几种天牛纤维素酶的来源

测定３种常见天牛：桑粒肩天牛（ＡｐｒｉｏｎａｇｅｒｍａｒｉＨｏｐｅ）、星天牛（ＡｎｏｐｌｏｐｈｏｒａｃｈｉｎｅｎｓｉｓＦｏｒｓｔｅｒ）和桔褐天牛（ＮｅｄｅｚｈｄｉｅｌｌａｃａｎｔｏｒｉＨｏｐｅ）消化道纤维素酶活性，发现几种天牛幼虫均有完整的纤维素酶体系，即Ｃ１酶、Ｃｘ酶和β－葡萄糖苷酶，能真正消化利用食物中的纤维素成分；以微晶纤维素（ＭＣ）和羧甲基纤维素钠（ＣＭＣ－Ｎａ）为碳源的选择性培养基培养几种天牛肠液，未发现纤维分解真菌及能分解ＭＣ的细菌；肠道解剖及肠组织切片未发现特殊的共生菌囊及含菌细胞；比较天牛的纤维素酶与常见真菌纤维素酶的性质相差甚远。因而认为所研究的几种天牛其纤维素酶均为其自身分泌的内源性酶而不是来源于其它生物。     

两种产纤维素酶菌株分离的初步研究

本研究对采自贵州林区中的响叶杨(Populusadenopoda Maxim)腐朽木和白蚁(Odontotermes formosanus(Shiraki))进行分离培养,分离自杨树腐朽木的产纤维素酶菌株为木霉属康宁木霉(Trichodermakoningii Oud);分离自白蚁肠的产纤维素酶菌株为芽孢杆菌属(Bacillus sp.)。对所分离的两个菌株进行滤纸降解观察与测定。所分离的菌株(Trichoderma koningii Oud.)滤纸培养30天后,有菌丝生长,培养基浑浊,滤纸呈絮状,培养7天后该菌对滤纸降解强度为7.13%(校正值)。刚果红法初步证明该菌能产纤维素酶。从白蚁肠分离的菌株(Bacillus sp.)滤纸培养15天后,滤纸呈絮状,培养基变浑浊,有细菌菌体沉积;培养7天后,对滤纸的降解强度为8.18%(校正值)。刚果红法初步证明该菌能产生纤维素酶。用五种培养基质培养芽孢杆菌属(Bacillus sp.)菌株,并测定其生长量,其中以滤纸为主要基质的培养液中,最高可检测到0.3×107cfu/ml菌株量。     

不同酶制剂对水稻秸秆和白酒糟混合青贮品质的影响

为研究不同酶制剂对水稻（Oryza sativa L.）秸秆和白酒糟（m︰m=1︰1）混合青贮品质的影响，设置CK（对照）、CEL（250 U·g^-1纤维素酶）和XYL（250 U·g^-1木聚糖酶）3个处理，青贮120 d后取样测定其青贮品质。结果表明：CEL处理的感官评定等级为良好；CEL和XYL处理的干物质、粗脂肪、钾和镁含量均显著高于CK（P<0.05），且CEL处理的粗蛋白和磷含量显著高于CK（P<0.05）；CEL和XYL处理的中性洗涤纤维、酸性洗涤纤维、木质素和纤维素含量均显著低于CK（P<0.05），且CEL处理显著低于XYL处理（P<0.05）；CEL和XYL处理30 h的养分消化率显著高于CK（P<0.05），且CEL处理48 h的养分消化率均显著高于CK（P<0.05）；CEL和XYL处理的饲用价值显著高于CK（P<0.05），且CEL处理显著高于XYL处理（P<0.05）。综上所述，添加纤维素酶、木聚糖酶能够提高水稻秸秆和白酒糟混合青贮的营养品质、养分消化率和饲用价值，且纤维素酶的添加效果优于木聚糖酶。     

Enzymatic activity of ectendomycorrhizal fungi

 Enzymatic (cellulolytic, pectolytic, proteolytic, chitinolytic and acid phosphatase) activities in the cultures of ten different ectendomycorrhizal strains (mainly Wilcoxina spp.) were studied. Although most of the fungi produced all these enzymes, their activities were low (milliunits per hour), with the exception of those of the pectolytic enzymes, which were detected at considerably higher levels (units per hour). Hebeloma crustuliniforme, an ectomycorrhizal fungus (positive control in our studies) synthesized ca. 2,000 times more exoglucanase than the ectendomycorrhizal fungi. Wilcoxina mikolae (ATCC 52685) had the lowest enzymatic activities of all 11 strains compared. Received: 3 December 1999     

Constraints on development of fungal biomass and decomposition processes during restoration of arable sandy soils

