嗜碱细菌降解木质素的复合碳源共代谢研究Ⅱ—营养条件调控机制的研究

文章研究了在蔗糖（第一碳源）＋麦草木质素（第二碳源）的复合碳源组合方式培养条件下,嗜碱木质素降解菌降解木质素的降解率及营养条件调控机制对其的影响。结果表明,蔗糖初始浓度为1g/L,添加0．3％的T－80和0.5mmol/L的ABTS并静置培养对菌株的产酶及降解能力有较大的促进作用。     

嗜碱细菌降解木质素的复合碳源共代谢研究Ⅰ——复合碳源组合方式及氮源的选择

研究在碱性液体培养条件 (pH =1 0 .5)下 ,不同碳源组合方式对嗜碱木质素降解菌产酶及降解麦草木质素的降解率的影响。研究结果表明 ,在蔗糖 (第一碳源 ) +麦草木质素 (第二碳源 )的组合方式下 ,并将硝酸铵作为氮源且碳氮比为 1 :1 .2时 ,该菌株的产酶及降解能力效果理想。     

汽爆麦草固态发酵木质素酶

木质素是地球上主要的可再生芳香族化合物 ,是地球上仅次于纤维素的第二丰富可再生天然资源 ,然而 ,对它的利用研究却很少 ,是天然高分子中未开发的领城[1] .80年代初发现了木质素过氧化物酶 (ＬｉＰ) [2 ,3] 和锰过氧化物酶 (ＭｎＰ) [4 ] 以后 ,木质素酶和木质素生物降解研究取得了一定进展 .但木质素微生物转化降解研究中仍存在许多问题 .( 1)由于木质素酶解是一种非专一性的、以自由基为基础的链反应过程[5] .因此 ,木质素酶在化学工业、煤化学和环境保护方面具有很大的开发前景 .而现在木质素酶的生产大多用合成培养基 ,必需添加昂贵的…     

Improved degradation of azo dyes by lignin peroxidase following mutagenesis at two sites near the catalytic pocket and the application of peroxidase-coated yeast cell walls

• Mutations in Lignin peroxidase Trp171 environment improved azo dyes degradation. • Expression on yeast cell surface and cell lysis allowed reusability of biocatalyst. • Aga2-LiP chimeric variants were characterized. The enzymatic degradation of azo dyes is a promising alternative to ineffective chemical and physical remediation methods. Lignin peroxidase (LiP) from Phanerochaete chrysosporium is a heme-containing lignin-degrading oxidoreductase that catalyzes the peroxide-dependent oxidation of diverse molecules, including industrial dyes. This enzyme is therefore ideal as a starting point for protein engineering. Accordingly, we subjected two positions (165 and 264) in the environment of the catalytic Trp171 residue to saturation mutagenesis, and the resulting library of 10⁴ independent clones was expressed on the surface of yeast cells. This yeast display library was used for the selection of variants with the ability to break down structurally-distinct azo dyes more efficiently. We identified mutants with up to 10-fold greater affinity than wild-type LiP for three diverse azo dyes (Evans blue, amido black 10B and Guinea green) and up to 13-fold higher catalytic activity. Additionally, cell wall fragments displaying mutant LiP enzymes were prepared by toluene-induced cell lysis, achieving significant increases in both enzyme activity and stability compared to a whole-cell biocatalyst. LiP-coated cell wall fragments retained their initial dye degradation activity after 10 reaction cycles each lasting 8 h. The best-performing mutants removed up to 2.5-fold more of each dye than the wild-type LiP in multiple reaction cycles.     

麦秸与奶牛场废水高固体混合厌氧发酵产甲烷研究

在实验室条件下,以麦秸和奶牛场废水为原料,设计麦秸与奶牛场废水质量比1∶4(T1)、1∶3(T2)和1∶2(T3)以及对照(麦秸与水质量比1∶4(T4))4个处理,研究发酵过程中日产气量、甲烷含量、发酵前后麦秸理化特性和结构的变化.结果表明,奶牛场废水与麦秸在中温、高固体条件下,厌氧发酵可以正常进行,且产气期延长2周以上,对甲烷含量的影响不大;厌氧发酵初始TS浓度对系统产气的影响较大,麦秸产气量随TS浓度的增加而降低,以T1的产气效果最好,麦秸TS产气量为0.41 L.g-1,较T4提高了17.14%,平均甲烷含量为48.78%;厌氧发酵后,麦秸半纤维素含量大幅降低,纤维素含量少许降低,降低幅度均为T1>T2>T4>T3,木质素含量稍有增加,各处理间无显著差异(P>0.05);FTIR和XRD的结果表明,厌氧发酵后,麦秸纤维素结晶区的相对含量增加,混合发酵可以促进厌氧微生物对麦秸纤维素结晶区的破坏.将麦秸与奶牛场废水(质量比)1∶4混合发酵产沼气是可行的,且促进了厌氧微生物对麦秸有机物的分解破坏,提高了麦秸产气量.     

