酶法降解壳聚糖工艺研究

采用非专一性酶(溶菌酶、纤维素酶)和专一性酶(壳聚糖酶)降解壳聚糖,探讨了不同条件对壳聚糖降解的影响.结果表明,溶菌酶降解壳聚糖的最佳条件为反应时间3.0 h、反应温度50℃、pH值4.0、酶用量40 U·mL-1;纤维素酶降解壳聚糖的最佳条件为反应时间1.5 h、反应温度55℃、pH值5.5、酶用量40 U·mL-1;壳聚糖酶降解壳聚糖的最佳条件为反应时间2.0 h、反应温度45℃、pH值5.0、酶用量30 U·mL-1.对壳聚糖酶酶解产物进行HPLC分析,发现得到了分子量分布较窄的壳寡糖.     

粗纤维素酶液对微晶纤维素酶解的影响因素

栗生灰黑孔菌多被用于产漆酶的研究,很少有利用其纤维素酶的报道。为了降低在纤维素水解中纤维素酶的使用成本,利用栗生灰黑孔菌发酵制备的粗纤维素酶液,以微晶纤维素为底物模型,研究粗纤维素酶液水解微晶纤维素的最佳pH、温度和最佳表面活性剂助剂种类及浓度,并对不同表面活性剂存在条件下的纤维素酶解动力学、紫外和荧光光谱进行了研究。结果表明,粗纤维素酶水解微晶纤维素的最佳条件为pH 4.8,温度50℃,最佳表面活性剂助剂为吐温80,添加剂量为1.12mg/g底物;吐温80的添加可提高粗纤维素酶解的最大反应速度常数V_max,降低米氏常数K_m;表面活性剂改变了纤维素酶的紫外和荧光最大吸收峰,酰胺Ⅰ带和酰胺Ⅲ带的谱峰,可能通过与纤维素酶中的氨基酸残基发生反应影响了纤维素酶的结构,进而影响了微晶纤维素的水解反应。该研究为进一步降低纤维素水解成本提供了理论指导。     

竹浆分批加酶工艺对长时间纤维素酶解性能的影响

优化纤维素酶水解工艺对于纤维素的高效利用具有重要意义。实验在保证纤维素酶相同的添加量时,采用多种方式分批添加纤维素酶水解竹浆,在长时间(7 d)的酶解过程中测定每天还原性糖产量,最后对长时间酶解过程中的影响因素进行了探究。实验结果表明:采用(0.35 mL+0.15 mL+0.1mL)添加工艺提高了酶解效率,酶解7d后糖产量提高了4.36%;纤维素酶随着培育时间的进行活性不断减小,培育1 d后的酶活性仅为原酶活性的53.87%;酶解糖产量随葡萄糖浓度的增加而降低,当葡萄糖浓度为30 mg/mL时,糖产量较无葡萄糖实验组降低了30.7%。酶解过程中酶活的丧失以及底物的抑制可能是影响纤维素酶长时间水解过程中的重要因素。     

纤维素高效酶水解方法的探讨

纤维素酶解可为生物质基的能源与化工品开发提供必需的水溶性还原糖。实验以浓磷酸解结晶处理结合纤维素酶-β-葡萄糖苷酶混合酶液的协同酶解,构建了一种高效酶解方法。实验结果表明,酶水解的最佳条件为:酶液质量浓度为0.062 5 mg/m L,底物质量浓度为12.5 mg/m L,p H为4.8缓冲溶液,酶解温度为50℃,转速为120 r/min。解结晶MCC结合协同酶解方法作用120 h和解结晶麦草结合协同酶解方法作用120 h,产物得率均得到大幅提高。     

固定化纤维素酶水解玉米秸秆的研究

以壳聚糖小球为载体固定纤维素酶,研究了固定化纤维素酶的酶学性质,并以经过蒸汽爆破预处理的玉米秸秆为底物,研究了固定化酶水解的最优条件.结果表明纤维索酶经过固定化后,其米氏常数为0.42 mg/mL,半衰期为162 h;酶解的最佳条件为固定化酶与底物的质量比为1:2.5、固液比为1:2.4(m/V)、温度为50℃,pH为5.0.秸秆水解率可达33%,纤维素水解率可达70%.     

