One‐step production of 2,3‐butanediol from starch by secretory over‐expression of amylase in Klebsiella pneumoniae

BACKGROUND: 2,3‐Butanediol (2,3‐BD) is a valuable chemical that can be biosynthesized from many kinds of substrates. For commercial biological production of 2,3‐BD, it is desirable to use cheap substrate without pretreatment, such as starch. However, there have been few reports on the production of 2,3‐BD directly from starch. RESULTS: In this work, gene malS coding for α‐amylase (EC 3.2.1.1) precursor was inserted into plasmid pUC18K, and secretory over‐expression of α‐amylase was achieved by engineered Klebsiella pneumoniae. The extracellular recombinant amylase accelerated the hydrolyzation of starch, and one‐step production of 2,3‐BD from starch was carried out by engineered K. pneumoniae. A 2,3‐BD concentration of 3.8 g L^?1 and yield of 0.19 g 2,3‐BD g^?1 starch were obtained after 24 h fermentation. CONCLUSION: The one‐step production of 2,3‐BD from starch was achieved by secretory over‐expression of amylase in K. pneumoniae. This would simplify the process and reduce the production cost considerably by enabling use of starch with minimal pretreatment. Copyright © 2008 Society of Chemical Industry     

Prophylaxis and Treatment against Klebsiella pneumoniae: Current Insights on This Emerging Anti-Microbial Resistant Global Threat

Klebsiella pneumoniae (Kp) is an opportunistic pathogen and the leading cause of healthcare-associated infections, mostly affecting subjects with compromised immune systems or suffering from concurrent bacterial infections. However, the dramatic increase in hypervirulent strains and the emergence of new multidrug-resistant clones resulted in Kp occurrence among previously healthy people and in increased morbidity and mortality, including neonatal sepsis and death across low- and middle-income countries. As a consequence, carbapenem-resistant and extended spectrum β-lactamase-producing Kp have been prioritized as a critical anti-microbial resistance threat by the World Health Organization and this has renewed the interest of the scientific community in developing a vaccine as well as treatments alternative to the now ineffective antibiotics. Capsule polysaccharide is the most important virulence factor of Kp and plays major roles in the pathogenesis but its high variability (more than 100 different types have been reported) makes the identification of a universal treatment or prevention strategy very challenging. However, less variable virulence factors such as the O-Antigen, outer membrane proteins as fimbriae and siderophores might also be key players in the fight against Kp infections. Here, we review elements of the current status of the epidemiology and the molecular pathogenesis of Kp and explore specific bacterial antigens as potential targets for both prophylactic and therapeutic solutions.     

以葡萄糖为辅助底物发酵生产1,3-丙二醇的研究

采用Klebsiellapneumoniae对葡萄糖作为辅助底物发酵生产 1 ,3 丙二醇进行了研究。结果表明葡萄糖单独作为底物发酵时不生成 1 ,3 丙二醇。以葡萄糖和甘油为混合底物时 ,则可以显著提高菌体浓度 ,但是 1 ,3 丙二醇浓度和甘油到 1 ,3 丙二醇转化率没有提高。在甘油为底物的批式发酵过程中 ,通过流加葡萄糖作为辅助底物可以提高甘油到 1 ,3 丙二醇的摩尔转化率 ,同时可缩短发酵时间。通过选择合适的葡萄糖流加速率 ,较以甘油为单一底物的发酵结果 ,1 ,3 丙二醇的摩尔转化率最高可达0 649,提高了 53 4% ;生产强度为 1 0 0 5g/ (L·h) ,提高了 1 39 9%。     

Klebsieila sp．601细菌漆酶的鉴定及性质

从森林树木根部土壤中分离得到一株具有2，6-二甲氧基酚（DMP）和2，2＇-连氮基-双（3-乙基苯丙噻唑啉-6磺酸（ABTS）氧化活性的细菌。Cu^2＋、Ca^2＋、Mn^2＋、Fe^2＋和Mg^2＋可诱导细菌表达活性的胞质可溶性单体酶蛋白。该酶进行DMP或ABTS氧化时需要铜离子的协助，在pH5．0～7．5范围内氧化DMP，在pH4．0～4．5范围内氧化ABTS，但不能氧化酪氨酸。SDS-PAGE估测酶蛋白分子量大约为55kD。经60℃处理4h或pH10条件下透析20h仍能保持酶活性。16S rDNA序列分析结合常规细菌形态学分析，表明该细菌属于克雷伯氏菌（Klebsiella）属，并命名为Klebsiella sp．60l。这是首例报道Klebsiella细菌菌株具有漆酶活性。     

改进的蒽酮法检测肺炎链球菌荚膜多糖结合物中多糖浓度

目的改进检测肺炎链球菌荚膜多糖结合物中多糖浓度的蒽酮法。方法分别对蒽酮法中蒽酮的浓度和处理温度进行优化,并对改进的方法进行验证和重复性检测。结果适宜的蒽酮浓度为0·3g/300ml,处理温度为40℃,在检测肺炎链球菌多糖结合物原液中的多糖浓度时,标准曲线回归系数高,方法稳定,重现性好。结论改进的蒽酮法可以有效、准确、稳定地检测肺炎链球菌结合物多糖原液中的多糖浓度。     

