食用菌菌糠综合利用研究进展

中国是食用菌生产第一大国,每年产生约1亿多t菌糠,菌糠中含有大量的粗纤维和多糖等物质,但大部分菌糠都被当作废弃物直接丢弃或焚烧,造成严重的环境污染和资源浪费,同时也不符合我国新时期的环保政策,如何变废为宝,科学、环保、经济、合理地利用菌糠成为食用菌产业健康发展的重要环节。本文通过对食用菌菌糠综合利用的方式和现状进行阐述,探讨菌糠利用存在的主要问题,对未来菌糠利用的方向和发展趋势进行展望,为食用菌菌糠的高效利用提供参考和理论依据。     

发酵菌糠饲料的开发

目的 充分利用资源,降低养殖成本。方法 利用菌糠为主要原料,添加适量麸皮和豆粕,以2%的接种量加入植物乳杆菌ST-III、干酪乳杆菌LC2W,15 d密闭发酵。结果 获得具有良好风味和质构、粗蛋白含量（以干物质计）可达到16%、pH为4.3的发酵饲料。动物实验结果表明,其72 h瘤胃降解率可达42%,经25 d饲喂后[1.5 kg/（头·天）],后备牛（9~10月龄）的25 d平均日增重可以达到0.95 kg。结论 发酵菌糠饲料性能优于市售同类产品,具有一定的应用前景。     

饲用微生物的分离鉴定及其对杏鲍菇菌糠发酵的效果

【目的】菌糠的营养素含量齐全,但纤维素含量过高是阻碍其饲料化利用的主要因素。故本研究筛选适合于发酵杏鲍菇菌糠的微生物菌株,以改善其饲用品质。【方法】首先,本研究采用纤维素-刚果红、苯胺蓝和MRS-Ca (De Man, Rogosa, Sharpe-Ca)筛选培养基,结合纤维素、木质素酶活力及抑菌活性的测定,从EM (effective microorganisms)原液发酵的杏鲍菇菌糠中分离筛选具有较强纤维素、木质素降解能力及抑菌能力的细菌/真菌。通过细菌16S rRNA和真菌18S rDNA基因序列分析确定菌株所属种属。其次,将筛选出的菌株菌液等体积混合制成复合菌剂用于固态发酵杏鲍菇菌糠。测定不同发酵时长菌糠营养成分含量以确定最佳发酵时间,并与相同工艺条件下EM原液发酵的杏鲍菇菌糠进行饲用品质比较。【结果】筛选并鉴定得到纤维素酶活性较高的特基拉芽孢杆菌(Bacillus tequilensis)菌株P11、发酵毕赤酵母(Pichia fermentans)菌株R8和马克斯克鲁维应变酵母(Kluyveromyces marxianus)菌株MU5;木质素酶活性较高的解淀粉芽孢杆菌(Bacillus amyloliquefaciens subsp.plantarum)菌株MU7;抑菌活性较高的类肠膜魏斯氏菌(Weissella paramesenteroides)菌株R4和乳酸片球菌(Pediococcus acidilactici)菌株R9。使用以上菌株复合发酵杏鲍菇菌糠7 d后,各项指标达到稳定。与EM原液发酵的杏鲍菇菌糠相比,复合菌剂发酵杏鲍菇菌糠的NDF和ADF分别显著降低了19.6%和21.44%(P0.05);CP (crude protein)、CA (crude ash)和EE (ether extract)含量分别显著提高了10.44%、5.26%和123.53%(P0.05)。【结论】本研究筛选得到的芽孢杆菌、酵母菌和乳酸菌优势菌株复合后用于发酵杏鲍菇菌糠可以很好地改善其饲用品质,效果优于生产中常用市售EM原液。     

金针菇意杨菌糠有机肥发酵的初步研究

通过筛选发酵配方和接种量,测定发芽指数(GI)、水分、pH、酶活性的变化,对金针菇意杨菌糠有机肥发酵进行初步研究。筛选得最佳发酵培养基配方(菌糠70%、米糠15%、麦麸15%),GI为94%。接种量为2%时,GI为92%,脲酶活性最大,为17 U,比接种量为10%(12U)和14%(13 U)高,而纤维素酶活性大小受接种量的影响较小。发酵过程中培养基的水分含量呈下降趋势,pH则呈上升趋势。脲酶最大活性为21 U,出现在第6天,在第8天降到最低值,仅为8 U。而纤维素酶活性最高值在第8天,为10 U,随之下降至8 U。这些研究结果将为金针菇意杨菌糠的肥料化研究提供一定的参考依据。     

