Effect of carbon,nitrogen sources and inducers on ligninolytic enzyme production by Morchella crassipes

The effect of different carbon, nitrogen sources and inducers on growth and ligninolytic activity by Morel mushroom Morchella crassipes was investigated. The maximum growth was observed in mineral salts broth containing glucose as the carbon source and sodium nitrate as the nitrogen source. Among the inducers, chemical inducers inhibited the growth whereas in natural substrates, growth was not affected much. Manganese peroxidase and lignin peroxidase activity were not detected in the medium with different carbon and nitrogen sources, whereas laccase activity varied depending on carbon source (0.7–3.48 U/ml). Among the inducers, natural inducers resulted in an increase in the enzyme activities. Maximum laccase activity was observed in rice straw (12. 6 U/ml) followed by ABTS (11.6 U/ml); Manganese peroxidase activity was maximum in rice straw (14.32 U/l) wheat straw (12.16 U/l) and phenol red (15 U/l) as the inducers, whereas for Lignin peroxidase activity, rice straw (22 U/l), wheat straw (16 U/l) and veratrylalcohol (20 U/l) served as the best inducers.     

Effect of biodelignification of rice straw on humification and humus quality by Phanerochaete chrysosporium and Streptomyces badius

The effect of biodelignification of rice straw by two different ligninolytic organisms, Phanerochaete chrysosporium (white-rot fungus) and Streptomyces badius (actinomycetes), on humus quality was investigated during a 56-day incubation at 30 °C. Lignin degradation, the release of humic extract (HE), humic acid (HA) and fulvic acid (FA), E4/E6 ratio of HA, and humification index (HI, HA/FA) were measured during the incubation. Lignin was degraded by both organisms, but to different extents. Lignin was degraded to 41% and 31% by P. chrysosporium and S. badius, respectively. HE released by P. chrysosporium and S. badius were, respectively, 2.10 and 2.13 times larger than that in the control at the maximum values. A significant correlation between lignin degradation and humus-related parameters involving HA fraction showed that both organisms are converting lignin to humic substances.     

Increased PCP removal by <Emphasis Type="Italic">Amylomyces rouxii</Emphasis> transformants with heterologous <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis> peroxidases supplementing their natural degradative pathway

Fungal peroxidases and phenoloxidases are widely used in aromatic toxic compounds degradation. Peroxidases, such as lignin peroxidase and manganese peroxidase, as well as laccases are mainly produced by basidiomycetes and to a lower extent by other fungi, such as ascomycetes. Peroxidase-encoding genes have been described and homologous expression has been achieved in basidiomycetes. Heterologous expression has also been achieved in some non-producing peroxidase ascomycetes, like Penicillium and Aspergillus. In this work, heterologous expression of peroxidase-encoding genes, lignin peroxidase, and manganese peroxidase was achieved in a zygomycete producing only phenoloxidases (Amylomyces rouxii), aimed at coupling two different pathways used in nature for PCP removal in only one microbial strain. The ability of PCP removal was assayed with one of the obtained transformants, resulting in increased activity with respect to the ability of the parental strain cultured free of the inducer tyrosine (95% and 45%, respectively, of the initial PCP (12.5 mg L⁻¹) in 120 h, or 100% and 49%, respectively, of the initial PCP after 144 h of liquid culture).     

Effect of surfactants on fluoranthene degradation by <Emphasis Type="Italic">Pseudomonas alcaligenes</Emphasis> PA-10

Two surfactants, Tween 80 and JBR, were investigated for their effect on fluoranthene degradation by a Pseudomonad. Both surfactants enhanced fluoranthene degradation by Pseudomonas alcaligenes PA-10 in shake flask culture. This bacterium was capable of utilising the synthetic surfactant and the biosurfactant as growth substrates and the critical micelle concentration of neither compound inhibited bacterial growth. The biosurfactant JBR significantly increased polycyclic aromatic hydrocarbon (PAH) desorption from soil. Inoculation of fluoranthene-contaminated soil microcosms with P. alcaligenes PA-10 resulted in the removal of significant amounts (45 ± 5%) of the PAH after 28 days compared to an uninoculated control. Addition of the biosurfactant increased the initial rate of fluoranthene degradation in the inoculated microcosm. The presence of a lower molecular weight PAH, phenanthrene, had a similar effect on the rate of fluoranthene removal.     

