Enzymatic and fungal treatments on sugarcane bagasse for the production of mechanical pulps

Crude ligninolytic enzyme extracts from Phanerochaete chrysosporium fungi were applied to sugarcane bagasse, prior to thermomechanical (TMP) and chemithermomechanical pulping (CTMP), and their properties were compared with the normal TMP and CTMP and also with TMP and CTMP pretreated with Ceriporiopsis subvermispora and P. chrysosporium fungi. The sugarcane bagasse was impregnated with the crude enzyme extract containing lignin peroxidase (LiP), manganese peroxidase (MnP), and laccase (Lac). The results show that pretreatment with enzyme crude extract is an advantageous way to produce TMP and CTMP from sugarcane bagasse, as compared with only fungal pretreatment. Enzymatic pretreatments need only hours to enhance pulping and paper properties, compared with the weeks necessary for fungal treatments. Higher pulp yields were obtained compared with the fungal pretreatments. Enzymatic pretreatment reduced the energy consumption in a proportion similar to that of C. subvermispora fungal pretreatment and increased the pulp tensile index compared with the normal TMP and CTMP pulps, although the tensile strength was somewhat lower than that for pulps from C. subvermispora fungal pretreatment before CTMP processing. An advantage of enzymatic pretreatment is that brightness is increased compared with normal TMP and CTMP processes, whereas fungal pretreatments reduce the brightness.     

甜高粱茎汁及茎渣同步糖化发酵工艺优化

为了提高甜高粱秸秆乙醇生产中茎汁和茎渣的利用,以甜高粱茎汁及其渣为发酵原料,对茎汁茎渣混合原料同步糖化乙醇发酵的工艺条件进行优化研究。采用Plackett-Burman(PB)筛选设计试验筛选出影响甜高粱茎秆渣汁同步糖化乙醇发酵的显著因素。采用响应面法建立了同步糖化发酵乙醇生产的乙醇产量数学模型。根据该模型进行了工艺参数的优化,以乙醇产量为指标,试验所得甜高粱茎秆渣汁同步糖化化乙醇发酵的优化工艺条件为:发酵温度36.58℃,混合纤维素酶添加量=23.5(FBU/m L)/35.25(CBU/m L),甜高粱渣汁质量体积比为8.2%,理论预测乙醇产量为89.2%,在此条件下进行验证试验,乙醇产量为88.98%,平均质量浓度,验证了数学模型的有效性,为提高甜高粱茎汁及茎渣混合原料同步糖化发酵产乙醇和提高发酵效率提供参考。     

Growth and anthraquinone production of Morinda elliptica cell suspension cultures in a stirred-tank bioreactor

The effects of medium strategy, number of impellers, aeration mode, and mode of operation on Morinda elliptica cell suspension cultures in a stirred-tank bioreactor are described. A lower number of impellers and continuous aeration contributed toward high cell growth rate, whereas a higher number of impellers reduced cell growth rate, although not anthraquinone yield. The semicontinuous mode could indirectly imitate the larger scale version of production medium strategy and improved anthraquinone production even with 0. 012% (v/v) antifoam addition. Production medium promoted both growth (maximum dry cell weight of 24.6 g/L) and anthraquinone formation (maximum content of 19.5 mg/g of dry cell weight), without any necessity for antifoam addition. Cultures in production medium or with higher growth rate and anthraquinone production were less acidic than cultures in growth medium or with lower growth rate and anthraquinone production. Using the best operating variables, growth of M. elliptica cells (24.6 g/L) and anthraquinone yield (0.25 g/L) were 45% and 140%, respectively, lower than those using a shake flask culture after 12 days of cultivation.     

Slow turnover and production of fungal hyphae during a Californian dry season

We used minirhizotrons to examine the production and turnover of fungal hyphae in situ during the dry season in a Californian grassland. Hyphae were produced relatively slowly throughout the season at rates that did not vary significantly over time, indicating that a portion of the fungal community was active even when soils were very dry. In addition, fungi displayed relatively long residence times, with half of the hyphae remaining in the soil for at least 145 days. Together, these results suggest that a contingent of active fungi may be capable of performing nutrient transformations when plants are otherwise dormant, while relatively long-lasting hyphae may immobilize nutrients for several months before turning over.     

