Comparison of surfactant production from kerosene by four species of Corynebacterium

Four species of Corynebacterium-C. fasciens, C. hydrocarboclastus, C. lepus and C. xerosis, were compared for growth and surfactant production from kerosene. Nitrate as sole nitrogen source resulted in a sharp peak in surfactant concentration during mid-exponential growth phase. Ammonium plus nitrate resulted in growth-associated surfactant production. Surfactant was produced at a concentration of 50–80 times the critical micelle concentration by these species; greatest concentrations were produced by C. lepus. All species were pleomorphic, with greatest cell length occurring in early exponential phase. C. hydrocarboclastus had distinctly longer cells (5 ) than the other species (3 ) when grown in shake-flask culture. Ammonium sulfate at concentrations greater than 0.1 % (w/v) increased cell length in stationary phase.This work was supported by The Department of Energy, Mines and Resources of Canada.     

Engineering Clostridium acetobutylicum for production of kerosene and diesel blendstock precursors

Processes for the biotechnological production of kerosene and diesel blendstocks are often economically unattractive due to low yields and product titers. Recently, Clostridium acetobutylicum fermentation products acetone, butanol, and ethanol (ABE) were shown to serve as precursors for catalytic upgrading to higher chain-length molecules that can be used as fuel substitutes. To produce suitable kerosene and diesel blendstocks, the butanol:acetone ratio of fermentation products needs to be increased to 2–2.5:1, while ethanol production is minimized. Here we show that the overexpression of selected proteins changes the ratio of ABE products relative to the wild type ATCC 824 strain. Overexpression of the native alcohol/aldehyde dehydrogenase (AAD) has been reported to primarily increase ethanol formation in C. acetobutylicum. We found that overexpression of the AAD^D485G variant increased ethanol titers by 294%. Catalytic upgrading of the 824(aad^D485G) ABE products resulted in a blend with nearly 50 wt%≤C₉ products, which are unsuitable for diesel. To selectively increase butanol production, C. beijerinckii aldehyde dehydrogenase and C. ljungdhalii butanol dehydrogenase were co-expressed (strain designate 824(Cb ald-Cl bdh)), which increased butanol titers by 27% to 16.9 g L⁻¹ while acetone and ethanol titers remained essentially unaffected. The solvent ratio from 824(Cb ald-Cl bdh) resulted in more than 80 wt% of catalysis products having a carbon chain length≥C₁₁ which amounts to 9.8 g L⁻¹ of products suitable as kerosene or diesel blendstock based on fermentation volume. To further increase solvent production, we investigated expression of both native and heterologous chaperones in C. acetobutylicum. Expression of a heat shock protein (HSP33) from Bacillus psychrosaccharolyticus increased the total solvent titer by 22%. Co-expression of HSP33 and aldehyde/butanol dehydrogenases further increased ABE formation as well as acetone and butanol yields. HSP33 was identified as the first heterologous chaperone that significantly increases solvent titers above wild type C. acetobutylicum levels, which can be combined with metabolic engineering to further increase solvent production.     

Flocculant and Chemical Properties of a Polysaccharide from Pullularia pullulans

An extracellular polysaccharide, PP-floc, was synthesized from glucose by Pullularia pullulans (or Aureobasidium pullulans) in a pilot plant batch fermentor containing 175 liters of culture medium. At 58 h of fermentation, the concentration of PP-floc was 1.03 g/100 ml, giving a 25.8% conversion of initial glucose to polysaccharide. The flocculant activity of the culture medium increased during the fermentation process and reached its maximum at 50 h of culture age. Less PP-floc (0.33 lb/ton of slimes [approximately 149.7 g/0.907 t]) was required to give the same flocculant activity as a synthetic polymer of acrylamide, Separan NP-10 (0.5 lb/ton of slimes [approximately 226.8 g/0.907 t]), at all temperatures from 25 to 100 C. The degree of inactivation of PP-floc and Separan NP-10 at elevated temperatures was almost identical, and they were completely inactivated at about the same temperature (80 C). PP-floc also gave better compaction of slimes than Separan NP-10 at all temperatures tested. PP-floc was soluble in water and its specific optical rotation was [alpha](D) (25) + 194 degrees in water (c, 0.4). PP-floc contained 83.3% carbohydrate, 3.2% protein, and 8.1% water. Glucose was found to be the principal sugar monomer with traces (>5%) of galactose and mannose present. Structural studies on the fractions of purified polysaccharide by methylation and by periodate oxidation techniques prove that PP-floc is linear and composed of alpha-(1 --> 4) and alpha-(1 --> 6) glucopyranosyl units in the approximate ratio of 2:1. The action of pullulanase on crude PP-floc suggested the ordered arrangement of two consecutive alpha-(1 --> 4)-linked glucopyranosyl units flanked by alpha-(1 --> 6)-linked glucopyranose residues.     