Isolation and screening of potential actinobacteria for rapid composting of rice straw

Rice straw is produced as a by-product from rice cultivation, which is composed largely of lignocellulosic materials amenable to general biodegradation. Lignocellulolytic actinobacteria can be used as a potential agent for rapid composting of bulky rice straw. Twenty-five actinobacteria isolates were isolated from various in situ and in vitro rice straw compost sources. Isolates A2, A4, A7, A9 and A24 were selected through enzymatic degradation of starch, cellulose and lignin followed by the screening for their adaptability on rice straw powder amended media. The best adapted isolate (A7) was identified as Micromonospora carbonacea. It was able to degrade cellulose, hemicelluloses and carbon significantly (P ≤ 0.05) over the control. C/N ratio was reduced to 18.1 from an initial value of 29.3 in 6 weeks of composting thus having the potential to be used in large scale composting of rice straw.     

Application of solid waste from anaerobic digestion of poultry litter in Agrocybe aegerita cultivation: mushroom production, lignocellulolytic enzymes activity and substrate utilization

The degradation and utilization of solid waste (SW) from anaerobic digestion of poultry litter by Agrocybe aegerita was evaluated through mushroom production, loss of organic matter (LOM), lignocellulolytic enzymes activity, lignocellulose degradation and mushroom nutrients content. Among the substrate combinations (SCs) tested, substrates composed of 10–20% SW, 70–80% wheat straw and 10% millet was found to produce the highest mushroom yield (770.5 and 642.9 g per 1.5 kg of substrate). LOM in all SCs tested varied between 8.8 and 48.2%. A. aegerita appears to degrade macromolecule components (0.6–21.8% lignin, 33.1–55.2% cellulose and 14–53.9% hemicellulose) during cultivation on the different SCs. Among the seven extracellular enzymes monitored, laccase, peroxidase and CMCase activities were higher before fruiting; while xylanase showed higher activities after fruiting. A source of carbohydrates (e.g., millet) in the substrate is needed in order to obtain yield and biological efficiency comparable to other commercially cultivated exotic mushrooms.     

Biodegradation of paddy straw obtained from different geographic locations by means of <Emphasis Type="Italic">Phlebia</Emphasis> spp. for animal feed

Various cereal straws are used as feed by supplementing the green forage or other feed stuffs. An experiment was designed to see the effect of different geographic locations and climatological conditions on biochemical constituents, fungal degradation and in vitro digestibility of paddy straw. Paddy straw (PS) obtained from three different geographic locations of India was subjected to solid state fermentation using four white rot fungi i.e. Phlebia brevispora, P. fascicularia, P. floridensis and P. radiata. Changes in the biochemical constituents like water soluble content, hemicellulose, cellulose, lignin, total organic matter, and in vitro digestibility of paddy straw was analyzed over a period of 60 days along with lignocellulolytic enzymes i.e. laccase, xylanase and carboxymethyl cellulase. All the fungi degraded the straw samples and enhanced the in vitro digestibility. The paddy straw, obtained from north western zone (NWZ) suffered a maximum loss (228 g/kg) of lignin by P. radiata, while a maximum enhancement of in vitro digestibility from 185 to 256 g/kg was achieved by P. brevispora, which also caused minimum loss in total organic matter (98 g/kg). In PS obtained from central eastern zone (CEZ) and north eastern zone (NEZ), a maximum amount of lignin (210 and 195 g/kg, respectively) was degraded by P. floridensis and resulted into a respective enhancement of in vitro digestibility from 172 to 246 g/kg and 188 to 264 g/kg. The study demonstrates that geographic locations not only affect the biochemical constituents of paddy straw but the fungal degradation of fibers, their in vitro digestibility and lignocellulolytic enzyme activity of the fungus may also vary.     

Production of Ligninolytic Enzymes by Phlebia Floridensis

Summary The present work reports the production of laccase, lignin peroxidase and manganese peroxidase by the little studied white-rot fungus Phlebia floridensis under a variety of nutritional and physicochemical conditions. Among the different media and supplements the highest yields of laccase, lignin peroxidase and manganese peroxidase were recorded in the presence of sugarcane bagasse, wheat straw and rice straw, respectively. Laccase and manganese peroxidase activities were best expressed at a pH of 4.5 while lignin peroxidase was optimally active at a lower pH. Laccase proved to be much more thermostable as compared to the other two enzymes.     

