Use of fungi for the bioconversion of rice straw into cellulase enzyme

Cellulase production was carried out by solid state bioconversion (SSB) method using rice straw, a lignocellulosic material and agricultural waste, as the substrate of three Trichoderma spp. and Phanerochaete chrysosporium in lab-scale experiments. The results were compared to select the best fungi among them for the production of cellulase. Phanerochaete chrysosporium was found to be the best among these species of fungi, which produced the highest cellulase enzyme of 1.43 IU/mL of filter paper activity (FPase) and 2.40 IU/mL of carboxymethylcellulose activity (CMCase). The “glucosamine” and “reducing sugar” parameters were observed to evaluate the growth and substrate utilization in the experiments. In the case of Phanerochaete Chrysosporium, the highest glucosamine concentration was 1.60 g/L and a high concentration of the release of reducing sugar was measured as 2.58 g/L obtained on the 4th day of fermentation. The pH values were also recorded. The range of the pH was about 5.15 to 5.56 in the case of Phanerochaete Chrysosporium.     

Phanerochaete chrysosporium吸附载体的选择及染料降解研究

研究了游离细胞与载体吸附培养、不同载体材料对Phanerochaete chrysosporium进行连续染料脱色及产酶能力的影响。结果表明,P.chrysosporium可在载体上良好生长,甚至生长到载体内部。木屑、玉米芯、花生壳3种载体材料中,以木屑载体吸附培养物的持续脱色和产酶效果最佳,该培养物经三轮连续脱色后对染料RB5仍能达到最高95%的脱色率,并产生596 U/L锰依赖过氧化物酶（MnP）和1 326 U/L木质素过氧化物酶（LiP）,对染料的持续脱色和产酶能力明显优于游离细胞培养物。     

纤维素高效降解菌NU-H的诱变选育及降解特性

以草酸青霉(Penicillium oxalicum)D1为出发菌株,经过紫外线、亚硝酸以及紫外与亚硝酸复合诱变处理,选育出1株高产纤维素酶突变株NU-H,与出发菌株相比,CMCase酶活提高69.8%,滤纸酶活提高75.28%,另外木聚糖酶活,还原糖得率也有显著地提高。通过单因子及正交实验,研究了突变株纤维素酶对稻草粉水解的最适宜条件。结果表明,该菌株酶解糖化的最佳条件为:温度50℃,时间28 h,纤维素酶浓度为60%,底物浓度3%,pH 4.8,还原糖得率为23.25%。     

Biodegradation of aromatic compounds by white rot and ectomycorrhizal fungal species and the accumulation of chlorinated benzoic acid in ectomycorrhizal pine seedlings

The capability of different white rot (WR, Heterobasidion annosum, Phanerochaete chrysosporium, Trametes versicolor) and ectomycorrhizal (ECM, Paxillus involutus, Suillus bovinus) fungal species to degrade different aromatic compounds and the absorption of 3-chlorobenzoic acid (3-CBA) by ECM pine seedlings was examined. The effect of aromatic compounds on the fungal biomass development varied considerably and depended on (a) the compound, (b) the external concentration, and (c) the fungal species. The highest effect on the fungal biomass development was observed for 3-CBA. Generally the tolerance of WR fungi against aromatic compounds was higher than that of the biotrophic fungal species. The capability of different fungi to degrade aromatic substances varied between the species but not generally between biotrophic and saprotrophic fungi. The highest degradation capability for aromatic compounds was detected for T. versicolor and H. annosum, whereas for Phanerochaete chrysosporium and the ECM fungi lower degradation rates were found. However, Paxillus involutus and S. bovinus showed comparable degradation rates at low concentrations of benzoic acid and 4-hydroxybenzoic acid. In contrast to liquid cultures, where no biodegradation of 3-CBA by S. bovinus was observed, mycorrhizal pines inoculated with S. bovinus showed a low capability to remove 3-CBA from soil substrates. Additional X-ray microanalytical investigations showed, that 3-CBA supplied to mycorrhizal plants was accumulated in the root cell cytoplasm and is translocated across the endodermis to the shoot of mycorrhizal pine seedlings.     