菌糠二次利用栽培黑木耳试验研究

为充分利用营养没有得到充分吸收的栽培过食用菌的废弃菌糠、明确在牡丹江地区再利用食用菌菌糠生产黑木耳的适宜比例,以灵芝和鲍鱼菇的废弃菌糠为试验材料,进行不同配方的对比试验。试验结果表明：黑木耳培养料中灵芝菌糠体积分数为30%时栽培效果最佳,显著优于参入鲍鱼菇菌糠的各种培养料配方（P〈0.01）。因此,可以利用灵芝菌糠废料作为替代料栽培黑木耳使用。     

阿魏菇菌糠提取液对四种食用菌菌丝生长的影响

目的:解决菌糠再利用技术问题.方法:采用平板培养法,对在PDA培养基中加入不同体积比的阿魏菇菌糠提取液对鸡腿菇、香菇、杏鲍菇和金针菇菌丝生长的影响进行了研究.结果:当PDA培养基中菌糠提取液加入量为10%～50%时,对香菇和金针菇菌丝生长有促进或明显促进作用;而加入量为10%的菌糠提取液对杏鲍菇菌丝生长无显著影响,但当阿魏菇菌糠提取液加入量大于10%时,明显抑制杏鲍菇和鸡腿菇的菌丝生长.结论:可利用阿魏菇菌糠作为栽培香菇和金针菇的培养料.     

蛹虫草MF27菌糠多糖对肝细胞氧化损伤的保护作用

蛹虫草是一种可利用大米、小麦等谷物培育的名贵药用真菌,其采收后的菌糠里仍富含许多生物活性物质。本研究立足于蛹虫草菌糠多糖,先分析其化学抗氧化活性,再以H₂O₂诱导氧化应激损伤的LO2细胞为模型,评价其对肝细胞氧化损伤的保护作用,进而解析活性多糖的单糖组分。结果显示,菌糠多糖能够有效清除DPPH自由基、羟基（?OH）自由基和ABTS自由基,EC₅₀分别为0.26mg/mL、1.03mg/mL、0.57mg/mL,提示其具有良好的抗氧化能力;在H₂O₂诱导氧化应激损伤的LO2细胞中,菌糠多糖能有效地保护细胞形态的完整性,并且随浓度梯度递增式地提高细胞存活率,当多糖浓度为5mg/mL时,细胞存活率可达91.83%;在分析其作用机制上,与模型组对比,菌糠多糖能通过调节细胞抗氧化酶SOD（提高4.91倍）和CAT（提高3.40倍）的表达来清除ROS含量（P<0.01）,降低氧化损害;经检测,虫草菌糠活性多糖主要含有葡萄糖、甘露糖、半乳糖、阿拉伯糖、葡萄糖醛酸、木糖、半乳糖醛酸、鼠李糖和岩藻糖等单糖。研究结果表明蛹虫草MF27菌糠多糖具有保护肝细胞氧化损伤的作用,为进一步开发和利用虫草菌糠提供了重要理论依据。     

香菇菌糠提取物处理对双孢蘑菇呼吸代谢的影响

为了延长双孢蘑菇Agaricus bisporus的保质期,本研究通过优化香菇Lentinula edodes菌糠提取物(LR-UE)采前喷施条件,探究其对双孢蘑菇贮藏过程中呼吸代谢的影响。研究表明,采前喷施的最佳条件是LR-UE的浓度为1.0mg/mL,喷施时间为采前1d,喷施量为200mL/m2。LR-UE处理能够有效降低双孢蘑菇的失重,延缓双孢蘑菇呼吸峰的出现;同时,它还能调节双孢蘑菇糖酵解途径、三羧酸循环和磷酸戊糖途径的酶活性,减少丙二醛的积累,减少电解质的泄漏,维持较高的可溶性蛋白质含量,有效保持双孢蘑菇的贮藏品质。     

