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Food intake, nutrient utilization and water turnover in the lesser mouse-deer (Tragulus javanicus) given lundai (Sapium baccatum)

Nolan JV, Liang JB, Abdullah N, Kudo H, Ismail H, Ho YW, et al.

Comp. Biochem. Physiol. A Physiol., 1995 May;111(1):177-82.
PMID: 7735907

Voluntary food intake, digestibility and water turnover were determined in adult Malaysian lesser mouse-deer (Tragulus javanicus) given unlimited access to lundai foliage (Sapium baccatum). Daily dry matter (DM) intake was 42.4 g/kg metabolic live mass (M0.73) or 3.7% M. Digestible energy intake was 853 kJ/day (571 kJ metabolisable energy per M0.73), calculated to be used with 79% efficiency. Apparent digestibility (%) of organic matter was 83.8, crude fibre 63.7, acid detergent fibre 60.5, neutral detergent fibre 72.1 and crude protein 65.0. Urinary excretion of the purine derivative, allantoin, was 0.05 mg/g digestible DM intake suggesting rumen microbial yield efficiency may be lower than in other ruminant species. Total water intake was 182 ml/M0.82. The body-water content of the fed mouse-deer, from tritiated water dilution, was 77% M, consistent with a very lean carcass. Turnover of body water was 17% per day. The mouse-deer produced relatively dry, well-defined faecal pellets.

Matched MeSH terms: Lignin/analysis
Comparison on pore development of activated carbon produced from palm shell and coconut shell

Daud WM, Ali WS

Bioresour Technol, 2004 May;93(1):63-9.
PMID: 14987722

A series of experiments were conducted to compare the pore development in palm-shell and coconut-shell-based activated carbons produced under identical experimental conditions. Carbonization and activation processes were carried out at 850 degrees C using a fluidized bed reactor. Within the range of burn-off studied, at any burn-off, the micropore and mesopore volumes created in palm-shell-based activated carbon were always higher than those of coconut-shell-based activated carbon. On macropore volume, for palm-shell-based activated carbon, the volume increased with increase in burn-off up to 30% and then decreased. However, for coconut-shell-based activated carbon, the change in macropore volume with burn-off was almost negligible but the absolute macropore volume decreased with burn-off.

Matched MeSH terms: Lignin/metabolism
Solid state bioconversion of oil palm biomass for ligninase enzyme production

Alam MZ, Mahmat ME, Muhammad N

Artif Cells Blood Substit Immobil Biotechnol, 2005;33(4):457-66.
PMID: 16317964

A laboratory-scale study of bioconversion of local lignocellulosic material, oil palm biomass (OPB) was conducted by evaluating the enzyme production through microbial treatment in solid state bioconversion (SSB). OPB in the form of empty fruit bunches (EFB) was used as a solid substrate and treated with the white-rot fungus, Phanerochaete chrysosporium, to produce ligninase. The results showed that the highest ligninase activity of 400.27 U/liter was obtained at day 12 of fermentation. While the optimum study indicated the enzyme production of 1472.8 U/liter with moisture content of 50%, 578.7 U/liter with 10% v/w of inoculum size, and 721.8 U/liter with co-substrate concentration of 1% (w/w) at days 9, 9 and 12 of fungal treatment, respectively. The parameters glucosamine and reducing sugar were observed to evaluate the growth and substrate utilization in the experiment.

Matched MeSH terms: Lignin/metabolism
Identification of differentially expressed genes in oil palms (Elaeis guineensis) related to infections by Ganoderma boninense

Mohamad Arif, A.M., Zetty Norhana, B.Y., Abdullah, F., Tan, Su.G., Ismail, S.

Buletin Persatuan Genetik Malaysia, 2005;11(2):0-0.
MyJurnal

Basal stem rot (BSR) caused by species Ganoderma, is one of the most serious disease of oil palm in Malaysia. As far as the disease problem to oil palm in Malaysia is concerned, BSR is the only disease requiring urgent solution. The BSR is not new to Malaysia, it has been known to attack oil palm since the early years when the crop was introduced into this country. There is an indication that there are differences in susceptibility to basal stem rot between germplasm materials from different genetic origin [2]. This provides hope in generating oil palm varieties with reduced level of susceptibility using existing genetic materials. There is also interest in developing diagnostic tools such as using PCR primers for detection of the pathogen in oil palms [1]. Altered expression of several classes of genes was observed in plants in response to fungal infection. These include genes associated with cell maintenance and development, genes involved in biosynthesis of lignin and phenolics and genes implicated in oxidative burst, programmed cell death or hypersensitive response [5].