林可霉素菌（Streptomyces lincolnensis）利用吡唑酮废水的研究

采用被吡唑酮废液驯化、分离、筛选后的林可霉素菌,并对其在摇瓶上利用吡唑酮废液中的硫酸铵发酵（7 d）生产林可霉素进行了研究。实验结果表明,废液加入培养基体积比都为1∶10,实验1中菌丝代谢和对照比正常,其中还原糖利用最快,在发酵后期为0.24 mg/L,林可霉素起步效价最低为2 100 IU/mL,与对照相比最后发酵效价降低了70 IU/mL;实验2发酵过程pH值偏低,全程为5.86～6.50,氨基氮代谢缓慢为40 mg/100 mL,最后林可霉素效价最低为4 480 IU/mL;实验3中废液在发酵进入48 h中后期的时候补入能促进菌丝体分泌,最后林可霉素为5 180 IU/mL,比对照发酵水平高出8.82%。可见实验3的实验设计有利于林可霉素菌利用吡唑酮废液生产林可霉素,为废物循环利用、变废为宝的可行性作了有意义尝试。     

林可霉素菌(Streptomyces lincolnensis)利用吡唑酮废水的研究

采用被吡唑酮废液驯化、分离、筛选后的林可霉素菌,并对其在摇瓶上利用吡唑酮废液中的硫酸铵发酵(7 d)生产林可霉素进行了研究。实验结果表明,废液加入培养基体积比都为1∶10,实验1中菌丝代谢和对照比正常,其中还原糖利用最快,在发酵后期为0.24 mg/L,林可霉素起步效价最低为2 100 IU/mL,与对照相比最后发酵效价降低了70 IU/mL;实验2发酵过程pH值偏低,全程为5.86～6.50,氨基氮代谢缓慢为40 mg/100 mL,最后林可霉素效价最低为4 480 IU/mL;实验3中废液在发酵进入48 h中后期的时候补入能促进菌丝体分泌,最后林可霉素为5 180 IU/mL,比对照发酵水平高出8.82%。可见实验3的实验设计有利于林可霉素菌利用吡唑酮废液生产林可霉素,为废物循环利用、变废为宝的可行性作了有意义尝试。     

白腐真菌F1对染料脱色特性的研究

本研究从我国生物资源中筛选出一株白腐真菌F1,确定了其生长条件 ,实现了扩大培养 ,并研究了F1与黄孢原毛平革菌对 4种难降解染料的脱色效果。试验结果表明 ,F1对中性深黄GRL、酸性媒介漂蓝B和刚果红的脱色效率都超过 90 % ;白腐真菌能在短时间内将有毒染料酸性媒介漂蓝B降低到较低浓度 ;白腐真菌对染料脱色的同时 ,自身能够繁殖生长。     

紫外诱变黄孢原毛平革菌染料脱色研究

对黄孢原毛平革菌孢子悬浮液进行了紫外诱变,并将筛选出的优势菌ZWYB1与未经诱变的菌作对比实验,结果表明:诱变筛选得到的ZWYB1不仅可缩短对染料的脱色时间,提高对杂环类染料蕃红花红及高浓度染料400 mg/L的孔雀石绿的脱色效果,而且诱变后的菌种具有一定的脱色能力稳定性.     

紫外诱变黄孢原毛平革菌染料脱色研究

对黄孢原毛平革菌孢子悬浮液进行了紫外诱变，并将筛选出的优势菌ZWYB1与未经诱变的菌作对比实验，结果表明：诱变筛选得到的ZWYB1不仅可缩短对染料的脱色时间，提高对杂环类染料蕃红花红及高浓度染料400mg／L的孔雀石绿的脱色效果，而且诱变后的菌种具有一定的脱色能力稳定性。     

Mycelial growth and solid-state fermentation of lignocellulosic waste by white-rot fungus Phanerochaete chrysosporium under lead stress

Lignocellulosic biomass is an abundant renewable resource difficult to degrade. Its bioconversion plays important roles in carbon cycles in nature, which may be influenced by heavy metals in environment. Mycelial growth and the degradation of lignocellulosic waste by lignin-degrading fungus Phanerochaete chrysosporium under lead stress were studied. It was shown that P. chrysosporium could grow in liquid media with 400 mg L?1 Pb(II), and mycelial dry weight was reduced by 54% compared to the control. Yellow mycelia in irregular short-strip shape formed in Pb-containing media, whereas the control showed ivory-white regular mycelial pellets. Two possible responses to Pb stress were: dense hyphae, and secretion from mycelia to resist Pb. During solid-state fermentation of straw, fungal colonization capability under Pb stress was positively correlated with the removal efficiency of soluble-exchangeable Pb when its content was higher than 8.2 mg kg?1 dry mass. Carboxymethyl cellulase activity and cellulose degradation were inhibited at different Pb concentrations, whereas low Pb concentrations increased xylanase and ligninolytic enzyme activities and the hemicellulose and lignin degradation. Cluster analyses indicated that Pb had similar effects on the different microbial indexes related to lignin and hemicellulose degradation. The present findings will advance the understandings of lignocellulose degradation by fungi under Pb pollution, which could provide useful references for developing metal-polluted waste biotreatment technology.     