稻草秆粉基质中白腐茵对三苯甲烷类染料的降解特性

研究了稻草秆粉基质中自腐菌对三苯甲烷类染料（孔雀绿、溴酚蓝和结晶紫）的降解特性．结果表明，白腐菌10d内对三苯甲烷类染料脱色率均可达到93％以上．染料加入对木素过氧化物酶分泌有抑制作用．随着培养过程的进行，染料脱色率增加，脱色降解以漆酶为主．紫外扫描发现，染料吸收波长有偏移，标志着染料结构发生变化，其中溴酚蓝能被完全矿化．图2表2参10     

Field evaluation and determination of four herbicides in a wheat ecosystem by a simple and versatile QuEChERS method with liquid chromatography-tandem mass spectrometry

Wheat products are consumed worldwide and every day; therefore, herbicides misuse in the wheat ecosystem has become a global food safety issue. Here, a simple and versatile method, QuEChERS (quick, easy, cheap, effective, rugged, and safe) with liquid chromatography-tandem mass spectrometry was developed to measure four herbicides in soil, wheat, and wheat straw – florasulam, carfentrazone-ethyl, fluroxypyr-meptyl, and fluroxypyr. By adjusting the amount of graphitized carbon black from 0 to 10 mg, the herbicides could be extracted with satisfactory recoveries of 80%–110%. Application of two water-dispersible granules showed first-order kinetics as well as half-lives between 1.2 and 5.1 d under open-field conditions. The dissipation kinetics of the four herbicides differed in the soil, wheat, and wheat straw, with residual concentrations in straw at 2 h after application being higher than in soil and faster degradation occurring in straw. The terminal residues in soil, wheat grain, and wheat straw were all below the maximum residue limits. The developed method was easy to handle and versatile; thus, it will facilitate regulation and inspection for possible misuse against Good Agriculture Practices. Moreover, the results of this study will contribute to global environmental protection as well as food safety issues.     

UV-Fenton法促进白腐菌处理草浆造纸蒸煮黑液

UV—Fenton法能够产生羟基自由基氧化草浆造纸蒸煮黑液中的有机质，白腐菌能够降解草浆造纸蒸煮黑液中的木质素，降低黑液COD，但是分别采用两种方法处理草浆造纸蒸煮黑液，效果都不明显．本研究初步探索了UV—Fenton法作为预处理对白腐菌处理草浆造纸蒸煮黑液体系的影响．与仅采用白腐菌处理黑液的效应相比，UV—Fenton法氧化黑液体系中易氧化的物质，改变了体系中难降解物的特性，降低了可溶性糖的含量，结果能提高白腐菌木质素降解酶系的分泌及酶的活性，增强白腐菌降解木质素及去除黑液COD的能力．图5表1参10     

氨化预处理对生物质秸秆厌氧发酵的影响

为探明氨化预处理对小麦秸秆干式厌氧发酵产气量影响的内在原因,采用傅立叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）、视频光学接触角测定仪（GBX）、热重分析（TGA）等方法,对氨化处理前后小麦秸秆中纤维结构的变化进行了多方面研究。结果表明,氨化预处理使小麦秸秆纤维部分官能团发生断裂,表层的亲水性增加,秸秆的热稳定性提高,并且氨化作用对纤维结构的破坏,有利于厌氧微生物接触到更多的可发酵物质,提高了秸秆的利用率,也表明小麦秸秆纤维结构的变化是厌氧发酵产气量提高的主要原因。     

