Effects of C/N Ratio on Lignocellulose Degradation and Enzyme Activities in Aerobic Composting

Lignocellulosic materials have a complex physicochemical composition and structure that reduces their decomposition rate and hinders the formation of humic substances during composting. Therefore, a composting experiment was conducted to evaluate the effects of different C/N ratios on lignocellulose (cellulose, hemicellulose and lignin) degradation and the activities of corresponding enzymes during aerobic composting. The study had five C/N ratios, namely, T1 (C/N ratio of 15), T2 (C/N ratio of 20), T3 (C/N ratio of 25), T4 (C/N ratio of 30) and T5 (C/N ratio of 35). The results showed that treatments T3 and T4 had the highest rate of degradation of cellulose and hemicellulose, while treatment T3 had the highest rate of degradation of lignin. Among the five treatments, treatment T3 enhanced the degradation of the lignocellulose constituents, indicating a degradation rate of 6.86–35.17%, 15.63–44.08% and 31.69–165.60% for cellulose, hemicellulose and lignin, respectively. The degradation of cellulose and lignin occurred mainly at the thermophilic and late mesophilic phases of composting, while hemicellulose degradation occurred at the maturation phase. Treatment T3 was the best C/N ratio to stimulate the activities of manganese peroxidase, lignin peroxidase, polyphenol oxidase and peroxidase, which in turn promoted lignocellulose degradation.

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Identification of enzymes for lignocellulose degradation in the genome and transcriptome of the aquatic hyphomycete Clavariopsis aquatica

10.1101/2020.06.18.151886 ◽

2020 ◽

Author(s):

Felix Heeger ◽

Elizabeth C. Bourne ◽

Christian Wurzbacher ◽

Elisabeth Funke ◽

Anna Lipzen ◽

...

Keyword(s):

Cytochrome P450 ◽

Lignin Degradation ◽

Carbon Sources ◽

Lignocellulose Degradation ◽

Cytochrome P450 Monooxygenases ◽

P450 Monooxygenases ◽

Lignin Modification ◽

Hemicellulose Degradation ◽

Differential Gene ◽

Carbohydrate Components

AbstractFungi are ecologically important decomposers of lignocellulose. Basidiomycetes use peroxidases, laccases, and enzymes of the cytochrome P450 superfamily for cometabolic lignin degradation in order to access cellulose and hemicellulose as carbon sources. Limited lignin modification capabilities have also been reported for some terrestrial ascomycetes. Here we newly sequenced the genome of an exclusively aquatic ascomycete, Clavariopsis aquatica, documented the presence of genes for the modification of lignocellulose and its constituents, and compared differential gene expression between C. aquatica cultivated on lignocellulosic and sugar-rich substrates. We identified potential peroxidases, laccases, and cytochrome P450 monooxygenases several of which were differentially expressed when experimentally grown on different substrates. Additionally, we found regulation of pathways for cellulose and hemicellulose degradation. Our results suggest that C. aquatica is able to modify lignin, detoxify aromatic lignin constituents, or both. This may facilitate the use of carbohydrate components of lignocellulose as carbon and energy sources.

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Effects of supplementation with phosphorus, calcium and manganese during oil palm frond fermentation by Phanerochaete chrysosporium on ligninase enzyme activity

Biodiversitas Journal of Biological Diversity ◽

10.13057/biodiv/d210509 ◽

2020 ◽

Vol 21 (5) ◽

Author(s):

Roni Pazla ◽

Novirman Jamarun ◽

Fauzia Agustin ◽

Mardiati Zain ◽

Arief Arief ◽

...

Keyword(s):

