氮源对里氏木霉木聚糖酶和纤维素酶生物合成的影响

研究了氮源种类和比例、碳氮比(C／N)等因素对里氏木霉木聚糖酶和纤维素酶生物合成的影响。在各种氮源中，蛋白胨是最好的氮源。复合氮源中当硫酸铵N和尿素N的比例为1：3时，木聚糖酶活力最高，达93．3IU／mL；当比例为1：1时，滤纸酶活力和羧甲基纤维素(CMC)酶活力达到最大值，分别为0．263FPIU／mL和0．026IU／mL。当控制培养基的C／N为8．0和6．0时，它们对木聚糖酶和纤维素酶的诱导作用最强，分别为95．1IU／mL和0．310FPIU／mL。     

β-聚糖酶的诱导合成(Ⅰ)--碳源、氮源及碳氮比的影响

以绿色木霉Trichoderma viride LH-0ll为产酶菌株,利用自制复合碳源,就碳源浓度、氮源种类及碳氮比等因素对β-聚糖酶的诱导合成的影响进行了研究.结果表明:以15 g/L预处理玉米芯Ⅰ为碳源,利用复合氮源((NNH4)2SO4 尿素 蛋白胨).控制C/N为6.4,调节初始pH值4.8,温度为28～30℃,150 r/min摇瓶培养72 h,纤维素酶活力和木聚糖酶活力分别达到了3.34 U/mL和13.2 U/mL.     

木聚糖相对分子质量分布对里氏木霉合成木聚糖酶的影响

以里氏木霉(Trichoderma reesei)Rut C-30为产酶茵,研究了相对分子质量(Mw)分布不同的木聚糖对木聚糖酶合成的影响。通过SephadexG一100凝胶过滤色谱分级分离发现木聚糖A中低Mw组分较多,木聚糖B中低Mw组分较少,木聚糖C中低Mw组分最少。分别以这3种木聚糖为碳源合成木聚糖酶,最高木聚糖酶活力分别为153．64、120．84和110.84IU／mL,产酶时间分别为60、72和96h。用这3种碳源合成的木聚糖酶酶解粗木聚糖,酶解2h时,产物中低聚木糖分别占总糖的80．70％、68．56％和66．92％。这表明低Mw组分较多的木聚糖不仅有利于促进木聚糖酶的诱导合成,而且有利于促进内切-1,4-木聚糖酶的合成。     

耐热型木聚糖酶制备工艺的研究

以木糖渣为碳源，利用耐热子囊菌（Thermoascus aurantiacus)液体发酵生产木聚糖酶，确立了最佳工艺参数为：木糖渣质量浓度4％，培养温度45摄氏度，转速180r/min，培养基初始pH值4．8，初始C／N为7．1，木聚糖酶活性可达900U/mL。酶学性质研究表明，该酶反应的最适温度为75摄氏度，最适pH值为4．8，该酶的耐热性强，在75摄氏度下温保1h，酶活力剩余85％以上。     

芽孢杆菌产纤维素酶的研究

对芽孢杆菌 (Bacillussp.ZU 0 4 )产纤维素酶的工艺参数进行了优化 ,研究结果表明 :木糖渣和豆饼粉分别是该菌合成纤维素酶的适宜碳源和氮源 ,NaCl和KH2 PO4对纤维素酶的合成具有重要作用 ,其适宜质量用量分别为 0 .5%～ 1.0 %及 0 .1% ,麸皮的添加可明显提高发酵液中的酶活力 ;3L发酵罐中的适宜发酵条件为 :搅拌速度 30 0r/min ,通气量 0 .3L/(L·min) ,培养温度 37℃ ,中性和碱性纤维素酶活力分别达到了2 57.6和 12 5.6U/mL ,显示了良好的工业应用前景。     

碱性纤维素酶发酵条件的研究

采用芽孢杆菌Bacillus sp.AC-2作为碱性纤维素酶的生产菌株，对其发酵条件进行了研究。适宜的种子茵龄为24h；适于产酶的培养温度为37℃，培养基初始pH值10．5；在碳源和氮源分别为1％的淀粉和2％的豆饼粉时，碱性纤维素酶活力可达到76．04U／mL。该酶反应的最适温度为60℃；酶液在70℃以下、pH值6．0～10．0范围内稳定。该酶制剂在棉织物洗涤和纸浆脱墨方面具有良好的应用前景。     

