scholarly journals Direct Ethanol Production from Xylan and Acorn Using the Starch-Fermenting Basidiomycete Fungus Phlebia acerina

Fermentation ◽  
2021 ◽  
Vol 7 (3) ◽  
pp. 116
Author(s):  
Kenji Okamoto ◽  
Takato Goda ◽  
Takeru Yamada ◽  
Masafumi Nagoshi
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Environmentally Friendly ◽  
White Rot ◽  
Starch Concentration ◽  
Wide Range ◽  
Mycelia Growth ◽  
Basidiomycete Fungus ◽  
Basidiomycete Fungi

During our search for ethanol-producing basidiomycete fungi for a wide range of substrates, we isolated Phlebia acerina, which is a white rot basidiomycete fungus. It favorably converted starch into ethanol with approximately 70% yield. Although the yield decreased as the starch concentration increased, growth and fermentation were observed even at 200 g/L of starch. P. acerina produced ethanol from glucose, galactose, mannose, xylose, cellobiose, and maltose with 93%, 91%, 86%, 72%, 92%, and 68% yields, respectively. Additionally, P. acerina, which secreted xylanase and xylosidase, was capable of assimilating xylan and directly converting it to ethanol with a yield of 63%. Furthermore, P. acerina produced ethanol directly from acorns, which are plant fruits containing starch and tannins, with a yield of 70%. Tannin delayed mycelia growth, thus prolonging ethanol production; however, this did not particularly affect the yield. These results were similar to those of fermentation in a medium with the same amounts of starch and tannin as the target crop acorn, thus suggesting that P. acerina could successfully produce environmentally friendly ethanol from starch-containing lignocellulosic biomass, unlike previously reported ethanol-producing basidiomycete fungi.

scholarly journals Purification and Characterization of Two Novel Laccases from Peniophora lycii

2020 ◽  
Vol 6 (4) ◽  
pp. 340
Author(s):  
Olga A. Glazunova ◽  
Konstantin V. Moiseenko ◽  
Olga S. Savinova ◽  
Tatyana V. Fedorova
Keyword(s):  
Phylogenetic Relationships ◽  
White Rot ◽  
Laccase Production ◽  
Purification And Characterization ◽  
Orthologous Genes ◽  
Basidiomycete Fungus ◽  
Scarce Data ◽  
Bromcresol Green ◽  
Basidiomycete Fungi

Although, currently, more than 100 laccases have been purified from basidiomycete fungi, the majority of these laccases were obtained from fungi of the Polyporales order, and only scarce data are available about the laccases from other fungi. In this article, laccase production by the white-rot basidiomycete fungus Peniophora lycii, belonging to the Russulales order, was investigated. It was shown that, under copper induction, this fungus secreted three different laccase isozymes. Two laccase isozymes—Lac5 and LacA—were purified and their corresponding nucleotide sequences were determined. Both purified laccases were relatively thermostable with periods of half-life at 70 °C of 10 and 8 min for Lac5 and LacA, respectively. The laccases demonstrated the highest activity toward ABTS (97 U·mg−1 for Lac5 and 121 U·mg−1 for LacA at pH 4.5); Lac5 demonstrated the lowest activity toward 2,6-DMP (2.5 U·mg−1 at pH 4.5), while LacA demonstrated this towards gallic acid (1.4 U·mg−1 at pH 4.5). Both Lac5 and LacA were able to efficiently decolorize such dyes as RBBR and Bromcresol Green. Additionally, phylogenetic relationships among laccases of Peniophora spp. were reconstructed, and groups of orthologous genes were determined. Based on these groups, all currently available data about laccases of Peniophora spp. were systematized.

scholarly journals Genomic Studies of White-Rot Fungus Cerrena unicolor SP02 Provide Insights into Food Safety Value-Added Utilization of Non-Food Lignocellulosic Biomass