Enzyme Activity ◽

Phanerochaete Chrysosporium ◽

Oil Palm ◽

Manganese Peroxidase ◽

Crude Protein ◽

Lignin Degradation ◽

Lignin Content ◽

Oil Palm Frond ◽

Range Test ◽

Palm Frond

Abstract. Pazla R, Jamarun N, Agustin F, Zain M, Cahyani NO. 2020. Effects of supplementation with phosphorus, calcium and manganese during oil palm frond fermentation by Phanerochaete chrysosporium on ligninase enzyme activity. Biodiversitas 21: 1833-1838. The objective of this study was to evaluate the effects of supplementation with phosphorus (P) in combination with calcium (Ca) and manganese (Mn) during oil palm frond (OPF) fermentation by Phanerochaete chrysosporium on ligninase enzyme activity and lignin degradation. This study was carried out using a randomized complete design with 3 treatments (addition of P, Ca and Mn) and 5 replicates. The following treatments were performed: T1 (P 1000 + Ca 2000 + Mn 150 ppm), T2 (P 1500 + Ca 2000 + Mn 150 ppm), and T3 (P 2000 + Ca 2000 +Mn 150 ppm). The data were subjected to an analysis of variance (ANOVA), and differences between treatment means were tested using Duncan's multiple range test (DMRT). The parameters measured were as follows: lignin peroxidase (LiP) activity (U/mL), manganese peroxidase (MnP) activity (U/mL), crude protein (CP) content (%), crude fiber (CF) content (%) and the decrease in lignin (%). The results revealed a significant increase in LiP activity and CP content and a decrease in the lignin content (p<0.05) by the addition of P in the T3 treatment. However, the treatment nonsignificantly increased (p>0.05) MnP activity and significantly decreased (P<0.05) the CF content. In conclusion, supplementation of the OPF fermentation process with P 2000, Ca 2000, and Mn 150 ppm resulted in the highest ligninase enzyme activity and in decreased lignin content.

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Linking Enzymatic Oxidative Degradation of Lignin to Organics Detoxification

International Journal of Molecular Sciences ◽

10.3390/ijms19113373 ◽

2018 ◽

Vol 19 (11) ◽

pp. 3373 ◽

Cited By ~ 9

Author(s):

Xiaolu Wang ◽

Bin Yao ◽

Xiaoyun Su

Keyword(s):

Redox Potential ◽

Lignin Peroxidase ◽

Manganese Peroxidase ◽

Lignin Degradation ◽

Oxidative Degradation ◽

Class Ii ◽

Substrate Preference ◽

Versatile Peroxidase

The major enzymes involved in lignin degradation are laccase, class II peroxidases (lignin peroxidase, manganese peroxidase, and versatile peroxidase) and dye peroxidase, which use an oxidative or peroxidative mechanism to deconstruct the complex and recalcitrant lignin. Laccase and manganese peroxidase directly oxidize phenolic lignin components, while lignin peroxidase and versatile peroxidase can act on the more recalcitrant non-phenolic lignin compounds. Mediators or co-oxidants not only increase the catalytic ability of these enzymes, but also largely expand their substrate scope to those with higher redox potential or more complicated structures. Neither laccase nor the peroxidases are stringently selective of substrates. The promiscuous nature in substrate preference can be employed in detoxification of a range of organics.

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Enhanced degradation of ammonium-pretreated wheat straw by lignocellulolytic Streptomyces spp.

Canadian Journal of Microbiology ◽

10.1139/m92-168 ◽

1992 ◽

Vol 38 (10) ◽

pp. 1022-1025 ◽

Cited By ~ 11

Author(s):

Marina Basaglia ◽

Giuseppe Concheri ◽

Stefano Cardinali ◽

Maria B. Pasti-Grigsby ◽

Marco P. Nuti

Keyword(s):

Weight Loss ◽

Wheat Straw ◽

Lignin Degradation ◽

Cellulose Degradation ◽

Lignocellulose Degradation ◽

Cellulose Content ◽

Streptomyces Spp ◽

Initial Substrate ◽

Streptomyces Chromofuscus ◽

Pretreated Wheat Straw

Eleven actinomycetes, isolated from the gut of worker termites (Macrotermes, Armitermes, Microcerotermes, Odontotermes), were identified as Streptomyces chromofuscus, S. chromogenus, S. diastaticus, and S. rochei. Their ability to grow on natural lignocellulosic substrates was tested in solid state fermentation experiments using wheat straw (C/N = 49.8) as a sole carbon source. Weight loss was 4.7–20.9% of the initial substrate, after 5 weeks at 30 °C; lignin and cellulose content decreased 2.0–16.1 and 3.5–32.9%, respectively. When the 11 Streptomyces were grown on wheat straw pretreated with (NH4)HCO3 (C/N = 28.2), weight loss was 9.3–29.9% of the initial substrate, indicating an overall enhancement of lignocellulose degradation. Weight, lignin, and cellulose losses were enhanced when S. chromofuscus (strain A2 and A11) and S. rochei A4 were grown on pretreated wheat straw instead of the untreated substrate. With S. rochei A10 the weight loss and lignin degradation were enhanced, while cellulolysis was slightly depressed. Weight loss and cellulose degradation were both enhanced when the remaining strains were grown on pretreated wheat straw. In this case, lignin degradation was depressed (S. chromofuscus A6 and A8, S. diastaticus A12, S. rochei A14) or remained essentially the same (S. diastaticus A3 and S. chromogenus A7). Key words: Streptomyces, wheat straw, degradation, lignin, cellulose.