剩余污泥合成PHAs的影响因素研究

利用序批式反应器，以乙酸钠为碳源驯化活性污泥，选择和富集具有积累聚羟基链烷酸酯（PHAs）能力的微生物菌群，探究了不同的碳源条件对PHAs合成产率、单体（PHB、PHV）组成的影响，寻求PHAs总量和PHAs单体产率最高的碳源组成和类型；考察了影响剩余污泥合成PHAs的因素，重点调控pH、碳氮比条件，实现剩余污泥贮存PHAs的工艺优化。实验结果表明：在微好氧循环体系中，相对于丙酸、丁酸等单一有机酸为碳源的培养条件，乙酸为单一碳源时可获得最高的PHAs合成量，达到了0.234mg／mg ，其中PHB的产率为0.225mg／mg ，占96％。不同比例的乙酸－丙酸和丁酸－丙酸等混合酸的合成条件下，以乙酸／丙酸＝1∶2的混合酸作为碳源时，单位质量干重污泥PHAs的产率可达到为0.384mg／mg ；对比C／N分别为20、100、500的培养条件，当 C／N ＝500时剩余污泥贮存 PHAs 的能力较强，最高产率约为0.356mg／mg ；分别控制pH值＝5、pH值＝7、pH值＝9的酸度条件，则中性环境更有利于剩余污泥贮存，可达0.39mg／mg的PHAs。     

纸浆漂白用木聚糖酶的选择性合成

以里氏木霉(Trichoderma reesei) Rut C-30为产酶菌,研究了碳源、培养温度、初始pH值、碳氮比对木聚糖酶和纤维素酶合成的影响.结果表明,粗木聚糖和亚硫酸盐纸浆混合作为碳源有利于木聚糖酶和纤维素酶的合成;低温有利于木聚糖酶和纤维素酶的合成,但产酶时间较长,高温对木聚糖酶的合成有一定的影响,对纤维素酶的合成能有效地抑制,且产酶时间较短;初始pH值低有利于纤维素酶的合成,初始pH值高则延长了木聚糖酶的合成时间,且强烈抑制纤维素酶的合成;低碳氮比有利于纤维素酶的合成,高碳氮比使得木聚糖酶的合成滞后,能够有效抑制纤维素酶的合成.以粗木聚糖和亚硫酸盐纸浆混合作为碳源,调控培养温度、初始pH值和碳氮比能有效地促进木聚糖酶的合成,抑制纤维素酶的合成,致使木聚糖酶活与纤维素酶活的比值提高,从而有利于选择性合成纸浆漂白用木聚糖酶,调控培养方式为:提高碳氮比(7.2)和初始pH值(6.0),在培养初期(1 d)培养温度为35～36 ℃,中后期培养温度25～26 ℃,调控6 d后,木聚糖酶酶活和纤维素酶酶活分别为186.93和0.156 IU/mL,酶活比为1 198.     

纤维素酶系组成对二次纤维形态与表面结构的影响

用不同滤纸酶活力和羧甲基纤维素酶活力比(F／C)值的纤维素酶系处理轻量涂布纸二次纤维，研究了纤维素酶系组成对二次纤维性能的影响。随着F／C值从0增到2．5，长度小于0．2mm的纤维分布密度由23．55％降低到17．50％。纤维加权平均长度由1．067mm增加到1．168mm，纤维的卷曲系数由0．103下降到0．087；纤维的扭结强度由0．78个／mm增加到0．84个／mm。随着F／C值的增加，纤维素酶系外切酶的酶解作用增强，细小组分逐渐减少，二次纤维平均长度呈上升趋势，卷曲程度下降，有利于浆料滤水性能的改善，但纤维扭结强度上升对单根纤维强度和纸张强度产生不利影响。     

响应面法优化芽孢杆菌产脂肪酶发酵条件

通过单因素试验、Plackett-Burrman设计及响应面法对芽孢杆菌T3—4的产脂肪酶条件进行优化。确定最适C、N源分别为麦芽糖、蛋白胨+硫酸铵,碳氮比为1∶3。其最适产酶条件为:七水硫酸镁0.05%,蓖麻油1.5%,蛋白胨0.75%,磷酸氢二钾0.1%,麦芽糖0.5%,硫酸铵0.3%,pH值6.5,接种量10%。在此条件下,发酵脂肪酶活力可达805.836 U/mL。     

里氏木霉选择性合成木聚糖酶的研究(Ⅰ)

