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Potential uses of spent mushroom substrate and its associated lignocellulosic enzymes

Phan CW, Sabaratnam V

Appl Microbiol Biotechnol, 2012 Nov;96(4):863-73.
PMID: 23053096 DOI: 10.1007/s00253-012-4446-9

Mushroom industries generate a virtually in-exhaustible supply of a co-product called spent mushroom substrate (SMS). This is the unutilised substrate and the mushroom mycelium left after harvesting of mushrooms. As the mushroom industry is steadily growing, the volume of SMS generated annually is increasing. In recent years, the mushroom industry has faced challenges in storing and disposing the SMS. The obvious solution is to explore new applications of SMS. There has been considerable discussion recently about the potentials of using SMS for production of value-added products. One of them is production of lignocellulosic enzymes such as laccase, xylanase, lignin peroxidase, cellulase and hemicellulase. This paper reviews scientific research and practical applications of SMS as a readily available and cheap source of enzymes for bioremediation, animal feed and energy feedstock.

Matched MeSH terms: Cellulose/metabolism
Fulltext Enhancement of Versatile Extracellular Cellulolytic and Hemicellulolytic Enzyme Productions by Lactobacillus plantarum RI 11 Isolated from Malaysian Food Using Renewable Natural Polymers

Mohamad Zabidi NA, Foo HL, Loh TC, Mohamad R, Abdul Rahim R

Molecules, 2020 Jun 03;25(11).
PMID: 32503356 DOI: 10.3390/molecules25112607

Lactobacillus plantarum RI 11 was reported recently to be a potential lignocellulosic biomass degrader since it has the capability of producing versatile extracellular cellulolytic and hemicellulolytic enzymes. Thus, this study was conducted to evaluate further the effects of various renewable natural polymers on the growth and production of extracellular cellulolytic and hemicellulolytic enzymes by this novel isolate. Basal medium supplemented with molasses and yeast extract produced the highest cell biomass (log 10.51 CFU/mL) and extracellular endoglucanase (11.70 µg/min/mg), exoglucanase (9.99 µg/min/mg), β-glucosidase (10.43 nmol/min/mg), and mannanase (8.03 µg/min/mg), respectively. Subsequently, a statistical optimization approach was employed for the enhancement of cell biomass, and cellulolytic and hemicellulolytic enzyme productions. Basal medium that supplemented with glucose, molasses and soybean pulp (F5 medium) or with rice straw, yeast extract and soybean pulp (F6 medium) produced the highest cell population of log 11.76 CFU/mL, respectively. However, formulated F12 medium supplemented with glucose, molasses and palm kernel cake enhanced extracellular endoglucanase (4 folds), exoglucanase (2.6 folds) and mannanase (2.6 folds) specific activities significantly, indicating that the F12 medium could induce the highest production of extracellular cellulolytic and hemicellulolytic enzymes concomitantly. In conclusion, L. plantarum RI 11 is a promising and versatile bio-transformation agent for lignocellulolytic biomass.

Matched MeSH terms: Cellulose/metabolism*
Draft genome sequence of Parvularcula flava strain NH6-79 T, revealing its role as a cellulolytic enzymes producer

Abdul Karim MH, Lam MQ, Chen SJ, Yahya A, Shahir S, Shamsir MS, et al.

Arch Microbiol, 2020 Nov;202(9):2591-2597.
PMID: 32607725 DOI: 10.1007/s00203-020-01967-z

To date, the genus Parvularcula consists of 6 species and no potential application of this genus was reported. Current study presents the genome sequence of Parvularcula flava strain NH6-79 T and its cellulolytic enzyme analysis. The assembled draft genome of strain NH6-79 T consists of 9 contigs and 7 scaffolds with 3.68 Mbp in size and GC content of 59.87%. From a total of 3,465 genes predicted, 96 of them are annotated as glycoside hydrolases (GHs). Within these GHs, 20 encoded genes are related to cellulosic biomass degradation, including 12 endoglucanases (5 GH10, 4 GH5, and 3 GH51), 2 exoglucanases (GH9) and 6 β-glucosidases (GH3). In addition, highest relative enzyme activities (endoglucanase, exoglucanase, and β-glucosidase) were observed at 27th hour when the strain was cultured in the carboxymethyl cellulose/Avicel®-containing medium for 45 h. The combination of genome analysis with experimental studies indicated the ability of strain NH6-79 T to produce extracellular endoglucanase, exoglucanase, and β-glucosidase. These findings suggest the potential of Parvularcula flava strain NH6-79 T in cellulose-containing biomass degradation and that the strain could be used in cellulosic biorefining process.