Matched MeSH terms: Lignin
Fulltext Isolation and characterization of pulp from sugarcane bagasse and rice straw

Saiful Azhar, S., Suhardy, D., Kasim, F.H., Nazry Saleh, M.

Journal of Nuclear and Related Technologies, 2007;2007(1):109-114.
MyJurnal

The amount of sugarcane bagasse and rice straw in the state of Perlis (Malaysia) is abundant while its utilization is still limited. One of the alternatives for the bagasse and straw utilization is as pulp raw material. This paper reviews on pulp from sugarcane bagasse and rice straw and its suitability for paper production. In this study, the pulp was extracted by the Soxhlet extraction method. The objective of this study was to investigate the cellulose, lignin and silica content of the pulp from sugarcane bagasse and rice straw. For rice straw, the presence of large amount of pentosanes in the pulp and black liquors, which also contain silica were decreased the using of straw in the paper industry. Therefore, formic acid pulping and NaOH treatment are studied to reduce or prevent silica. The isolated pulp samples were further characterized by Scanning Electron Microscope (SEM) to investigate their fiber dimensions.

Matched MeSH terms: Lignin
Physicochemical properties of hydrothermally treated hemicellulose from oil palm frond

Fazilah A, Azemi MN, Karim AA, Norakma MN

J Agric Food Chem, 2009 Feb 25;57(4):1527-31.
PMID: 19166335 DOI: 10.1021/jf8028013

Hemicelluloses from oil palm frond (OPF) were extracted using 3 M potassium hydroxide (KOH) for 4 h at 40 degrees C with stirring at 400 rpm to obtain hemicelluloses A and B. The total yield of the hemicellulose isolated from OPF was 33% (dry weight). Both hemicelluloses A and B were then subjected to hydrothermal treatment at 121 degrees C and 1.03 x 10(5) Pa for 10, 30, and 50 min. Physicochemical characterizations of hydrothermally treated hemicelluloses, such as Klason lignin content and reducing sugar content, were performed to study the effect of autohydrolysis processing on OPF-derived hemicelluloses. It was shown that Klason lignin content in hemicellulose A was higher than that in hemicellulose B and decreased after hydrothermal treatment. Hydrothermal treatment enhanced the solubility of hemicelluloses, which reflects their higher reducing sugar content. Monosaccharide analysis using HPLC showed that xylose was the predominant monosaccharide for both hemicelluloses A and B.

Matched MeSH terms: Lignin/analysis
Pretreatment of empty palm fruit bunch for production of chemicals via catalytic pyrolysis

Misson M, Haron R, Kamaroddin MF, Amin NA

Bioresour Technol, 2009 Jun;100(11):2867-73.
PMID: 19232488 DOI: 10.1016/j.biortech.2008.12.060

The effect of chemical pretreatments using NaOH, H(2)O(2), and Ca(OH)(2) on Empty Palm Fruit Bunches (EPFB) to degrade EPFB lignin before pyrolysis was investigated. Spectrophotometer analysis proved consecutive addition of NaOH and H(2)O(2) decomposed almost 100% of EPFB lignin compared to 44% for the Ca(OH)(2), H(2)O(2) system while NaOH and Ca(OH)(2) used exclusively could not alter lignin much. Next, the pretreated EPFB was catalytically pyrolyzed. Experimental results indicated the phenolic yields over Al-MCM-41 and HZSM-5 catalysts were 90 wt% and 80 wt%, respectively compared to 67 wt% yield for the untreated sample under the same set of conditions. Meanwhile, the experiments with HY zeolite yielded 70 wt% phenols.