Effect of cysteinyl caffeic acid, caffeic acid, and L-dopa on the oxidative cross-linking of feruloylated arabinoxylans by a fungal laccase

To study a way to covalently link arabinoxylans and proteins using a fungal laccase from the fungus Pycnoporus cinnabarinus, the effect of cysteinyl caffeic acid on the cross-linking of wheat arabinoxylans was investigated by means of capillary viscometry and RP-HPLC of alkali labile phenolic compounds. Cysteinyl caffeic acid provoked a delay in gelation and in the consumption of the esterified ferulic acid on arabinoxylans. When reacting free ferulic acid and cysteinyl caffeic acid with laccase, the ferulic acid consumption and the dehydrodimers production were also diminished. These results suggest that cysteinyl caffeic acid is oxidized while reducing the semiquinones of ferulic acid produced by laccase. Thus, ferulic acid could not be oxidized into dimers until all cysteinyl caffeic acid was consumed, preventing the cross-linking of feruloylated arabinoxylan chains. A similar mechanism is proposed in the case of caffeic acid and of L-Dopa.     

Optimization of simultaneous saccharification and fermentation for the production of ethanol from lignocellulosic biomass

Simultaneous saccharification and fermentation (SSF) of alkaline hydrogen peroxide pretreated Antigonum leptopus (Linn) leaves to ethanol was optimized using cellulase from Trichoderma reesei QM-9414 (Celluclast from Novo) and Saccharomyces cerevisiae NRRL-Y-132 cells. Response surface methodology (RSM) and a three-level four-variable design were employed to evaluate the effects of SSF process variables such as cellulase concentration (20-100 FPU/g of substrate), substrate concentration (5-15% w/v), incubation time (24-72 h), and temperature (35-45 degrees C) on ethanol production efficiency. Cellulase and substrate concentrations were found to be the most significant variables. The optimum conditions arrived at are as follows: cellulase = 100 FPU/g of substrate, substrate = 15% (w/v), incubation time = 57.2 h, and temperature = 38.5 degrees C. At these conditions, the predicted ethanol yield was 3.02% (w/v) and the actual experimental value was 3.0% (w/v).     

Transformation of catechol in the presence of a laccase and birnessite

The transformation of naturally occurring phenols to humic polymers through oxidative coupling reactions may involve oxidoreductive enzymes and soil minerals as catalysts. There is limited information on the possible inhibitory or synergistic interactions between oxidoreductases and mineral catalysts as they participate in oxidative coupling of phenolic substrates. In this study, a ternary system was investigated, in which a fungal enzyme (Trametes villosa laccase), birnessite (δ-MnO₂), and a naturally occurring phenolic compound (catechol) were reacted together to model soil processes. Binary systems (catechol/laccase and catechol/birnessite) were included for comparison. In the absence of the mineral, T. villosa laccase (950 katal ml⁻¹) transformed 31% of catechol, whereas birnessite (1 mg ml⁻¹) in the absence of the enzyme showed a 24% catechol transformation. The percentages of catechol transformation in the binary systems did not accumulate in the ternary system; instead, birnessite and laccase tested together transformed only 36% of catechol. This suggested that birnessite had an inhibitory effect on substrate transformation by laccase catalysis. Enzyme assays indicated that inhibition was a result of enzyme deactivation by humic-like polymers produced by birnessite, and by Mn²⁺ ions released from the mineral. These observations underscore the importance of considering enzyme-soil mineral-organic matter interactions in studies of humus formation and contaminant removal.     

Impact of cultivation and sugar-cane green trash management on carbon fractions and aggregate stability for a Chromic Luvisol in Queensland, Australia

