Continuous, high level production and excretion of a plasmid-encoded protein by Escherichia coli in a two-stage chemostat

The stable continuous overproduction of a plasmidencoded protein, beta-lactamase, for at least 50 days by Escherichia coli K-12, RB791(pKN), with release into the culture medium has been demonstrated in two-stage chemostats. The second-stage culture was continuously induced with 0.1 mM IPTG. Continuous expression of beta-lactamase could not be sustained with this strain in a single-stage chemostat because of cell death and selection for lac(-1) cells. beta-Lactamase production in the second stage was sensitive to the second-stage dilution rate and the distribution of the limiting substrate (i.e., glucose) between the first and second stages. The fraction of viable, excreting cells and the average copy number in the induced culture was measurably higher under those conditions of dilution rate and substrate distribution which yielded high beta-lactamase levels. The best operating conditions found at 20 degrees C were a first-stage dilution rate of 0.12 h(-1), a second-stage dilution rate of 0.03 h(-1), and equal glucose feed supplied to each stage. Enzymatically active beta-lactamase was produced at a level of 25% of total cellular protein with 90% excretion yielding 300 mg beta-lactamase/L that was 50% pure at an OD(600) < 6. (c) 1993 Wiley & Sons, Inc.     

大鼠脑突触质膜糖皮质激素受体的纯化

本文利用抗大鼠肝细胞内糖皮质激素受体的单克隆抗体制备的免疫亲和层析柱,将大鼠脑突触质膜糖皮质激素受体纯化了约1150倍,SDS聚丙烯酰胺簿层梯度凝胶电泳显示,在约67kD处有一较明显的染色条带。     

花生种子活力与贮藏蛋白质降解的关系

花生种子吸胀2d后,子叶中肽链内切酶活性上升,贮藏蛋白质开始降解。高活力种子肽链内切酶活性在吸胀2d后迅速上升,至4d时达到高峰,而中等活力种子的肽链内切酶活性上升速度绶慢。高活力种子萌发时贮藏蛋白质降解速度高于中等活力种子。中等活力种子经PEG和PUT处理可提高种子活力,也促进了种子贮藏蛋白质降解能力的提高。     

花生种子老化相关蛋白质的初步研究

本文以粤油 116花生(Arachis hypogaea L.)为材料,对不同处理种子的除子叶“种胚”(以下简称“种胚”)的蛋白质进行了研究.实验结果表明,当花生种子活力下降到一定程度时,其“种胚”内出现一种新蛋白质( pI6.2、MW 10 KD),随种子老化程度加深,含量逐渐增多.我们认为该蛋白质与花生种子老化存在着一定的相关关系,可作为该种子老化的标志.     

鼎湖山自然林豆科固氮植物资源的调查研究

本文在调查鼎湖山自然林木本豆科植物结瘤固氮的基础上,参阅了国内外有关豆科植物结瘤固氮的主要文献,研究了鼎湖山自然林木本豆科植物的固氮资源。结果得出鼎湖山自然林中常见的木本豆科植物共有41种,其中乔木15种,灌木6种,木质藤本20种;有结瘤固氮特性的26种,其中乔木11种,灌木5种,木质藤本10种;经初步调查未见根瘤的6种,其中乔木2种,灌木1种,木质藤本3种;未调查的9种,其中乔木2种,木质藤本7种。本研究结果为鼎湖山木本豆科固氮植物资源的保护、管理和开发利用提供了科学论据,在理论和应用方面均有重要意义。     

粟穗螟滞育的形成和解除与环境条件的关系

粟穗螟Manpava bipunctella Ragonot在川南地区为二化性兼性滞育的昆虫。光周期是诱发滞育的主导因素,在中位温度下,滞育与否主要取决于幼虫发育期间的每日光照时数。在2s℃恒温下,临界光周期为14小时38分。幼虫对光照刺激反应的敏感期为低龄期。 温度和食料效应只发生在每天14小时以上的长光照下,低温有抵销长光照抑制滞育的作用,高温影响不显著;取食玉米的幼虫滞育率比高粱的高,并随寄主生育阶段的发展而增高。该虫滞育解除必需每天14-15小时的长光照;不利于滞育发育和解除,适宜温度为10一25℃。本文最后讨论了该虫滞育形成和解除的特点对发生规律的作用及在测报上的意义。     

