固态发酵的参数周期变化及对微生物发酵的影响

研究了压力脉动固态发酵反应器内环境参数的周期性变化以及这些周期性的环境刺激对于固态培养的斜卧青霉发酵的影响。研究结果显示:在这种反应器中,在空气压力脉动的带动下,反应器内的温度和空气湿度也会发生较大幅度的周期性变化,变化的周期和空气压力脉动的周期相同,变化的幅度随着压力脉动的幅度增大而增加;在外界周期刺激的条件下,较未加外界周期刺激斜卧青霉的生物量增加了104% ,二氧化碳的产生总量增加了229%和纤维素酶的产量增加了320 % ,数据显示外界周期刺激不仅增加了菌体的生物量同时增加了其代谢活性。     

极低频率（ELF）磁场对细胞生长周期分布谱的影响及其机理的计算机模拟分析

测定在各种温度条件下和在ＥＬＦ（极低频率）磁场作用下细胞生长周期分布谱的变化。实验结果表明温度不仅能使周期分布谱的离散性发生改变,而且也能使谱的峰值产生位移,而ＥＬＦ磁场只能使周期分布谱的离散性发生变化,对谱的峰值没有显著影响。这一差异是由于温度和ＥＬＦ磁场对细胞生长产生影响的机理不同。用计算机模拟不同机理对细胞生长周期分布谱的影响与我们早先提出的机理相吻合,即温度影响细胞内各种生长因子、生物离子的活性,而ＥＬＦ磁场则可能通过对细胞膜的影响使细胞内的细胞生长必不可少的生物离子的浓度发生变化。     

微小型生物反应器及其在生物医药领域的应用展望

微小型生物反应器体积微小但在线分析检测和过程控制功能媲美台式装备。其核心支撑技术包括一次性材料及微加工技术、非接触式光学传感器、自动化以及实验设计 (DOE)、数据分析软件与过程控制的整合。由于体积微小、湍流程度和单位能耗较低,微小型反应器内的混合、传质、剪切特性与工业规模设备有一定的区别。现阶段微小型生物反应器主要用于菌株和细胞系筛选和工艺优化,在实现高通量工艺的同时确保了数据的丰度,对缩短研发周期和加速产品上市,尤其是在应对突发性传染性疾病方面有着重要的意义。未来,精准医疗概念的落实也依赖功能柔性化的微小型生物反应器系统。     

生物脱硫的研究新进展*

化石燃料的脱硫形势日益严峻。生物技术为脱有机硫提供了一条经济有效的可行之路。阐述了近几年生物脱硫在许多方面的重大进展 ,主要包括 :新菌种的分离 ,生物脱硫机制的研究 ,应用直接进化技术提高酶的催化效率 ,新型反应器的设计及有价值的化学副产品的生产等。     

生物反应器工程

本文在外界周期刺激强化生物反应和细胞膜传质速率新理论指导下,根据生态学的思维方式,提出了生物反应器工程的新概念,阐明了它的发展过程,并提出了生物反应器工程研究的新方法。     

生物反应器工程

本文在外界周期刺激强化生物反应和细胞膜传质速率新理论指导下,根据生态学的思维方式,提出了生物反应器工程的新概念,阐明了它的发展过程,并提出了生物反应器工程研究的新方法。     

由微分方程所描述的微生物连续培养动力系统(Ⅰ)

陆续介绍微生物连续培养(Chemostat)的基本原理,以单种微生物连续培养模型为基础,较详细地介绍几类由微分方程所描述的微生物连续培养动力系统模型,涉及的问题有解的稳定性,系统的持久性,周期解和Hopf分支等．     

rBTI通过上调PTEN的表达抑制Hep G2细胞增殖

磷酸酶及张力蛋白的同源基因(PTEN) 是一种抑癌基因,可以调控细胞的增殖,与癌症的发生和发展息息相关。本研究采用MTT法和流式细胞术分别检测了重组荞麦胰蛋白酶抑制剂(rBTI)对人肝癌细胞株Hep G2细胞的增殖以及周期的影响。免疫荧光及Western印迹法检测了PTEN和p PTEN的亚细胞定位及蛋白表达的变化。采用qRT-PCR及Western印迹法检测了周期相关蛋白的表达。旨在探究PTEN和p PTEN在rBTI抑制Hep G2细胞增殖和周期阻滞中的作用。结果表明,rBTI能显著抑制Hep G2细胞增殖,将细胞周期阻滞在G0/G1期,并呈时间和剂量依赖性;rBTI作用于Hep G2后,可显著上调PTEN和p-PTEN的表达。同时发现,p-PTEN主要分布于细胞核中,能与核仁发生共定位;周期相关蛋白检测表明,细胞内p53、p21转录水平和蛋白水平均增加。综上所述,rBTI通过上调PTEN的表达,使得细胞周期阻滞于G0/G1期,进而抑制Hep G2细胞的增殖。     