2021 ◽  
Vol 7 (10) ◽  
pp. 835
Author(s):  
Zichen Zhang ◽  
Aabid Manzoor Shah ◽  
Hassan Mohamed ◽  
Yao Zhang ◽  
Nino Tsiklauri ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Molecular Mechanisms ◽  
Genetic Material ◽  
Value Added ◽  
White Rot ◽  
White Rot Fungus ◽  
Industrial Biotechnology ◽  
Lignocellulolytic Enzymes ◽  
Cerrena Unicolor ◽  
Wide Range

Cerrena unicolor is an ecologically and biotechnologically important wood-degrading basidiomycete with high lignocellulose degrading ability. Biological and genetic investigations are limited in the Cerrena genus and, thus, hinder genetic modification and commercial use. The aim of the present study was to provide a global understanding through genomic and experimental research about lignocellulosic biomass utilization by Cerrena unicolor. In this study, we reported the genome sequence of C. unicolor SP02 by using the Illumina and PacBio 20 platforms to obtain trustworthy assembly and annotation. This is the combinational 2nd and 3rd genome sequencing and assembly of C. unicolor species. The generated genome was 42.79 Mb in size with an N50 contig size of 2.48 Mb, a G + C content of 47.43%, and encoding of 12,277 predicted genes. The genes encoding various lignocellulolytic enzymes including laccase, lignin peroxidase, manganese peroxidase, cytochromes P450, cellulase, xylanase, α-amylase, and pectinase involved in the degradation of lignin, cellulose, xylan, starch, pectin, and chitin that showed the C. unicolor SP02 potentially have a wide range of applications in lignocellulosic biomass conversion. Genome-scale metabolic analysis opened up a valuable resource for a better understanding of carbohydrate-active enzymes (CAZymes) and oxidoreductases that provide insights into the genetic basis and molecular mechanisms for lignocellulosic degradation. The C. unicolor SP02 model can be used for the development of efficient microbial cell factories in lignocellulosic industries. The understanding of the genetic material of C. unicolor SP02 coding for the lignocellulolytic enzymes will significantly benefit us in genetic manipulation, site-directed mutagenesis, and industrial biotechnology.

Ethanol Production from Lignocellulosic Biomass Using Xylotrophic Basidiomycetes

2015 ◽  
Vol 51 (5) ◽  
pp. 516-525 ◽  
Author(s):  
N. R. Al’myasheva ◽  
A. A. Novikov ◽  
E. Yu. Kozhevnikova ◽  
A. V. Golyshkin ◽  
A. V. Barkov ◽  
...  
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Xylotrophic Basidiomycetes

scholarly journals Protein Engineering Approaches to Enhance Fungal Laccase Production in S. cerevisiae

2021 ◽  
Vol 22 (3) ◽  
pp. 1157
Author(s):  
Pablo Aza ◽  
Felipe de Salas ◽  
Gonzalo Molpeceres ◽  
David Rodríguez-Escribano ◽  
Iñigo de la Fuente ◽  
...  
Keyword(s):  
Protein Engineering ◽  
Directed Evolution ◽  
Signal Peptide ◽  
Laccase Activity ◽  
Laccase Production ◽  
Wide Range ◽  
Mating Factor ◽  
Conserved Amino Acids ◽  
Basidiomycete Fungi

Laccases secreted by saprotrophic basidiomycete fungi are versatile biocatalysts able to oxidize a wide range of aromatic compounds using oxygen as the sole requirement. Saccharomyces cerevisiae is a preferred host for engineering fungal laccases. To assist the difficult secretion of active enzymes by yeast, the native signal peptide is usually replaced by the preproleader of S. cerevisiae alfa mating factor (MFα1). However, in most cases, only basal enzyme levels are obtained. During directed evolution in S. cerevisiae of laccases fused to the α-factor preproleader, we demonstrated that mutations accumulated in the signal peptide notably raised enzyme secretion. Here we describe different protein engineering approaches carried out to enhance the laccase activity detected in the liquid extracts of S. cerevisiae cultures. We demonstrate the improved secretion of native and engineered laccases by using the fittest mutated α-factor preproleader obtained through successive laccase evolution campaigns in our lab. Special attention is also paid to the role of protein N-glycosylation in laccase production and properties, and to the introduction of conserved amino acids through consensus design enabling the expression of certain laccases otherwise not produced by the yeast. Finally, we revise the contribution of mutations accumulated in laccase coding sequence (CDS) during previous directed evolution campaigns that facilitate enzyme production.

