各种化合物,是解决人类面临的不可再生资源和能源危机的重要途径,这激发了人们对木质纤维素水解液为原料和环保节能型浓醪发酵技术的极度关注。然而高浓度底物、产物、渗透压、木质纤维素水解液中抑制性物质、发酵过程温度的提高均会抑制微生物生长代谢及发酵性能,这是发酵行业"瓶颈"问题。本文简述了渗透压、温度及抑制性物质对酵母细胞生长的危害,并从胞内稳态平衡、分子水平等方面着重叙述工业酵母对渗透压、温度及抑制性物质的抗逆机制研究进展。     

分别利用产甘油假丝酵母抗逆转录因子CgSTB5和CgSEF1对酿酒酵母菌株糠醛耐受改造

【背景】纤维素是生物转化解决能源问题的主要原料之一,其水解物中存在严重影响抑制菌株生长的糠醛,需脱毒才可应用于发酵,提高菌株耐受性是解决纤维素水解液实际生产应用的关键。【目的】酿酒酵母(Saccharomyces cerevisiae)是主要的纤维素水解液发酵工业菌株,但糠醛耐受性较低,通过分子改造获得具有高糠醛耐受性的菌株。【方法】利用新获得的产甘油假丝酵母(Candidaglycerinogenes)的相关抗逆转录因子CgSTB5、CgSEF1和CgCAS5,通过分子技术进行S.cerevisiae改造,考察其对酿酒酵母糠醛耐受性的影响,并尝试应用于未脱毒纤维素乙醇发酵。【结果】单个表达CgSTB5和CgSEF1的酿酒酵母,通过菌株点板实验表明菌株的糠醛耐受性提高25%以上,并且摇瓶发酵结果显示糠醛降解性能明显提高,生长延滞期明显缩短,S.cerevisiae W303/p414-CgSTB5的未脱毒纤维素乙醇发酵生产效率提高12.5%左右。【结论】转录因子CgSTB5和CgSEF1均能对提高酿酒酵母糠醛耐受性起到重要作用,并且有助于提高酿酒酵母菌株未脱毒纤维素乙醇发酵性能。     

重组大肠杆菌利用废纸水解液发酵产L-乳酸

前期通过基因工程手段,构建了一株大肠杆菌工程菌E.coli WL204,该菌株可以有效利用木糖为底物发酵产L-乳酸。以废纸为发酵原料,研究该菌株利用木质纤维素发酵产乳酸的特性。原料以稀硫酸预处理后,经纤维素酶酶解,得到的水解液用Ca(OH)2脱毒后,接种E.coli WL204,在7L发酵罐中发酵72h,每100g废纸可以产生31g乳酸,糖酸转化率为79%。结果表明,E.coli WL204可以木质纤维素原料为底物发酵生产L-乳酸,具有一定的工业化开发潜力。     

Role for dopamine in malonate-induced damage in vivo in striatum and in vitro in mesencephalic cultures

Defects in mitochondrial energy metabolism have been implicated in the pathology of several neurodegenerative disorders. In addition, the reactive metabolites generated from the metabolism and oxidation of the neurotransmitter dopamine (DA) are thought to contribute to the damage to neurons of the basal ganglia. We have previously demonstrated that infusions of the metabolic inhibitor malonate into the striata of mice or rats produce degeneration of DA nerve terminals. In the present studies, we demonstrate that an intrastriatal infusion of malonate induces a substantial increase in DA efflux in awake, behaving mice as measured by in vivo microdialysis. Furthermore, pretreatment of mice with tetrabenazine (TBZ) or the TBZ analogue Ro 4-1284 (Ro-4), compounds that reversibly inhibit the vesicular storage of DA, attenuates the malonate-induced DA efflux as well as the damage to DA nerve terminals. Consistent with these findings, the damage to both DA and GABA neurons in mesencephalic cultures by malonate exposure was attenuated by pretreatment with TBZ or Ro-4. Treatment with these compounds did not affect the formation of free radicals or the inhibition of oxidative phosphorylation resulting from malonate exposure alone. Our data suggest that DA plays an important role in the neurotoxicity produced by malonate. These findings provide direct evidence that inhibition of succinate dehydrogenase causes an increase in extracellular DA levels and indicate that bioenergetic defects may contribute to the pathogenesis of chronic neurodegenerative diseases through a mechanism involving DA.     

