Effect of steam treatment for the enzymic saccharification of waste mushroom medium after cultivation of shiitake mushroom (Lentinula edodes) and enokitake mushroom (Flammulina velutipes)

Waste mushroom medium of shiitake and enokitake (WS and WE, respectively) was steamed (StmWS and StmWE, respectively) and saccharized with cellulase to obtain glucose which can be feedstock of bioethanol or other bioproducts. Mushroom medium before cultivation of shiitake and enokitake (MS and ME, respectively) was also steamed (StmMS and StmME, respectively) and saccharized to compare with WS and WE. WS was stored at 15 °C for 1 month (SS), and SS was steamed (StmSS) and saccharized. The amount of glucose from StmSS was more than that from StmWS. The cellulose contained in StmSS and StmWE was saccharified earlier than that in StmMS and StmME, respectively. When StmSS and StmWE-added Meicelase of 5 and 3 FPU/g substrate (9.7 and 5.8 mg Meicelase/g substrate), respectively, was saccharized for 120 h, the saccharification ratio (the amount of glucose saccharized from cellulose/theoretical amount of glucose from cellulose) of them exceeded 80 %. The amount of glucose obtained from StmSS and StmWE was 311.0 and 207.7 mg/g substrate under the condition. It was found that the cellulose contained in mushroom medium was saccharized with less amount of cellulase by the influence of cultivation of shiitake or enokitake, storage of shiitake, and steaming.     

Evaluation of waste mushroom medium from cultivation of shiitake mushroom (<Emphasis Type="Italic">Lentinula edodes</Emphasis>) as feedstock of enzymic saccharification

The feasibility of using waste mushroom medium (WMM) as a substrate for enzymic saccharification to produce bioethanol or other bioproducts was investigated. WMM was broken up or left unbroken and stored at constant temperatures (5°, 15°, and 25°C) for 1 or 2 month(s) to accelerate lignin degradation by shiitake, which is a white rot fungus, and to increase the saccharification ratio. When the physicochemical properties of WMM and stored WMM were investigated, it was evident that the mushroom medium (MM) lignocellulose had decomposed during shiitake cultivation and its subsequent storage at a constant temperature. WMM and stored WMM were more susceptible to attack by cellulase than MM. When the unbroken WMM that was stored at 25°C for 2 months was saccharized with meicelase at 5 FPU/g substrate, its saccharification ratio was higher than that of unstored WMM. The maximum glucose yield of the stored WMM was 200.5 mg/g substrate, approximately 4.1 times as large as the value for autoclaved MM under the same saccharification condition. The saccharification ratio of the constituent cellulose was approximately 60%.     

Enzymatic saccharification of spent wood-meal media made of 5 different tree species after cultivation of edible mushroom <Emphasis Type="Italic">Auricularia polytricha</Emphasis>

The chemical characteristics and the suitability for enzymatic saccharification in the spent culture media (SCM) of Auricularia polytricha were examined in order to investigate the utilization of the SCM as a biomass resource for alternative energy production. Wood meals from 3 hardwood species (Quercus serrata, Betula platyphylla var. japonica, and Alnus japonica) and 2 softwood species (Pinus densiflora and Cryptomeria japonica) were used as basal culture media. Dry weight of fruiting bodies were higher in the cultural media made of B. platphylla var. japonica and A. japonica. Amount of weight loss in media showed almost the same value among the cultural media made of 5 species, except for media made of C. japonica. The amounts of the main chemical components (Klason lignin, holocellulose, and α-cellulose) in SCM showed lower values than those in wood meals (WM) and fresh media (FM). After saccharification of the media by Meiselase for 48 h, the hydrolysis weight decrease dramatically increased in SCM. The amount of glucose in SCM ranged from 10.9 to 19.2 g/100 g dry biomass. The highest amount of glucose was obtained in the SCM from B. platyphylla var. japonica. These results indicate that SCM of A. polytricha is a suitable biomass material to produce fermentable sugars for bio-ethanol production.     