scholarly journals Media selection for mycelia growth, antifungal activity against wood-degrading fungi, and GC-MS study by Pycnoporus sanguineus

BioResources ◽  
2011 ◽  
Vol 6 (3) ◽  
pp. 2719-2731 ◽  
Author(s):  
Yi P. Teoh ◽  
Mashitah M. Don ◽  
Salmiah Ujang
Keyword(s):  
Antifungal Activity ◽  
Water Extract ◽  
White Rot ◽  
White Rot Fungus ◽  
Gas Chromatography Mass Spectrometry ◽  
Malt Extract ◽  
Decay Fungi ◽  
Pycnoporus Sanguineus ◽  
Mycelia Growth ◽  
Property Changes

Wood-decaying fungi present a serious threat to items made from rubberwood (Hevea brasiliensis). Though conventional chemical control has been a successful method for preserving wood against stain and decay fungi growth, the effects of these chemicals are of concern because they create problems for the environment and public health. Pycnoporus sanguineus (P. sanguineus), is a white-rot fungus that invades wood during its growth, storage, or use, causing decay or other property changes. It was considered in this work as a potential source of bioactive compounds and investigated for its natural antifungal activity using a minimum inhibitory concentration assay against wood-degrading fungi. It was found that media consisting of 10.0 g/L malt extract, yeast extract, dextrose, and maltose, respectively at pH 4.7±0.2 provided the highest biomass production by P. sanguineus. Results showed that the antifungal properties of methanol and water extract of P. sanguineus mycelia and supernatant ranged from MIC values of 0.1 to 5.0 µg/µL. 4H-Pyran-4-one,2,3-dihydro-3,5-dihydroxy-6-methyl- (DDMP) was found to be the major component in the extract of this fungus, based on analysis using gas chromatography – mass spectrometry.

scholarly journals Bio-ethanol (2nd Generation Ethanol): A Solution to Ever Polluting Gasoline to Climate in India

2020 ◽  
pp. 1-15
Author(s):  
Shruti Mohapatra ◽  
Raj Kishore Mishra ◽  
Khitish K. Sarangi
Keyword(s):  
Ethanol Production ◽  
Lignocellulosic Biomass ◽  
Fossil Fuels ◽  
Economic Benefits ◽  
Policy Implications ◽  
Current Status ◽  
Environmentally Sustainable ◽  
2Nd Generation ◽  
Production Technologies ◽  
Rapid Pace

Environmentally sustainable energy sources are called for due to contemporaneous development in industries along with the rapid pace of urbanization. Ethanol produced from biomass can be deliberated as a clean and safest liquid fuel and an alternative to fossil fuels as they have provided unique environmental, strategic economic benefits. For the past decade, it has been noticed that there is an increasing trend found in bio ethanol production which has created a stimulus to go for advancement in bio ethanol production technologies. Several feed stocks have been used for the bio ethanol production but the second generation bio ethanol has concentrated on the lignocellulosic biomass. Plenteous lignocellulosic biomass in the world can be tapped for ethanol production, but it will require significant advances in the ethanol production process from lignocellulosic because of some technical and economic hurdles found in commercial scale. This review will encompass the current status of bio ethanol production in terms of their economic and environmental viability along with some research gaps as well as policy implications for the same.

scholarly journals A comprehensive study of the promoting effect of manganese on white rot fungal treatment for enzymatic hydrolysis of woody and grass lignocellulose

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Xiao Fu ◽  
Jialong Zhang ◽  
Xiangyu Gu ◽  
Hongbo Yu ◽  
Shulin Chen
Keyword(s):  
Wheat Straw ◽  
Lignocellulosic Biomass ◽  
Quantum Coherence ◽  
Plant Cell Wall ◽  
White Rot ◽  
Yield Enhancement ◽  
Biomass Recalcitrance ◽  
Fungal Treatment ◽  
Ether Linkage ◽  
Glucose Yield