在一定条件下,里氏木霉可选择性地合成木聚糖酶,选择性合成程度与碳源种类、碳源浓度及碳氮比大小有关。以低纤维素污染的木聚糖为碳源,适当降低碳源的浓度,提高碳氮比可以提高木聚糖酶选择性合成的程度。以玉米芯粗木聚糖为碳源,采用添加碳源和氮源的方法控制发酵过程中的碳源浓度在５ｇ／Ｌ下,碳氮比在１１２以上时,产生的木聚糖酶和纤维素酶酶活分别为３５５Ｕ／ｍＬ、０．２Ｕ／ｍＬ,两种酶活的比值为１１７     

Effects of nitrogen source and defoliation on growth and biological dinitrogen fixation of Gliricidia sepium seedlings

Effects of four N sources and two defoliation treatments on growth and nitrogenase activity of Gliricidia sepium (Jacq.) Walp seedlings were studied in a greenhouse. All nutrients were supplied in irrigation water to the sterile growing medium. The N sources were: (1) 100 mg l(-1) of N supplied as NO(3) (-) (high-NO(3) (-)), (2) 50 mg l(-1) of N supplied as NO(3) (-) and inoculation with Rhizobium spp. medium-NO(3) (-)), (3)100 mg l(-1) of N supplied as NH(4)NO(3), and (4) inoculation with Rhizobium spp without mineral N (N(2)). At 35 weeks after sowing, mean total biomass was 130.5, 50.5, 22.9 and 17.4 g seedling(-1) in the NH(4)NO(3), N(2), medium-NO(3) (-) and high-NO(3) (-) treatments, respectively. The root/shoot ratio was high in all of the N treatments (1.73-2.77) because the seedlings had big taproots. The medium-NO(3) (-) treatment completely inhibited nodulation, whereas seedlings in the N(2) treatment were profusely nodulated. At 32 weeks after sowing, groups of seedlings in the N(2) and high-NO(3) (-) treatments were subjected to 50 or 100% defoliation. Closed-chamber acetylene reduction assays of intact root systems were conducted to compare nitrogenase activity at 7, 14 and 28 days after defoliation (DAD). At 7 and 14 DAD, nitrogenase activity of completely and partially defoliated seedlings was about 10 and 60%, respectively, of that of undefoliated controls. At 28 DAD, nitrogenase activity of completely defoliated seedlings was twice the predefoliation value, whereas nitrogenase activity of partially defoliated seedlings was only 87% of the predefoliation value. Recovery of nitrogenase activity was strongly correlated with foliage regrowth in the completely defoliated seedlings, but not in the partially defoliated seedlings. Abundant belowground C and N reserves in the large taproot probably contributed to the rapid recovery from defoliation. Accumulation of belowground biomass may also improve defoliation tolerance of mature trees.     

来源于解酯菌的嗜热耐碱脂肪酶的表达纯化及其酶学性质研究

脂肪酶广泛应用于食品加工、生物柴油制备等领域。为了有效提高微生物脂肪酶的可利用度,将来源于解酯嗜热菌(Thermosyntropha lipolytica DSM 11003)的脂肪酶基因(tll2)克隆到大肠杆菌表达载体pET28a中,获得重组质粒pET28a TLL2,将pET28a TLL2转入大肠杆菌BL21(DE3)宿主菌中进行高效表达。通过热处理和镍柱亲和层析得到电泳纯的蛋白TlLip2,对其进行酶学性质表征。结果显示,TlLip2的亚基分子质量为57 ku,比酶活为31 U/mg,最适反应pH及温度分别为8.0和60℃。在60℃以下、pH 6.0~11.0范围内,TlLip2的稳定性较好,相对酶活均维持在60%以上。金属离子K+和Na+对TlLip2有较大激活作用,而Co^2+、Zn^2+、SDS和Tween 80对TlLip2酶活力抑制作用明显。TlLip2在极性小的有机溶剂中具有较好的耐受性。TlLip2对中等碳链长度的对硝基苯酯显示出更高的活性,其中以对硝基苯己酯为底物时,TlLip2的酶活最高;以对硝基苯棕榈酸酯为底物时,该酶的动力学常数Km为0.29 mmol/L,Vmax为2.28 mmol/(L·min^-1),kcat为6.19 s^-1。此外,将其应用到油酸乙酯的合成研究中,以油酸和乙醇为底物,研究TlLip2催化制备油酸乙酯的转化条件,结果表明在60℃、加酶量为100 U/g的TlLip2,催化摩尔比为1∶2的油酸和乙醇,12 h后油酸乙酯的转化率达到62.1%,表明TlLip2在催化脂肪酸酯化方面具有较大的应用潜能。     