微生物絮凝剂及其絮凝微生物的研究进展

介绍了近几年来国内外微生物絮凝剂和絮凝微生物的一些发展概况,列举了近几年发现的一些微生物絮凝剂的物质属性和组成,重点讨论了胞外絮凝剂和絮凝酵母的絮凝机理,详细综述了絮凝微生物的遗传学方面的研究进展,分析讨论微生物絮凝剂的应用概况,提出微生物絮凝剂的发展趋势和研究方向。     

微生物絮凝剂及其在污水处理中的应用

微生物絮凝剂是一种具有广泛应用前景的天然高分子絮凝剂 ,因其安全、高效、无二次污染等优点受到人们的广泛关注 ,已应用于污水处理、食品和发酵工业等。综述了微生物絮凝剂的主要组成、特点、絮凝机理及应用现状。     

新型复合型生物絮凝剂的制备与应用研究

目的:为了得到高效实用的复合型微生物絮凝剂.方法:利用甘蔗渣作为廉价底物,进行复发酵培养:前期为3d,后期为1d,温度为30℃;初始pH值为7.2;同时以珠江河水作为实验水源,对其性能进行研究.结果:复合型微生物絮凝剂对水的浊度、色度去除最佳投加量为12mL/L,同时考虑到其实际使用,制成含水率为80%左右的干粉,试验其对水中TP、邻苯二甲酸脂的去除效果,其最佳投加量为0.21 mg/L.     

变功率微波法制备壳聚糖絮凝剂的研究

研究了变功率下微波加热对甲壳素脱乙酰化反应和粘度的影响。结果表明:在4 80W下反应4min、然后在16 0W下反应6min的最佳功率组合下,可制得脱乙酰度77.3%、粘度4 6mPa·s的壳聚糖产品;与恒功率微波法相比,变功率微波法制备壳聚糖的脱乙酰度稍低,但粘度高出31% ,反应时间缩短三分之一,能耗降低6 0 %     

微生物絮凝剂产生菌的筛选与特性研究

从活性污泥中筛选到1株絮凝活性很高的菌株MY-88。研究结果表明,MY-88的初始培养pH为6.5~7.0,最适温度为25~35℃,最适碳源为葡萄糖;在摇床速度为180 r/min时,其絮凝率达100%。Na 、K 、Ca2 、Fe3 均有较好的促絮凝作用,其中Ca2 尤为显著,使絮凝率达99.6%。MY-88在较大的pH、温度、摇床速度、搅拌速度、水样pH等范围内均具有较高的絮凝活性,显示出良好的应用前景。     

Ethylene production from methionine

1. A new reaction is described in which ethylene is formed from the Cu(+)-catalysed breakdown of methionine in phosphate buffer at 30 degrees in air. Some of the other products of the reaction are methionine sulphone, methionine sulphoxide, homocysteic acid, homocystine, acrolein, dimethyl disulphide, methanethiol, ethyl methyl sulphide, methane and ethane. These are considered to be produced in different reaction pathways. 2. Hydrogen peroxide is an intermediate in this reaction and can support ethylene production in the model system in anaerobic atmospheres. Cuprous copper is the active form that catalyses the formation of ethylene from an oxidized intermediate. The initial reaction is probably a Strecker degradation, but the aldehyde product is further degraded to ethylene and other products. 3. Methional (CH(3).S.CH(2).CH(2).CHO) is the most effective producer of ethylene in the model system and appears to be an intermediate in the reaction. 4. The evidence, from both tracer studies and from other precursors of ethylene in the reaction, indicates that ethylene is derived from the -CH(2).CH(2)- group of methionine.     