Establishment of the white rot fungus Phanerochaete chrysosporium on unsterile straw in solid substrate fermentation systems intended for degradation of pesticides

The effects of different inoculum-loading rates and pre-treatment of wheat straw with formic acid and hot water (50 °C) on the establishment of Phanerochaete chrysosporium on unsterile straw were studied in laboratory scale and in a 1.5-m³ bioreactor. The establishment of P. chrysosporium on unsterile straw was satisfactory. Phanerochaete chrysosporium and other fungi, which developed simultaneously, were able to produce the activity necessary to degrade two herbicides, bentazon and MCPA (4-chloro-2-methylphenoxyacetic acid) in 20 days (65 and 75%, respectively). The decrease of both herbicides coincided with the presence of the activity of the lignin-degrading enzymes lignin peroxidase and manganese peroxidase/laccase. Extensive growth of P. chrysosporium or other lignin-degrading fungi on unsterile straw would be excellent for inexpensive solid substrate systems intended for degradation of pesticides.     

Bioconversion and biotransformation of coir pith for economic production of Pleurotus florida: chemical and biochemical changes in coir pith during the mushroom growth and fructification

Coir pith represents ∼50% of the waste from the coir industries and was tested for its potential in serving as a growth substrate for the production of species of oyster mushroom, Pleurotus florida. Due to its high lignin (∼48%) content and amorphous powdery nature, coir pith supported poor mushroom mycelial growth and yields were considerably low (∼25% bioconversion efficiency). Pre-treating coir pith with hot water did not prove economical to produce the mushroom yields. Acid swelling and alkali delignification of coir pith though served to change the structure of coir pith; the mushroom yields were not improved. Amendment of coir pith with rice (Oryza sativa) straw and horse gram (Dolichos biflorus) plant residue tended to greatly modify the physical characteristics of the inoculated mushroom bed. Such a supplementation of coir pith growth substrate resulted in production of mushroom yields with 110–125% bioconversion efficiency. Implications of supplementing coir pith with rice straw/horse gram plant residue in terms of holocellulose:lignin ratio are discussed. Sensorially, the mushrooms so produced did not differ from that on rice straw, the economic growth substrate recommended for production of the mushroom yields on commercial scale. Changes in cellulose, hemicellulose and lignin contents of coir pith amended with rice straw were studied. Cellulase, hemicellulase and protease enzyme activities in the amended coir pith substrate showed a continuous increase from inoculation till the end of fructification, whereas laccase activity decreased during fructification, in consonance with decreased lignin degradation during fructification.     

木质素降解菌BYL-7的筛选及降解条件优化

【背景】微生物降解木质素因其具有降解效率高和环保等特点而备受关注。【目的】筛选高效木质素降解真菌,并对其降解条件进行优化。【方法】通过愈创木酚-马铃薯葡萄糖琼脂(potato dextroseagar,PDA)和苯胺蓝平板法筛选高效木质素降解菌株,利用单因素筛选及响应面试验对培养条件进行优化。【结果】筛选到一株高效木质素降解菌BYL-7,经形态和多序列分析初步确定为Trametes versicolor。单因素试验证明初始pH、温度和接种量为降解木质素显著影响因子,响应面试验确定降解木质素最优条件为:初始pH 6.7,温度25°C,接种量8%。在此条件下,碱性木质素降解率为36.5%,比未优化前提高54.0%;水稻秸秆木质素、半纤维素和纤维素降解率分别为32.8%、21.5%、13.2%,其中木质素降解率比未优化前提高36.1%;漆酶活性在第6天达到峰值120.0 U/L,比未优化前提高25.0%;木质素过氧化物酶活性在第6天达到峰值1343.8U/L,比未优化前提高36.0%;锰过氧化物酶活性在第5天达到峰值463.8U/L,比未优化前提高31.7%。【结论】研究结果为木质素的降解提...     

黄孢原毛平革菌对松木、稻草和芦苇去木质化作用的研究

为确定黄孢原毛平革菌对不同植物材料的去木质化作用,以pH、干物质重、半纤维素、纤维素和木质素为主要技术指标,比较黄孢原毛平革菌对松木、稻草和芦苇降解能力的差异。松木、芦苇在发酵过程中pH呈下降趋势,稻草呈上升趋势。在干物质重、半纤维素、纤维素降解率三个指标上皆为松木〈芦苇〈稻草,在木质素降解率上则为松木〈稻草〈芦苇,且差异显著。表明黄孢原毛平革菌对不同植物材料去木质化能力有较大差异,其中芦苇的木质素降解率为13％,是三种材料中最易于被去木质化的。     

Screening of a fungus capable of powerful and selective delignification on wheat straw