Matched MeSH terms: Lignin/chemistry*
Exploring the antioxidant potential of lignin isolated from black liquor of oil palm waste

Bhat R, Khalil HP, Karim AA

C. R. Biol., 2009 Sep;332(9):827-31.
PMID: 19748457 DOI: 10.1016/j.crvi.2009.05.004

This study was conducted to evaluate the potential antioxidant activity of lignin obtained from black liquor, a hazardous waste product generated during the extraction of palm oil. Antioxidant potential of the extracted lignin was evaluated by dissolving the extracted samples in 2 different solvent systems, namely, 2-methoxy ethanol and DMSO. Results revealed high percent inhibition of the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical in the lignin sample dissolved in 2-methoxy ethanol over DMSO (concentration range of 1-100 microg/ml). Lignin extracted in 2-methoxy ethanol exhibited higher inhibition percentage (at 50 microg/ml, 84.2%), whereas a concentration of 100 microg/ml was found to be effective in the case of the DMSO solvent (69.8%). Fourier transform infrared (FTIR) spectrometry revealed that the functional groups from the extracted lignin and commercial lignin were highly similar, indicating the purity of the lignin extracted from black liquor. These results provide a strong basis for further applications of lignin in the food industry and also illustrate an eco-friendly approach to utilize oil palm black liquor.

Matched MeSH terms: Lignin/pharmacology*
Optimization study of ethanolic fermentation from oil palm trunk, rubberwood and mixed hardwood hydrolysates using Saccharomyces cerevisiae

Chin KL, H'ng PS, Wong LJ, Tey BT, Paridah MT

Bioresour Technol, 2010 May;101(9):3287-91.
PMID: 20056407 DOI: 10.1016/j.biortech.2009.12.036

Ethanolic fermentation using Saccharomyces cerevisiae was carried out on three types of hydrolysates produced from lignocelulosic biomass which are commonly found in Malaysia such as oil palm trunk, rubberwood and mixed hardwood. The effect of fermentation temperature and pH of hydrolysate was evaluated to optimize the fermentation efficiency which defined as maximum ethanol yield in minimum fermentation time. The fermentation process using different temperature of 25 degrees Celsius, 30 degrees Celsius and 40 degrees Celsius were performed on the prepared fermentation medium adjusted to pH 4, pH 6 and pH 7, respectively. Results showed that the fermentation time was significantly reduced with the increase of temperature but an adverse reduction in ethanol yield was observed using temperature of 40 degrees Celsius. As the pH of hydrolysate became more acidic, the ethanol yield increased. Optimum fermentation efficiency for ethanolic fermentation of lignocellulosic hydrolysates using S. cerevisiae can be obtained using 33.2 degrees Celsius and pH 5.3.

Matched MeSH terms: Lignin/metabolism
Bio-ethanol from lignocellulose: Status, perspectives and challenges in Malaysia

Goh CS, Tan KT, Lee KT, Bhatia S

Bioresour Technol, 2010 Jul;101(13):4834-41.
PMID: 19762229 DOI: 10.1016/j.biortech.2009.08.080

The present study reveals the perspective and challenges of bio-ethanol production from lignocellulosic materials in Malaysia. Malaysia has a large quantity of lignocellulosic biomass from agriculture waste, forest residues and municipal solid waste. In this work, the current status in Malaysia was laconically elucidated, including an estimation of biomass availability with a total amount of 47,402 dry kton/year. Total capacity and domestic demand of second-generation bio-ethanol production in Malaysia were computed to be 26,161 ton/day and 6677 ton/day, respectively. Hence, it was proven that the country's energy demand can be fulfilled with bio-ethanol if lignocellulosic biomass were fully converted into bio-ethanol and 19% of the total CO(2) emissions in Malaysia could be avoided. Apart from that, an integrated national supply network was proposed together with the collection, storage and transportation of raw materials and products. Finally, challenges and obstacles in legal context and policies implementation were elaborated, as well as infrastructures shortage and technology availabilities.

Matched MeSH terms: Lignin/chemistry*
Second-generation bio-ethanol (SGB) from Malaysian palm empty fruit bunch: energy and exergy analyses

Tan HT, Lee KT, Mohamed AR

Bioresour Technol, 2010 Jul;101(14):5719-27.
PMID: 20223656 DOI: 10.1016/j.biortech.2010.02.023

Recently, second-generation bio-ethanol (SGB), which utilizes readily available lignocellulosic biomass has received much interest as another potential source of liquid biofuel comparable to biodiesel. Thus the aim of this paper is to determine the exergy efficiency and to compare the effectiveness of SGB and palm methyl ester (PME) processes. It was found that the production of bio-ethanol is more thermodynamically sustainable than that of biodiesel as the net exergy value (NExV) of SGB is 10% higher than that of PME. Contrarily, the former has a net energy value (NEV) which is 9% lower than the latter. Despite this, SGB is still strongly recommended as a potential biofuel because SGB production can help mitigate several detrimental impacts on the environment.