NaOH-Fenton试剂预处理桑木的纤维素酶分段酶水解研究

采用分段酶水解木质纤维原料的方法,以NaOH-Fenton试剂预处理桑木为原料,通过在反应过程中及时移除葡萄糖和纤维二糖,减轻产物的抑制作用,最终达到提高酶水解得率和缩短酶解反应时间的目的。实验结果表明：纤维素酶用量为15FPIU/g（以纤维素计,下同）时,在三段（8+8+8h）水解过程中,经第一段水解,纤维素酶反应速率从1.25g/（L·h）提高到2.21g/（L·h）,第二段水解后,酶反应速率为1.54g/（L·h）,比未分段水解的酶反应速率提高了73%;当纤维素酶用量为40FPIU/g时,三段（8+8+8h）水解得率增至88.08%;三段（8+8+8h）水解充分利用了酶解残渣上的结合酶进行后续水解。对纤维素酶在预处理桑木上的吸附情况进行研究,发现桑木经NaOH-Fenton试剂预处理后,对纤维素酶的最大吸附量为8.08mg/g,预处理增加了纤维素酶与桑木间的吸附位点。     

玉米秸秆蒸爆渣的氨基酸辅助纤维素酶水解

以商品纤维素酶C2730酶解玉米秸秆蒸汽爆破渣,研究了不同氨基酸、氨基酸浓度、温度对水洗蒸汽爆破渣纤维素酶水解的影响,优化纤维素酶水解条件,提高纤维素酶水解得率。实验结果表明,纤维素酶水解蒸汽爆破渣的优化氨基酸为苯丙氨酸,优化水解条件为每克纤维素酶用量15FPIU,苯丙氨酸质量浓度为1.5 g/L,温度为50℃,水解时间为48 h,还原糖和葡萄糖得率分别为51.38%和36.78%。     

蔗渣酶法水解在非缓冲溶剂体系中的反应机理探讨

通过测定蔗渣酶法水解在水系介质中的反应过程的pH随各种反应条件的变化,探讨了酶解液的各种特性,并测定了甘蔗渣、纤维素及纤维素酶相互作用的红外图谱。通过分析认为,纤维素在酶水解过程中,纤维素和纤维素酶相互作用形成了中间产物,使纤维素酶的立体构象发生变化而放出H^ ,使酶解液的pH发生变化,并形成了一种微弱的缓冲体系,致使甘蔗渣酶解反应在采用普通纯水代替缓冲液时,酶解反应的还原糖得率不变。     

甘露聚糖酶促进纤维素酶水解转化马尾松聚糖至可发酵单糖

由于形态结构和化学组成等原因,针叶材原料利用纤维素酶水解法转化聚糖至可发酵单糖一直存在着难水解、总糖转化率低的问题。采用硫酸盐(KP)蒸煮联合氧脱木素和机械打浆对马尾松木片进行预处理,基于针叶材原料中含有较多的甘露聚糖的特点,研究在纤维素酶水解预处理后的浆料过程中添加甘露聚糖酶对酶解效率的影响。研究结果显示,添加甘露聚糖酶对纤维素酶水解具有促进作用,其作用效果因原料预处理方法和程度的不同而不同。KP蒸煮后再经氧脱木素或打浆处理,则甘露聚糖酶对纤维素酶水解的促进作用比单独KP蒸煮效果更加明显。当KP蒸煮至样品木素含量相对于原料为4.1%时,再对原料进行氧脱木素,将木素含量降至相对于原料为1.9%时,分别用10 FPU/g和15FPU/g的复合纤维素酶CTec2对预处理后的样品进行酶解,酶解总糖得率分别为66.0%和83.6%;添加5U/g甘露聚糖酶,上述样品酶解总糖得率分别提高至70.6%和89.2%;相对于原料中聚糖含量,上述条件下酶解总糖转化率分别为53.9%和68.1%。纤维素酶水解至3小时左右再添加甘露聚糖酶,其促进纤维素酶水解效果更好。     

纤维素酶预处理花生壳工艺条件优化

用纤维素酶预处理花生壳,以乙醇提取木犀草素。结果表明,纤维素酶预处理花生壳的最佳工艺条件为:酶解液pH值5.2,酶解温度50℃,酶解时间1.5 h,酶用量0.10%。在最佳预处理条件下预处理后的花生壳木犀草素的提取率可达2.831 0 mg/g,而未经纤维素酶预处理的花生壳木犀草素的提取率仅为1.518 0 mg/g。     

Direct observation of freeze-thaw instability of latex coatings via high pressure freezing and cryogenic SEM

Despite its industrial importance, the subject of freeze-thaw (F/T) stability of latex coatings has not been studied extensively. There is also a lack of fundamental understanding about the process and the mechanisms through which a coating becomes destabilized. High pressure (2100 bar) freezing fixes the state of water-suspended particles of polymer binder and inorganic pigments without the growth of ice crystals during freezing that produce artifacts in direct imaging scanning electron microscopy (SEM) of fracture surfaces of frozen coatings. We show that by incorporating copolymerizable functional monomers, it is possible to achieve F/T stability in polymer latexes and in low-VOC paints, as judged by the microstructures revealed by the cryogenic SEM technique. Particle coalescence as well as pigment segregation in F/T unstable systems are visualized. In order to achieve F/T stability in paints, latex particles must not flocculate and should provide protection to inorganic pigment and extender particles. Because of the unique capabilities of the cryogenic SEM, we are able to separate the effects of freezing and thawing, and study the influence of the rate of freezing and thawing on F/T stability. Destabilization can be caused by either freezing or thawing. A slow freezing process is more detrimental to F/T stability than a fast freezing process; the latter actually preserves suspension stability during freezing. Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, October 27–29, 2004 in Chicago, IL. Tied for first place in The John A. Gordon Best Paper Competition.     