SSTR5 ablation in islet results in alterations in glucose homeostasis in mice

Somatostatin (SST) peptide is a potent inhibitor of insulin secretion and its effect is mediated via somatostatin receptor 5 (SSTR5) in the endocrine pancreas. To investigate the consequences of gene ablation of SSTR5 in the mouse pancreas, we have generated a mouse model in which the SSTR5 gene was specifically knocked down in the pancreatic beta cells (betaSSTR5Kd) using the Cre-lox system. Immunohistochemistry analysis showed that SSTR5 gene expression was absent in beta cells at three months of age. At the time of gene ablation, betaSSTR5Kd mice demonstrated glucose intolerance with lack of insulin response and significantly reduced serum insulin levels. Insulin tolerance test demonstrated a significant increase of insulin clearance in vivo at the same age. In vitro studies demonstrated an absence of response to SST-28 stimulation in the betaSSTR5Kd mouse islet, which was associated with a significantly reduced SST expression level in betaSSTR5Kd mice pancreata. In addition, betaSSTR5Kd mice had significantly reduced serum glucose levels and increased serum insulin levels at 12 months of age. Glucose tolerance test at an older age also indicated a persistently higher insulin level in betaSSTR5Kd mice. Further studies of betaSSTR5Kd mice had revealed elevated serum C-peptide levels at both 3 and 12 months of age, suggesting that these mice are capable of producing and releasing insulin to the periphery. These results support the hypothesis that SSTR5 plays a pivotal role in the regulation of insulin secretion in the mouse pancreas.     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2018年中国植物科学若干领域重要研究进展

2018年中国植物科学继续呈现快速发展的态势, 我国科学家在国际植物科学主流学术刊物发表论文数量大幅增加, 取得了多项具有重要影响的成果。调控植物生长-代谢平衡实现可持续农业发展入选2018年度中国科学十大进展; 中国被子植物区系进化历史研究入选2018年度中国生命科学十大进展。以水稻为代表的农作物和果蔬等经济作物研究在国际上已呈现出明显的优势, 若干领域已从“追赶”状态跨越到“领跑”地位。该文对2018年中国科学家在植物科学若干领域取得的重要研究成果进行了概括性评述, 旨在全面追踪和报道当前中国植物科学领域的发展前沿和热点, 展示中国科学家所取得的杰出成就。     

2017年中国植物科学若干领域重要研究进展

2017年中国植物科学继续保持高速发展态势, 重大成果频出, 具体表现在中国植物学家在国际顶级学术期刊发表的文章数量平稳上升。中国植物科学领域的研究工作者成果精彩纷呈, 如新型广谱抗病机制的发现、水稻广谱抗病遗传基础及机制和疫霉菌诱发病害成灾机制研究等。2017年中国生命科学领域十大进展评选中, 有两项植物科学领域的研究成果入选。水稻生物学、进化与基因组学和激素生物学等领域学科发展突出。另外, 值得一提的是, 长期从事高等植物与代谢途径调控分子网络研究和水稻品种设计育种的李家洋院士的研究成果“水稻高产优质性状形成的分子机理及品种设计”荣获2017年国家自然科学一等奖。这一具有重大国际影响的开创性贡献标志着中国植物科学在该领域的国际科学前沿居于引领和卓越地位。该文对2017年中国本土科学家在植物科学若干领域取得的重要研究成果进行了系统梳理, 旨在全面追踪和报道当前中国植物科学领域发展的最新前沿动态, 与广大读者共同分享我国科学家所取得的辉煌成就。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义。综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望。     

我国葫芦科植物离体培养研究进展

葫芦科植物包括多种瓜类蔬菜,对其进行离体培养研究具有重要的理论和实践意义.综述了国内在葫芦科植物器官培养、体细胞胚胎发生、花药培养、原生质体培养和体细胞杂交及离体遗传转化等方面取得的研究进展,并对葫芦科植物离体培养、遗传转化与育种的前景作了展望.     