The smell and odorous components of dried shiitake mushroom, Lentinula edodes III: substances that increase the odorous compound content

This study formed part of an effort to improve the quality of dried shiitake mushroom [Lentinula edodes (Berk.) Pegler], in accordance with consumer preferences, and deals with the search for substances that increase the odorous component content. From analysis of sulfur and sulfur-containing substances in the culture substrate, rice bran was found to be the main source of sulfur, and 75% of this was present as cysteine and methionine. The sulfur-containing substances were added to a sawdust medium containing only rice bran and sawdust as a substrate; shiitake mushrooms were cultivated in the medium, the fruiting bodies were dried, and the 1,2,4-trithiolane content in the fruiting bodies was measured as an indicator of the odorous compounds. Of the sulfur-containing substances, those that increased odorous compounds the most were cysteine and methionine. The efficiency of cysteine in this regard was higher than that of methionine. It was also noted that the amount of glutamic acid increased the odorous compounds in combination with cysteine and methionine. Furthermore, the addition of both amino acids and glutamic acid had no negative effect on the yield. These results showed that it is possible to produce dried shiitake mushrooms with a smell suitable for particular consumer preferences.     

The smell and odorous components of dried shiitake mushroom, Lentinula edodes VI: increase in odorous compounds of dried shiitake mushroom cultivated on bed logs

Odor is one of the most important characteristics affecting consumer preference for dried shiitake mushrooms [Lentinula edodes (Berk.) Pegler]. In our previous studies, we found that the odor content of commercial dried products was too weak for most people, and that the odorous compound content could be increased by adding amino acids to sawdust media. Currently, however, bed-log cultivation is used to produce fruiting bodies for dried products. The purpose of this study was to fi nd a method to increase the content of odorous compounds in dried products cultivated on bed logs. Pressure injection of amino acids from the side of the bed log was the most effi cient method, but it had some problems. Hence, a simpler and less troublesome method was developed, i.e., injecting amino acid solution from small bottles set in deep holes bored in the sides of the bed logs. In fruiting bodies cultivated on bed logs injected with amino acid solution by the improved method, the mean contents of lentinic acid, a precursor of the odorous compound lenthionine, approximately doubled compared to that in the untreated logs, although the infi ltration area of the solution injected by the improved method was smaller than that by the former method.     

Production and investment evaluation of oyster mushroom cultivation on the waste dendromass: a case study on aspen wood in Slovakia

This paper addresses the processes and investment evaluation of utilizing available dendromass in an easy and close-to-nature way under Central European conditions in Slovakia. Wood of the European aspen (Populus tremula L.) that has generally been considered of little economic value was efficiently used as a growth medium for the oyster mushroom (Pleurotus ostreatus (Jacq.) P. Kumm.). The production of the fruiting bodies of this mushroom were evaluated for 5 years under natural forest conditions. The evaluation focused solely on aspen wood, specifically, the most valuable part – the trunk. The investment assessment revealed the high economic value of the fruiting bodies of the oyster mushroom inoculated into aspen wood blocks for the period of five years. In particular, the results of the net present value (NPV?=?347.78 €/m³) as well as the profitability index (PI?=?1.63) and discounted payback period (DPP ≈ 2 years and 8 months) point to the high profitability of such a project under the given conditions. The results could be helpful and inspiring for forest and garden owners when deciding how to utilize low-value waste dendromass, low-quality aspen wood and other tree species in small-scale forest management.     

Recycled ionic liquid 1-ethyl-3-methylimidazolium acetate pretreatment for enhancing enzymatic saccharification of softwood without cellulose regeneration