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New and different lignocellulosic materials from Turkey for laccase and manganese peroxidase production byTrametes versicolor

Engineering in Life Sciences ◽

10.1002/elsc.200700025 ◽

2009 ◽

Vol 9 (1) ◽

pp. 60-65 ◽

Cited By ~ 11

Author(s):

Emre Erden ◽

Meryem Cigdem Ucar ◽

Yasin Kaymaz ◽

Nurdan Kasikara Pazarlioglu

Keyword(s):

Manganese Peroxidase ◽

Lignocellulosic Materials

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Pretreated Sugarcane Bagasse Result in More Efficient Degradation by Streptomyces sp S2

10.21203/rs.3.rs-1059961/v1 ◽

2021 ◽

Author(s):

Stanislaus Aditya Agung ◽

Dede Heri Yuli Yanto ◽

Anja Meryandini ◽

Titi Candra Sunarti

Keyword(s):

Hydrogen Peroxide ◽

Sugarcane Bagasse ◽

Ligninolytic Enzymes ◽

Lignocellulose Degradation ◽

Lignocellulosic Materials ◽

Lignocellulosic Material ◽

Chemical Treatments ◽

Material Condition ◽

Pretreated Sugarcane Bagasse ◽

Guaiacyl Lignin

Abstract Streptomyces genera plays important role in lignocellulose degradation. Many research founds Streptomyces has cellulolytic and ligninolytic enzymes that sufficient to degrade lignocellulosic materials. However, minimum lignocellulosic material condition that can efficiently degraded by Streptomyces sp. has not been fully understood. In this research, three pretreament conditions (physical, alkaline-hydrotermal, and hydrogen-peroxide chemical treatments) of sugarcane bagasse used as lignocellulosic material, to further degraded by Streptomyces sp. S2. Lignocellulose component measurement conclude that raw (physical treated only) bagasse wasn’t efficiently degraded by Streptomyces sp S2. Hydrogen-peroxide was effective on reducing both syringil and guaiacyl lignin, meanwhile alkaline-hydrotermal pretreatment was very effective on reducing syringil lignin. This study suggest that hydrogen-peroxide pretreatment can be used in many type of lignocellulosic material, which can be further degraded by Streptomyces sp. S2. Alkaline-hydrotermal preteatment on the other hand is best suited to degrade lignocellulosic material that have high percentage of syringil lignin.

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Physiological Characteristics and Comparative Secretome Analysis of Morchella importuna Grown on Glucose, Rice Straw, Sawdust, Wheat Grain, and MIX Substrates

Frontiers in Microbiology ◽

10.3389/fmicb.2021.636344 ◽

2021 ◽

Vol 12 ◽

Author(s):

YingLi Cai ◽

XiaoLong Ma ◽

QianQian Zhang ◽

FuQiang Yu ◽

Qi Zhao ◽

...

Keyword(s):

