Purification of Aspergillus fumigatus (Pdf1) Poly(β-hydroxybutyrate) (PHB) Depolymerase Using a New, Single-Step Substrate Affinity Chromatography Method: Characterization of the PHB Depolymerase Exhibiting Novel Self-Aggregation Behavior

The extracellular poly(-hydroxybutyrate) (PHB) depolymerase of Aspergillus fumigatus Pdf1 was purified by a new, simple, one-step affinity chromatography method using the substrate PHB. The purified enzyme was glycosylated, with the molecular mass of 40 KD, and exhibited a novel self-aggregation behavior by means of hydrophobic interaction that was resolved by Triton X-100 (TX-100) pretreatment of enzyme and also TX-100 incorporation in the native gel. The apparent K _m value of purified enzyme for PHB was 119 g/mL and 3-hydroxybutyrate was detected as the main endproduct of PHB hydrolysis. The depolymerase was insensitive to phenylmethyl sulfonyl fluoride (PMSF), sodium azide, ethylenediaminetetraacetic acid (EDTA), and para-chloromercuric benzoic acid (PCMB), but was inactivated by dithioerythritol (DTT) and showed specificity for short chain-length poly(-hydroxyalkanoates) (PHAs) such as PHB, poly(hydroxyvalerate) (PHV), and copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Medium-chain-length PHA failed to get hydrolyzed. The enzyme, however, exhibited strong cross reactivity with the Comamonas sp. PHB depolymerase antibodies, but not with PHV depolymerase antibodies of Pseudomonas lemoignei. Southern hybridization and dot blot analysis of A. fumigatus Pdf1 genomic DNA with alkaline phosphatase labeled probes of P. lemoignei PHB and PHV depolymerase genes revealed no homology, although the enzyme hydrolyzed both PHB and PHV.     

曲霉WZ-2对染料脱色研究

从废水池污泥中分离到一株染料脱色曲霉ＷＺ－２,它对不同染料均有较好的脱色效果。在３３￣４０℃温度范围内ＷＺ－２对活性桃红ＫＤ－８Ｂ的脱色率为８４％￣９８％,最佳脱色温度为３３℃,ｐＨ值在４．０￣７．０范围内ＷＺ－２对直接耐晒翠蓝ＧＬ的脱色率为８３％￣９６％,最佳脱色ｐＨ为６．０,碳源、氮源及接种量对其脱色率均有影响,染料脱色物的紫外－可见光谱分析表明,可见光区５２０ｎｍ处的吸收峰完全消失,而紫外光     

Bioremediation of metal contamination

A study was initiated to evaluate the use of the fungus Aspergillus niger for bioleaching and then todetermine the effect of process steps, the tailingsconcentration and type of substrate. An oxidized miningtailing containing mainly copper (7240 mg kg^-1 tailings) was studied. A sucrose and mineral salts medium was initially used to produce citric and gluconic acids by A. niger atvarious concentrations of tailings (1, 5, 7, 10 and 15% w/v).Maximal removal of up to 60% of the copper was obtained forthe 5% tailings when the organic acid supernatant was addedto the tailings. In a single step process, A. niger wasthen grown in the presence of mining tailings at variousconcentrations. Maximum copper solubilization (63%) occurredwith 10% mining tailings using sucrose as the substrate.Other substrates were then evaluated including molasses, corncobs and brewery waste (10% mining tailings). Sucrose gavethe best results for copper removal, followed by molasses,corn cobs and brewery waste. Other experiments usingultrasound as a pretreatment showed that 80% removal of thecopper could be obtained for a 5% tailings concentration. Inconclusion, leaching of copper from mining tailings istechnically feasible using A. niger but furtherresearch will be required to increase the economic feasibilityof the process.     

氧化乐果曲霉降解特性和中间产物的研究

在氧化乐果和葡萄糖共基质条件下,曲霉G21优先利用葡萄糖作为碳源,葡萄糖耗尽后开始快速降解氧化乐果,高浓度氧化乐果抑制了曲霉G21的生物降解活性.GC-MS分析表明,曲霉G21降解氧化乐果的主要中间产物为O,O,S-三甲基磷酸酯,氧化乐果降解率提高不会导致该有毒中间产物的累积效应.     