In this work, pretreatment of wood meals using a recycled ionic liquid (IL), 1-ethyl-3-methylimidazolium acetate ([Emim]Ac), enhanced glucose liberation by enzymatic saccharification, without dissolution of cellulose and lignin. In contrast, previous studies on IL pretreatment have mostly focused on lignocellulosic dissolution to regenerate cellulose and removing lignin. Softwood (Cryptomeria japonica) was pretreated with [Emim]Ac at 60–100 °C for 2–8 h without collecting regenerated cellulose. The pretreatment did not have a strong effect on wood component dissolution (weight of residues: 91.7–98.8%). The residues contained relatively high amounts of lignin (26.6–32.6%) with low adsorption of [Emim]Ac (0.9–2.7%). Meanwhile, the crystallinity index (C _r I) of cellulose in the wood was significantly reduced by pretreatment, from 50.9% to 28.4–37.1%. In spite of the high lignin contents in the residues, their glucose liberation values by enzymatic saccharification using a cellulase mixture were 3–16 times greater than that of untreated wood. A good correlation was found between the saccharification effectiveness of pretreated samples and the C _r I. Although lignin dissolved in [Emim]Ac continued to accumulate after repeated use of [Emim]Ac, the pretreatment was found to be effective for three consecutive cycles without the need to remove the dissolved materials.     

The effects of chemical components and particle size on the mechanical properties of binderless boards made from oak (<Emphasis Type="Italic">Quercus</Emphasis> spp.) logs degraded by shiitake fungi <Emphasis Type="Italic">(Lentinula edodes</Emphasis>)

Binderless boards are composite boards that rely on self-bonding mechanisms for inter-fibre bonding. Quercus acutissima and Quercus serrata logs degraded by Lentinula edodes (shiitake fungi) were used in this study to investigate whether physical and chemical changes induced by shiitake fungi can enhance board mechanical properties. Binderless boards were manufactured with 0.8 g/cm³ target density, 220 °C pressing temperature, 5 MPa pressure, and pressing duration of 10 min. Boards made from logs degraded for ≥?26 months were stronger than control boards and met modulus of rupture (MOR) and internal bonding (IB) requirements for fibreboards. Chemical composition and particle size distribution of the wood powder used to make the boards were determined to elucidate the drivers of board mechanical properties. The proportion of small particles (<?150 µm) showed a strong positive correlation with MOR for both species and hot water extractives showed a strong positive correlation with IB for Q. acutissima boards. Introduction of shiitake fungi pre-treatment to the production process may enhance the mechanical strength of binderless boards.     

Storage of maitake mushroom (Grifola frondosa) culture medium after harvesting fruit bodies is an effective pretreatment for ethanol conversion

The storage of spent maitake culture medium (SMCM) under various conditions was investigated as a potential pretreatment of SMCM for increased ethanol conversion. When SMCM was stored at 25°C for 12?weeks, the glucose yield by enzymatic saccharification increased from 22 to 52%. Selective degradation of lignin and hemicellulose occurred in SMCM during storage. The optimal storage temperature of SMCM was at the active growing temperature of maitake mycelium, which is between 25 and 30°C. Storage of SMCM under anaerobic conditions did not increase the glucose yield compared to non-stored conditions. The glucose yield from the SMCM stored for 4?weeks at 25°C was increased by about 30% with either NaOH or vibrating ball milling pretreatment. After 12?weeks of storage, the glucose yield from SMCM without any other pretreatment was higher than that of non-stored SMCM with additional pretreatments. An ethanol yield of 42.1% was obtained from SMCM stored for 12?weeks by simultaneous saccharification and fermentation, which was comparable to yields after NaOH (41.3%) or vibrating ball milling (44.1%) pretreatments. Therefore, storage of SMCM is a very useful pretreatment for bioethanol production.     

Growth,flowering and fruiting of stecklings,grafts and seedlings of <Emphasis Type="Italic">Allanblackia floribunda</Emphasis> Oliver (Clusiaceae)

A study was carried out to assess the growth, flowering and fruiting of stecklings (rooted cuttings), grafts and seedlings of Allanblackia floribunda in Cameroun. Thirty-one individuals of each plantlet type were planted in May 2006. Height, collar and crown diameters were recorded in September and October, while flowering and fruiting were recorded each month from January to December. In 2014, the trees with the greatest height were those from seeds (6.98 ± 0.25 m), followed by those from grafts (1.65 ± 0.23 m) and cuttings (1.09 ± 0.21 m). Grafts started flowering and fruiting 3 years after planting, while seedlings started flowering and fruiting after four and 6 years, respectively. Stecklings still had not flowered 8 years after planting. Fruit weight and length from seed-origin trees are about seven times higher than grafts. Correlations between height and crown diameter were significant and positive irrespective of the nature of the planting material. In conclusion, grafts fruited earlier, while seedlings grew faster and had greater yields than grafts and stecklings of A. floribunda.     