Butanol production from lignocellulosics

Clostridium spp. produce n-butanol in the acetone/butanol/ethanol process. For sustainable industrial scale butanol production, a number of obstacles need to be addressed including choice of feedstock, the low product yield, toxicity to production strain, multiple-end products and downstream processing of alcohol mixtures. This review describes the use of lignocellulosic feedstocks, bioprocess and metabolic engineering, downstream processing and catalytic refining of n-butanol.     

Solvent production from xylose

Xylose is the second most abundant sugar derived from lignocellulose; it is considered less desirable than glucose for fermentation, and strategies that specifically increase xylose utilization in wild type or engineered cells are goals for biofuel production. Issues arise with xylose utilization because of carbohydrate catabolite repression, which is the preferential utilization of glucose relative to xylose in fermentations with both pure and mixed cultures. Taken together the low substrate utilization rates and solvent yields with xylose compared to glucose, many industrial fermentations ignore the xylolytic portion of the reaction in lieu of methods to maintain high glucose. This is shortsighted given the massive potential for xylose generation from a number of sustainable biomass feedstocks, based on utilization of the hemicellulose fraction(s) that enter pretreatment. A number of strategies have been developed in recent years to address xylose utilization and solvent production from xylose in systems with just xylose, or in systems with mixtures of glucose plus xylose, which are more typical of pretreated lignocellulose. The approaches vary in terms of complexity, stability, and ease of introduction to existing fermentation infrastructure (i.e., so-called drop-in fermentation strategies). Some approaches can be considered traditional engineering approaches (e.g., change the reaction conditions), while others are more subtle cellular approaches to eliminate the impacts of catabolite repression. Finally, genetic engineering has been used to increase xylose utilization, although this can be considered a relatively nascent approach compared to manipulations completed to date for glucose utilization.

     

Ethylene production from propanal

Tracer studies using a model system consisting of Cu²⁺ and ascorbate indicate that carbons 2 and 3 of propanal are converted to ethylene, and carbon 1 is converted to formic acid and CO₂. A mechanism accounting for this reaction is described. In apple tissue, methionine but not propanal is readily incorporated into ethylene. It is therefore concluded that propanal is not a precursor of ethylene in this fruit.     

Utilization of kerosene by a hydrocarbon-assimilating strain of Fusarium oxisporum

Fusarium oxysporum IFO 6384 was grown in three different media, one containing glucose and two containing kerosene as carbon source. One of the kerosene media contained also Tween 60. The harvested dry mats were weighed at various stages of growth. Substitution of glucose with kerosene led to a sharp drop in the production of fungal mycelia. Analysis of the mat samples representing maximum yields indicated that the mycelia grown in the kerosene media were richer in free amino acid and poorer in protein as well as in polymeric and low-molecular weight carbohydrates than those grown in the glucose medium. Treatment of the culture filtrates with three volumes of acetone led to the separation of two fractions. In the first, acetone soluble fraction the free sugars and amino acids were determined, whereas the second, acetone precipitable one, was examined for its proteolytic activity. The gelatinase activity of the acetone precipitable material (APM) obtained from the kerosene media was lower than that from the glucose medium. The former activity, in contrast to the latter reached its maximum and minimum values at pHs 4 and 9, respectively.     

Determination of kerosene and light oil components in blood.

A sensitive and rapid method to analyse fuel components in blood from rats exposed to kerosene or light oil vapour was developed by making use of capillary gas chromatography/mass spectrometry. The aliphatic hydrocarbons with carbon numbers 8-10 and aromatics such as toluene, xylene, 3- and 4-ethyltoluene and trimethylbenzenes were clearly detected in blood from rats exposed to kerosene or light oil vapour, using the head-space method combined with the salting-out technique. The concentration ratio of pseudocumene to toluene in blood exposed to light oil was higher than that in the case of exposure to kerosene. The lower limits of detection were 50 pg and 1 ng in toluene and pseudocumene, respectively. Our suggestion is that this method is useful in forensic investigations to detect fuel components in blood and for the purposes of differentiating kerosene and light oil in blood tissues.     