麦季稻秆还田方式对后续稻季CH4排放的影响

通过田间试验研究了麦季4种稻秆还田方式（不还田、表面覆盖、均匀混施和原位焚烧）对后续稻季CH4排放的影响,以探讨稻．麦轮作系统中秸秆还田对稻田温室气体排放的后续效应。试验于2007年小麦播种期将4．8t·hm^-2水稻秸秆分别以不同方式还田（不还田处理除外）,利用静态箱／气相色谱法对2008年后续稻季CH4排放进行观测。结果表明,不同麦季稻秆还田方式显著影响后续稻季的CH4排放。与不施稻秆处理相比,表面覆盖和均匀混施处理后续稻季CH4排放量增加了75％和40％,且CH4排放量差异主要体现在水稻生长前期（0．60d）;原位焚烧处理CH4排放量与稻秆不还田处理相比无显著差异（P〉0．05）;与不施稻秆处理相比,均匀混施处理显著增加稻季开始前土壤全C质量分数6％和全N质量分数12％（P〈0．05）;各处理水稻（Oryza sativa L．）产量无显著差异（P〉0．05）;稻秆麦季均匀混施与表面覆盖相比能在一定程度上抑制后续稻季CH4排放．同时避免了秸秆焚烧导致的C、N、P等元素的大量损失,是较为合理的麦季稻秆还田方式。     

三相鼓泡塔生物反应器培养黄孢原毛平革菌合成木素过氧化物酶系

利用三相鼓泡塔反应器固定化培养黄孢原毛平革菌，可以高效地合成木素过氧化物酶系，固定化载体为聚氨酯泡沫塑料．实验表明，合成木素过氧化物酶和锰过氧化物酶的最佳通气量均是1．0vvm。在此通气量下，最大木素过氧化物酶的酶活达367U/L，最大锰过氧化物酶的酶活达4．72U／mL。在使用相同的培养基和固定化载体单位体积用量条件下，与摇瓶培养相比，酶活分别增大1倍和1．2倍．一定条件下，在三相鼓泡塔中可以进行重复间歇培养生产木素过氧化物酶，连续进行了5批培养，每批最大木素过氧化物酶的活力均在250U／L以上，最高酶活出现在第二批为480U／L，总培养时间达22d．图9参15     

不同作物原料热裂解生物质炭对溶液中Cd2+和Pb2+的吸附特性

选择由小麦秸秆、玉米秸秆和花生壳经350~500℃热裂解制成的生物质炭,研究生物黑炭对水溶液中Cd2+和Pb2+的吸附特性,分析了pH值、吸附时间、溶液初始质量浓度、生物质炭粒径和投加量对吸附效果的影响。结果表明:生物质炭对Cd2+和Pb2+的吸附约10 min即达平衡;3种生物质炭对Cd2+和Pb2+的等温吸附均可用Langmuir方程和Freundlich方程拟合,玉米秸秆炭对Cd2+和Pb2+的最大吸附量远大于小麦秸秆炭和花生壳炭;在生物黑炭投加量为150 mg(6 g.L-1)时,3种生物黑炭对溶液Cd2+的去除率均在90%以上,玉米秸秆炭对溶液Pb2+的去除率达90.30%,而小麦秸秆炭和花生壳炭的去除率仅为52%和47%,玉米秸秆炭有望成为处理重金属污染废水的新型吸附材料。     

不同作物原料热裂解生物质炭对溶液中Cd2＋和Pb2＋的吸附特性

选择由小麦秸秆、玉米秸秆和花生壳经350-500℃热裂解制成的生物质炭,研究生物黑炭对水溶液中Cd2＋和Pb2＋的吸附特性,分析了pH值、吸附时间、溶液初始质量浓度、生物质炭粒径和投加量对吸附效果的影响。结果表明：生物质炭对Cd2＋和Pb2＋的吸附约10 min即达平衡;3种生物质炭对Cd2＋和Pb2＋的等温吸附均可用Langmuir方程和Freundlich方程拟合,玉米秸秆炭对Cd2＋和Pb2＋的最大吸附量远大于小麦秸秆炭和花生壳炭;在生物黑炭投加量为150 mg（6 g.L-1）时,3种生物黑炭对溶液Cd2＋的去除率均在90%以上,玉米秸秆炭对溶液Pb2＋的去除率达90.30%,而小麦秸秆炭和花生壳炭的去除率仅为52%和47%,玉米秸秆炭有望成为处理重金属污染废水的新型吸附材料。     