Rice Straw ◽

Lignin Degradation ◽

Extracellular Enzymes ◽

Plant Biomass ◽

Growth Potential ◽

Glycoside Hydrolases ◽

Lignocellulose Degradation ◽

Wheat Grain ◽

Predominant Component ◽

Morchella Importuna

Morels (Morchella sp.) are economically important edible macro-fungi, which can grow on various synthetic or semi-synthetic media. However, the complex nutritional metabolism and requirements of these fungi remain ill-defined. This study, based on the plant biomass commonly used in the artificial cultivation of morels, assessed and compared the growth characteristics and extracellular enzymes of Morchella importuna cultivated on glucose, rice straw, sawdust, wheat grain, and a mixture of equal proportions of the three latter plant substrates (MIX). M. importuna could grow on all five tested media but displayed significant variations in mycelial growth rate, biomass, and sclerotium yield on the different media. The most suitable medium for M. importuna was wheat and wheat-containing medium, followed by glucose, while rice straw and sawdust were the least suitable. A total of 268 secretory proteins were identified by liquid chromatography coupled with tandem mass spectrometry detection. Functional classification and label-free comparative analysis of these proteins revealed that carbohydrate-active enzyme (CAZYme) proteins were the predominant component of the secretome of M. importuna, followed by protease, peptidase, and other proteins. The abundances of CAZYme proteins differed among the tested media, ranging from 64% on glucose to 88% on rice straw. The CAZYme classes of glycoside hydrolases and carbohydrate-binding module were enriched in the five secretomes. Furthermore, the enzyme activities of CMCase, lignase, amylase, xylase, pNPCase, and pNPGase were detected during the continuous culture of M. importuna in MIX medium, and the relative expression of the corresponding genes were detected by quantitative real-time PCR. The combined data of growth potential, secretome, extracellular enzyme activity, and gene expression on different substrates inferred that M. importuna was weak in lignocellulose degradation but a good starch decomposer. Specifically, in terms of the degradation of cellulose, the ability to degrade cellulose into oligosaccharides was weaker compared with further degradation into monosaccharides, and this might be the speed-limiting step of cellulose utilization in M. importuna. In addition, M. importuna had a strong ability to decompose various hemicellulose glycosidic bonds, especially α- and β-galactosidase. Only a very few lignin-degradation-related proteins were detected, and these were in low abundance, consistent with the presence of weak lignin degradation ability. Furthermore, the presence of lipase and chitinase implied that M. importuna was capable of decomposition of its own mycelia in vitro. The study provides key data that facilitates a further understanding of the complex nutritional metabolism of M. importuna.

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Isolation and Characterization of a Novel Laccase LacZ1 for Lignin Degradation

Applied and Environmental Microbiology ◽

10.1128/aem.01355-21 ◽

2021 ◽

Author(s):

Weiran Zhang ◽

Weiwei Wang ◽

Jinghong Wang ◽

Guinan Shen ◽

Yuan Yuan ◽

...

Keyword(s):

Enzyme Activity ◽

High Temperature ◽

Lignin Degradation ◽

Microbial Consortium ◽

Organic Polymer ◽

Lignocellulose Degradation ◽

Bacterial Laccase ◽

Isolation And Characterization ◽

Acid Pathway ◽

Insight Into

Lignin is a complex natural organic polymer and is one of the primary components of lignocellulose. The efficient utilization of lignocellulose is limited because it is difficult to degrade lignin. In this study, we screened a lacz1 gene fragment encoding laccase from the macro transcriptome data of a microbial consortium WSC-6, which can efficiently degrade lignocellulose. The RT-qPCR results demonstrated that the expression level of the lacz1 gene during the peak period of lignocellulose degradation by WSC-6 increased by 30.63 times compared to the initial degradation period. Phylogenetic tree analysis demonstrated that the complete lacz1 gene is derived from Bacillus sp. and encoded laccase. The corresponding protein LacZ1 was expressed and purified by Ni-chelating affinity chromatography. The optimum temperature was 75°C, the optimum pH was 4.5, and the highest enzyme activity reached 16.39 U/mg. We found that Cu 2+ was an important cofactor needed for LacZ1 to have enzyme activity. The molecular weight distribution of lignin was determined by Gel Permeation Chromatography (GPC) and changes in the lignin structure were determined by 1H Nuclear Magnetic Resonance Spectra (1H NMR). The degradation products of lignin by LacZ1 were determined by Gas Chromatography and Mass Spectrometry (GC-MS), and three lignin degradation pathways (the gentian acid pathway, benzoic acid pathway, and protocatechuic acid pathway) were proposed. This study provides insight into the degradation of lignin and new insights into high-temperature bacterial laccase. IMPORTANCE Lignin is a natural aromatic polymer that is not easily degraded, hindering the efficient use of lignocellulose-rich biomass resources, such as straw. Biodegradation is a method of decomposing lignin that has recently received increasing attention. In this study, we screened a gene encoding laccase from the lignocellulose-degrading microbial consortium WSC-6, purified the corresponding protein LacZ1, characterized the enzymatic properties of laccase LacZ1, and speculated that the degradation pathway of LacZ1 degrades lignin. This study identified a new, high-temperature bacterial laccase that can degrade lignin, providing insight into lignin degradation by this laccase.