海军航材仓库中曲霉属的耐药性研究

黄曲霉是航材仓库中常见的霉菌之一,其适应性强易变异,容易产生耐药性,在航材仓库中很难防治;通过分析其特性和几种药物对其防治效果以及同一种药剂对几种霉菌防治效果的比较,初步确定了黄曲霉的耐药性并给出了初步的治理建议。     

Inhibitory properties of Enterococcus spp. isolated from faeces of healthy dogs

The aim of the present study was to evaluate the inhibitory effect by the cross-streak method of nine Enterococcus faecium strains isolated from faeces of healthy dogs and their treated and non-treated cell-free supernatant (CFS) by the well-diffusion test on the growth of potentially pathogenic bacteria isolated from clinical cases and aflatoxigenic Aspergillus section Flavi and the consequent aflatoxin B₁ (AFB₁) production. Results obtained from the cross-strake assay showed that E. faecium MF1, GJ18 and GJ40 presented the major inhibitory activity against all pathogenic strains assayed; E. faecium GJ40 produced the larger inhibitory zones (26–27 mm). Well-diffusion test results showed that the majority of the enterococci strains CFS had antimicrobial activity against the pathogenic microorganisms, especially on Gram negative indicators. Cell-free supernatant of E. faecium GJ40 was the one that produced the largest inhibition zones (14 to 21 mm) in the majority of the indicator microorganisms assayed. All supernatants treated with 10 N NaOH (pH6) showed no inhibitory effect on the indicator strain assayed. With respect to fungal inhibition, any of the CFS assayed significantly inhibited the Aspergillus strains growth. But, in general, all CFS reduced AFB₁ production from 8 to 87%. The results demonstrate that enterococci isolated from healthy dog feaces produce substances with the capacity to inhibit some potential pathogenic bacteria growth and the capacity of inhibiting or reducing the AFB₁ production in vitro.     

黑曲霉死菌与活性炭对直接耐晒翠蓝FBL的吸附性能

采用批式实验,系统考察了黑曲霉死菌和活性炭的粉剂投加量,染料初始浓度,pH和反应时间对酞菁染料FBL脱色效果的影响;并采用扫描电镜图像,分析吸附剂的结构变化。结果表明,对于FBL染料的吸附处理,黑曲霉死菌粉剂与活性炭粉剂适宜的吸附条件为:酸性至弱碱性pH下,投加量为8 g/L;黑曲霉死菌粉剂比活性炭粉剂的吸附速度快、脱色性能高、抗染料浓度负荷冲击能力强。扫描电镜图像分析显示,黑曲霉死菌粉剂所具有的多层纤维结构为吸附染料分子提供较大的比表面。     

连续培养酱油曲霉(Aspergillus sojae)产微生物絮凝剂的研究

为了利用廉价材料规模化生产微生物絮凝剂,以味精废水作为廉价培养基质,对酱油曲霉的摇瓶连续培养和发酵罐连续培养进行了研究。摇瓶中以5%的接种量进行连续培养,最适温度在30~33℃之间,每6 h替换一次新鲜废水培养基,5次替换新鲜废水培养基后最大絮凝率仍达到97.8%。在发酵罐扩大连续培养中以5%的接种量接种后经过9 h预培养达到稳定生产絮凝剂后,以4 mL/min的补料流量进行连续培养,生产的絮凝剂产量达到2.392 g/L,且最大絮凝率为98.1%。生产36 h后对发酵系统中菌体进行稀释,使菌体量保持在50~200 g/L之间可持续进行生产。     

柠檬酸中和废水回用技术研究

本文研究了柠檬酸中和废水回用工艺技术。中和废水经碱（ＮａＯＨ）沉淀和酸（Ｈ２ＳＯ４）中和预处理措施,直接用于发酵配料。实验用菌株Ｃ－９８自黑曲霉ＣＢＸ－１２分离获得。最佳发酵条件：总还原糖１４％～１６％,麦麸２．５％,初始ｐＨ６．５,乙醇１．５％。同传统的柠檬酸废水处理工艺比较,该工艺在投资和运转成本方面具有潜在的优势。     

黑曲霉吸附弱酸性艳蓝RAWL机理研究

考察了黑曲霉对染料吸附性能的pH响应模式,红外光谱表征了菌体表面的主要官能团组成,化学修饰定量研究了不同官能团的染料吸附贡献.结果显示,在低pH条件下,菌体表现出最优的染料吸附特性:随着pH值由2.0升至6.0,染料吸附量由39.6mg/g降至9.3mg/g.菌体表面含有氨基、羧基和磷酸根.高pH下,羧基和磷酸根电离导致菌体表面带负电并与染料发生静电斥力,使得染料吸附量下降;低pH下,氨基的质子化使得菌体带正电并通过静电吸引提高染料吸附.甲基化氨基在酸性条件下仍然可以质子化,故氨基甲基化修饰后染料吸附性能不变;乙酰化氨基在酸性体系中失去质子化能力,乙酰化修饰菌体染料吸附性能下降51.6%.氨基质子化引起的菌体正电性和染料负电性之间的静电引力是染料吸附的重要机制.图6参10