Matched MeSH terms: Cellulose/metabolism
Simultaneous production of cellulase and reducing sugar through modification of compositional and structural characteristic of sugarcane bagasse

Yoon LW, Ngoh GC, Chua AS

Enzyme Microb Technol, 2013 Sep 10;53(4):250-6.
PMID: 23931690 DOI: 10.1016/j.enzmictec.2013.05.005

This study examined the potential of untreated and alkali-pretreated sugarcane bagasse (SCB) in cellulase, reducing sugar (RS) and fungal biomass production via solid state fermentation (SSF) using Pycnoporus sanguineus. The impact of the composition, structure and cellulase adsorption ability of SCB on the production of cellulase, RS and fungal biomass was investigated. From the morphological and compositional analyses, untreated SCB has relatively more structural changes with a higher percentage of depolymerisation on the cellulose, hemicellulose and lignin content compared to alkali-pretreated SCB. Thus, untreated SCB favoured the production of cellulase and fungal biomass whereas alkali-pretreated SCB yielded a higher amount of RS. The composition and morphology of untreated SCB did not encourage RS production and this suggested that RS produced during SSF might be consumed in a faster rate by the more abundantly grown fungus. Besides that, alkali-pretreated SCB with higher cellulase adsorption ability could have adsorbed the cellulase produced and resulted in a lower cellulase titre. In short, the production of specific bioproducts via SSF is dependent on the structure and composition of the substrate applied.

Matched MeSH terms: Cellulose/metabolism*
A review of bacterial cellulose-based drug delivery systems: their biochemistry, current approaches and future prospects

Abeer MM, Mohd Amin MC, Martin C

J Pharm Pharmacol, 2014 Aug;66(8):1047-61.
PMID: 24628270 DOI: 10.1111/jphp.12234

The field of pharmaceutical technology is expanding rapidly because of the increasing number of drug delivery options. Successful drug delivery is influenced by multiple factors, one of which is the appropriate identification of materials for research and engineering of new drug delivery systems. Bacterial cellulose (BC) is one such biopolymer that fulfils the criteria for consideration as a drug delivery material.

Matched MeSH terms: Cellulose/metabolism*
Inhibition and kinetic studies of cellulose- and hemicellulose-degrading enzymes of Ganoderma boninense by naturally occurring phenolic compounds

Surendran A, Siddiqui Y, Ali NS, Manickam S

J Appl Microbiol, 2018 Jun;124(6):1544-1555.
PMID: 29405525 DOI: 10.1111/jam.13717

AIM: Ganoderma sp, the causal pathogen of the basal stem rot (BSR) disease of oil palm, secretes extracellular hydrolytic enzymes. These play an important role in the pathogenesis of BSR by nourishing the pathogen through the digestion of cellulose and hemicellulose of the host tissue. Active suppression of hydrolytic enzymes secreted by Ganoderma boninense by various naturally occurring phenolic compounds and estimation of their efficacy on pathogen suppression is focused in this study.
METHODS AND RESULTS: Ten naturally occurring phenolic compounds were assessed for their inhibitory effect on the hydrolytic enzymes of G. boninense. The enzyme kinetics (Vmax and Km ) and the stability of the hydrolytic enzymes were also characterized. The selected compounds had shown inhibitory effect at various concentrations. Two types of inhibitions namely uncompetitive and noncompetitive were observed in the presence of phenolic compounds. Among all the phenolic compounds tested, benzoic acid was the most effective compound suppressive to the growth and production of hydrolytic enzymes secreted by G. boninense. The phenolic compounds as inhibitory agents can be a better replacement for the metal ions which are known as conventional inhibitors till date. The three hydrolytic enzymes were stable in a wide range of pH and temperature.
CONCLUSION: These findings highlight the efficacy of the applications of phenolic compounds to control Ganoderma.
SIGNIFICANCE AND IMPACT OF THE STUDY: The study has proved a replacement for chemical controls of G. boninense with naturally occurring phenolic compounds.