Matched MeSH terms: Lignin/chemistry*
Hot compressed water pretreatment of oil palm fronds to enhance glucose recovery for production of second generation bio-ethanol

Goh CS, Lee KT, Bhatia S

Bioresour Technol, 2010 Oct;101(19):7362-7.
PMID: 20471249 DOI: 10.1016/j.biortech.2010.04.048

This work presents the pretreatment of oil palm fronds (OPF) using hot compressed water (HCW) to enhance sugar recovery in enzymatic hydrolysis. A central, composite rotatable design was used to optimize the effect of reaction temperature, reaction time and liquid-solid ratio on the pretreatment process. All variables were found to significantly affect the glucose yield. A quadratic polynomial equation was used to model glucose yield by multiple regression analysis, using response surface methodology (RSM). Using a 10 bar pressurized reactor, the optimum conditions for pretreatment of OPF were found at 178 degrees C, 11.1 min and a liquid-solid ratio of 9.6. The predicted glucose yield was 92.78 wt.% at the optimum conditions. Experimental verification of the optimum conditions gave a glucose yield in good agreement with the estimated value of the model.

Matched MeSH terms: Lignin/analysis
Influence of humic acids on radium adsorption by coir pith in aqueous solution

Zalina Laili, Muhamad Samudi Yasir, Muhamat Omar, Mohd Zaidi Ibrahim, Philip E

Sains Malaysiana, 2010;39:333-336.

This study examines the influence of humic acids (HA) on adsorption of radium (Ra) ions onto coir pith (CP) in aqueous solution. The adsorption behaviours of Ra ions onto CP under the influence of HA in aqueous solution were investigated in the series of batch mode adsorption experiments. The effects of various experimental conditions such as pH, contact time, adsorbent dosage and initial concentration of Ra ions have been studied. The results revealed that the presence of HA in aqueous solution enhanced the adsorption of Ra ions onto CP. The adsorption results showed that the percentage of Ra adsorbed was increased with an increase in the pH or alkalinity of aqueous solutions. Time dependence of the batch studies showed that a contact time of one day was sufficient to reach equilibrium. The result also showed that there was no significant difference on the effect of adsorbent dose on adsorption of radium onto CP. It was shown that the equilibrium data could be fitted by Freundlich equation.

Matched MeSH terms: Lignin
Kesan mangkin ke atas resin fenolik biopolimer dengan menggunakan kaedah pencecairan lignin soda

Sharifah Nabihah Syed Jaafar, Sarani Zakaria, Rozaidi Rasid, Nurul Ain Zulkifli, Ali Ahmadzadeh

Sains Malaysiana, 2010;39.

Proses pencecairan lignin soda telah dilakukan dengan menggunakan fenol dengan nisbah 1:3. Proses dilakukan selama 90 minit pada suhu 130oC dalam keadaan refluk. Hasil pencecairan iaitu pemfenolan lignin (PL) telah dianalisis dengan Spektrometer Inframerah Transformasi Fourier (FTIR), reometer dan analisis termogravimetri (TGA). Sampel yang disintesis dengan menggunakan asid hidroklorik sebagai mangkin memberikan keputusan yang terbaik. Analisis FTIR menunjukkan kehadiran kumpulan berfungsi yang signifikan seperti gelang aromatik, alkohol dan karbonil. Kesemua sampel PL mematuhi persamaan Arhenius dan bersifat pseudo-plastik. Peratus kehilangan berat sampel dan kadar penguraian sampel PL dipengaruhi oleh jenis mangkin yang digunakan. Sampel PL yang disintesis dikelaskan sebagai biopolimer resin fenolik.