不同氮素水平对红厚壳幼苗生长及光合特性的影响

采用温室盆栽方法,设置0(对照)、50、100、150、200、300、400、600 mg.株-18个氮素处理,研究氮素供应对红厚壳幼苗生长及光合特性的影响。结果表明:随供氮量的增加,红厚壳幼苗高、地径、叶面积及整株生物量均呈现出先增加后降低的趋势,均在200 mg.株-1时达到峰值,分别为22.06 cm、0.65 cm、327.27 cm2和10.43 g.株-1,是对照的1.59、1.25、2.58、1.90倍;地下生物量N5显著高于N7、N8处理,但与其余处理无显著性差异,根冠比随供氮量的增加呈现单调递减趋势;红厚壳幼苗叶绿素a、b及总量随供氮量的增加而增加;红厚壳幼苗的净光合速率、气孔导度、胞间CO2浓度以及蒸腾速率随供氮量的增加呈现出先增加后减小的趋势,均在200 mg.株-1时达最大值,分别是7.29μmol.m-2.s-1、0.071 mol.m-2.s-1、220.22μmol.mol-1和1.34 mmol.m-2.s-1,分别是对照的2.26、1.92、1.37、1.79倍,不同氮素处理间红厚壳光合气体交换参数差异均显著。结果表明:施氮量200 mg.株-1左右为红厚壳幼苗温室栽培的最佳施肥量。     

Soil nitrate dynamics in relation to nitrogen source and landscape position in Malawi

Nitrogen is normally the nutrient most limiting production of maize (Zea mays) — the main staple food crop — in southern Africa. We conducted a field study to determine the effect of N sources on soil nitrate dynamics at three landscape positions in farmers' fields in southern Malawi. The landscape positions were dambo valley or bottomland, dambo margin, and steep slopes. The N sources were calcium ammonium nitrate fertilizer applied at 120 kg N ha^–1, biomass from Sesbania sesban, and no added N. Sesbania biomass was produced in situ in the previous season from sesbania relay cropped with maize. Nitrate in the topsoil (0 to 15 cm depth) increased to 85 days after maize planting (mean = 48 kg N ha^–1) and then decreased markedly. Application of N fertilizer and sesbania biomass increased soil nitrate, and nitrate-N in topsoil correlated positively with amount of incorporated sesbania biomass. The strongest correlation between sesbania biomass added before maize planting and topsoil nitrate was observed at 85 days after maize planting. This suggests that the sesbania biomass (mean N content = 2.3%) mineralized slowly. Inorganic N accumulated in the subsoil at the end of the maize cropping season when N fertilizer and sesbania were applied. This study demonstrated the challenges associated with moderate quality organic N sources produced in smallholder farmer's fields. Soil nitrate levels indicated that N was released by sesbania residues in the first year of incorporation, but relay cropping of sesbania with maize may need to be supplemented with appropriately timed application of N fertilizer.This revised version was published online in November 2005 with corrections to the Cover Date.     

不同N源对兴安落叶松苗木生长,养分吸收及离子平衡的影响(英文)

本文研究了不同N源对落叶松苗木干物重、养分吸收及离子平衡的影响。结果表明:施铵态N的苗木干物重小于对照处理;施硝铵态N的苗木生长与对照和施硝态N的苗木差别不大,但较施铵态N的苗木生长要好。铵态N与硝铵态N处理苗木地上部的阳离子总浓度受N水平的影响很小,而硝态N处理时则阳离子浓度增加;各处理对地上部阴离子浓度的影响均不大。在根部除硫酸根与钙离子外,阳离子与阴离子浓度均变化不大。硝态N处理的苗木具有较高的C-A值;△(C-A)/△Norg值在硝态N处理的苗木中随N供应量的增加而略有变化,但铵态N处理则明显下降;硝铵态N处理苗木的△(C-A)/△Norg值界于铵态N处理与硝态N处理之间。     