Matched MeSH terms: Cellulose/metabolism*
Effect of feeding Pleurotus pulmonarius-treated empty fruit bunch on nutrient digestibility and milk fatty acid profiles in goats

Zailan MZ, Salleh SM, Abdullah S, Yaakub H

Trop Anim Health Prod, 2023 Nov 10;55(6):402.
PMID: 37950132 DOI: 10.1007/s11250-023-03817-8

This study aimed to evaluate the effect of feeding P. pulmonarius-treated empty fruit bunch (FTEFB) on the nutrient intakes, digestibility, milk yield and milk profiles of lactating Saanen goats. A total of nine lactating Saanen goats were used in an incomplete cross-over experimental design. The balanced dietary treatments contain different replacement levels of Napier grass with FTEFB at 0% (0-FT), 25% (25-FT) and 50% (50-FT). The FTEFB contained crude protein (CP), neutral detergent fibre (NDF), acid detergent fibre (ADF) and acid detergent lignin (ADL) at 4.10, 94.6, 70.8 and 19.4% DM, respectively. The replacement of FTEFB in 25-FT did not alter dry matter, NDF, hemicellulose, ADL, ether extract and gross energy intakes when compared to the control fed group (0-FT). The ADF and cellulose intake was higher in 25-FT than in the others (P 0.05). There are no differences in milk fatty profiles between dietary treatments (P > 0.05), except for OCFA. Goat fed with 25-FT had the lowest OCFA (P

Matched MeSH terms: Cellulose/metabolism
Fulltext Comparative Study of Extracellular Proteolytic, Cellulolytic, and Hemicellulolytic Enzyme Activities and Biotransformation of Palm Kernel Cake Biomass by Lactic Acid Bacteria Isolated from Malaysian Foods

Lee FH, Wan SY, Foo HL, Loh TC, Mohamad R, Abdul Rahim R, et al.

Int J Mol Sci, 2019 Oct 09;20(20).
PMID: 31600952 DOI: 10.3390/ijms20204979

Biotransformation via solid state fermentation (SSF) mediated by microorganisms is a promising approach to produce useful products from agricultural biomass. Lactic acid bacteria (LAB) that are commonly found in fermented foods have been shown to exhibit extracellular proteolytic, β-glucosidase, β-mannosidase, and β-mannanase activities. Therefore, extracellular proteolytic, cellulolytic, and hemicellulolytic enzyme activities of seven Lactobacillus plantarum strains (a prominent species of LAB) isolated from Malaysian foods were compared in this study. The biotransformation of palm kernel cake (PKC) biomass mediated by selected L. plantarum strains was subsequently conducted. The results obtained in this study exhibited the studied L. plantarum strains produced versatile multi extracellular hydrolytic enzyme activities that were active from acidic to alkaline pH conditions. The highest total score of extracellular hydrolytic enzyme activities were recorded by L. plantarum RI11, L. plantarum RG11, and L. plantarum RG14. Therefore, they were selected for the subsequent biotransformation of PKC biomass via SSF. The hydrolytic enzyme activities of treated PKC extract were compared for each sampling interval. The scanning electron microscopy analyses revealed the formation of extracellular matrices around L. plantarum strains attached to the surface of PKC biomass during SSF, inferring that the investigated L. plantarum strains have the capability to grow on PKC biomass and perform synergistic secretions of various extracellular proteolytic, cellulolytic, and hemicellulolytic enzymes that were essential for the effective biodegradation of PKC. The substantial growth of selected L. plamtraum strains on PKC during SSF revealed the promising application of selected L. plantarum strains as a biotransformation agent for cellulosic biomass.