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Research on the Efficiency of Acid-Producing and Gas-Producing Based on the Enhancement of High-Performance Composite Strains

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.807-809.1232 ◽

2013 ◽

Vol 807-809 ◽

pp. 1232-1235

Author(s):

Guo Xiang Zheng ◽

Tian Fu Liu ◽

Li Li Yin ◽

Yuan Bei Zhang ◽

Jie Yang

Keyword(s):

High Performance ◽

Degradation Rate ◽

Anaerobic Fermentation ◽

Lignocellulose Degradation ◽

Cow Dung ◽

Volatile Acid ◽

Two Phase ◽

Fermentation Products ◽

High Performance Composite ◽

Acidogenic Phase

A batch test was carried out to investigate the effect of acid-producing fermentative characteristic of a new complex strains LZF-12 with high lignocellulose-degradation ability, which straw was the only carbon source in acidogenic phase. Simultaneity, the biogas-producing performance was also researched by shifting the liquid end products from acidongenic phase to methane phase, which fermentative substrate was the mixture of cow dung and kitchen waste. The results indicated that major fermentation products of acidogenic phase was acetic acid accounted for 70% of total volatile acid, which would be in favor of microorganisms growth of methanogenic phase. In two-phase anaerobic fermentation system, the straw degradation rate of composite strains LZF-12 reached the maximum at 72 hour, among which the degradation rate of cellulose, hemicellulose and lignin reached 55%, 67% and 13% respectively. The methane percentage of biogas-producing phase was more than 50% and 134.7mL/g VS of methane production rate was obtained.

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Wood-feeding termite gut symbionts as an obscure yet promising source of novel manganese peroxidase-producing oleaginous yeasts intended for azo dye decolorization and biodiesel production

Biotechnology for Biofuels ◽

10.1186/s13068-021-02080-z ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Rania Al-Tohamy ◽

Jianzhong Sun ◽

Maha A. Khalil ◽

Michael Kornaros ◽

Sameh Samir Ali

Keyword(s):

Lipid Accumulation ◽

Manganese Peroxidase ◽

Azo Dye ◽

Dye Decolorization ◽

Biodiesel Production ◽

Lignocellulose Degradation ◽

Oleaginous Yeasts ◽

Hydroxymethyl Furfural ◽

Termite Gut ◽

Gut Symbionts

Abstract Background The ability of oxidative enzyme-producing micro-organisms to efficiently valorize organic pollutants is critical in this context. Yeasts are promising enzyme producers with potential applications in waste management, while lipid accumulation offers significant bioenergy production opportunities. The aim of this study was to explore manganese peroxidase-producing oleaginous yeasts inhabiting the guts of wood-feeding termites for azo dye decolorization, tolerating lignocellulose degradation inhibitors, and biodiesel production. Results Out of 38 yeast isolates screened from wood-feeding termite gut symbionts, nine isolates exhibited high levels of extracellular manganese peroxidase (MnP) activity ranged between 23 and 27 U/mL after 5 days of incubation in an optimal substrate. Of these MnP-producing yeasts, four strains had lipid accumulation greater than 20% (oleaginous nature), with Meyerozyma caribbica SSA1654 having the highest lipid content (47.25%, w/w). In terms of tolerance to lignocellulose degradation inhibitors, the four MnP-producing oleaginous yeast strains could grow in the presence of furfural, 5-hydroxymethyl furfural, acetic acid, vanillin, and formic acid in the tested range. M. caribbica SSA1654 showed the highest tolerance to furfural (1.0 g/L), 5-hydroxymethyl furfural (2.5 g/L) and vanillin (2.0 g/L). Furthermore, M. caribbica SSA1654 could grow in the presence of 2.5 g/L acetic acid but grew moderately. Furfural and formic acid had a significant inhibitory effect on lipid accumulation by M. caribbica SSA1654, compared to the other lignocellulose degradation inhibitors tested. On the other hand, a new MnP-producing oleaginous yeast consortium designated as NYC-1 was constructed. This consortium demonstrated effective decolorization of all individual azo dyes tested within 24 h, up to a dye concentration of 250 mg/L. The NYC-1 consortium's decolorization performance against Acid Orange 7 (AO7) was investigated under the influence of several parameters, such as temperature, pH, salt concentration, and co-substrates (e.g., carbon, nitrogen, or agricultural wastes). The main physicochemical properties of biodiesel produced by AO7-degraded NYC-1 consortium were estimated and the results were compared to those obtained from international standards. Conclusion The findings of this study open up a new avenue for using peroxidase-producing oleaginous yeasts inhabiting wood-feeding termite gut symbionts, which hold great promise for the remediation of recalcitrant azo dye wastewater and lignocellulosic biomass for biofuel production. Graphical Abstract

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