Technological advances in sugar-cane harvesting and processing is bringing about rapid changes in production systems which could impact on soil physical conditions. An increasing incidence of soil structural decline and depletion of soil carbon levels has increased the risk of soil erosion and crop yield reductions. Soil carbon (C) and aggregate stability were studied on a sugar-cane (Saccharum officinarum L.) green trash blanket trial that had been established on a Chromic Luvisol soil at Mackay, Qld, Australia in 1992. The experiment consisted of blocks with two blocks being harvested early and the remaining two blocks harvested late in the crushing season. Within each block, treatment combinations of trash burnt or green trash blanket, which are either cultivated between rows or not cultivated after harvest, were included. Cropping and cultivation of the soil reduced the different C fractions in the surface 0–100 mm layer by 66–67% when compared to an adjacent uncropped reference soil. The labile C (C_L) concentration was 11% lower in the burnt treatment compared to the trash returned treatment but the opposite was found for total C (C_T). After four years, the no cultivation treatment had higher concentrations of all C fractions measured, compared to the cultivated treatment. When compared to the uncropped reference soil, cropping resulted in marked reductions in aggregate mean weight diameter (MWD) and aggregates >250 μm and an increase in aggregates <125 μm determined by both immersion and tension wetting. The return of the green trash resulted in a 30% greater MWD and a 28% increase in aggregates >250 μm and an 18% reduction in aggregates <125 μm compared to the burnt treatment when immersion wetting was used. Four years of cultivation reduced the MWD, as determined by immersion wetting, by 26% compared to the no cultivation treatment. No significant correlations were found between any measured C fraction and aggregate stability. This study indicates that sustainable sugar-cane cropping systems will likely be those where cultivation is kept to a minimum and trash is retained in the system.     

Enterotoxigenicity and cytotoxicity of Bacillus thuringiensis strains and development of a process for Cry1Ac production

Bacillus thuringiensis is indistinguishable from Bacillus cereus except for the production of insecticidal crystal proteins (ICPs). B. thuringiensis strains may show enterotoxin profiles and toxin levels similar to those of B. cereus strains isolated from food-poisoning cases. It is important for the food industry and farmers to consider that with the application of B. thuringiensis strains to crops, their spores may be introduced into the human food chain. In this study, 59 B. thuringiensis strains were assayed for their hemolysin BL (HBL) using a BCET-RPLA kit and their cytotoxicity to Chinese hamster ovary (CHO) cells. The enterotoxin titer was as high as that of B. cereus diarrheal-type strain ATCC 49064. In an attempt to obtain a food safety strain for bioinsecticide use, in this study, a 3.5-kb cry1Ac DNA fragment was amplified with PCR from the total DNA of B. thuringiensis subsp. kurstaki CCRC 11502 and cloned into the Bacillus expression vector pHY300PLK. The alpha-amylase promoter, amyE, was then introduced into the promoter region and, afterward, the recombinant plasmid pHYe1Ac35 was introduced into a non-enterotoxigenic and non-cytotoxic B. thuringiensis subsp. kurstaki Tt14 strain. The transformant, without any detectable enterotoxigenicity or cytotoxicity, produced Cry1Ac toxin properly, and its insecticidal activity against Trichoplusia ni larvae was found to be satisfactory.     

Nitrous oxide production in soils and the ratio of the fungal to bacterial biomass

The proportion between the fungal and bacterial biomass, the potential activity of denitrification, and the intensity of N₂O production were determined in the soils (chernozem and soddy-podzolic) of secondary biocenoses formed upon the abandoning of agricultural lands. The substitution of meadow and forest vegetation for agrocenoses has led to an increase in the percentage of the fungal biomass in the upper soil horizons. The rate of the net N₂O production after the soil moistening positively correlated with the content of nitrates. In the soddy-podzolic soil (pH 3.7–5.6), the rate of nitrous oxide production was higher than that in the chernozem (pH 6.1–6.8). The rate of N₂O production was inversely proportional to the bacterial biomass in the soils.     

Diacylglycerol and triacylglycerol as responses in a dual response surface-optimized process for diacylglycerol production by lipase-catalyzed esterification in a pilot packed-bed enzyme reactor

Diacylglycerol (DAG) and triacylglycerol (TAG) as responses on optimization of DAG production using a dual response approach of response surface methodology were investigated. This approach takes the molecular equilibrium of DAG into account and allows for the optimization of reaction conditions to achieve maximum DAG and minimum TAG yields. The esterification reaction was optimized with four factors using a central composite rotatable design. The following optimized conditions yielded 48 wt % DAG and 14 wt % TAG: reaction temperature of 66.29 degrees C, enzyme dosage of 4 wt %, fatty acid/glycerol molar ratio of 2.14, and reaction time of 4.14 h. Similar results were achieved when the process was scaled up to a 10 kg production in a pilot packed-bed enzyme reactor. Lipozyme RM IM did not show any significant activity losses or changes in fatty acid selectivity on DAG synthesis during the 10 pilot productions. However, lipozyme RM IM displayed higher selectivity toward the production of oleic acid-enriched DAG. The purity of DAG oil after purification was 92 wt %.     