白腐菌应用于堆肥处理含木质素废物的研究

白腐菌对木质素具有较强的降解能力。通过在含大量木质素的模拟垃圾堆肥中采取添加白腐菌菌剂与不添加该菌剂2组实验对比,在相同堆肥处理条件下,接种菌剂的堆肥中木质素总量由274988mg降至15438mg,降解率达4386%,远高于未接种菌剂的堆肥中木质素的降解率,表明白腐菌可有效用于含木质素废弃物的堆肥处理,并有望于加速堆肥腐熟,提高堆肥效率。     

Harnessing dark fermentative hydrogen from pretreated mixture of food waste and sewage sludge under sequencing batch mode

Food waste and sewage sludge are the most abundant and problematic organic wastes in any society. Mixture of these two wastes may provide appropriate substrate condition for dark fermentative biohydrogen production based on synergistic mutual benefits. This work evaluates continuous hydrogen production from the cosubstrate of food waste and sewage sludge to verify mechanisms of performance improvement in anaerobic sequencing batch reactors. Volatile solid concentration and mixing ratio of food waste and sludge were adjusted to 5 % and 80:20, respectively. Five different hydraulic retention times (HRT) of 36, 42, 48, 72, and 108 h were tested using anaerobic sequencing batch reactors to find out optimal operating condition. Results show that the best performance was achieved at HRT 72 h, where the hydrogen yield, the hydrogen production rate, and hydrogen content were 62.0 mL H₂/g VS, 1.0 L H₂/L/day, and ~50 %, respectively. Sufficient solid retention time (143 h) and proper loading rate (8.2 g COD/L/day as carbohydrate) at HRT 72h led to the enhanced performance with better hydrogen production showing appropriate n-butyrate/acetate (B/A) ratio of 2.6. Analytical result of terminal-restriction fragment length polymorphism revealed that specific peaks associated with Clostridium sp. and Bacillus sp. were strongly related to enhanced hydrogen production from the cosubstrate of food waste and sewage sludge.     

“氧化镁/活性炭”新型吸附剂的制备及其对Cr(VI)的吸附研究

以氯化镁和造纸草浆黑液为原料，采用物理活化法制得“氧化镁/活性炭”新型吸附剂，其比表面积（BET）为388.96 m²/g、总孔容积为0.40 mL/g。测定了这种吸附剂对水中Cr(VI)的吸附性能，考察了吸附时间、pH值、吸附剂投加量、初始浓度等因素对Cr(VI)的吸附量和脱除率的影响，研究所得吸附水溶液中Cr(VI)的最佳条件为：吸附时间为120 min，吸附剂投加量为2 g/L，pH值为2。“氧化镁/活性炭”新型吸附剂对Cr(VI)的吸附过程符合Freundlich等温式。     

混凝法深度处理废纸造纸废水实验研究

按照烧杯实验方法,重点考察了pH值、混凝剂种类和投加量等因素对生化处理后废纸造纸废水混凝处理效果的影响。实验结果表明:PAC作为混凝剂,PAM作为助凝剂联合处理该废水时,能够取得较好的去浊率、SS、色度和COD去除率。混凝沉淀最佳运行条件为:废水pH为6.5,含铝量10%的PAC和2 g/L的PAM投加量分别为1 mL/L、0.5 mL/L,浊度从35 NTU降低到1 NTU,去除率达97.1%,SS从30 mg/L降低到7 mg/L,去除率达76.7%,色度从64倍降低到18倍,去除率达71.9%,COD从95 mg/L降低到44.8 mg/L,去除率可达52.8%,取得了较好的去除效果,达到国家造纸废水新排放标准限值。     

Fumaric acid production using renewable resources from biodiesel and cane sugar production processes

The microbial production of fumaric acid by Rhizopus arrhizus NRRL 2582 has been evaluated using soybean cake from biodiesel production processes and very high polarity (VHP) sugar from sugarcane mills. Soybean cake was converted into a nutrient-rich hydrolysate via a two-stage bioprocess involving crude enzyme production via solid state fermentations (SSF) of either Aspergillus oryzae or R. arrhizus cultivated on soybean cake followed by enzymatic hydrolysis of soybean cake. The soybean cake hydrolysate produced using crude enzymes derived via SSF of R. arrhizus was supplemented with VHP sugar and evaluated using different initial free amino nitrogen (FAN) concentrations (100, 200, and 400 mg/L) in fed-batch cultures for fumaric acid production. The highest fumaric acid concentration (27.3 g/L) and yield (0.7 g/g of total consumed sugars) were achieved when the initial FAN concentration was 200 mg/L. The combination of VHP sugar with soybean cake hydrolysate derived from crude enzymes produced by SSF of A. oryzae at 200 mg/L initial FAN concentration led to the production of 40 g/L fumaric acid with a yield of 0.86 g/g of total consumed sugars. The utilization of sugarcane molasses led to low fumaric acid production by R. arrhizus, probably due to the presence of various minerals and phenolic compounds. The promising results achieved through the valorization of VHP sugar and soybean cake suggest that a focused study on molasses pretreatment could lead to enhanced fumaric acid production.     