Abstract Background The efficiency of biological systems as an option for pretreating lignocellulosic biomass has to be improved to make the process practical. Fungal treatment with manganese (Mn) addition for improving lignocellulosic biomass fractionation and enzyme accessibility were investigated in this study. The broad-spectrum effect was tested on two different types of feedstocks with three fungal species. Since the physicochemical and structural properties of biomass were the main changes caused by fungal degradation, detailed characterization of biomass structural features was conducted to understand the mechanism of Mn-enhanced biomass saccharification. Results The glucose yields of fungal-treated poplar and wheat straw increased by 2.97- and 5.71-fold, respectively, after Mn addition. Particularly, over 90% of glucose yield was achieved in Mn-assisted Pleurotus ostreatus-treated wheat straw. A comparison study using pyrolysis gas chromatography mass spectrometry (Py-GC/MS) and two-dimensional 1H–13C heteronuclear single quantum coherence (2D HSQC) nuclear magnetic resonance (NMR) spectroscopy was conducted to elucidate the role of Mn addition on fungal disruption of the cross-linked structure of whole plant cell wall. The increased Cα-oxidized products was consistent with the enhanced cleavage of the major β-O-4 ether linkages in poplar and wheat straw lignin or in the wheat straw lignin–carbohydrate complexes (LCCs), which led to the reduced condensation degree in lignin and decreased lignin content in Mn-assisted fungal-treated biomass. The correlation analysis and principal component analysis (PCA) further demonstrated that Mn addition to fungal treatment enhanced bond cleavage in lignin, especially the β-O-4 ether linkage cleavage played the dominant role in removing the biomass recalcitrance and contributing to the glucose yield enhancement. Meanwhile, enhanced deconstruction of LCCs was important in reducing wheat straw recalcitrance. The findings provided not only mechanistic insights into the Mn-enhanced biomass digestibility by fungus, but also a strategy for improving biological pretreatment efficiency of lignocellulose. Conclusion The mechanism of enhanced saccharification of biomass by Mn-assisted fungal treatment mainly through Cα-oxidative cleavage of β-O-4 ether linkages further led to the decreased condensation degree in lignin, as a result, biomass recalcitrance was significantly reduced by Mn addition. Graphic abstract

scholarly journals A simple enzymatic assay for the quantification of C1-specific cellulose oxidation by lytic polysaccharide monooxygenases

2019 ◽  
Vol 42 (1) ◽  
pp. 93-102 ◽  
Author(s):  
M. B. Keller ◽  
C. Felby ◽  
C. A. Labate ◽  
V. O. A. Pellegrini ◽  
P. Higasi ◽  
...  
Keyword(s):  
Lignocellulosic Biomass ◽  
Oxidation Product ◽  
Gluconic Acid ◽  
Enzymatic Assay ◽  
Cellulose Oxidation ◽  
Metal Chelator ◽  
Pure Cellulose ◽  
Lytic Polysaccharide Monooxygenases ◽  
Wide Range ◽  
Polysaccharide Monooxygenases

Abstract Objective The development of an enzymatic assay for the specific quantification of the C1-oxidation product, i.e. gluconic acid of cellulose active lytic polysaccharide monooxygenases (LPMOs). Results In combination with a β-glucosidase, the spectrophotometrical assay can reliably quantify the specific C1- oxidation product of LPMOs acting on cellulose. It is applicable for a pure cellulose model substrate as well as lignocellulosic biomass. The enzymatic assay compares well with the quantification performed by HPAEC-PAD. In addition, we show that simple boiling is not sufficient to inactivate LPMOs and we suggest to apply a metal chelator in addition to boiling or to drastically increase pH for proper inactivation. Conclusions We conclude that the versatility of this simple enzymatic assay makes it useful in a wide range of experiments in basic and applied LPMO research and without the need for expensive instrumentation, e.g. HPAEC-PAD.

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