Cultivation of shiitake in sugi wood meal II: effects of seasoning treatment for wood meal on mycelial growth

The effects of seasoning treatment for fresh sugi wood meal on the mycelial growth of shiitake was investi-gated. The mycelial growth of shiitake in sugi wood meal increased to the same level as that in extract-free sugi wood meal on the 12th day and to approximately 70% of that in konara wood meal on the 28th day of the seasoning treatment. A drastic decrease in extracts of fresh sugi wood meal occurred during the early stage of treatment, and the neutral fraction of methanol extracts decreased to less than 2%. The drainage of sugi wood meal media significantly increased during the later stage of treatment. No changes in the chemical components of sugi wood meal were detected during the seasoning treatment. The mycelial growth of shiitake in the fully seasoned sugi wood meal, which was prepared for commercial mushroom cultivation, was at almost the same level as that in the extract-free sugi wood meal. All of the seasoned sugi wood meals contained a neutral fraction of less than 1%. These results suggested that seasoning treatment can promote mycelial growth of shiitake in sugi wood meal by eliminating inhibitors and improving the physical properties of sugi wood.Part of this paper was presented at the 49th Annual Meeting of the Japan Wood Research Society, Tokyo, April 1999     

乌梅良种单株选育试验初报

为了加快乌梅产业发展,提升乌梅品质,通过对全国乌梅情况进行了解,以四川省达川区为选育点,通过观察、对比等方法从500株单株中筛选出2株优株,分别在3个试验点139hm2区域进行试验研究。4a试验结果表明:选育出的"达梅-1号"鲜果平均单果质量23.20g,可食率为85%左右,平均单位冠幅面积鲜果产量3.31kg/m2,在同类型中花期相对较早,果实均匀,适应性较广;"达梅-2号"鲜果平均单果质量25.70g,可食率在86%左右,平均单位冠幅面积鲜果产量3.33kg/m2,花期最早,果实成熟期早,果实大,果面颜色美观、清洁,适应性强。区域试验证明,乌梅适宜沙质土壤,pH值5.9~6.3,喜温暖湿润气候,"达梅-1号"2月13日进入初花期,7d后进入展叶期,2月26日进入幼果期,果实6月8日开始成熟;"达梅-2号"初花期、展叶期和幼果期时间较"达梅-1号"早16~20d,挂果时间长,一般在6月下旬成熟。初步认为,二者均可在全区范围内进行推广。     

Enzymatic hydrolysis of wood with alkaline treatment

Pulverized samples of wood, cedar and eucalyptus were treated with 5 N NaOH solutions at 25–150 °C. Hemicellulose and lignin content in the samples decreased with increasing treatment temperatures, while the recovery of glucose was maintained at nearly 90 %. X-ray diffraction analysis showed that the content of the original cellulose I structure in the samples decreased with increasing temperature, and most of the cellulose in the sample treated at 150 °C was converted to cellulose II by mercerization. Enzymatic hydrolysis of the alkaline-treated samples was carried out at 37 °C using solutions comprising a mixture of cellulase and β-glucosidase. The samples treated at higher temperatures showed better enzymatic degradability. Treatment with an alkaline solution of lower concentration (1 N NaOH) at 150 °C was also used. Despite significant quantities of hemicellulose and lignin being removed, mercerization was not induced. The enzymatic degradability was much lower than that of the sample treated with a 5 N NaOH solution at 150 °C. Thus, treatment with concentrated alkaline solution at high temperature led to not only the removal of hemicellulose and lignin, but also to modification of the cellulose structure, which resulted in high efficiency of enzymatic saccharification of the wood samples.     