Reusing colored industrial wastewaters in a photofermentation for enhancing biohydrogen production by using ultrasound stimulated <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis>

One-time ultrasonication pre-treatment of Rhodobacter sphaeroides was evaluated for improving biohydrogen production via photofermentation. Batch experiments were performed by varying ultrasonication amplitude (15, 30, and 45%) and duration (5, 10, and 15 min) using combined effluents from palm oil as well as pulp and paper mill as a single substrate. Experimental data showed that ultrasonication at amplitude 30% for 10 min (256.33 J/mL) achieved the highest biohydrogen yield of 9.982 mL H₂/mL_medium with 5.125% of light efficiency. A maximum COD_total removal of 44.7% was also obtained. However, when higher ultrasonication energy inputs (>256.33 J/mL) were transmitted to the cells, biohydrogen production did not improve further. In fact, 20.6% decrease of biohydrogen yield (as compared to the highest biohydrogen yield) was observed using the most intense ultrasonicated inoculum (472.59 J/mL). Field emission scanning electron microscope images revealed the occurrence of cell damages and biomass losses if ultrasonication at 472.59 J/mL was used. The present results suggested that moderate ultrasonication pre-treatment was an effective technique to improve biohydrogen production performances of R. sphaeroides.     

“氧化镁/活性炭”新型吸附剂的制备及其对Cr（Ⅵ）的吸附研究

以氯化镁和造纸草浆黑液为原料,采用物理活化法制得＂氧化镁/活性炭＂新型吸附剂,其比表面积（BET）为388.96 m2/g、总孔容积为0.40 mL/g。测定了这种吸附剂对水中Cr（Ⅵ）的吸附性能,考察了吸附时间、pH值、吸附剂投加量、初始浓度等因素对Cr（VI）的吸附量和脱除率的影响,研究所得吸附水溶液中Cr（Ⅵ）的最佳条件为：吸附时间为120min,吸附剂投加量为2 g/L,pH值为2。＂氧化镁/活性炭＂新型吸附剂对Cr（Ⅵ）的吸附过程符合Freundlich等温式。     

Comparative studies of fungal degradation of single or mixed bioaccessible reactive azo dyes

A screening using several fungi (Phanerochaete chrysosporium, Pleurotus ostreatus, Trametes versicolor and Aureobasidium pullulans) was performed on the degradation of syringol derivatives of azo dyes possessing either carboxylic or sulphonic groups, under optimized conditions previously established by us. T. versicolor showed the best biodegradation performance and its potential was confirmed by the degradation of differently substituted fungal bioaccessible dyes. Enzymatic assays (lignin peroxidase, manganese peroxidase, laccase, proteases and glyoxal oxidase) and GC-MS analysis were performed upon the assay obtained using the most degraded dye. The identification of hydroxylated metabolites allowed us to propose a possible metabolic pathway. Biodegradation assays using mixtures of these bioaccessible dyes were performed to evaluate the possibility of a fungal wastewater treatment for textile industries.     

烟草下脚料发酵制取乙醇

通过单因素实验考察了硫酸浓度、固液比和水解时间对硫酸水解的影响。结果显示最优条件为:硫酸浓度为50%(w/w),固液比为10%(w/v),时间为100 min。烟草下脚料在最佳硫酸水解条件下,经5倍稀释,中和pH值至5～6。取经过滤后的水解液(FH)用酿酒酵母(Sacchharomyces cerevisiae)发酵产生乙醇,最大的乙醇浓度和乙醇产量分别为1.09g/L和54.5 g/kg。未过滤水解液(UFH,包括水解残渣)加入纤维素酶(70 U/100 mL)和酿酒酵母(Sacchharomyces cerevisiae)进行发酵,最大的乙醇浓度和乙醇产量分别为1.23 g/L和61.5 g/kg。     

Fenton氧化法深度处理抗生素废水二级出水

采用Fenton氧化法对青霉素和土霉素混合废水二级处理出水进行深度处理,通过正交和单因素实验研究了废水初始反应pH值、H2O2投加量、Fe2+/H2O2摩尔比及反应时间等因素对废水处理效果的影响。实验结果表明,Fenton氧化法处理的最佳反应条件为:初始pH值4、H2O2(30%)投加量50 mL/L、Fe2+/H2O2摩尔比1/20和反应时间60 min,处理后出水COD小于120 mg/L,COD去除率在75%以上,急性毒性(HgCl2毒性当量)小于0.07 mg/L,满足《发酵类制药工业水污染物排放标准》(GB21903-2008)表2标准要求。