银耳菌糠黑色素的理化性质及抗氧化和抑菌活性

银耳菌糠中存在一种银耳的伴生菌——炭团菌(俗称香灰菌),其菌丝生长到一定阶段后会产生大量的黑色素,具有广泛的应用价值。本研究从银耳菌糠中提取黑色素,研究其理化性质、抗氧化活性及抑菌作用。通过紫外-可见光谱、傅里叶红外光谱对提取的黑色素进行鉴定,表明银耳菌糠黑色素具有黑色素的典型特征。通过对银耳菌糠黑色素理化性质的研究,表明银耳菌糠黑色素是一种趋于黑色并略带红色和黄色的粉末;该黑色素易溶于碱性溶液;具有较好的热稳定性和光稳定性,其稳定性受氧化剂和还原剂的影响较小,受Ca²⁺、Cu²⁺的影响较明显。通过总抗氧化能力(FRAP法)、自由基清除能力检测银耳菌糠黑色素的抗氧化活性,研究表明黑色素具有较高的抗氧化活性,羟自由基、ABTS自由基清除的EC₅₀值分别为0.429 mg/mL和0.016 mg/mL。本研究还检测了黑色素对革兰氏阳性、革兰氏阴性细菌的抑菌能力,结果表明该黑色素在浓度为3.2 mg/mL时对供试菌株的抑菌率超过90%,且对革兰氏阳性菌会较敏感。本研究为银耳菌糠的有效利用及其黑色素产品的开发提供了理论基础,具有较高的经济价值。     

可培养盐碱菌多样性的研究进展

存在于高盐强碱极端环境的微生物因其独特的生命方式,引起了广泛的关注。根据盐碱环境所含的可溶性盐成分,可分为"NaCl型"和"苏打型(Na_2CO_3/NaHCO_3)"两大类,前者的碱性pH值较低而后者碱性pH值较高。本文总结了盐碱菌适宜生长条件在盐度0.5 mol/L和碱性pH 9.0之上且有效发表的标准菌株,并对这些菌株的生物多样性及生理特性进行了阐述;可培养盐碱细菌的数量及其多样性远远大于盐碱古菌,但是盐碱细菌对高盐度和强碱性p H依赖程度相对较低。盐碱细菌主要组成依次为芽孢杆菌纲(Bacilli,占总数约40%)、γ-变形菌纲(γ-Proteobacteria,30%)、梭菌纲(Clostridia,11%)、δ-变形菌纲(δ-Proteobacteria,6%)和放线菌纲(Actinobacteria,6%),而盐碱古菌主要组成为盐古菌纲(Halobacteria,92%)和甲烷微菌纲(Methanomicrobia,8%)。这些极端微生物在生物地球化学过程中或生态循环中扮演着重要的角色和功能,挖掘和利用盐碱菌具有重要意义。     

Bioremediation of Fungicides by Spent Mushroom Substrate and Its Associated Microflora

Experiments were conducted both under in vitro and in situ conditions to determine the biodegradation potential of button mushroom spent substrate (SMS) and its dominating microbes (fungi and bacteria) for carbendazim and mancozeb, the commonly used agricultural fungicides. During 6 days of incubation at 30 ± 2°C under broth culture conditions, highest degradation of carbendazim (17.45%) was recorded with B-1 bacterial isolate, while highest degradation of mancozeb (18.05%) was recorded with Trichoderma sp. In fungicide pre-mixed sterilized SMS, highest degradation of carbendazim (100.00–66.50 μg g⁻¹) was recorded with mixed inoculum of Trichoderma sp. and Aspergillus sp., whereas highest degradation of mancozeb (100.00–50.50 μg g⁻¹) was with mixed inoculum of Trichoderma sp., Aspergillus sp. and B–I bacterial isolate in 15 days of incubation at 30 ± 2°C. All these microbes both individually as well as in different combinations grew well and produced extracellular lignolytic enzymes on SMS, which helped in fungicides degradation. Under in situ conditions, among three different proportions of SMS (10, 20 and 30%, w/w) mixed with fungicide pre-mixed soil (100 μg g⁻¹ of soil), the degradation of carbendazim was highest in 30% SMS treatment, while for mancozeb it was in 20% SMS treatment. The residue levels of both fungicides decreased to half of their initial concentration after 1 month of SMS mixing.     

Utilization of Brewery Spent Grain Liquor by Aspergillus niger

Aspergillus niger was found capable of rapidly converting about 97% of the sugar from brewery spent grain liquor to fungal mass. The yield of dry mycelium, based on the sugar consumed, was approximately 57%. This fungus produced 1.10% titratable acid calculated as citric acid and reduced the biochemical oxygen demand by 96%.     