Matched MeSH terms: Cellulose/metabolism*
Bioconversion of oil palm frond by Aspergillus niger to enhances it's fermentable sugar production

Lim SH, Ibrahim D

Pak J Biol Sci, 2013 Sep 15;16(18):920-6.
PMID: 24502148

The aim of this study was to develop an economical bioprocess to produce the fermentable sugars at laboratory scales Using Oil Palm Frond (OPF) as substrate in Solid State Fermentation (SSF). OPF waste generated by oil palm plantations is a major problem in terms of waste management. However, this lignocellulosic waste material is a cheap source of cellulose. We used OPF as substrate to produce fermentable sugars. The high content of cellulose in OPF promises the high fermentable sugars production in SSF. Saccharification of OPF waste by A. niger USMAI1 generates fermentable sugars and was evaluated through a solid state fermentation. Physical parameters, e.g., inoculum size, initial substrate moisture, initial pH, incubation temperature and the size of substrate were optimized to obtain the maximum fermentable sugars from oil palm fronds. Up to 77 mg of fermentable sugars per gram substrate was produced under the optimal physical parameter conditions. Lower productivity of fermentable sugars, 32 mg fermentable sugars per gram substrate was obtained under non optimized conditions. The results indicated that about 140.6% increase in fermentable sugar production after optimization of the physical parameters. Glucose was the major end component amongst the fermentable sugars obtained. This study indicated that under optimum physical parameter conditions, the OPF waste can be utilized to produce fermentable sugars which then convert into other products such as alcohol.

Matched MeSH terms: Cellulose/metabolism
Investigating process parameters to enhance (hemi)cellulolytic enzymes activity produced by Trichoderma reesei RUT-C30 using deoiled oil palm mesocarp fiber in solid-state fermentation

Al-Qassab AA, Zakaria MR, Yunus R, Salleh MAM, Mokhtar MN

Int J Biol Macromol, 2024 Sep;276(Pt 2):134030.
PMID: 39038578 DOI: 10.1016/j.ijbiomac.2024.134030

This study investigates the synthesis of (hemi)cellulolytic enzymes, including endoglucanase (CMCase), xylanase, and β-glucosidase, employing Trichoderma reesei RUT-C30 and deoiled oil palm mesocarp fiber (OPMF) through solid-state fermentation (SSF). The objective was to determine the optimal process conditions for achieving high enzyme activities through a one-factor-at-a-time approach. The study primarily focused on the impact of the solid-to-liquid ratio, incubation period, initial pH, and temperature on enzyme activity. The effects of OPMF pretreatment, particularly deoiling and fortification, were explored. This approach significantly improved enzyme activity levels compared to the initial conditions, with CMCase increasing by 111.6 %, xylanase by 665.2 %, and β-Glucosidase by 1678.1 %. Xylanase and β-glucosidase activities, peaking at 1346.75 and 9.89 IU per gram dry substrate (GDS), respectively, under optimized conditions (1:4 ratio, pH 7.5, 20 °C, 9-day incubation). With lower moisture levels, CMCase reached its maximum activity of 227.84 IU/GDS. The study highlights how important it is for agro-industrial byproducts to support environmentally sustainable practices in the palm oil industry. It also emphasizes how differently each enzyme reacts to changes in process parameters.

Matched MeSH terms: Cellulose/metabolism
Inhibition by toxic compounds in the hemicellulosic hydrolysates on the activity of xylose reductase from Candida tropicalis

Rafiqul IS, Sakinah AM, Zularisam AW

Biotechnol Lett, 2015 Jan;37(1):191-6.
PMID: 25214231 DOI: 10.1007/s10529-014-1672-5

Xylose reductase (XR) is an oxidoreductase having potential applications in the production of various specialty products, mainly xylitol. It is important to screen for compounds that can decrease XR activity and consequently can decrease xylitol production. We have identified the byproducts in the hemicellulosic hydrolysate that inhibit XR from Candida tropicalis and measured their effects. XR inhibitory activities of byproducts, glucose, acetic acid, arabinose, lignin-degradation products (LDPs), furfural and hydroxymethylfurfural (HMF), were evaluated by measuring the MIC and IC50 values. XR activity was 11.2 U/ml. Acetic acid, LDPs, furfural and HMF significantly inhibited XR with IC50 values of 11, 6.4, 2.3 and 0.4 g/l, respectively. This is the first report on the inhibitory activities of several byproducts for XR.