Growth kinetics for shelf-life prediction: Theory and practice

Summary Predictive microbiology can be used to determine and predict the shelf-life of perishable foods under commercial distribution conditions based on microbial growth kinetics. This paper presents general microbial growth kinetics with the Monod model and the Gompertz function. Additional models are given to describe effects of food composition (e. g.a _w) and environmental conditions (e.g. temperature, gas atmosphere) as well as their interaction on the growth kinetic parameters (lag time and specific growth rate). These models can be used to predict the time to reach a critical level under any constant conditions within the range tested. A combination of microbial kinetics with an engineering accumulation approach can be used to predict the final microbial level in a food, or the loss of shelf-life, for any known time-temperature sequence, if there is no history effect or the history effect is negligible. A time-temperature indicator, could be used for predicting the remaining shelf-life of perishable foods under any distribution condition based on microbial growth kinetics.Mention of brand or firm names does not constitute an endorsement by the US Department of Agriculture over others of a similar nature not mentioned.     

Avicularin alleviates acute liver failure by regulation of the TLR4/MyD88/NF-κB and Nrf2/HO-1/GPX4 pathways to reduce inflammation and ferroptosis

Acute liver failure (ALF) is an inflammation-mediated hepatocyte death process associated with ferroptosis. Avicularin (AL), a Chinese herbal medicine, exerts anti-inflammatory and antioxidative effects. However, the protective effect of AL and the mechanism on ALF have not been reported. Our in vivo results suggest that AL significantly alleviated lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced hepatic pathological injury, liver enzymes, inflammatory cytokines, reactive oxygen species and iron levels and increased the antioxidant enzyme activities (malondialdehyde and glutathione). Our further in vitro experiments demonstrated that AL suppressed inflammatory response in LPS-stimulated RAW 264.7 cells via blocking the toll-like receptor 4 (TLR4)/myeloid differentiation protein-88 (MyD88)/nuclear factor kappa B (NF-κB) pathway. Moreover, AL attenuated ferroptosis in D-GalN-induced HepG2 cells by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1)/glutathione peroxidase 4 (GPX4) pathway. Therefore, AL can alleviate inflammatory response and ferroptosis in LPS/D-GalN-induced ALF, and its protective effects are associated with blocking TLR4/MyD88/NF-κB pathway and activating Nrf2/HO-1/GPX4 pathway. Moreover, AL is a promising therapeutic option for ALF and should be clinically explored.     

鸭血清胆碱酯酶的纯化及性质研究

首次采用新技术双水相萃取方法作为鸭血清胆碱酯酶(EC.3.1.1.8 CHE) 纯化的第一步,后经 DEAE-Sephadex A50,sephadex G200 柱层析,获得电泳纯鸭血清胆碱酯酶,提纯倍数1018倍,酶活力回收43.4%,比活274.9U/mg。鸭血清胆碱酯酶性质研究表明:此酶是糖蛋白和酸性蛋白水解酶,等电点 4.2 左右,最适 pH7.5 左右;对底物碘化硫代丁酰胆碱的 K_m=9.8×10^-5mol/L;SDS-PAGE 电泳和聚丙烯酰胺梯度电泳表明,鸭血清胆碱酯酶以相同亚基组成的不同聚合体形式存在,亚基分子量 78000,具有完整的酶活性.不同聚合体带电状态相同.     

闽南渔场主要游泳动物时空生态位特征及其影响因素

根据2019—2021年在闽南渔场进行的秋季、冬季、春季和夏季四个航次定点底拖网调查资料,利用相对重要性指数、种群聚集强度、生态位宽度、生态位重叠及冗余分析对主要游泳动物时空生态位特征及其影响因素进行研究。结果表明,(1)调查海域共鉴定出游泳动物214种,主要优势种有18种,优势种存在明显的季节更替现象;夏季优势种的丛生指数和平均拥挤度较高,春季较低;(2)在时间维度上,须赤虾(Metapenaeposis barbata De Haan)生态位宽度最大(0.99), 7组种对时间生态位重叠值等于1.00;在空间维度上,带鱼(Trachurus japonicus Temminck&Schlegel)生态位宽度最大(2.57),空间生态位重叠值超过0.6的种类占71.3%;在时空维度上,带鱼生态位宽度最大(2.45),鹿斑仰口鲾(Leiognathus ruconius Hamilton)与赤鼻棱鳀(Thrissa kammalensis Bleeker)时空生态位重叠值最大(0.94);(3)冗余分析表明,底层温度和底层盐度是影响闽南渔场主要游泳动物时空生态位特征的重要环境因...