New technique for rose production in soilless culture system and disease reduction

Rose flower is one of the most important cut flowers, with the world's highest production rate. Soilless culture has been increasingly popular in plant production worldwide in the past few decades. In two separate trials, the efficacy of Trichoderma harzianum, Anearobacillus migululanas, slow filtration, and UV ultraviolet (UV) treatment and sand filtration in eliminating Fusarium oxyporum, artificially added in the recirculating nutrient solution, was evaluated with Rosea hybrid plants grown in closed soilless systems. The dynamics of these fungi in the recirculating nutrient solution and in the sand filter was also investigated. T. harzianum, A. migululanas, UV treatment and slow sand filtration were effective in reducing Fusarium root rot. However, slow sand filtration may be a more feasible disinfection method than UV because of lower costs of installation and maintenance and for its adaptability to a wide range of production systems. Moreover, these disinfection techniques can be successfully combined with the application of antagonistic microorganisms.     

NaCl浓度对SBBR同步脱氮及N2O释放的影响

盐度是影响生物脱氮过程的重要因素。盐度增加会导致生物硝化和反硝化过程中N_2O的产生并释放。该文以添加NaCl的生活污水为研究对象,采用固定填料序批式生物膜反应器(sequencing batch biofilm reactor,SBBR),考察了不同NaCl浓度(0、5、10、15和20g/L)对SBBR脱氮性能及N_2O释放的影响。结果表明,试验NaCl浓度范围内,SBBR出水COD稳定在40～60mg/L。硝化过程NO_2~-/NO_3~-随NaCl浓度增加而增加。NaCl浓度≤10g/L时,NH_4~+-N去除率大于95%,N_2O产率由4.08%(NaCl浓度为0)增至6.72%(NaCl浓度为10 g/L)。NaCl浓度为20 g/L时,驯化后SBBR内平均NH_4~+-N去除率为70%,平均N_2O产率为13.60%。无添加NaCl时,N_2O主要产生于硝化阶段的AOB好氧反硝化过程,SBBR内缺氧区有助于减少N_2O释放;高NaCl浓度条件下,N_2O主要产生于AOB好氧反硝化过程和内源同步反硝化过程,高盐度加剧内源反硝化阶段NO_2~-和N_2O之间电子竞争,抑制N_2O还原,其活性抑制性能与电子受体和初始C/N有关。与硝态氮还原速率和亚硝态氮还原速率相比,氧化亚氮还原速率受NaCl抑制最为明显,是导致高盐度条件下N_2O释放量增加的重要因素。     

New volatile sulfur-containing constituents in a simultaneous distillation-extraction extract of red bell peppers (Capsicum annuum)

An extract of red bell peppers ( Capsicum annuum) was prepared by simultaneous distillation-extraction (SDE, Likens-Nickerson). In addition to the already known (3 E)-3-hepten-2-one ( 1), the unsaturated C9-ketones 1-nonen-4-one ( 2), (2 E)-2-nonen-4-one ( 3), and (2 E,5 E)-2,5-nonadien-4-one ( 4), 2-methoxy-3-isobutylpyrazine ( 5), and heptane-2-thiol ( 6), we identified 19 new thiols (the aliphatic saturated and unsaturated thiols 14- 16, and 22- 27, the mercapto-ketones 12 and 13, the mercapto-alcohols 17, 18, and 30, the dithiols 19 and 28, the methylthio-thiols 20 and 21, and the thiophene-thiol 31) and the two new dithiolanes 10 and 29. All of them are structurally related to the unsaturated C7- and C9-ketones 1- 4. The free thiols were enriched using Affi-Gel 501 ( p-aminophenyl-mercuric acetate grafted on an agarose gel). The new compounds were confirmed by syntheses and were organoleptically evaluated.     

