短梗霉多糖研究进展

短梗霉多糖是一种微生物发酵产物,具有极好的成膜性,无色、无味,且不透气,易生物降解,对人体和环境无毒无害,受到国际上广泛关注,是一种极具研究潜力和经济价值的新型生物环保材料。该文综合了国内外众多学者的研究成果,从发酵底物(包括碳源、氮源、二价离子等)、发酵条件(包括pH、温度、通气量、接种量、种龄等)、多糖性质、应用研究(包括传统应用和最新应用成果)等方面进行阐述,为短梗霉多糖的进一步研究和应用提供参考。     

环境pH诱导出芽短梗霉细胞多形化

出芽短梗霉具有酵母状细胞、膨大细胞、菌丝、厚垣孢子、念珠状菌丝和分生组织状结构。在最适pH条件下,出芽短梗霉生长繁殖以酵母状细胞（CBS100225等4菌株）或膨大细胞（CBS249.65等4菌株）为主。pH 2.2或pH 7.0诱导全部8株出芽短梗霉形成分生组织状结构。酵母状细胞转变成膨大细胞受低pH值诱导的占75%,还受高pH诱导的占50%。膨大细胞是多形性细胞转变的中心环节,可以转变成菌丝、厚垣孢子或分生组织状结构。     

短梗霉多糖发酵条件的研究

在摇瓶发酵条件研究的基础上。于16L自控发酵罐上进行了罐上发酵条件优化研究。发现以10％淀粉水解物为碳源时,淀粉水解物的最适DE值为40－50,发酵培养基中的硫酸铵最适用量不同于摇瓶发酵时的量,种龄和接种量、通气量、罐压、搅拌速度和搅拌叶轮挡数等均对多糖的产生有较大的影响。另外还进行了发酵过程的动力学的研究。     

出芽短梗霉胞外酸性漆酶

通过愈创木酚法平板检测10株出芽短梗霉,发现5株菌能够分泌胞外多酚氧化酶,反应最适pH在2.0左右,均属于酸性多酚氧化酶。菌株NG的酶活最高,达110 U/mL。添加H2O2、EDTA以及过氧化氢酶不显著影响菌株NG胞外酶活,表明NG分泌的多酚氧化酶中不含有锰过氧化物酶（MnP）和不依赖Mn2＋的过氧化物酶（MiP）,属于漆酶（Lac）。     

表面活性剂对出芽短梗霉多糖生产影响的研究

研究了表面活性剂对出芽短梗霉细胞培养过程中多糖释放的影响。在摇瓶中,比较添加0.05%(w/v)的Tween 80、Tween 60、Tween 40,结果显示几种表面活性剂均能促进细胞释放多糖,其中以Tween 80的效果最佳。在5L发酵罐中,以100g/L玉米粉水解液做碳源的出芽短梗霉细胞培养液中分别添加了表面活性剂Tween 80 0.01%、0.05%、0.1%,其中以添加Tween 800.05%时的效果最好,与不添加表面活性剂相比多糖产量提高25%左右,发酵周期缩短了将近2d。     

几株出芽短梗霉在不同发酵条件下产生多糖的比较

将已有的４株出芽短梗霉在摇瓶中于不同发酵条件下进行比较,考察了它们的生长情况,不同的碳源、氮源、磷酸盐、初始ｐＨ和通气量等对短梗霉多糖合成的影响,获得一株产短梗霉多糖的高产菌株,为以后工作打下良好基础。     

短梗霉胞外多糖发酵及其发酵动力学

对短梗霉（Aureobasidiumpullulans)胞外多糖（EPS）的发酵及其发酵动力学进行了研究。短梗霉菌体前期生长速度较快,到48b生长趋于稳定期,其胞外多糖合成随菌体生长的不断上升,到84b多糖的产量达到最高,为14.24(g/L)发酵实验基于Logitic方程和Luedeking-Piret方程,得到了描述发酵过程的动力学数学模型和模型参数,同时对实验数据与模型进行了验证比较。     

出芽短梗霉的研究进展

出芽短梗霉是一类类酵母真菌,具有酵母样和真菌菌丝体两种形态,影响其形态的因素有碳源,氮源,离子种类及浓度和pH值等,出芽短梗霉的发酵产物多种多样,如多聚糖,酶,抗真菌素等,通过选育优良菌株可提高发酵产物的产量。     

出芽短梗霉发酵过程溶氧控制的研究

在搅拌罐式生物反应器中,通过控制DO（溶氧浓度）的变化,对出芽短梗霉（Aureobasidium pullulans）发酵过程的控制进行了研究。以100g/L玉米粉水解液做碳源,比较了不同溶氧控制条件下发酵参数的变化及其对出芽短梗霉发酵结果的影响。结果表明,过低的DO对菌体生长和多糖生产都不利,过高的DO使培养液中糖大部分消耗在菌体的生长上,也不利于多糖的生产,通过控制搅拌速度和通气量能将DO维持在较合适的水平。     

出芽短梗霉色素变异菌株R45的普鲁蓝糖发酵研究

用正交实验确定了出芽短梗霉（Aureobasidiumpullulans）色素变异菌株R45的发酵优化条件。在此条件下,摇瓶培养的普鲁蓝糖产量最高可达82．4g／L,转化率为54．9％。实验表明,CaCO3是多糖发酵的重要影响因素,多糖的合成与发酵pH值及细胞形态密切相关。     