In an earlier study we reported the apparent stabilization of a low fungal biomass in ex-arable lands during the first decades after abandonment. It was hypothesized that the lack of increase in fungal biomass was due to constraints on development of fungi with persistent hyphae such as lignocellulolytic basidiomycetes and ericoid mycorrhizal fungi. With respect to the former group, the slow increase of the pool of lignocellulose-rich organic matter was expected to be the major constraint for their development. To study this, we enriched soil samples of one arable land, of two recently abandoned arable lands, of one older abandoned arable land and of heathland with carbon substrates that differed in composition (glucose, cellulose and sawdust). In addition, we combined the effect of carbon addition on fungal biomass development in arable and recently abandoned lands with inoculation of 1% of soil from the older abandoned site and the heathland. All treatments induced a fast increase and a subsequent rapid decline in fungal biomass in the arable and ex-arable fields. Denaturing Gradient Gel Electrophoresis (DGGE) band patterns and enzyme activities did show differences between the carbon treatments but not between the recent and older abandoned field sites, indicating a similarly responding fungal community even after three decades of land abandonment and irrespective of soil inoculation. Identification of fungi by sequencing and culturing confirmed that decomposition processes were mostly dominated by opportunistic fungi in arable and ex-arable fields. In the heathland, only a very slow increase of microbial activity was observed after addition of carbon and sequencing of DGGE bands showed that ericoid mycorrhiza (ERM) fungi were responsible for carbon decomposition. We conclude that an increase of enduringly present fungal hyphae in ex-arable land may only be possible when a separate litter layer develops and/or when suitable host plants for ERM fungi become established.     

产纤维素酶菌群的筛选及其酶学特性研究

从常年堆放的腐质豆秆中分离到1组产纤维素酶菌群MO,并在50℃、pH8.0条件下培养,90h时达到最高酶活,活性为1.3611IU。该酶最适反应温度为60℃,pH为7.0,在60℃以下和pH3~8范围内具有良好的热稳定性和pH稳定性。在最适反应条件下,该酶的最高活性可达2.13IU。通过生长动力学研究了菌群内部不同生长阶段pH、酶活和失重率的相互关系,并通过非变性电泳对酶谱进行初步研究,得到7条活性条带,说明MO中具有多种产酶菌株。     

RNA干扰技术和家蚕杆状病毒表达系统在纤维素酶研究中的应用前景

植物通过光合作用产生的纤维素是地球上最丰富、最廉价的可再生资源。纤维素酶是糖苷水解酶的一种,能有效地将农作物秸杆等富含纤维素的物质水解为葡萄糖,进而发酵产生生物乙醇,从而解决农业、再生能源以及环境污染等问题。随着生物化学、分子生物学以及基因工程等多种交叉学科的快速发展,获得适合工业化的高活力的纤维素酶指日可待。RNA干扰是利用双链RNA特异性地降解相应序列的mRNA,从而特异性地阻断相应基因的表达,是后基因组时代的一种强有力的调控目的基因表达的实验工具。家蚕杆状病毒表达系统是一种快速、高效表达外源基因的技术手段,目前该系统的发展和应用已经非常成熟。本文综述了纤维素酶的特性以及近年来国内外对纤维素酶、RNA干扰和家蚕杆状病毒表达系统的研究进展,并对该两种实验技术手段在今后纤维素酶研究中的应用前景作了预测和展望。     

酶解法提取海藻酸钠研究

采用正交实验研究了酶解法提取海藻酸钠的工艺及影响因素,并分析了单因素对酶解反应的影响,从而得出最佳酶解反应条件:缓冲液用量55 ml,pH值5.0,温度45 ℃,反应时间18 h,加酶量90 U/g,此时海藻酸钠浸出量可达2.448 7 g.     

稻瘟菌的胞外纤维素酶和果胶酶活性的研究

液体培养5种稻瘟菌生理小种.随着培养天数增加,ZC_3、ZD_1和ZG_1培养液中纤维素酶活性增高较快,而ZE_1和ZF_1增高较慢.培养液中纤维素酶活性增高快的,葡萄糖含量增加也快.培养8天的稻瘟菌培养液中各生理小种果胶酶活性高低顺序,果胶甲酯酶为:ZD_1>ZC_3>ZG_1>ZE_1>ZF_1;多聚半乳糖醛酸酶活性为:ZD_1>ZC_3>ZE_1>ZF_1>ZG_1.研究表明,胞外纤维素酶和果胶酶活性高的稻瘟菌生理小种,其致病性也强.     

咪唑型离子液体溶液中纤维素酶荧光活性测定

为考查纤维素酶在离子液体溶液中的酶活,采用荧光光谱分析法对离子液体中纤维素酶的荧光活性测定,研究离子液体的浓度、类型对纤维素酶的活性影响。结果表明:纤维素酶在咪唑类离子液体溶液中荧光活性随离子液体浓度的升高而降低。离子液体为[C4 mim]Cl、[C4 mim]Br,且浓度为0.1 mol/L使纤维素酶的荧光强度减低最小,即对酶活性影响最小。