IGF1和IGF1R在小鼠第一个毛囊生长周期皮肤的表达

毛囊生长周期中,真皮乳头和毛基质间的基质 上皮信号调控细胞的增殖和分化。多功能细胞调控因子胰岛素样生长因子1（IGF1)是该信号路径的成员之一。第1个毛囊生长周期决定着毛囊的正常生长和发育,但IGF1在此期的作用未见报道。实时荧光定量PCR结果显示,IGF1在生长期皮肤中的相对表达量最低,在退化期表达量最高,在静止期表达量又降低。与生长初期相比,IGF1在退化期和静止期的表达量呈差异极显著（P＜0.01）;胰岛素样生长因子1受体（IGF1R）在生长期皮肤中的相对表达量最高,在退化期表达量最低,而在静止期表达量又升高。与生长初期相比,IGF1R在退化期和静止期的表达量呈差异极显著（P＜0.01）。Western 印迹结果显示,IGF1和IGF1R蛋白在小鼠皮肤第1个毛囊生长周期各阶段的表达趋势分别与其mRNA的表达趋势一致;免疫组织化学结果表明,IGF1主要分布在小鼠表皮,而IGF1R免疫阳性在小鼠毛囊毛球部、内外根鞘和毛乳头均有分布。以上实验结果揭示,IGF1和IGF1R在小鼠皮肤第1个毛囊生长周期的各阶段的差异性表达,可能在毛囊生长周期各阶段的转化过程中参与了黑色素的形成。然而,IGF1和IGF1R表达趋势不一致,提示IGF1在小鼠皮肤中发挥作用时,并非只与IGF1R结合才能发挥作用。     

板状电泳在苏芸金芽孢杆菌亚种鉴定上的应用

作者用聚丙烯酰胺凝胶板状电泳技术,对1986年以前发表的苏芸金杆菌22个血清型29个亚种进行了酯酶分析,获得了较圆盘电泳更为准确、迅速、易区分的酮酶分析结果。其中H_1—H_(14)的20个亚种与圆盘电泳的结果相同。本文报道了1978—1986年间所发表的苏芸金杆菌9个亚种板状电泳的酮酶分析和生化测定的结果。从而确立了以酮酶型为主结合生化特性对苏芸金杆菌未知菌株进行鉴定的可行方法。     

Enhanced production of poly(3-hydroxybutyrate) in a novel airlift reactor with in?situ cell retention using Azohydromonas australica

Economic production of biodegradable plastics is a challenge particularly because of high substrate and energy cost inputs for its production. Research efforts are being directed towards innovations to minimize both of the above costs to economize polyhydroxybutyrate (PHB) production. A novel airlift reactor (ALR) with outer aeration and internal settling was utilized in this investigation. Although it featured no power consumption for agitation, it facilitated increased oxygen transfer rate and better cell retention than stirred tank reactor (STR), thereby resulting in enhanced PHB productivity. ALR with in?situ cell retention demonstrated a significant improvement in biomass concentration and biopolymer accumulation. The total PHB production rate, specific biomass, and product yield in the ALR were observed to be 0.84?g/h, 0.43?g/g, and 0.32?g/g, respectively. The studies revealed that the volumetric oxygen mass transfer rate and mixing time for ALR were 0.016?s(-1) and 3.73?s, respectively, at 2.0?vvm as compared with corresponding values of 0.005?s(-1) and 4.95?s, respectively, in STR. This demonstrated that ALR has better oxygen mass transfer and mixing efficiency than STR. Hence, ALR with cell retention would serve as a better bioreactor design for economic biopolymer production than STR, particularly due to its lower cost of operation and simplicity along with its enhanced oxygen and heat transfer rates.     

Comparison of air-lift and stirred tank reactors for itaconic acid production by Aspergillus terreus

In a 2-l stirred tank reactor (STR), maximum production rate ofitaconic acid was 0.48g/l.h , for an agitation rate of 400 rpm andan aeration rate of 0.5 vvm. In an air-lift reactor (ALR) themaximum production rate was 0.64 g/l.h at an O supply rate of0.41 l O /l. min. Power input per unit volume which gave themaximum production rates for STR and ALR were 1180 and 542 W/m 3,respectively. If O -enriched air was used in place of air for ALR,the corre-sponding power input per unit volume was decreased to 34W/m 3 . ALR requires less power input per unit volume in comparisonwith that of STR whether therefore air or O -enriched air is used.ALR would be a suitable bioreactor for a large production of itaconicacid.     