麦秸还田与土壤耕作对稻季CH_4和N_2O排放的影响

2008年在大田试验条件下,设置麦秸还田旋耕、麦秸不还田旋耕、麦秸还田翻耕、麦秸不还田翻耕4个处理,采用静态暗箱-气相色谱法田间原位观测稻麦两熟制农田水稻生长季CH_4和N_2O排放通量,研究小麦秸秆全量还田与土壤耕作两项技术措施对稻季CH_4和N_2O排放的影响及其温室效应,以期为稻麦两熟制农田温室气体减排提供对策.结果表明:麦秸还田对稻季CH_4排放总量的影响达极显著水平,麦秸还田与耕作方式的互作效应对CH_4排放总量有显著影响,麦秸还田和耕作方式对N_2O排放总量的影响均达极显著水平;不同麦秸还田与土壤耕作处理稻季OH4排放总量为:麦秸还田旋耕>麦秸还田翻耕>麦秸不还田翻耕>麦秸不还田旋耕,N_2O的排放总量为:麦秸不还田翻耕>麦秸不还田旋耕>麦秸还田翻耕>麦秸还田旋耕;与麦秸不还田相比,相同耕作措施下麦秸还用排放CH_44和N_2O所产生的全球增温潜势(GWP)明显提高.麦秸还田条件下旋耕处理的GWP高于翻耕处理,而"单位产量的GWP"无明显差异,麦秸不还田条件下采用旋耕措施较翻耕可减轻温室效应.     

Influence of glucose feeding on the ligninolytic enzyme production of the white-rot fungus Phanerochaete chrysosporium

The present work studied the influence of glucose feeding on the ligninolytic enzyme production of Phanerochaete chrysosporium in a nitrogen-limited (C/N ratio is 56/8.8 mmol/L) medium. Several sets of shaking flask experiments were conducted. The results showed that 2 g/L glucose feeding on the first day of the culture (24 h after the inoculation) simulated both fungal biomass growth and enzyme production. The manganese peroxidase (MnP) activity was 2.5 times greater than that produced in cultures without glucose feeding. Furthermore, the glucose feeding mode in fed-batch culture was also investigated. Compared to cultures with glucose feeding every 48 h, cultures with glucose feeding of 1.5 g/L (final concentration) every 24 h produced more enzymes. The peak and total yield of MnP activity were 2.7 and 3 times greater compared to the contrast culture, respectively, and the enzyme was kept stable for 4 days with an activity of over 200 U/L.     

秸秆腐解剂在秸秆还田中的效果研究初报

在大田、微区和盆栽条件下,研究了秸秆腐解剂对小麦、水稻生长及产量的影响,同时研究了秸秆腐解剂对小麦、水稻秸秆腐解速率及对土壤肥力的影响。结果表明,稻、麦秸秆还田时施用秸秆腐解剂对提高稻、麦产量具有明显的增产效果,增产的原因是穗数和粒数增加;稻、麦秸秆还田量不同时,还田量大且配施秸秆腐解剂的效果较还田量小好;麦秸秆还田方式不同时,麦秸以栽稻前耕翻还田且配施秸秆腐解剂的效果较好,上水沤制的效果较差;秸秆腐解剂能促进稻、麦秸秆较快腐解,减轻和防止多量秸秆还田给作物生长带来不利影响,并可稳定和提高土壤养分含量。     

配施猪粪对麦季CH4和N2O排放及温室效应的影响

采用遮光密闭箱和气相色谱法研究配施猪粪条件下麦季CH4和N2O的排放特征,并运用全球增温潜势(PGW)对麦田CH4和N2O排放的温室效应进行估算。结果表明,常规施肥、秸秆还田、50%猪粪和100%猪粪替代化肥处理CH4平均排放通量分别为6.10、10.26、5.32和2.35μg.m-2.h-1,N2O平均排放通量分别为24.25、38.24、12.21和16.06μg.m-2.h-1。CH4排放通量在拔节后随温度升高而增加,N2O排放主要发生在苗期灌溉或降水后。与常规施肥相比,100%猪粪和50%猪粪替代化肥处理麦季CH4和N2O排放产生的总PGW分别降低34.3%和48.9%,单位产量的PGW分别降低26.0%和48.9%,秸秆还田措施的PGW及单位产量的PGW分别提高57.9%和52.0%。然而,与常规施肥与秸秆还田处理相比,100%猪粪处理的小麦产量显著降低(P<0.05)。试验结果表明,在作物高产、稳产要求下,50%猪粪替代化肥措施的减排效果较好。     