Harvest date affects colour and soluble solids concentrations (SSC) of <Emphasis Type="Italic">Uapaca kirkiana</Emphasis> (Muell. Arg.) fruits from natural woodlands

Uapaca kirkiana (Muell. Arg.) is a woodland fruit tree that is native to parts of eastern, central and southern Africa. Unripe fruits are harvested from wild and semi-wild sources by dislodging them from trees. Ripe fruits are gathered from the ground after abscission. Utilization of the fruits is affected by problems such as variability of fruit quality and high perishability. Quality attributes of fruits harvested on 1 and 17 November, and 3 December 2003 and kept at 25–30°C, have been evaluated. On the second and third harvests, fruits were also stored in polythene bags to investigate a common incubation method to hasten ripening. Fruit weight and colour before and after ripening, and the soluble solids concentrations (SSC) of ripe fruits were measured. The redness (a*) values at harvest increased from 3.5 to 5.3 units from 1 November to 3 December. Over this period there was a significant decline in both lightness (L*; 63.8 to 58.9 units) and yellowness (b*, 39.6 to 36.4 units). Skin colour became darker during storage as reflected by the decreasing lightness (L* values from 63.8 to 44.5 units), yellowness (b* values from 36.6 to 20.1 units) and increasing redness (a* values from 3.5 to 8.8 units). Fruits harvested on 17 November and 3 December did not darken during storage as much as fruits harvested on 1 November. Fruits harvested on 3 December lost less weight (13.7%) during storage than those harvested on 1 November (34.0%), while the respective SSC measured 6 days after harvest were 18.1% and 9.9%. Fruits kept in polythene bags had 5.3% lower SSC than those kept on plates. The results demonstrate the benefits of delaying fruit harvest to improve quality attributes such as fruit skin colour at harvest and during storage, to reduce weight loss and to obtain higher SSC during storage.     

Ethanol production from sugi pulp under simultaneous saccharification and fermentation using a cocktail enzyme of T. reesei and A. tubingensis produced by solid-state fermentation

The process of solid-state fermentation was used to produce a cocktail enzyme of Trichoderma reesei ATCC 66587 and Aspergillus tubingensis KRCF 700-33. Wheat bran, corncob, and sugi pulp were supplemented with ammonium sulfate as an enzyme-producing medium using T. reesei and A. tubingensis. The corncob blend ratio, duration of incubation, and ammonium sulfate concentration were optimized for enhancing cellulase production from T. reesei using a Box-Behnken design. Filter paper degrading activity more than tripled when T. reesei was grown in the optimized medium, as compared with the initial medium. The highest activity of 4.03 FPU/ml (about 29 FPU/g of dry material) was obtained with a cocktail enzyme having a 25 % content of A. tubingensis and 75 % of T. reesei. The sugi pulp was then fermented to ethanol with the cocktail enzyme and thermotolerant yeast (Saccharomyces cerevisiae BA-11) under simultaneous saccharification and fermentation (SSF) at 40 °C. An ethanol concentration of 4.48 % (w/v) was achieved using the cocktail enzyme (4 FPU/g-pulp) that was produced on-site with a substrate loading level of 12.5 wt %, which achieved an ethanol yield of 76 % after 72 h.     

蒸汽预处理玉米秸秆酶水解速率下降的因素分析

为评价导致纤维素酶水解速率下降的因素,以底物质量浓度为50 g/L葡聚糖及酶用量为20 FPIU/g滤纸酶活和10 IU/g β-葡萄糖苷酶活的蒸汽预处理玉米秸秆酶水解为研究对象,探讨了影响酶水解速率的潜在因素,包括物料反应性能、纤维素酶非特异性吸附、酶失活及终产物抑制。结果表明：酶用量40 FPIU/g条件下酶水解6 h及12 h后,蒸汽预处理玉米秸秆的物料反应性能分别下降了16.0%及23.7%,然而,在酶用量为20 FPIU/g时,物料反应性能的下降对酶水解速率的影响极其有限;酶解木质素的添加使得1 h酶解上清液中酶蛋白浓度降低了20.8%,但初始酶水解速率并未显著降低,即木质素对纤维素酶的非特异性吸附对酶水解速率影响不大;两段酶水解中纤维素酶的更新使得7h酶水解速率由一段酶水解中的1.30 g/(L·h)提高至1.83 g/(L·h);两段酶水解中终产物的去除则使得7 h酶水解速率提高至4.76 g/(L·h),是一段酶水解中7 h酶水解速率的3.66倍。综合而言,酶失活及终产物抑制对酶水解速率影响较大,其中终产物抑制是导致蒸汽预处理玉米秸秆酶水解速率降低的关键因素。     