Insect antifeedant properties of anthranoids from the genusVismia

Vismiones and ferruginins, representatives of a new class of lypophilic anthranoids from the genusVismia were found to inhibit feeding in larvae of species ofSpodoptera, Heliothis, and inLocusta migratoria.     

Ethanol and (−)-α-Pinene: Attractant Kairomones for Bark and Ambrosia Beetles in the Southeastern US

In 2002–2004, we examined the flight responses of 49 species of native and exotic bark and ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) to traps baited with ethanol and/or (−)-α-pinene in the southeastern US. Eight field trials were conducted in mature pine stands in Alabama, Florida, Georgia, North Carolina, and South Carolina. Funnel traps baited with ethanol lures (release rate, about 0.6 g/day at 25–28°C) were attractive to ten species of ambrosia beetles (Ambrosiodmus tachygraphus, Anisandrus sayi, Dryoxylon onoharaensum, Monarthrum mali, Xyleborinus saxesenii, Xyleborus affinis, Xyleborus ferrugineus, Xylosandrus compactus, Xylosandrus crassiusculus, and Xylosandrus germanus) and two species of bark beetles (Cryptocarenus heveae and Hypothenemus sp.). Traps baited with (−)-α-pinene lures (release rate, 2–6 g/day at 25–28°C) were attractive to five bark beetle species (Dendroctonus terebrans, Hylastes porculus, Hylastes salebrosus, Hylastes tenuis, and Ips grandicollis) and one platypodid ambrosia beetle species (Myoplatypus flavicornis). Ethanol enhanced responses of some species (Xyleborus pubescens, H. porculus, H. salebrosus, H. tenuis, and Pityophthorus cariniceps) to traps baited with (−)-α-pinene in some locations. (−)-α-Pinene interrupted the response of some ambrosia beetle species to traps baited with ethanol, but only the response of D. onoharaensum was interrupted consistently at most locations. Of 23 species of ambrosia beetles captured in our field trials, nine were exotic and accounted for 70–97% of total catches of ambrosia beetles. Our results provide support for the continued use of separate traps baited with ethanol alone and ethanol with (−)-α-pinene to detect and monitor common bark and ambrosia beetles from the southeastern region of the US.     

Many Drugs of Abuse May Be Acutely Transformed to Dopamine,Norepinephrine and Epinephrine In Vivo

It is well established that a wide range of drugs of abuse acutely boost the signaling of the sympathetic nervous system and the hypothalamic–pituitary–adrenal (HPA) axis, where norepinephrine and epinephrine are major output molecules. This stimulatory effect is accompanied by such symptoms as elevated heart rate and blood pressure, more rapid breathing, increased body temperature and sweating, and pupillary dilation, as well as the intoxicating or euphoric subjective properties of the drug. While many drugs of abuse are thought to achieve their intoxicating effects by modulating the monoaminergic neurotransmitter systems (i.e., serotonin, norepinephrine, dopamine) by binding to these receptors or otherwise affecting their synaptic signaling, this paper puts forth the hypothesis that many of these drugs are actually acutely converted to catecholamines (dopamine, norepinephrine, epinephrine) in vivo, in addition to transformation to their known metabolites. In this manner, a range of stimulants, opioids, and psychedelics (as well as alcohol) may partially achieve their intoxicating properties, as well as side effects, due to this putative transformation to catecholamines. If this hypothesis is correct, it would alter our understanding of the basic biosynthetic pathways for generating these important signaling molecules, while also modifying our view of the neural substrates underlying substance abuse and dependence, including psychological stress-induced relapse. Importantly, there is a direct way to test the overarching hypothesis: administer (either centrally or peripherally) stable isotope versions of these drugs to model organisms such as rodents (or even to humans) and then use liquid chromatography-mass spectrometry to determine if the labeled drug is converted to labeled catecholamines in brain, blood plasma, or urine samples.     

Novel polyurethane coating technology through glycidyl carbamate chemistry

Glycidyl carbamate chemistry combines the excellent properties of polyurethanes with the crosslinking chemistry of epoxy resins. Glycidyl carbamate functional oligomers were synthesized by the reaction of polyfunctional isocyanate oligomers and glycidol. The oligomers were formulated into coatings with several amine functional crosslinkers at varying stoichiometric ratios and cured at different temperatures. Properties such as solvent resistance, hardness, and impact resistance were dependent on the composition and cure conditions. Most coatings had an excellent combination of properties. Studies were carried out to determine the kinetics of the curing reaction of the glycidyl carbamate functional oligomers with multifunctional and model amines. Detailed kinetic analysis of the curing reactions was also undertaken. The results indicated that the glycidyl carbamate functional group is more reactive than a glycidyl ether group. Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, on October 27–29, 2004, in Chicago, IL.