Model‐based high cell density cultivation of Rhodospirillum rubrum under respiratory dark conditions

The potential of facultative photosynthetic bacteria as producers of photosynthetic pigments, vitamins, coenzymes and other valuable products has been recognized for decades. However, mass cultivation under photosynthetic conditions is generally inefficient due to the inevitable limitation of light supply when cell densities become very high. The previous development of a new cultivation process for maximal expression of photosynthetic genes under semi‐aerobic dark conditions in common bioreactors offers a new perspective for utilizing the facultative photosynthetic bacterium Rhodospirillum rubrum for large‐scale applications. Based on this cultivation system, the present study aimed in determining the maximal achievable cell density of R. rubrum in a bioreactor, thereby providing a major milestone on the way to industrial bioprocesses. As a starting point, we focus on aerobic growth due to higher growth rates and more facile process control under this condition, with the option to extend the process by an anaerobic production phase. Process design and optimization were supported by an unstructured computational process model, based on mixed‐substrate kinetics. Key parameters for growth and process control were determined in shake‐flask experiments or estimated by simulation studies. For fed‐batch cultivation, a computer‐controlled exponential feed algorithm in combination with a pH‐stat element was implemented. As a result, a maximal cell density of 59 g cell dry weight (CDW) L^?1 was obtained, representing so far not attainable cell densities for photosynthetic bacteria. The applied exponential fed‐batch methodology therefore enters a range which is commonly employed for industrial applications with microbial cells. The biochemical analysis of high cell density cultures revealed metabolic imbalances, such as the accumulation and excretion of tetrapyrrole intermediates of the bacteriochlorophyll biosynthetic pathway. Biotechnol. Bioeng. 2010. 105: 729–739. © 2009 Wiley Periodicals, Inc.     

重组基因工程菌在不对称还原羰基化合物中的应用

手性醇是药物合成的重要手性砌块,利用生物催化剂不对称还原羰基化合物是手性醇制备的重要方法。介绍了生物催化还原羰基化合物的反应原理及特点,综述了重组基因工程菌的构建及其在不对称还原羰基化合物中的应用情况,展望了今后研究发展的方向。     

不同碱度条件下中华水韭昼夜CO2气体交换的特征

 运用pH-drift的方法研究了在不同碱度条件下中华水韭(Isoetes sinensis)的沉水叶片昼夜CO₂吸收的特征。结果表明中华水韭的沉水叶片具有昼夜吸收水中CO₂的能力, 而不具备利用水中的HCO ₃^–的能力, 进一步证明了水生植物中华水韭的光合碳同化途径具有景天酸代谢(CAM)的特征。中华水韭沉水叶片光照条件下对水中CO₂的吸收速率在一定的浓度范围内正相关于水中的CO₂浓度。光照条件下, 中华水韭的pH-drift实验的pH补偿点分别为(8.1±0.3)和(7.9±0.1) mmol·L^–1, 最终[CT]/Alk值为(1.009±0.01)和(1.022±0.004)。碱度对中华水韭夜晚CO2的吸收速率有显著的影响(F = 38.73, p < 0.000 1)。总碱度1.70 mmol·L^–1溶液中的中华水韭沉水叶片在相对较低的CO₂浓度(0.04±0.001 mmol·L^–1)水平下即表现出对CO₂的净吸收。调查了野外一处中华水韭沉水种群的生境pH值及CO₂浓度的昼夜变化, 发现水体碱度约为1.59 mmol·L^–1, 一昼夜的pH值波动不大, 平均为(6.1±0.04), 昼夜CO₂浓度存在波动, 午夜水中的CO₂浓度是午后的近3倍。     

Constitutive inhibition of plasma CETP by apolipoprotein C1 is blunted in dyslipidemic patients with coronary artery disease

Plasma cholesteryl ester transfer protein (CETP) promotes the cholesterol enrichment of apoB-containing lipoproteins (VLDL and LDL) at the expense of HDL. Recent studies demonstrated that apoC1 is a potent CETP inhibitor in plasma of healthy, normolipidemic subjects. Our goal was to establish whether the modulation of CETP activity by apoC1 is influenced by dyslipidemia in patients with documented coronary artery disease (CAD). In the total CAD population studied (n = 240), apoC1 levels correlated negatively with CETP activity, independently of apoE-epsilon, CETP-Taq1B, and apoC1-Hpa1 genotypes. In multivariate analysis, the negative relationship was observed only in normolipidemic patients, not in those with hypercholesterolemia, hypertriglyceridemia, or combined hyperlipidemia. In the normolipidemic subjects, apoC1 levels were positively associated with higher HDL- to LDL-cholesterol ratio (r = 0.359, P < 0.001). It is concluded that apoC1 as a CETP inhibitor no longer operates on cholesterol redistribution in high-risk patients with dyslipidemia, probably due to increasing amounts of VLDL-bound apoC1, which is inactive as a CETP inhibitor. Patients with dyslipidemia could experience major benefits from treatment with pharmacological CETP inhibitors, which might compensate for blunted endogenous inhibition.     