Matched MeSH terms: Lignin
Lignin biodegradation and ligninolytic enzyme studies during biopulping of Acacia mangium wood chips by tropical white rot fungi

Liew CY, Husaini A, Hussain H, Muid S, Liew KC, Roslan HA

World J Microbiol Biotechnol, 2011 Jun;27(6):1457-68.
PMID: 25187145 DOI: 10.1007/s11274-010-0598-x

White rot fungi are good lignin degraders and have the potential to be used in industry. In the present work, Phellinus sp., Daedalea sp., Trametes versicolor and Pycnoporus coccineus were selected due to their relatively high ligninolytic enzyme activity, and grown on Acacia mangium wood chips under solid state fermentation. Results obtained showed that manganese peroxidase produced is far more compared to lignin peroxidase, suggesting that MnP might be the predominating enzymes causing lignin degradation in Acacia mangium wood chips. Cellulase enzyme assays showed that no significant cellulase activity was detected in the enzyme preparation of T. versicolor and Phellinus sp. This low cellulolytic activity further suggests that these two white rot strains are of more interest in lignin degradation. The results on lignin losses showed 20-30% of lignin breakdown at 60 days of biodegradation. The highest lignin loss was found in Acacia mangium biotreated with T. versicolor after 60 days and recorded 26.9%, corresponding to the percentage of their wood weight loss recorded followed by P. coccineus. In general, lignin degradation was only significant from 20 days onwards. The overall percentage of lignin weight loss was within the range of 1.02-26.90% over the biodegradation periods. Microscopic observations conducted using scanning electron microscope showed that T. versicolor, P. coccineus, Daedalea sp. and Phellinus sp. had caused lignin degradation in Acacia mangium wood chips.

Matched MeSH terms: Lignin
Dissolution and delignification of bamboo biomass using amino acid-based ionic liquid

Muhammad N, Man Z, Bustam MA, Mutalib MI, Wilfred CD, Rafiq S

Appl Biochem Biotechnol, 2011 Oct;165(3-4):998-1009.
PMID: 21720837 DOI: 10.1007/s12010-011-9315-y

In the present work, the dissolution of bamboo biomass was tested using a number of ionic liquids synthesized in laboratory. It was observed that one of the synthesized amino acid-based ionic liquids, namely 1-ethyl-3-methylimidazolium glycinate, was capable of dissolving the biomass completely. The dissolved biomass was then regenerated using a reconstitute solvent (acetone/water) and was characterized using Fourier transform infrared spectroscopy, X-ray diffraction, and scanning electron microscopy. The results were compared to preconditioned bamboo biomass. The regenerated biomass was found to have a more homogenous macrostructure, which indicates that the crystalline form and structure of its cellulose has changed from type Ι to type ΙΙ during the dissolution and regeneration process.

Matched MeSH terms: Lignin/metabolism*; Lignin/chemistry
Comparison of the carbon-sequestering abilities of pineapple leaf residue chars produced by controlled combustion and by field burning

Leng LY, Husni MH, Samsuri AW

Bioresour Technol, 2011 Nov;102(22):10759-62.
PMID: 21958525 DOI: 10.1016/j.biortech.2011.08.131

This study was undertaken to compare the chemical properties and yields of pineapple leaf residue (PLR) char produced by field burning (CF) with that produced by a partial combustion of air-dried PLR at 340 °C for 3 h in a furnace (CL). Higher total C, lignin content, and yield from CL as well as the presence of aromatic compounds in the Fourier Transform Infrared spectra of the char produced from CL suggest that the CL process was better in sequestering C than was the CF process. Although the C/N ratio of char produced from CL was low indicating a high N content of the char, the C in the char produced from CL was dominated by lignin suggesting that the decomposition of char produced from CL would be slow. To sequester C by char application, the PLR should be combusted in a controlled process rather than by burning in the field.

Matched MeSH terms: Lignin/analysis
Fulltext Expression profile of small RNAs in Acacia mangium secondary xylem tissue with contrasting lignin content - potential regulatory sequences in monolignol biosynthetic pathway

Ong SS, Wickneswari R

BMC Genomics, 2011 Nov 30;12 Suppl 3(Suppl 3):S13.
PMID: 22369296 DOI: 10.1186/1471-2164-12-S3-S13