Matched MeSH terms: Cellulose/metabolism*
Isolation, screening, and identification of potential cellulolytic and xylanolytic producers for biodegradation of untreated oil palm trunk and its application in saccharification of lemongrass leaves

Ang SK, Yahya A, Abd Aziz S, Md Salleh M

Prep Biochem Biotechnol, 2015;45(3):279-305.
PMID: 24960316 DOI: 10.1080/10826068.2014.923443

This study presents the isolation and screening of fungi with excellent ability to degrade untreated oil palm trunk (OPT) in a solid-state fermentation system (SSF). Qualitative assay of cellulases and xylanase indicates notable secretion of both enzymes by 12 fungal strains from a laboratory collection and 5 strains isolated from a contaminated wooden board. High production of these enzymes was subsequently quantified in OPT in SSF. Aspergillus fumigates SK1 isolated from cow dung gives the highest xylanolytic activity (648.448 U g(-1)), generally high cellulolytic activities (CMCase: 48.006, FPase: 6.860, beta-glucosidase: 16.328 U g(-1)) and moderate lignin peroxidase activity (4.820 U/g), and highest xylanolytic activity. The xylanase encoding gene of Aspergillus fumigates SK1 was screened using polymerase chain reaction by a pair of degenerate primers. Through multiple alignment of the SK1 strain's xylanase nucleotide sequences with other published xylanases, it was confirmed that the gene belonged to the xylanase glycoside hydrolase family 11 (GH11) with a protein size of 24.49 kD. Saccharification of lemongrass leaves using crude cellulases and xylanase gives the maximum reducing sugars production of 6.84 g/L with glucose as the major end product and traces of phenylpropanic compounds (vanillic acid, p-coumaric acid, and ferulic acid).

Matched MeSH terms: Cellulose/metabolism*
Cellulose acetate phthalate microencapsulation and delivery of plasmid DNA to the intestines

Hanafi A, Nograles N, Abdullah S, Shamsudin MN, Rosli R

J Pharm Sci, 2013 Feb;102(2):617-26.
PMID: 23192729 DOI: 10.1002/jps.23389

Cellulose acetate phthalate (CAP) microcapsules were formulated to deliver plasmid DNA (pDNA) to the intestines. The microcapsules were characterized and were found to have an average diameter of 44.33 ± 30.22 μm, and were observed to be spherical with smooth surface. The method to extract pDNA from CAP was modified to study the release profile of the pDNA. The encapsulated pDNA was found to be stable. Exposure to the acidic and basic pH conditions, which simulates the pH environment in the stomach and the intestines, showed that the release occurred in a stable manner in the former, whereas it was robust in the latter. The loading capacity and encapsulation efficiency of the microcapsules were low but the CAP recovery yield was high which indicates that the microcapsules were efficiently formed but the loading of pDNA can be improved. In vitro transfection study in 293FT cells showed that there was a significant percentage of green-fluorescent-protein-positive cells as a result of efficient transfection from CAP-encapsulated pDNA. Biodistribution studies in BALB/c mice indicate that DNA was released at the stomach and intestinal regions. CAP microcapsules loaded with pDNA, as described in this study, may be useful for potential gene delivery to the intestines for prophylactic or therapeutic measures for gastrointestinal diseases.

Matched MeSH terms: Cellulose/metabolism
Solid acid catalysts pretreatment and enzymatic hydrolysis of macroalgae cellulosic residue for the production of bioethanol

Tan IS, Lee KT

Carbohydr Polym, 2015 Jun 25;124:311-21.
PMID: 25839825 DOI: 10.1016/j.carbpol.2015.02.046

The aim of this study is to investigate the technical feasibility of converting macroalgae cellulosic residue (MCR) into bioethanol. An attempt was made to present a novel, environmental friendly and economical pretreatment process that enhances enzymatic conversion of MCR to sugars using Dowex (TM) Dr-G8 as catalyst. The optimum yield of glucose reached 99.8% under the optimal condition for solid acid pretreatment (10%, w/v biomass loading, 4%, w/v catalyst loading, 30min, 120°C) followed by enzymatic hydrolysis (45FPU/g of cellulase, 52CBU/g of β-glucosidase, 50°C, pH 4.8, 30h). The yield of sugar obtained was found more superior than conventional pretreatment process using H2SO4 and NaOH. Biomass loading for the subsequent simultaneous saccharification and fermentation (SSF) of the pretreated MCR was then optimized, giving an optimum bioethanol yield of 81.5%. The catalyst was separated and reused for six times, with only a slight drop in glucose yield.