Mycotoxin production from fungi isolated from grapes

AIMS: In order to assess the potential for producing mycotoxins, fungi were isolated from wine producing grapes. METHODS AND RESULTS: The isolates were identified and Penicillium expansum, the most well recognized mycotoxin producer, was analysed for mycotoxin production by TLC. Many of the strains produced patulin and/or citrinin, often depending on whether they were grown on a grape or yeast extract sucrose media. CONCLUSION: Citrinin was produced by all strains grown in the yeast extract sucrose medium, but only one strain (from 51) was able to produce this compound in grape juice medium. Patulin was produced in the yeast extract medium by 20 strains and in grape juice medium by 33 strains. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of mycotoxins in wine producing grapes is discussed. Grapes contamination with patulin seems not to contribute to wine contamination, and no ochratoxin producing fungi was identified.     

基于航空煤油细菌检测的触变显色培养基的制备

利用正交试验测定了三种凝胶在不同配比下的触变性,并添加营养物质葡萄糖、蛋白胨、NaCl和显色剂TTC(四氮唑红),得到具有触变性的显色培养基,可直接用于航空煤油样品的检测。结果表明:黄原胶0.9%、琼脂粉0.4%和卡拉胶0.2%配伍使用,触变效果最好,其中影响因素顺序依次为黄原胶、卡拉胶、琼脂;培养基中TTC的最佳浓度范围为0.0008%～0.002%,能使细菌染成红色,并对菌的生长影响很小。     

Purification and Chemical Properties of a Flocculant Produced by Paecilomyces

A new flocculant for microbial cells was purified by ethanol precipitation and gel chromatography from the culture fluid of Paecilomyces sp. I-1. Isoelectric focusing of the flocculant (PF-101) showed a single band at pH 8.5, and its molecular weight was estimated to be over 300,000 daltons by the ultra-filtration method. The results of elemental analysis, the IR spectrum and investigation of the acid hydrolysate by gas and liquid chromatography and colorimetrie analysis suggested that PF-101 was a polysaccharide composed of galactosamine. About 80% of the galactosamine residues were N-unsubstituted and 8% were N-acetylated. Studies on deaminative cleavage, periodate oxidation and Smith degradation suggested that the galactosamine residues were mainly linked by α (→4)-linkaees.     

Bacterial nanocellulose production from naphthalene

Polycyclic aromatic compounds (PAHs) are toxic compounds that are released in the environment as a consequence of industrial activities. The restoration of PAH-polluted sites considers the use of bacteria capable of degrading aromatic compounds to carbon dioxide and water. Here we characterize a new Xanthobacteraceae strain, Starkeya sp. strain N1B, previously isolated during enrichment under microaerophilic conditions, which is capable of using naphthalene crystals as the sole carbon source. The strain produced a structured biofilm when grown on naphthalene crystals, which had the shape of a half-sphere organized over the crystal. Scanning electron microscopy (SEM) and GC-MS analysis indicated that the biofilm was essentially made of cellulose, composed of several micron-long nanofibrils of 60 nm diameter. A cellulosic biofilm was also formed when the cells grew with glucose as the carbon source. Fourier transformed infrared spectroscopy (FTIR) confirmed that the polymer was type I cellulose in both cases, although the crystallinity of the material greatly depended on the carbon source used for growth. Using genome mining and mutant analysis, we identified the genetic complements required for the transformation of naphthalene into cellulose, which seemed to have been successively acquired through horizontal gene transfer. The capacity to develop the biofilm around the crystal was found to be dispensable for growth when naphthalene was used as the carbon source, suggesting that the function of this structure is more intricate than initially thought. This is the first example of the use of toxic aromatic hydrocarbons as the carbon source for bacterial cellulose production. Application of this capacity would allow the remediation of a PAH into such a value-added polymer with multiple biotechnological usages.     

Uncoupling chitosanase production from chitosan

There is a growing interest in chitosanases as enzymatic tools to hydrolyze chitosan into bioactive forms: low molecular weight chitosan (LMWC) or chitosan oligosaccharides (CHOS). However chitosanases are still expensive and methods of large-scale production of these enzymes are not yet established. The article reviews the approaches used for chitosanase production in various bacterial hosts, pointing out the difficulties resulting from the necessity to include chitosan into the medium composition. A mutated Streptomyces host allows for the efficient production of several chitosanases originating from actinobacteria in the absence of chitosan as inducer.