N-acetylaspartate in the vertebrate brain: metabolism and function

N-Acetyl-l-aspartate (NAA) is an amino acid that is present in the vertebrate brain. Its concentration is one of the highest of all free amino acids and, although NAA is synthesized and stored primarily in neurons, it cannot be hydrolyzed in these cells. Furthermore, neuronal NAA is dynamic and turns over more than once each day by virtue of its continuous efflux, in a regulated intercompartmental cycling via extracellular fluids, between neurons and a second compartment in oligodendrocytes. The metabolism of NAA, between its anabolic compartment in neurons and its catabolic compartment in oligodendrocytes, and its possible physiological role in the brain has been the subject of much speculation. There are two human inborn errors in metabolism of NAA. One is Canavan disease (CD), in which there is a buildup of NAA (hyperacetylaspartia) and associated spongiform leukodystrophy, caused by a lack of aspartoacylase activity. The other is a singular human case of lack of NAA (hypoacetylaspartia), where the enzyme that synthesizes NAA is apparently absent. There are two animal models currently available for studies of CD. One is a rat with a natural deletion of the catabolic enzyme, and the other a gene knockout mouse. In addition to the presence of NAA in neurons, its prominence in ¹H nuclear magnetic resonance spectroscopic studies has led to its wide use in diagnostic human medicine as both an indicator of brain pathology and of disease progression in a variety of CNS diseases. In this review, various hypotheses regarding the metabolism of NAA and its possible role in the CNS are evaluated. Based on this analysis, it is concluded that although NAA may have several functions in the CNS, an important role of the NAA intercompartmental system is osmoregulatory, and in this role it may be the primary mechanism for the removal of intracellular water, against a water gradient, from myelinated neurons.     

日粮添加复合饲用益生菌对肉牛生长性能及肠道微生物的影响

[目的]为研究添加饲用益生菌对肉牛生长性能、血液生理生化指标及肠道微生物区系的影响.[方法]在青海地区选取西门塔尔牛与荷斯坦牛的杂交1代18头,按每组平均体重相近的原则随机分为2组,每组9头,试验组饮食添加地衣芽孢杆菌、枯草芽孢杆菌和酿酒酵母的复合饲用益生菌.[结果]试验结果表明:试验组显著地提高了胸围日增长量(P=0...     

gacS基因插入失活对绿针假单胞菌G-05的两种胞外次生代谢物合成的影响

一株来自大棚温室甜椒根际的绿针假单胞菌Pseudomonas chlororaphis G-05,可分泌抗生物质吩嗪-1-羧酸,并具有抑制辣椒疫霉的生物防治功效。【目的】为了系统研究该菌株的生物防治功能及抗生物质合成与分泌机制。【方法】首先通过生化法和16S rDNA同源比对法对该菌株进行系统分类的初步鉴定,再根据基因的同源性从G-05基因组DNA中克隆长1.4 kb的gacS基因的部分保守区段,采用抗庆大霉素基因（gentamycin resistance cassette, aacC1）插入失活的策略构建了该基因突变株G-05S。【结果】在King’ s B（KMB）或PPM培养基中,突变株G-05S合成吩嗪-1-羧酸的能力受到明显抑制。然而,突变株G-05S分泌的吲哚乙酸与野生株相比无显著差异。互补实验表明,gacS基因的表达可以使突变株G-05S的吩嗪-1-羧酸的合成恢复到野生株水平。【结论】由此推测,GacS(Global activator sensor )对不同次生代谢物的调控具有特异性。     

The relationship between hydrogen metabolism,sulfate reduction and nitrogen fixation in sulfate reducers

Summary Hydrogenase and nitrogenase activities of sulfate-reducing bacteria allow their adaptation to different nutritional habits even under adverse conditions. These exceptional capabilities of adaptation are important factors in the understanding of their predominant role in problems related to anaerobic metal corrosion. Although the D₂–H⁺ exchange reaction indicated thatDesulfovibrio desulfuricans strain Berre-Sol andDesulfovibrio gigas hydrogenases were reversible, the predominant activity in vivo was hydrogen uptake. Hydrogen production was restricted to some particular conditions such as sulfate or nitrogen starvation. Under diazotrophic conditions, a transient hydrogen evolution was followed by uptake when dinitrogen was effectively fixed. In contrast, hydrogen evolution proceeded when acetylene was substituted as the nitrogenase substrate. Hydrogen can thus serve as an electron donor in sulfate reduction and nitrogen metabolism.     

水果中抗病原真菌物质成分的提取分离方法及应用前景

诱发因子如草酸,拮抗菌产生的抗素等,以及采后热处理,紫外线处理可以诱导水果产生抗病性,此外有的水果具有先天性抗病成分,对这些活性成分进行分析,并用之于水果采后病害的生物防治,为水果的防腐保鲜提供了新的方法,追踪这些抗病物质的产生和分布,需要采用一些专门的分析方法,目前采用的方法是将果实组织破碎,离心,通过萃取,层析或电泳技术进行纯化,然后用核磁共振,X－射线衍射及各种波谱分析技术进行结构鉴定,抗真菌物质的活性可以用薄层层层析生物学方法,孢子萌发或纸碟法进行鉴定,而抗真菌活性物质的定位则用组织化学法,电子显微术,放射自显著技术和免疫印迹技术也广泛用物于组织化学分析。