Matched MeSH terms: Cellulose/metabolism*
Fulltext Microbial diversity of thermophiles with biomass deconstruction potential in a foliage-rich hot spring

Lee LS, Goh KM, Chan CS, Annie Tan GY, Yin WF, Chong CS, et al.

Microbiologyopen, 2018 12;7(6):e00615.
PMID: 29602271 DOI: 10.1002/mbo3.615

The ability of thermophilic microorganisms and their enzymes to decompose biomass have attracted attention due to their quick reaction time, thermostability, and decreased risk of contamination. Exploitation of efficient thermostable glycoside hydrolases (GHs) could accelerate the industrialization of biofuels and biochemicals. However, the full spectrum of thermophiles and their enzymes that are important for biomass degradation at high temperatures have not yet been thoroughly studied. We examined a Malaysian Y-shaped Sungai Klah hot spring located within a wooded area. The fallen foliage that formed a thick layer of biomass bed under the heated water of the Y-shaped Sungai Klah hot spring was an ideal environment for the discovery and analysis of microbial biomass decay communities. We sequenced the hypervariable regions of bacterial and archaeal 16S rRNA genes using total community DNA extracted from the hot spring. Data suggested that 25 phyla, 58 classes, 110 orders, 171 families, and 328 genera inhabited this hot spring. Among the detected genera, members of Acidimicrobium, Aeropyrum, Caldilinea, Caldisphaera, Chloracidobacterium, Chloroflexus, Desulfurobacterium, Fervidobacterium, Geobacillus, Meiothermus, Melioribacter, Methanothermococcus, Methanotorris, Roseiflexus, Thermoanaerobacter, Thermoanaerobacterium, Thermoanaerobaculum, and Thermosipho were the main thermophiles containing various GHs that play an important role in cellulose and hemicellulose breakdown. Collectively, the results suggest that the microbial community in this hot spring represents a good source for isolating efficient biomass degrading thermophiles and thermozymes.

Matched MeSH terms: Cellulose/metabolism
Sulfated membrane adsorbers for economic pseudo-affinity capture of influenza virus particles

Opitz L, Lehmann S, Reichl U, Wolff MW

Biotechnol Bioeng, 2009 Aug 15;103(6):1144-54.
PMID: 19449393 DOI: 10.1002/bit.22345

Strategies to control outbreaks of influenza, a contagious respiratory tract disease, are focused mainly on prophylactic vaccinations in conjunction with antiviral medications. Currently, several mammalian cell culture-based influenza vaccine production processes are being established, such as the technologies introduced by Novartis Behring (Optaflu) or Baxter International Inc. (Celvapan). Downstream processing of influenza virus vaccines from cell culture supernatant can be performed by adsorbing virions onto sulfated column chromatography beads, such as Cellufine sulfate. This study focused on the development of a sulfated cellulose membrane (SCM) chromatography unit operation to capture cell culture-derived influenza viruses. The advantages of the novel method were demonstrated for the Madin Darby canine kidney (MDCK) cell-derived influenza virus A/Puerto Rico/8/34 (H1N1). Furthermore, the SCM-adsorbers were compared directly to column-based Cellufine sulfate and commercially available cation-exchange membrane adsorbers. Sulfated cellulose membrane adsorbers showed high viral product recoveries. In addition, the SCM-capture step resulted in a higher reduction of dsDNA compared to the tested cation-exchange membrane adsorbers. The productivity of the SCM-based unit operation could be significantly improved by a 30-fold increase in volumetric flow rate during adsorption compared to the bead-based capture method. The higher flow rate even further reduced the level of contaminating dsDNA by about twofold. The reproducibility and general applicability of the developed unit operation were demonstrated for two further MDCK cell-derived influenza virus strains: A/Wisconsin/67/2005 (H3N2) and B/Malaysia/2506/2004. Overall, SCM-adsorbers represent a powerful and economically favorable alternative for influenza virus capture over conventional methods using Cellufine sulfate.