Degradation of Polycyclic Aromatic Hydrocarbons by Crude Extracts from Spent Mushroom Substrate and its Possible Mechanisms

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by pure laccase has been reported, but the high cost limited its application in environmental bioremediation. Here, we reported a study about PAHs degradation by crude extracts (CEs) containing laccase, which were obtained by extracting four spent mushroom (Agaricus bisporus, Pleurotus eryngii, Pleurotus ostreatus, and Coprinus comatus) substrates. The results showed that anthracene, benzo[a]pyrene, and benzo[a]anthracene were top three degradable PAHs by CEs while naphthalene was most recalcitrant. The PAHs oxidation was enhanced in the presence of 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). Laccase included in CE might play a major role in PAHs degradation. The maximum degradation rate of anthracene and benzo[a]pyrene was observed by using crude extracts from P. eryngii while the highest laccase activities were found in crude extracts from A. bisporus, moreover, crude extracts from P. eryngii, which contained less laccase activities, degraded more anthracene and benzo[a]pyrene than pure laccase with higher laccase activities. The lack of correlation between laccase activity and PAHs degradation rate indicated that other factors might also influence the PAHs degradation. Boiled CEs were added to determine the effect on PAHs degradation by laccase. The results showed that all four boiled CEs had improved the PAHs oxidation. The maximum improvement was observed by adding CEs from P. eryngii. It suggested that some mediators indeed existed in CEs and CEs from P. eryngii contained most. As a result, CEs from P. eryngii has the most application potential in PAHs bioremediation.     

苏云金杆菌ZLL13晶体蛋白分析及其菌糠液态培养基的优化

食用菌栽培废料,简称菌糠(spent mushroom substrate, SMS)是食用菌栽培和生产的残留物,其含有丰富的甲壳素、木质纤维和蛋白质等,可为苏云金芽孢杆菌(Bacillus thuringiensis, Bt)的生长提供所需的营养物质。本研究以优化后的前处理条件制备的菌糠浸提液(2%硫酸, 121℃, 1 h)作为主要碳源,通过单因素试验、Plackett-Burman设计、最陡爬坡和响应面分析等方法来优化最佳培养基组分,结果表明,54%SMS浸提液,31.9 g/L大豆饼粉、0.88 g/L CaCO3、0.4 g/L MnSO4、0.5 g/L K2HPO4和0.4 g/L吐温100为最佳培养基配方,且优化后培养基(1.8×108/mL)产生的孢子数是原始SMS培养基(0.065×108/mL)的27倍,这不仅为菌糠的二次利用提供一种新的有效方法,而且也可以大大降低生产Bt所需要的发酵成本,具有良好的应用前景。     

Differential Utilization of Carbon Substrates by Bacteria and Fungi in Tundra Soil