肺炎链球菌荚膜多糖-破伤风类毒素结合疫苗两种免疫途径对小鼠抗体应答的比较

目的比较肺炎链球菌荚膜多糖-破伤风类毒素结合疫苗两种免疫途径对小鼠的抗体应答。方法选NIH小鼠,分别经皮下和腹腔途径免疫3次同等剂量的结合疫苗,每次间隔2周,用ELISA方法检测两种途径免疫小鼠的血清抗体滴度。结果结合疫苗经腹腔免疫后的抗体滴度优于皮下免疫。小鼠经腹腔免疫3次后,血清抗体的几何平均滴度分别是第1次免疫的6·6倍和3·5倍。结论结合疫苗经腹腔免疫较皮下免疫获得的抗体应答水平高。     

Separation of 1,3-propanediol from glycerol-based fermentations of Klebsiella pneumoniae by alcohol precipitation and dilution crystallization

The separation of 1,3-propanediol from the glycerol-based fermentation broth of Klebsiella pneumoniae plays an important role during the microbial production of 1,3-propanediol. In this paper, the separation of 1,3-propanediol from fermentative broth by a combination of ultrafiltration and alcohol dilution crystallization was investigated. The broth was first filtered by ultrafiltration, and 99% of cells, 89.4% of proteins and 69% of nucleic acids were removed. The obtained broth was further condensed by vacuum distillation, and then alcohol was added. The macromolecular impurities, such as nucleic acids, polysaccharides and proteins, were precipitated, and inorganic and organic salts were crystallized. The optimal volume ratio of alcohol added to the condensed fermentation broth was determined to be 2:1. As a result, proteins, nucleic acids and electric conductivity decreased by 97.4%, 89.7% and 95.8%, respectively, compared with the fermentative broth. The influences of pH and water content in condensed broth on alcohol precipitation and dilution crystallization were also investigated. The experimental results indicated that alcohol precipitation and dilution crystallization was feasible and effective for the separation of 1,3-propanediol from actual fermentation broth. Translated from The Chinese Journal of Process Engineering, 2006, 6(3): 454–457 [译自: 过程工程学报]     

有氧条件下Klebsiella pneumoniae发酵生产1,3-丙二醇的研究

探讨了有氧条件下利用Klebsiella pneumoniae发酵生产1,3-丙二醇的可能性,研究了通气量、初始底物浓度、pH和3-羟基丙醛等因素对发酵过程的影响.当空气通量为0.25vvm时,1,3-丙二醇的得率和生产强度与厌氧时相近.有氧条件下,当初始甘油质量浓度大于50g/L时,发酵中后期出现3-羟基丙醛的长期积累,甘油不能消耗完全.控制pH为7.75～8.00可以促进3-羟基丙醛的转化使甘油完全消耗.     

1,3-丙二醇发酵过程中底物抑制及其对策的研究

研究了底物甘油浓度对Klebsiellapneumoniae发酵生产 1 ,3 丙二醇的影响 ,并通过实验确定了最佳的初始甘油浓度和甘油开始对产物生成产生抑制作用的浓度。针对底物抑制现象 ,提出了菌种驯化和流加甘油发酵两种解决途径。结果表明 :采用原始菌株发酵 ,适宜的甘油初始浓度为 642 .4mmol/L ,此时 1 ,3 丙二醇浓度和转化率分别可达 2 60mmol/L和 0 .492mol/mol;在较高甘油初始浓度 (789mmol/L)的情况下 ,经驯化培养获得的耐底物抑制菌株 ,可将最终 1 ,3 丙二醇浓度和转化率分别提高到 30 0 .9mmol/L和 0 .530mol/mol;采用流加甘油发酵工艺 ,1 ,3 丙二醇浓度和转化率分别可提高到 397.7mmol/L和 0 .62 5mol/mol。     

Microbial Precipitation of Pb(II) with Wild Strains of Paraclostridium bifermentans and Klebsiella pneumoniae Isolated from an Industrially Obtained Microbial Consortium

The study focused on determining the microbial precipitation abilities of bacterial strains that were isolated from an industrially obtained Pb(II)-resistant microbial consortium. Previous research has demonstrated the effectiveness of the consortium on the bioprecipitation and adsorption of Pb(II) from solution. The bioremediation of Pb(II) using microbial precipitation provides an alternative option for Pb(II) removal from wastewater. Both strains, Klebsiella pneumoniae and Paraclostridium bifermentans, were successfully isolated from the consortium obtained from a battery recycling plant in South Africa. The experiments were conducted over both 30 h and 5 d, providing insight into the short- and long-term precipitation abilities of the bacteria. Various initial concentrations of Pb(II) were investigated, and it was found that P. bifermentans was able to remove 83.8% of Pb(II) from solution with an initial Pb(II) concentration of 80 mg L−1, while K. pneumoniae was able to remove 100% of Pb(II) with the same initial Pb(II) concentration after approximately 5 d. With the same initial Pb(II) concentration, P. bifermentans was able to remove 86.1% of Pb(II) from solution, and K. pneumoniae was able to remove 91.1% of Pb(II) from solution after 30 h. The identities of the precipitates obtained for each strain vary, with PbS and Pb0 being the main species precipitated by P. bifermentans and PbO with either PbCl or Pb3(PO4)2 precipitated by K. pneumoniae. Various factors were investigated in each experiment, such as metabolic activity, nitrate concentration, residual Pb(II) concentration, extracellular and intracellular Pb(II) concentration and the precipitate identity. These factors provide a greater understanding of the mechanisms utilised by the bacteria in the bioprecipitation and adsorption of Pb(II). These results can be used as a step towards applying the process on an industrial scale.