Aims: To screen and characterize a novel fungus with powerful and selective delignification capability on wheat straw. Methods and Results: A fungus capable of efficient delignification under solid‐state fermentation (SSF) conditions on wheat straw was screened. After 5 days of incubation, 13·07% of the lignin was removed by fungal degradation, and 7·62% of the holocellulose was lost. Furthermore, 46·53% of the alkali lignin was removed after 2 days of liquid fermentation. The fungus was identified as Fusarium concolor based on its morphology and an analysis of its 18S rDNA gene sequence. The molecular weight distribution of lignin was evaluated by gel permeation chromatography. Enzyme assay indicated that the fungus produced laccase, cellobiose dehydrogenase, xylanase and cellulase during the incubation period. Intracellular lignin peroxidase, manganese peroxidase and laccase were produced during liquid fermentation. Conclusions: We have successfully screened a fungus, F. concolor, which can efficiently degrade the lignin of wheat straw, with slight damage to the cellulose, after 5 days of SSF. Significance and Impact of the Study: The newly isolated strain could be used in pretreatment of lignocellulose materials prior to biopulping, bioconversion into fuel and substrates for the chemical industry.     

Roles of apoplastic peroxidases, laccases, and lignification in the manganese tolerance of hyperaccumulator Phytolacca americana

We investigated the response of Mn-hyperaccumulator Phytolacca americana L. to manganese excess as well as the relationships between lignin deposition in the plant’s leaves, peroxidase and laccase activities in the leaf apoplast, and Mn toxicity. The exceptionally high tolerance of P. americana to Mn, both in solution and in tissue, was confirmed. No visible brown spot was observed in the leaves of plants treated with ≤10,000 μM Mn for 10 days. Mn treatment significantly increased lignin content and laccase activity in the apoplastic washing fluid (AWF) of P. americana leaves. In contrast, an increase in the Mn supply was paralleled by a significant decrease in the concentration of total phenolic compounds (TPCs) and in water-soluble guaiacol peroxidase (SPOD) activity in leaf AWF. This result suggested that an increase in lignin deposition decreased the concentration of apoplastic TPCs that are available to generate potentially toxic intermediates by acting as peroxidase substrates. Thus, data of the present study indicate that lignin formation by laccase activities reduces Mn toxicity and increases Mn tolerance of P. americana by depressing SPOD-mediated formation of toxic intermediates from TPCs.     

Lignin and manganese peroxidase activity in extracts from straw solid substrate fermentations

Manganese and lignin peroxidase (MnP, LiP) activities were measured in straw extracts from cultures of Phanerochaete chrysosporium. Out of six MnP substrates, the MBTH/DMAB (3-methyl-2-benzothiazolinone hydrazone/3-(dimethylamino)benzoic acid), gave the highest MnP activity. Detection of LiP activity as veratryl alcohol oxidation was inhibited by phenols in the straw culture extracts. Appropriate levels of veratryl alcohol and peroxide (4 mM and 0.4 mM, respectively), and a restricted sample volume (not larger than 10%) were necessary to detect activity.     

不同木质纤维素基质上白腐菌降解特性的研究

通过测定木质素、纤维素、半纤维素和漆酶分泌的变化,研究白腐菌在稻草、木屑、粗纤维素、滤纸、黑液木素基质上的降解特性。结果表明,除黑液木素上白腐菌不能生长外,在前25d,各基质中纤维素、半纤维素和木质素含量呈持续下降趋势,之后,降解速率减少,其中木质素的降解速率大于纤维素和半纤维素的降解速率。漆酶分泌在生长初期呈快速上升趋势,第10d酶活达到最大,第10～20d快速下降,其后基本不变,基质中酶活大小顺序为稻草基质、木屑基质、粗纤维和滤纸基质,显示了木质素存在对漆酶分泌的诱导作用。     

Comparative study on the growth and yield of<Emphasis Type="Italic"> Pleurotus ostreatus</Emphasis> mushroom on different lignocellulosic by-products

Eight lignocellulosic by-products were evaluated as substrates for cultivation of the oyster mushroom, Pleurotus ostreatus (Jacq. ex. fr) Kummer. The yields of mushroom on the different substrates were 183.1, 151.8, 111.5, 87.8, 49.5, 23.3, 13.0 and 0.0 g for composted sawdust of Triplochiton scleroxylon, rice straw, banana leaves, maize stover, corn husk, rice husk, fresh sawdust, and elephant grass, respectively. The biological efficiency (BE) followed the same pattern and ranged from 61.0% for composted sawdust to 0.0% for elephant grass. The yield of mushroom was positively correlated to cellulose (r ² =0.6), lignin (r ² =0.7) and fibre (r ² =0.7) contents of the substrates. Based on the yield and BE of the substrates tested, rice straw appeared to be the best alternate substrate for growing oyster mushrooms. Electronic Publication     