Little is known about the contribution of bacteria and fungi to decomposition of different carbon compounds in arctic soils, which are an important carbon store and possibly vulnerable to climate warming. Soil samples from a subarctic tundra heath were incubated with ¹³C-labeled glucose, acetic acid, glycine, starch, and vanillin, and the incorporation of ¹³C into different phospholipid fatty acids (PLFA; indicative of growth) and neutral lipid fatty acids (NLFA; indicative of fungal storage) was measured after 1 and 7 days. The use of ¹³C-labeled substrates allowed the addition of substrates at concentrations low enough not to affect the total amount of PLFA. The label of glucose and acetic acid was rapidly incorporated into the PLFA in a pattern largely corresponding to the fatty acid concentration profile, while glycine and especially starch were mainly taken up by bacteria and not fungi, showing that different groups of the microbial community were responsible for substrate utilization. The ¹³C-incorporation from the complex substrates (starch and vanillin) increased over time. There was significant allocation of ¹³C into the fungal NLFA, except for starch. For glucose, acetic acid, and glycine, the allocation decreased over time, indicating use of the storage products, whereas for vanillin incorporation into fungal NLFA increased during the incubation. In addition to providing information on functioning of the microbial communities in an arctic soil, our study showed that the combination of PLFA and NLFA analyses yields additional information on the dynamics of substrate degradation.Bacteria and fungi comprise more than 90% of the soil microbial biomass and are the main agents for decomposition of organic matter in soil. Until recently it was thought that these two organism groups could be lumped together in this respect, and total microbial biomass or total activity (respiration) was often the only variable included in soil microbiology studies of decomposition and soil organic matter turnover (39). However, there is increasing evidence suggesting that whether decomposition is performed by bacteria or fungi, thereby channeling energy through the bacterial or the fungal food web, has profound effects on the ecosystem. Such effects can have direct influence on the higher trophic levels in the food web (30) or indirect effects on nutrient mineralization rates (14) and nutrient transfer (19, 20), and they can even determine the extent of carbon sequestration in the soil (37). The situation becomes even more complex when the impact of changes in climate, nitrogen availability, and litter input on the balance between bacteria and fungi is taken into account. The Arctic region has been identified as an area that will be especially vulnerable to these changes (3).Little is known about the contribution of bacteria and fungi to the utilization of plant-derived carbon substrates in arctic soils. Differentiation of the bacterial and fungal contributions to decomposition has hitherto relied to a large extent on changes in bacterial and fungal biomasses, for example, by analysis of patterns of phospholipid fatty acids (PLFA) (40). PLFA are components of the cell membrane, and some of the PLFA extracted from the soil are characteristic for a certain microbial group in the environment. However, for changes in PLFA concentrations after the addition of substrates to be detected, substrates often have to be added at unrealistically large amounts. Even then only small changes in the PLFA concentrations will often be detected (35).One way of overcoming these problems is to follow the incorporation of ¹³C label from added substrates into specific fatty acids (8, 17). This approach adds a new dimension—metabolic function—to the study of soil microbial communities without the need of cultivation. It also increases the sensitivity in tracing responses of organism groups to different substrates as the addition of substrates at low and more realistic concentrations with high specific ¹³C label will induce large changes in the ¹³C concentration of the PLFA without changing the total amount of PLFA.Carbon-13 labeling has been used to follow uptake of recent photosynthates (11, 13, 27), pure substrates (10, 12, 32, 33, 41), and complex labeled plant material (28, 41, 43, 44) into PLFA although seldom in arctic soils. However, microorganisms incorporate carbon not only into phospholipids (indicating growth) but also into storage products, for example, when a nutrient other than carbon is limiting growth or under growth-restricting conditions. Thus, with excess carbon both bacteria and fungi will store carbon for later need, for example, as polyhydroxyalkanoate or glycogen (bacteria) and triacylglycerols (fungi). Thus, neutral lipid fatty acids (NLFA) of fungal origin can be used to indicate storage in fungi (4). Degraded PLFA, resulting in diacylglycerols, will also end up in the corresponding NLFA fraction, and NLFA has thus been suggested as an indicator of recently dead bacterial biomass (42). Therefore, the NLFA/PLFA ratio serves two purposes: for fungal lipids a higher NLFA/PLFA ratio would indicate allocation of lipids to energy storage while for bacterial lipids it would indicate turnover of this bacterial group. However, the latter will probably be of minor importance during short incubations. As far as we know, no studies on soil microorganisms have used incorporation of ¹³C from substrates to indicate both effects on growth (incorporation into PLFA) and storage (incorporation into NLFA).We assessed the uptake of ¹³C-labeled substrates into lipid biomarkers of different microbial groups in a laboratory incubation experiment using soil from an arctic tundra heath. The selected substrates represented carbon sources present in soil. Glucose, acetic acid, and glycine are simple compounds common in plant root exudates, and glycine is also a nitrogen source. Starch is a very common polysaccharide in plant residues. Vanillin is a common product of lignin depolymerization (18) containing a phenol ring and is often used as a model substance to indicate lignin degradation. Starch and vanillin are therefore examples of more complex substrates and are supposedly more difficult to decompose. We followed the incorporation of the label into different PLFA and NLFA over time. We hypothesized that ¹³C from the simple compounds would be more rapidly incorporated into microbial PLFA than ¹³C from the more complex substrates (more rapid growth), and thus we expected ¹³C emanating from the complex substrates to increase in concentration in the PLFA and NLFA over time. We also hypothesized that bacteria would be better than fungi in utilizing simple compounds while the label from the more complex substrates would preferentially be incorporated into PLFA, indicating fungi (6, 29). We also expected ¹³C from the C-rich substrates to be incorporated into NLFA (fungal storage) to a larger extent than C from glycine, which also serves as a nitrogen source (4). However, with time the carbon in storage structures would decrease as it would be used for growth or maintenance energy.     

甲烷氧化菌及甲烷单加氧酶的研究进展

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