Level of sugars in Scots pine trees of different sensitivity to fluoride and sulphur dioxide

On the average there is in the needles of the studied pines 1.9 and 4.9% of reducing sugars in the fresh and dry weight of wintertime needles respectively. There is 1.4% glucose of needle fresh weight (3.0% of needle dry weight) while for fructose the corresponding values are 0.8 and 2.1%. The relative ratio of glucose to fructose was on the average 1.33. Trees more tolerant to fluorides and sulphur dioxide emitted from the phosphate fertilizers factory have a statistically significant lower level of directly reducing sugars in the dry weight of needles (37.1 mg/g) compared to sensitive trees (50.5 mg/g). The control (more tolerant trees) accumulate more glucose and fructose than sensitive ones.     

容器规格、肥料种类和育苗基质对桢楠幼苗生长的影响

以半年生桢楠（Phoebe zhennan）幼苗为试材,采用正交试验设计,初步研究了容器规格、肥料种类和育苗基质对桢楠容器苗的苗高、地径、根系长、全株鲜重、全株干重影响。结果表明,不同的肥料种类对幼苗的苗高、地径、全株鲜重、全株干重均有显著影响（P<0.05）,而容器规格和育苗基质对容器苗的生长无显著影响。对不同因素、不同水平的各个组合处理进行分析,半年生桢楠的容器育苗最佳组合为8 cm×12 cm的容器规格,施用5%有机肥作为基肥,采用80%森林土+20%的火烧土作为育苗基质,其次是10 cm×14 cm的容器规格,施用5%有机肥作为基肥,采用100%森林土的育苗基质。     

The smell and odorous components of dried shiitake mushroom, <Emphasis Type="Italic">Lentinula edodes</Emphasis> I: relationship between sensory evaluations and amounts of odorous components

The smell of food is one of the most important factors in assessing its quality. Concerning the smell of dried shiitake mushrooms [Lentinula edodes (Berk.) Pegler], 1,2,3,5,6-pentathiepane, commonly known as lenthionine, has been reported as a key compound. However, other compounds have not been studied sufficiently in connection with smell. From the results of sensory intensity studies and sensory evaluations of dried shiitake mushrooms, a positive significant correlation at 1% risk was observed between sensory intensity and sulfur perception. This showed that the smell of dried shiitake mushrooms was characterized by a sulfurous smell. Also, comparing the sensory intensity with the amounts of volatile components showed positive significant correlations at 1% risk between sensory intensity and three compounds: 1,2,4-trithiolane, 1,2,4,6-tetrathiepane, and lenthionine. Furthermore, significant correlations at 5% risk were obtained between the amounts of these three compounds and sensory intensity by multiple regression analysis. This showed that the smell of dried shiitake mushrooms depended on these compounds. The partial regression coefficient of 1,2,4-trithiolane was larger than those of the others, and so it was proposed that 1,2,4-trithiolane could serve as an indicator to estimate the smell of dried shiitake mushroom.     

光皮树饼粕用于杏鲍菇营养基质的效果初探

本文尝试采用杏鲍菇菌丝处理光皮树提油后的饼粕,以期使光皮树饼粕微生物资源化。结果表明:以水酶法提取光皮树油的固体残渣为基料,适当添加部分配料,杏鲍菇的菌丝能正常生长,且菌丝洁白、浓密、粗壮,结块性强。菌丝满瓶后,粗放管理都可长出子实体。不同配方的光皮树饼粕培养料栽培杏鲍菇,生物转化率均在50%以上,单个子实体鲜重均在15 g以上。无论是从菌丝生长还是单个子实体均重和生物转化率角度评估,光皮树饼粕都是栽培杏鲍菇的良好基质。说明利用食用菌转化技术使光皮树饼粕资源化是一条可行的途径。