Effect of cultivation conditions on spore production from Bacillus amyloliquefaciens B128 and its antagonism to Botrytis elliptica

Aims:  To maximize spore production by Bacillus amyloliquefaciens B128, and its antagonism to the fungal pathogen Botrytis elliptica B061. Methods and Results:  In the 5-l stirred-tank bioreactor (STR), with the 0·5 vvm aeration rate, an agitation rate of 200 rev min⁻¹ significantly enhanced the spore yield compared to the same in 300 rev min⁻¹ cultivations. In a 20-l airlift bioreactor (ALR) the maximal spore production was further increased with a controlled aeration rate of 2·5 vvm operated in a 24-mesh net-draft tube mode, and no pH control cultivation. This spore yield in the 20-l ALR was five- and eightfold higher; in addition the cultivation period was 19 h shorter, compared to that obtained from shaker flask and in the 5-l STR cultivations respectively. Conclusions:  Although culture conditions are still to be optimized, by using an ALR with net-draft tube, a scaling up from shaker flasks and STR to ALR of spore production by the strain B128 is technically feasible. Significance and Impact of the Study:  The spore yields obtained using bioreactors were much higher than those previously reported. The freshly produced spore preparations from the B128 strain significantly antagonized the grey mould pathogen B. elliptica.     

Exploring antidiabetic potential of adamantyl-thiosemicarbazones via aldose reductase (ALR2) inhibition

The role of aldose reductase (ALR2) in diabetes mellitus is well-established. Our interest in finding ALR2 inhibitors led us to explore the inhibitory potential of new thiosemicarbazones. In this study, we have synthesized adamantyl-thiosemicarbazones and screened them as aldehyde reductase (ALR1) and aldose reductase (ALR2) inhibitors. The compounds bearing phenyl 3a, 2-methylphenyl 3g and 2,6-dimethylphenyl 3m have been identified as most potent ALR2 inhibitors with IC₅₀ values of 3.99 ± 0.38, 3.55 ± 0.26 and 1.37 ± 0.92 µM, respectively, compared with sorbinil (IC₅₀ = 3.14 ± 0.02 μM). The compounds 3a, 3g, and 3m also inhibit ALR1 with IC₅₀ value of 7.75 ± 0.28, 7.26 ± 0.39 and 7.04 ± 2.23 µM, respectively. Molecular docking was also performed for putative binding of potent inhibitors with target enzyme ALR2. The most potent 2,6-dimethylphenyl bearing thiosemicarbazone 3m (IC₅₀ = 1.37 ± 0.92 µM for ALR2) and other two compound 3a and 3g could potentially lead for the development of new therapeutic agents.     

Modulation of aldose reductase activity through S-thiolation by physiological thiols

The glutathionyl-modified aldose reductase (GS-ALR2) is unique, among different S-thiolated enzyme forms, in that it displays a lower specific activity than the native enzyme (ALR2). Specific interactions of the bound glutathionyl moiety (GS) with the ALR2 active site, were predicted by a low perturbative molecular modelling approach. The outcoming GS allocation, involving interactions with residues relevant for catalysis and substrate allocation, explains the rationale behind the observed differences in the activity between GS-ALR2 and other thiol-modified enzyme forms. The reversible S-glutathionylation of ALR2 observed in cultured intact bovine lens undergoing an oxidative/non oxidative treatment cycle is discussed in terms of the potential of ALR2/GS-ALR2 inter-conversion as a response to oxidative stress conditions.     

Modulation of aldose reductase activity through S-thiolation by physiological thiols

The glutathionyl-modified aldose reductase (GS-ALR2) is unique, among different S-thiolated enzyme forms, in that it displays a lower specific activity than the native enzyme (ALR2). Specific interactions of the bound glutathionyl moiety (GS) with the ALR2 active site, were predicted by a low perturbative molecular modelling approach. The outcoming GS allocation, involving interactions with residues relevant for catalysis and substrate allocation, explains the rationale behind the observed differences in the activity between GS-ALR2 and other thiol-modified enzyme forms. The reversible S-glutathionylation of ALR2 observed in cultured intact bovine lens undergoing an oxidative/non oxidative treatment cycle is discussed in terms of the potential of ALR2/GS-ALR2 inter-conversion as a response to oxidative stress conditions.     