Changes in aquatic vegetation in Rhine floodplain streams in Alsace in relation to disturbance

Abstract. Changes are described in aquatic vegetation in oligotrophic, groundwater-fed Rhine floodplain streams in Alsace (eastern France), resulting from disturbance. Disturbance factors include changes in nutrients, either permanent ones - effluent from a waste water treatment plant or trout hatcheries - or periodic ones: flooding. Regular inputs of high levels of phosphate and ammonia modified the macrophyte vegetation in these streams. The floristic composition, which was characteristic of oligotrophic waters upstream of the eutrophicated sector, changed to that of a eutrophic situation as originally found downstream. Periodic disturbance by floods which normally occur once a year, irregularly eutrophicates the small streams, causing the development of a mixture of eutrophic and oligotrophic species. Six macrophyte communities are distinguished, indicating different trophic levels. The aquatic vegetation is adapted to the variations of phosphate and ammonia levels. Hence, aquatic macrophytes can be used as bio-indicators of fluctuations in water nutrient levels in relation to the type of disturbance.     

Morphine-potentiated platelet aggregation in in vitro and platelet plug formation in in vivo experiments

The detailed mechanisms underlying morphine-signaling pathways in platelets remain obscure. Therefore, we systematically examined the influence of morphine on washed human platelets. In this study, washed human platelet suspensions were used for in vitro studies. Furthermore, platelet thrombus formation induced by irradiation of mesenteric venules with filtered light in mice pretreated with fluorescein sodium was used for an in vivo thrombotic study. Morphine concentration dependently (0.6, 1, and 5 microM) potentiated platelet aggregation and the ATP release reaction stimulated by agonists (i.e., collagen and U46619) in washed human platelets. Yohimbine (0.1 microM), a specific alpha(2)-adrenoceptor antagonist, markedly abolished the potentiation of morphine in platelet aggregation stimulated by agonists. Morphine also potentiated phosphoinositide breakdown and intracellular Ca(2+) mobilization in human platelets stimulated by collagen (1 microg/ml). Moreover, morphine (0.6-5 microM) markedly inhibited prostaglandin E(1) (10 microM)-induced cyclic AMP formation in human platelets, while yohimbine (0.1 microM) significantly reversed the inhibition of cyclic AMP by morphine (0.6 and 1 microM) in this study. The thrombin-evoked increase in pH(i) was markedly potentiated in the presence of morphine (1 and 5 microM). Morphine (2 and 5 mg/g) significantly shortened the time require to induce platelet plug formation in mesenteric venules. We concluded that morphine may exert its potentiation in platelet aggregation by binding to alpha(2)-adrenoceptors in human platelets, with a resulting inhibition of adenylate cyclase, thereby reducing intracellular cyclic AMP formation followed by increased activation of phospholipase C and the Na(+)/H(+) exchanger. This leads to increased intracellular Ca(2+) mobilization, and finally potentiation of platelet aggregation and of the ATP release reaction.     

Apoptosis in experimental myocardial infarction in situ and in the perfused heart in vitro

We report the appearance of apoptotic cells in experimental myocardial infarction (rabbit heart) in in situ and in vitro preparations. Apoptosis was recognized by intravital staining with Hoechst 33342 (Ho342), by nick-end labeling (TUNEL) and by DNA laddering. A steady rise in the relative number of apoptotic cardiomyocytes (apoptotic index) was noted in in situ preparations. Apoptosis was first noted 6 h after the onset of ischemia with its highest value occurring after 72 h. Apoptotic nuclei were absent in remote areas of the left and right ventricles. Apoptotic nuclei within the infarcted area showed diminished intensity of Ho342 fluorescence. Three days after ischemia, a border zone adjacent to the infarcted area consisting of apoptotic macrophages was recognized. A novel finding was the appearance of apoptotic cardiomyocytes in the isolated perfused ischemic heart. Occurring as early as 50 min after the onset of ischemia, a high apoptotic index was present adjacent to the ligature placed around the coronary artery. This observation provides the opportunity to selectively examine factors leading to apoptosis in the ischemic heart under controlled experimental conditions.