通气提高Klebsiella oxytoca HP1发酵产氢

研究了Klebsiella oxytoca HP1在不同通气条件下的生长和放氢特性．批式发酵实验结果表明：通氩气放氢效果最好，通二氧化碳放氢效果最差．提高氩气通气速率能显著提高产氢量。0．25L／Lmin条件下的产氢量达到1155mL是不通气时的2．41倍，但通气也影响延迟期的长短。氩气通量应该与微生物生长阶段相匹配．代谢产物分析表明：K．oxytoca HP1呈混合酸发酵特性，主要代谢产物有乙酸、乙醇、乳酸和2，3-丁二醇，氩气通气速率明显影响丙酮酸节点的代谢途径和通量，从高通气条件到不通气条件，代谢主要途径按乙酸途径→乙醇途径→乳酸途径的次序转变，通气速率是K．oxytoca HP1发酵产氢的重要调控因子．     

产酸克雷伯氏菌的吸附固定及其产氢研究

利用曲霉(Aspergillussp.XF101)所形成的菌丝球吸附产氢细菌产酸克雷伯氏菌(Klebsiella oxytocaHP1),当曲霉培养时间为50 h,菌丝球直径为1.3~1.8 mm,菌悬液pH值为5.0~6.0,吸附温度为30℃,吸附时间为1.5 h时,可获得最佳吸附效果,吸附率可达99%以上.利用菌丝球固定产氢菌可进行连续产氢,其最大产氢速率为18 mmol/L.h,平均产氢速率为1.7 mmol/L.h,持续产氢时间为15 d.     

Enhanced hydrosen production by insertional inactivation of adhE gene in Klebsiella oxytoca HP1

Ethanol is the main byproduct of anaerobic H2-producing fermentation in Klebsiella oxytoca HP1. Two moles of NAD（P）H are consumed to yield one mole of ethanol that may decrease bacterial hydrogen production. In this article the adhE gene that codes for acetaldehyde dehydrogenase was disrupted for the first time. A homologous recombination vector pTA-Str was constructed in which the adhE gene was disrupted by inserting an aminoglycoside-3＇-adenyltransferase （aadA） gene. As expected, the vector includes the insertion 5＇-adhE-aadA-adhE-3＇. The amplified DNA fragment 5＇-adhE-aadA-adhE-3＇ from pTA-Str was transformed into K. oxytoca HP1 and one recombinant was obtained. PCR analysis of the resulting genomic DNA indicated the appropriate deletion and insertion. Compared with the H2-production of wild type K. oxytoca HP1, the hydrogen yield of the mutant increased by 16.07% and ethanol concentration decreased by 77.47%, suggesting that inactivation of the adhE gene in K. oxytoca HP1 is a potential method for enhancing bacterial H2-production.     

Enhanced hydrogen production by insertional inactivation of adhE gene in Klebsiella oxytoca HP1

Ethanol is the main byproduct of anaerobic H2-producing fermentation in Klebsiella oxytoca HP1. Two moles of NAD(P)H are consumed to yield one mole of ethanol that may decrease bacterial hydrogen production. In this article the adhE gene that codes for acetaldehyde dehydrogenase was disrupted for the first time. A homologous recombination vector pTA-Str was constructed in which the adhE gene was disrupted by inserting an aminoglycoside-3'-adenyltransferase (aadA) gene. As expected, the vector includes the insertion 5′-adhE-aadA-adhE-3′. The amplified DNA fragment 5′-adhE-aadA-adhE-3′ from pTA-Str was transformed into K. oxytoca HP1 and one recombinant was obtained. PCR analysis of the resulting genomic DNA indicated the appropriate deletion and insertion. Compared with the H2-production of wild type K. oxytoca HP1, the hydrogen yield of the mutant increased by 16.07% and ethanol concentration decreased by 77.47%, suggesting that inactivation of the adhE gene in K. oxy- toca HP1 is a potential method for enhancing bacterial H2-production.     

PTD-Tat之C端融合在活体体内的跨膜递送作用

报道了Tat蛋白转导区域(PTD-Tat)的C端融合蛋白在线虫和小鼠体内的跨膜递送作用.将重组表达质粒pGEX-GFP-Tat在大肠杆菌中高效表达出的融合蛋白GST-GFP-Tat对小鼠腹腔注射(ip)17h后,荧光显微镜观察其心、肝和肾组织,均检测到强烈的绿色荧光,甚至该蛋白跨越了血脑屏障(BBB).此外用融合蛋白喂食线虫发现蛋白分布于线虫消化管道及其原体腔,且表现出的跨膜递送活性与喂食时间和蛋白浓度呈现正相关.该研究结果拓宽了PTD-Tat在蛋白药物递送方面的应用范围,为蛋白质疗法开辟了新的视野,并为其有效应用提供理论指导.