内蒙古耕地资源变化过程与粮食生产安全问题研究

内蒙古自治区地域辽阔,水热等基本条件相差悬殊,土地资源分异非常显著,耕地资源也表现出较明显的地域差异。本文利用内蒙古自治区50多年的统计数据和近9年的土地利用变更调查数据,分析了内蒙古自治区耕地面积、人均耕地面积变化过程,耕地面积变化过程的地域差异及耕地面积变化与粮食产量的关系。结果表明,内蒙古人均粮食产量的变化趋势随人均耕地的增减有较大起伏,两者的变化基本同步,并随社会经济的发展和人口的不断增加,人均耕地面积有减少的趋势,故从粮食生产安全角度考虑,保护耕地成为粮食生产安全的根本保障。基于此,从珍惜和保护好现有耕地,大力改善农业基础设施建设、特别是水利工程建设,严格控制非农业建设侵占耕地,切实保护耕地资源,加强基本农田的行政管理和社会监督及建立耕地利用考核体系,缩减土地征用利益空间等方面提出了耕地保护与保障粮食生产安全的对策。     

Decomposition of willow-leaf litter in a short-rotation forest in relation to fungal colonization and palatability for earthworms

Summary The influence of leaf litter from three Salix spp. on fungal growth and microbial decomposition was studied using 1-mm-mesh litter-bags, and the effect on additional soil macrofaunal activity was studied by measuring litter disappearance from 4-mm-mesh bags and under 4-mm-mesh nets. Mineral macro-elements, water-and ethanol-extractable substances, lignin, and protein-precipitating substances (astringency) in the litter were determined, taking contaminating of the litter with soil particles into account. As expected, the litter disappeared more quickly from the large-mesh bags than from the small-mesh bags, which was attributed to earthworm activity. During the 1st year, the rate of leaf disappearance from both types of bags and under the nets was much higher for S. daphnoides than for S. viminalis and S. fragilis. The lower initial astringency, related to the tannin content, of the S. daphnoides litter might account for this difference. Tannin metabolites probably hampered both microbial decomposition and earthworm acceptability for some time also after the astringency was lost. Neither the content of macronutrients nor that of the other organic fractions studied can be assumed to have had any effect on weight losses due to microbial decomposition. Although, the S. daphnoides leaves initially contained the least amount of fungal mycelium (m g^-1 dry weight), the increase after contact with soil was most pronounced in this litter. The species composition of Fungi Imperfecti in the leaves of S. viminalis and S. daphnoides differed only for fresh litter, whereas the number of isolates was somewhat higher for S. daphnoides throughout the study. Similar seasonal variations in fungal composition occurred in both the S. viminalis and the S. daphnoides litter.     

Development of a biocatalytic process for the production of c6-aldehydes from vegetable oils by soybean lipoxygenase and recombinant hydroperoxide lyase

Volatile C6- and C9-aldehydes and alcohols are widely used as food flavors to reconstitute the "fresh green" odor of fruits and vegetables lost during processing. To meet the high demand for natural flavors, an efficient, cheap, and versatile biocatalytic process was developed to produce C6-aldehydes on a large scale. Vegetable oils were converted by soybean lipoxygenase and recombinant hydroperoxide lyase into hexanal and (2E)- or (3Z)-hexenal. In contrast to plant extracts, generally used as enzyme sources, high molar conversions were obtained with recombinant hydroperoxide lyase (50% for hexanal and 26% for hexenal formation), and no side products were formed. Furthermore, recombinant hydroperoxide lyase lacks isomerase activity, allowing production of (3Z)-hexenal, which could not be obtained in previously described processes. Recombinant hydroperoxide lyase is stable and can be stored at 4 degrees C for 1 month without significant loss of activity.     

New organic substrates and boron fertilizing for production of yellow passion fruit seedlings

The identification of alternative substrates suitable for seedling production is very important, while boron nutrition is also relevant due to plant requirement and its difficult management. Thus, four experiments were carried out from October 2012 to April 2013 to evaluate the effect of substrates using decomposed buriti (Mauritia vinifera Mart.) stem (DBS) and boron fertilizing for seedling production of yellow passion fruit (Passiflora edulis Sims). The experimental design was completely randomized in factorial scheme 6 × 2 + 1, referring to the substrates [S1: DBS, S2: soil and sand 1:1 (20%) + DBS (80%), S3: soil and sand 1:1 (40%) + DBS (60%), S4: soil and sand 1:1 (60%) + DBS (40%), S5: soil and sand 1:1 (80%) + DBS (20%), S6: commercial substrate (additional treatment) and S7: soil, sand and manure 1:1:2] with and without boron. The seedling emergence, emergence rate, plant height, stem diameter, root length, root volume and dry mass of roots and shoots were recorded. The substrates S2 (without B fertilizing) and S4 (B fertilized with 0.5 mg dm^?3) produced seedlings with more than 3.0 g of shoot dry mass and at least 30 cm in height, and can be used for production of high-quality yellow passion fruit seedlings.