Substrate supply, fine roots, and temperature control proteolytic enzyme activity in temperate forest soils

Temperature and substrate availability constrain the activity of the extracellular enzymes that decompose and release nutrients from soil organic matter (SOM). Proteolytic enzymes are the primary class of enzymes involved in the depolymerization of nitrogen (N) from proteinaceous components of SOM, and their activity affects the rate of N cycling in forest soils. The objectives of this study were to determine whether and how temperature and substrate availability affect the activity of proteolytic enzymes in temperate forest soils, and whether the activity of proteolytic enzymes and other enzymes involved in the acquisition of N (i.e., chitinolytic and ligninolytic enzymes) differs between trees species that form associations with either ectomycorrhizal or arbuscular mycorrhizal fungi. Temperature limitation of proteolytic enzyme activity was observed only early in the growing season when soil temperatures in the field were near 4 degrees C. Substrate limitation to proteolytic activity persisted well into the growing season. Ligninolytic enzyme activity was higher in soils dominated by ectomycorrhizal associated tree species. In contrast, the activity of proteolytic and chitinolytic enzymes did not differ, but there were differences between mycorrhizal association in the control of roots on enzyme activity. Roots of ectomycorrhizal species but not those of arbuscular mycorrhizal species exerted significant control over proteolytic, chitinolytic, and ligninolytic enzyme activity; the absence of ectomycorrhizal fine roots reduced the activity of all three enzymes. These results suggest that climate warming in the absence of increases in substrate availability may have a modest effect on soil-N cycling, and that global changes that alter belowground carbon allocation by trees are likely to have a larger effect on nitrogen cycling in stands dominated by ectomycorrhizal fungi.     

Organochlorine pesticide residues in fodder from rural areas of Haryana,India

Residues of organochlorine pesticides (OCPs) namely 1,2,3,4,5,6-hexachlorocyclohexane isomers (HCH-isomers), 1,1,1-trichloro-2,2-bis(p-chlorophenyl) ethane (DDT) and its metabolites, and endosulfan stereoisomers were analyzed in dry and green fodder samples from rural areas of Ambala, Gurgaon, and Hisar districts of Haryana, India during winter, summer, and post-monsoon seasons. The HCH isomers γ-HCH and β-HCH, and DDT and its metabolites p,p′-DDD, p,p′-DDE, and p,p′-DDT had more traceability in test samples as compared to other isomers and metabolites studied. Total OCPs (ΣOCPs), i.e., ΣHCH, ΣDDT, and Σendosulfan were found to be the highest in wheat straw (1.1–1.2?mg?kg^?1) from Ambala and Gurgaon, followed by that in sorghum straw (0.46?mg?kg^?1) from Hisar. Dry fodder samples were found to have relatively higher residue levels than green fodders. In case of green fodder samples, maximum ΣOCP residues of 0.44?mg?kg^?1 were found in whole plant samples of sorghum from Gurgaon district followed by that in pearl millet (0.40?mg?kg^?1) from Ambala. The findings indicate highly significant differences (p?>?0.0001) in ΣOCPs and ΣDDT in wheat straw between different districts and reveal the persistence of OCP residues in both dry and green fodder samples in the study area.     

Isolation and identification of fungi tolerant to polycyclic aromatic hydrocarbons and coal tar from different habitats in Lithuania

A group of 36 fungal strains, belonging to the Lithuanian mycobiota, was collected and isolated from different locations, habitats, and matrices, including creosote-treated wood in storage yards for crosstie wastes. The eight most perspective strains selected according to preliminary assessment of tolerance to coal tar were subsequently identified combining taxonomical evaluation and molecular techniques. The tolerance of the eight identified fungal species (five basidiomycetes and three ascomycetes) to the presence of various concentrations of coal tar, and for the four most perspective fungal strains (Pleurotus sp., Schizophyllum sp., Irpex lacteus, Bjerkandera adusta) to polycyclic aromatic hydrocarbons was evaluated. The ligninolytic enzymatic activity assay of the isolated strains resulted in a good correspondence between the tolerance to pollutants and the capability to produce ligninolytic enzymes indicating that this group of white-rot fungi is perspective for further investigation and eventual usage for mycoremediation of polycyclic aromatic hydrocarbons polluted substrates.