川金丝猴食源地衣长松萝生长、存活与生理学特征对模拟氮沉降的响应

【目的】探讨氮沉降对长松萝生长、存活的影响及其生理响应特征,为理解氮沉降增加背景下神农架地区树生地衣群落的演替趋势、科学管理川金丝猴的地衣食物供应提供理论支撑。【方法】在野外对长松萝个体和繁殖体模拟施加不同强度的氮沉降,测定其个体生长速率和繁殖体存活率;同时开展室内试验,测定地衣体对铵态氮和硝酸盐氮吸收速率、地衣体的氮磷含量及其比值以及氮磷代谢关键酶(谷氨酰胺合成酶、硝酸还原酶和酸性磷酸酶)活性等指标,综合分析氮沉降影响长松萝生长、存活的规律及其生理学指标响应机制。【结果】低强度氮沉降(<20.94 kgN ·hm -2 a -1 )能促进长松萝个体生长,高强度氮沉降(>32.99 kgN ·hm -2 a -1 )则对长松萝生长与繁殖体存活均有显著不利影响( P ≤0.05);与硝态氮相比,长松萝偏向于吸收铵态氮,且吸收量随氮沉降强度增加而增加;长松萝地衣体氮磷比、酸性磷酸酶活性与氮沉降强度显著正相关( P ≤0.05);随氮沉降强度增加,长松萝谷氨酰胺合成酶和硝酸还原酶活性呈先增加后降低趋势。【结论】低强度氮沉降(<20.94 kgN ·hm -2 a -1 )促进长松萝生长和存活,而高强度氮沉降(>32.99 kgN ·hm -2 a -1 )抑制长松萝生长和存活。过量氮沉降损伤长松萝地衣健康是多种生理机制综合作用的结果,包括选择性吸收铵态氮导致的长松萝体内铵态氮的积累、铵态氮积累引起谷氨酰胺合成酶和硝酸还原酶的活性下降以及铵态氮积累导致地衣磷限制等方面。     

Intsia palembanica wood extracts and its isolated compounds as Propionibacterium acnes lipase inhibitor

Potential compounds from Intsia palembanica methanol extracts were isolated. Three isolated compounds as well as 7 reported compounds from Merbau were analyzed for their ability to inhibit the lipase activity of Propionibacterium acnes. Lipase was isolated from P. acnes and used as enzyme for activity analysis. The lipase activity test was performed using the 2,3-dimercapto-1-propanol tributyrate (BALB) method. The results showed that methanol extract and water fraction were active to inhibit lipase activity but n-hexane fraction and EtOAc fraction did not reach 50 % inhibition until 500 μg/ml. EtOAc fraction consisted of flavonoid, naringenin, robinetin, and (+)-epirobidanol accelerated the lipase activity. The other 7 compounds showed inhibitory activity of lipase in a concentration-dependent manner. In conclusion, fustin is the most active compound to inhibit the P. acnes lipase activity.     

拟南芥悬浮细胞生长特性及其继代培养条件

研究了拟南芥悬浮细胞的生长特性及其继代培养条件。结果表明:在每个7d一次的继代周期中,拟南芥悬浮细胞的生长表现出明显的S型生长曲线。正常培养条件下,继代后2-4 d出现快速生长,此后生长量迅速下降,但接种量不同,其继代后悬浮细胞生物量及快速增长发生时期不同。维持7 d继代周期的理想接种量为起始悬浮液按10 mL／50 mL稀释。接种量小,生长迟缓,甚至不能增殖。培养基中不同激素及不同蔗糖含量均会对其生长产生极明显的影响,在B5 0．2 mg／L NAA条件下反复继代后的细胞,再加入外源NAA时,反而会抑制其生长。蔗糖含量以30 g／L最好,蔗糖含量低,悬浮生物量及细胞活性明显下降。     

Comparison of two coniferous plantations in central Japan with respect to forest productivity, growth phenology and soil nitrogen dynamics

Japanese cypress (Chamaecyparis obtusa Endl.) and Japanese cedar (Cryptomeria japonica D. Don) are common species for plantation forestry in Japan. Cypress is conventionally planted on sites of low fertility whereas for cedar high fertility sites are used. Objectives of this study were to compare the productivities of cypress and cedar plantations grown on adjacent sites where common properties of soils, such as pH values and C and N contents, were similar, and to relate the N cycling at their site with productivities. The stem diameter of trees, aboveground litter production and fine root biomass were measured as indices of forest productivity. Parameters of N cycling included pools of total N and mineral N (ammonium + nitrate), annual N leaching, and potentially mineralizable N. The radial stem increment of the two tree species was similar. However, cedar site had higher total basal area and annual basal increment than cypress site reflecting higher tree density on the cedar site. Aboveground litter, fine root biomass, soil organic matter, and N turnover were higher on the cedar site than on the cypress site. However, litter production and fine root biomass per unit basal area was greater at the cypress site. Phenological pattern of stem growth and periodical litter production were similar for both species during the study period (1992–2000), but showed distinct annual variations caused by the fluctuation in the ambient temperature and precipitation. Mineral N content and the N mineralization potential were greater on the cedar site, indicating greater N availability and higher total tree productivity at the cedar site than those at the cypress site. When provided with more space in the canopy to expand more needles and in the soil to develop more fine roots to exploit sufficient resources, the individual cypress trees have the potential to grow faster. On fertile site and at lower tree density, thicker logs of cypress might be yielded.