BACKGROUND: Lignin, after cellulose, is the second most abundant biopolymer accounting for approximately 15-35% of the dry weight of wood. As an important component during wood formation, lignin is indispensable for plant structure and defense. However, it is an undesirable component in the pulp and paper industry. Removal of lignin from cellulose is costly and environmentally hazardous process. Tremendous efforts have been devoted to understand the role of enzymes and genes in controlling the amount and composition of lignin to be deposited in the cell wall. However, studies on the impact of downregulation and overexpression of monolignol biosynthesis genes in model species on lignin content, plant fitness and viability have been inconsistent. Recently, non-coding RNAs have been discovered to play an important role in regulating the entire monolignol biosynthesis pathway. As small RNAs have critical functions in various biological process during wood formation, small RNA profiling is an important tool for the identification of complete set of differentially expressed small RNAs between low lignin and high lignin secondary xylem.
RESULTS: In line with this, we have generated two small RNAs libraries from samples with contrasting lignin content using Illumina GAII sequencer. About 10 million sequence reads were obtained in secondary xylem of Am48 with high lignin content (41%) and a corresponding 14 million sequence reads were obtained in secondary xylem of Am54 with low lignin content (21%). Our results suggested that A. mangium small RNAs are composed of a set of 12 highly conserved miRNAs families found in plant miRNAs database, 82 novel miRNAs and a large proportion of non-conserved small RNAs with low expression levels. The predicted target genes of those differentially expressed conserved and non-conserved miRNAs include transcription factors associated with regulation of the lignin biosynthetic pathway genes. Some of these small RNAs play an important role in epigenetic silencing. Differential expression of the small RNAs between secondary xylem tissues with contrasting lignin content suggests that a cascade of miRNAs play an interconnected role in regulating the lignin biosynthetic pathway in Acacia species.
CONCLUSIONS: Our study critically demonstrated the roles of small RNAs during secondary wall formation. Comparison of the expression pattern of small RNAs between secondary xylem tissues with contrasting lignin content strongly indicated that small RNAs play a key regulatory role during lignin biosynthesis. Our analyses suggest an evolutionary mechanism for miRNA targets on the basis of the length of their 5' and 3' UTRs and their cellular roles. The results obtained can be used to better understand the roles of small RNAs during lignin biosynthesis and for the development of gene constructs for silencing of specific genes involved in monolignol biosynthesis with minimal effect on plant fitness and viability. For the first time, small RNAs were proven to play an important regulatory role during lignin biosynthesis in A. mangium.

Matched MeSH terms: Lignin/biosynthesis*; Lignin/chemistry
Fulltext Identification of lignin genes and regulatory sequences involved in secondary cell wall formation in Acacia auriculiformis and Acacia mangium via de novo transcriptome sequencing

Wong MM, Cannon CH, Wickneswari R

BMC Genomics, 2011;12:342.
PMID: 21729267 DOI: 10.1186/1471-2164-12-342

Acacia auriculiformis × Acacia mangium hybrids are commercially important trees for the timber and pulp industry in Southeast Asia. Increasing pulp yield while reducing pulping costs are major objectives of tree breeding programs. The general monolignol biosynthesis and secondary cell wall formation pathways are well-characterized but genes in these pathways are poorly characterized in Acacia hybrids. RNA-seq on short-read platforms is a rapid approach for obtaining comprehensive transcriptomic data and to discover informative sequence variants.

Matched MeSH terms: Lignin/biosynthesis; Lignin/metabolism*
Fulltext Overliming effects on xylitol production from sago trunk hydrolysate

Mohamad, N.L., Mustapa Kamal, S.M., Abdullah, A.G.L.

Pertanika Journal of Science & Technology, 2011;19(2):415-422.
MyJurnal

Xylitol can be obtained from lignocellulosic materials containing xylose. However, the fraction of lignocellulose converted through dilute acid hydrolysis contains compounds that inhibit the fermenting micro-organisms. These inhibitors can be removed from the hydrolysate by detoxification method, prior to fermentation. This study describes effectiveness of overliming process to reduce the toxicity of hydrolysates generated from pre-treatment of sago trunk for xylitol production. The overliming pH 9 and 10 was studied and the results showed that pH 9 was showed 20% of sugar loss, which is low compared to pH 10. Candida tropicalis strain was used to evaluate the fermentability of overlimed sago trunk hydrolysate at pH 9 and non-overlimed hydrolysate medium. Meanwhile, Xylitol accumulation and productivity in the overlimed medium was found to be higher than the non-treated medium. The maximum production of xylitol was increased up to 74% and converted within 76 h. The results obtained improved the fermentation process when compared with the nontreated medium.

Matched MeSH terms: Lignin

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