Augmenter of liver regeneration protects the kidney from ischaemia‐reperfusion injury in ferroptosis

Acute kidney injury (AKI) is a common and severe clinical condition with high morbidity and mortality. Ischaemia‐reperfusion (I/R) injury remains the major cause of AKI in the clinic. Ferroptosis is a recently discovered form of programmed cell death (PCD) that is characterized by iron‐dependent accumulation of reactive oxygen species (ROS). Compelling evidence has shown that renal tubular cell death involves ferroptosis, although the underlying mechanisms remain unclear. Augmenter of liver regeneration (ALR) is a widely distributed multifunctional protein that is expressed in many tissues. Our previous study demonstrated that ALR possesses an anti‐oxidant function. However, the modulatory mechanism of ALR remains unclear and warrants further investigation. Here, to elucidate the role of ALR in ferroptosis, ALR expression was inhibited using short hairpin RNA lentivirals (shRNA) in vitro model of I/R‐induced AKI. The results suggest that the level of ferroptosis is increased, particularly in the shRNA/ALR group, accompanied by increased ROS and mitochondrial damage. Furthermore, inhibition of system xc‐ with erastin aggravates ferroptosis, particularly silencing of the expression of ALR. Unexpectedly, we demonstrate a novel signalling pathway of ferroptosis. In summary, we show for the first time that silencing ALR aggravates ferroptosis in an in vitro model of I/R. Notably, we show that I/R induced kidney ferroptosis is mediated by ALR, which is linked to the glutathione‐glutathione peroxidase (GSH‐GPx) system.     

Fermentative production of 1,3-propanediol from glycerol by Clostridium butyricum up to a scale of 2m3

Summary The conversion of glycerol to 1,3-propanediol (PD) by Clostridium butyricum DSM 5431 was studied in anaerobic culture. Growth and product formation were optimal at pH = 7.0 and T = 35° C, while aeration rate and stirrer speed were found to have no significant influence. As increasing amounts of initial glycerol led to inhibition of growth, cultivations were done in fed-batch operation. Comparative cultivations were carried out in an air-lift (ALR) and a stirred-tank reactor (STR) having equal working volumes (V _L = 30 l) and no difference in product formation was found. The process was scaled up to reactor sizes of 1.2 m³ (ALR) and 2.0 m³ (STR). The same results were obtained irrespective of reactor volume as well as reactor type (STR/ALR). PD concentrations of approximately 50–58 g·l^–1 and overall productivities of 2.3–2.9 g·l^–1 ·h^–1 could be reached. Offprint requests to: W.-D. Deckwer     

Immunoquantitation of aldose reductase in human tissues

Rabbit antibodies raised against bovine kidney aldose reductase (ALR2) were shown to be monospecific for human ALR2 by Western blot analysis of human muscle homogenates. The human enzyme was detected, by reaction with the antiserum (alpha-BKALR2), in homogenates of adrenal gland, muscle, lens, brain, testes, kidney, and placenta, but not in erythrocytes or leukocytes. The amount of enzyme in each tissue was determined by densitometric analysis of autoradiographs of Western blots probed with alpha-BKALR2 and [125I]protein A. Standard curves of radiographic intensity versus amount of purified human muscle ALR2 were linear in the 20 to 200-ng range; a similar sensitivity was seen in tissue homogenates containing up to 675 micrograms total protein. The results presented here for the ALR2 level in human tissues (adrenal greater than muscle greater than lens approximately brain approximately testes greater than kidney approximately placenta) are in agreement with literature values for those tissues from which the enzyme has previously been purified. A notable exception was the absence of detectable ALR2 in human erythrocytes. A quantitative comparison of immunoradiographic response showed that bovine kidney ALR2 was about sevenfold more reactive with a alpha-BKALR2 compared to the human muscle enzyme.     

人脑醛糖还原酶的纯化与性质

联合采用ＤＥＡＥ－纤维素层析、色谱聚焦、ＮＡＤＰ亲和层析与ＳｅｐｈａｄｅｘＧ－１００的凝胶过滤,对人脑醛糖还原酶（ＥＣ１．１．１．２１;ＡＬＲ）进行纯化．现测得该酶的等电点ｐＨ值为５．８５．经聚丙烯酰胺凝胶盘状电泳和Ｗｅｓｔｅｒｎ印迹证实,获得了满意的酶纯度．同葡萄糖,葡糖－６－磷酸与ＮＡＤＰＨ保温后,人脑ＡＬＲ纯品的活性与对照酶组相似,且不被糖酵解途径的一些磷酸化中间产物抑制．苯基硼酸琼脂糖柱层析洗脱谱峰和氢硼化钠还原反应提示,当同葡萄糖保温时,人脑ＡＬＲ（特别是其均一态）可能未被进一步糖基化．在糖尿病并发症和按结构完成药物设计的研究工作中,纯品ＡＬＲ的应用可发挥重要作用