灵芝胞外生物活性多糖的pH控制发酵

灵芝深层发酵过程中 ｐＨ值对灵芝胞外多糖形成的影响。结果表明起始ｐＨ为５．５时,有利于胞外多糖的形成,找到了一种有效的ｐＨ控制发酵策略,当发酵过程中控制ｐＨ４．０时,胞外多糖的产量最大,达到２．３２９／Ｌ,较未控制提高了２４％。另外,还对胞外多糖的生物活性进行了检测,结果表明抗肿瘤活性平均达到５１．２％,且能明显提高小鼠的免疫力。     

SIGNIFICANT IMPROVEMENT OF QUALITY FOR LONG OLIGONUCLEOTIDES BY USING CONTROLLED PORE GLASS WITH LARGE PORES

A key factor influencing the quality of long oligonucleotides is the choice of controlled pore glass (CPG) which is used as a solid support during oligonucleotide synthesis. We studied the influence of CPG pore size on the quality of 75-mer oligonucleotides. Using electrophoresis and HPLC, we demonstrated failure modes that can occur at certain oligo lengths with 1000A pore size, and compared yield and purity of 75-mer oligos using 1000A and larger pore size CPG. We showed that oligonucleotides with much better quality are obtained using CPG with pore sizes of 1400A and larger. We also identified the key characteristics for CPG selection that lead to the best CPG performance.     

CALCIFICATION BY COCCOLITHOPHORIDS: EFFECTS OF pH AND Sr1

Cells of Coccolithus huxleyi which fail to deposit CaCO₃ and form coccoliths often occur as unwanted components in cultures used for studies of calcification. Non-calcified cells generally cannot be made to recalcify, but they can be removed from cultures by treatment at elevated pH or by a method based on faster sinking of calcified cells. Lowering the concentrations of nitrate, phosphate, or trace metals in the medium did not restore calcifying ability of non-calcified cells. However, addition of strontium did promote recalcification of decalcified Cricosphaera carterae grown under calcium limitation. Strontium seemed to promote coccolith attachment to cells rather than to affect calcium uptake or coccolith formation itself.     

EFFECTS OF CERTAIN LIMITING CONDITIONS ON THE SYNTHESIS OF B VITAMINS BY YEAST

In yeast crops which were grown in the presence of various inhibitors, there was considerable variation in content of the various B vitamins. A higher degree of parallelism in variation in content was found to exist between thiamine and niacin than between any other pair of vitamins; this has been interpreted as indicating that the predominant functions of these two vitamins are their established rôles in fermentation. Values for inositol indicate that it may be involved in fermentation processes, but this is not the case for other members of the B complex. Biotin appears to be unique since in no case did the biotin content of yeast grown in the presence of an inhibitor fall below that of the control yeast. There was some evidence of synthesis of biotin, or a material with biotin activity, in the presence of certain inhibitors, the most striking instance being with sulfaguanidine. An exogenous supply of biotin was essential for extensive proliferation of F. B. yeast, and yeast grown in a medium to which biotin was the only added vitamin contained the B vitamins in amounts very similar to those found in the control yeast, the most marked differences being in increased vitamin B₆ and p-aminobenzoic acid contents. In the absence of biotin, significant amounts of all of the B vitamins except biotin were synthesized, both in the presence and absence of certain other members of the B complex. The addition of thiamine, pyridoxine, inositol, and β-alanine to the culture medium caused a reduction in the amounts of vitamin B₆ and p-aminobenzoic acid synthesized. F. B. yeast was able to grow in a xylose medium only when certain of the B vitamins were present, and even then growth was limited. Evidence was obtained for some synthesis of all of the vitamins investigated except biotin and vitamin B₆. The most significant differences in vitamin content between galac yeast and the parent F. B. strain were in folic acid and vitamin B₆, the former being considerably reduced in amount, the latter being increased.     

SYNTHESIS OF A PRECURSOR TRIPEPTIDE Z-Asp-Val-Tyr-OH OF THYMOPENTIN BY CHEMO-ENZYMATIC METHOD

The precursor tripeptide of thymopentin was synthesized by a combination of chemical and enzymatic methods. First, Val-Tyr-OH dipeptide was synthesized by a novel chemical method in two steps involving preparation of NCA-Val. Second, the linkage of the third amino acid Z-Asp-OMe to Val-Tyr-OH was completed by an enzymatic method under kinetic control. An industrial alkaline protease alcalase was used in water–organic cosolvent systems. The synthesis reaction conditions were optimized by examining the effects of several factors including organic solvents, water content, temperature, pH, and reaction time on the yield of Z-Asp-Val-Tyr-OH. The optimum condition is of pH 10.0, 35°C, acetonitrile/Na₂CO₃-NaHCO₃ buffer system (85:15, v/v), and reaction time of 2.5 hr, which achieves tripeptide yield of more than 70%.     

低pH对鱼类胚胎发育、鱼苗生长及鳃组织损伤影响的研究

本文研究了低pH水平对鱼类的胚胎,鱼苗和鱼种的影响以及鱼鳃的组织学观察。在硬水环境,pH≤4.5时,对泥鳅胚胎发育有严重影响。胚胎在低pH水平下,发育进程明显地迟缓。pH≤5.5时,泥鳅幼苗的生长受到抑制;在软水环境,pH≤4.5时,影响草鱼苗和幼鱼的存活率。低pH水平加上铝则对鱼类呈现出协同毒性。低pH使鱼鳃直接遭受严重的损害:出现大量的粘液、渗血、鳃上皮肿胀和脱落,组织增生和融合。     

抗癌药物紫杉醇的全合成—Holton合成紫杉醇路线的剖析

从全合成战略与路线,关键反应,保护基应用和立体化学方面对Holton的线性全合成紫杉醇路线进行剖析归纳。