Optimization of lignin peroxidase production and stability by <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis> using sewage-treatment-plant sludge as substrate in a stirred-tank bioreactor

A laboratory-scale study was carried out to produce lignin peroxidase (ligninase) by white rot fungus (Phanerochaete chrysosporium) using sewage-treatment-plant (STP) sludge as the major substrate. The optimization was done using full-factorial design (FFD) with agitation and aeration as the two parameters. Nine experiments indicated by the FFD were fermented in a stirred-tank bioreactor for 3 days. A second-order quadratic model was developed using the regression analysis of the experimental results with the linear, quadratic, and interaction effects of the parameters. Analysis of variance (ANOVA) showed a high coefficient of determination (R ²) value of 0.972, thus indicating a satisfactory fit of the quadratic model with the experimental data. Using statistical analysis, the optimum aeration and agitation rates were determined to be 2.0 vvm and 200 rpm, respectively, with a maximum activity of 225 U l⁻¹ in the first 3 days of fermentation. The validation experiment showed the maximum activity of lignin peroxidase was 744 U l⁻¹ after 5 days of fermentation. The results for the tests of the stability of lignin peroxidase showed that the activity was more than 80% of the maximum for the first 12 h of incubation at an optimum pH of 5 and temperature of 55°C.     

Emerging biotechnological potentials of DyP-type peroxidases in remediation of lignin wastes and phenolic pollutants: a global assessment (2007–2019)

Dye decolourizing peroxidase (DyP) is an emerging biocatalyst with enormous bioremediation and biotechnological potentials. This study examined the global trend of research related to DyP through a bibliometric analysis. The search term ‘dye decolourizing peroxidase’ or ‘DyP-type peroxidase’ was used to retrieve published articles between 2007 and 2019 from the Web of Science (WoS) and Scopus databases. A total of 62 articles were published within the period, with an annual growth rate of 17·6%. The highest research output was observed in 2015, which accounted for about 13% of the total output in 12 years. Germany published the highest number of articles (n = 10, 16·1%) with a total citation of 478. However, the lowest number of published articles among the top 10 countries was observed in India and Korea (n = 2, 3·2%). Research collaboration was low (collaboration index = 4·08). In addition to dye decolourizing peroxidase(s) and DyP-type peroxidase(s) (n = 33, 53·23%), the top authors keywords and research focus included lignin and lignin degradation (n = 10, 16·1 %). More so, peroxidase (n = 59, 95·2%), amino acid sequence (n = 27, 46·8%), lignin (n = 24, 38·7%) and metabolism (n = 23, 37·1%) were highly represented in keywords-plus. The most common conceptual framework from this study include characterization, lignin degradation and environmental proteomics. Apart from the inherent efficient dye-decolourizing properties, this study showed that DyP has emerging biotechnological potentials in lignin degradation and remediation of phenolic environmental pollutants, which at the moment are under explored globally.     

Radiation and fermentation treatment of cellulosic wastes

The effect of radiation pasteurization of sugar cane bagasse and rice straw and fermentation using various strains of fungi were studied for upgrading of cellulosic wastes. The initial contamination by fungi and aerobic bacteria both in bagasse and straw was high. The doses of 30 kGy for sterilization and 8 kGy for elimination of fungi were required. Irradiation effect showed that rice straw contained comparatively radioresistant microorganisms. It was observed that all the fungi (Hericium erinacium, Pleurotus djamor, Ganoderma lucidum, Auricularia auricula, Lentinus sajor-caju, Coriolus versicolor, Polyporus arcularius, Coprinus cinereus) grow extending over the entire substrates during one month after inoculation in irradiated bagasse and rice straw with 3% rice bran and 65% moisture content incubated at 30°C. Initially, sugar cane bagasse and rice straw substrates contained 39.4% and 25.9% of cellulose, 22.9% and 26.9% of hemicellulose, and 19.6% and 13.9% of lignin + cutin, respectively. Neutral detergent fibre (NDF) values decreased significantly in sugar cane bagasse fermented byG. lucidum, A. auricula andP. arcularius, and in rice straw fermented by all the 8 strains of fungi. Acid detergent fibre (ADF) values also decreased in bagasse and rice straw fermented by all the fungi.P. arcularius, H. erinacium, G. lucidum andC. cinereus were found to be the most effective strains for delignification of sugar cane bagasse.