Matched MeSH terms: Cellulose/metabolism
Identification patterns of Trichoderma strains using morphological characteristics, phylogenetic analyses and lignocellulolytic activities

Asis A, Shahriar SA, Naher L, Saallah S, Fatihah HNN, Kumar V, et al.

Mol Biol Rep, 2021 Apr;48(4):3285-3301.
PMID: 33880673 DOI: 10.1007/s11033-021-06321-0

Trichoderma is a genus of soil-borne fungus with an abundance of reports of its economic importance in the agriculture industry. Thus, the correct identification of Trichoderma species is necessary for its commercial purposes. Globally, Trichoderma species are routinely identified from micro-morphological descriptions which can be tedious and prone to errors. Thus, we emphasize that the accurate identification of Trichoderma strains requires a three-pronged approach i.e. based on its morphological characteristics, multilocus gene sequences of the rDNA [internal transcribed spacer (ITS) 1 and 2 regions], translation elongation factor 1-α (TEF-1α), Calmodulin (CAL) and its lignocellulolytic activities. We used this approach to identify a total of 53 Trichoderma strains which were isolated from a wet paddy field located at Tuaran, Sabah, Malaysia. The 53 strains were positively identified as belonging to three Trichoderma species, namely T. asperellum (43 strains), T. harzianum (9 strains), and T. reesei (one strain) on the basis of its morphological characteristics and multilocus gene sequences. Phylogenetic trees constructed based on the UPGMA method of the ITS 1 and 2 regions of the rDNA, TEF-1α and CAL revealed three distinct groups with the T. asperellum, T. harzianum and T. reesei strains placed under the section of Trichoderma, Pachybasium and Longibrachiatum, respectively. In addition, the lignocellulolytic activities of the isolates were measured based on the diameters of the halo zones produced when degrading cellulose, lignin, and starch, respectively. This diagnostic assay can be used to identify Trichoderma as it produces polyphenol oxidase when Tannic Acid Media is used for the lignin test, endoglucanases when Jensen media is used for cellulose, and it hydrolyzes starch to glucose when the modified Melin-Nokrans media is used for the starch test. Accurate identification of Trichoderma species is needed as these strains can potentially be used as a biocontrol agent to prevent diseases and to increase yield in agriculture crops.

Matched MeSH terms: Cellulose/metabolism
Fulltext Prospecting Agro-waste Cocktail: Supplementation for Cellulase Production by a Newly Isolated Thermophilic B. licheniformis 2D55

Kazeem MO, Shah UKM, Baharuddin AS, AbdulRahman NA

Appl Biochem Biotechnol, 2017 Aug;182(4):1318-1340.
PMID: 28176140 DOI: 10.1007/s12010-017-2401-z

Bacteria isolated from thermophilic environment that can produce cellulase as well as utilise agro-waste biomass have a high potential for developing thermostable cellulase required in the biofuel industry. The cost for cellulase represents a significant challenge in converting lignocellulose to fermentable sugars for biofuel production. Among three potential bacteria examined, Bacillus licheniformis 2D55 (accession no. KT799651) was found to produce the highest cellulolytic activity (CMCase 0.33 U/mL and FPase 0.09 U/mL) at 18-24 h fermentation when grown on microcrystalline cellulose (MCC) as a carbon source in shake flask at 50 °C. Cellulase production process was further conducted on the untreated and NaOH pretreated rice straw (RS), rice husk (RH), sugarcane bagasse (BAG) and empty fruit bunch (EFB). Untreated BAG produced the highest FPase (0.160 U/mL), while the highest CMCase (0.150 U/mL) was supported on the pretreated RH. The mixture of untreated BAG and pretreated RH as agro-waste cocktail has remarkably improved CMCase (3.7- and 1.4-fold) and FPase (2.5- and 11.5-fold) compared to the untreated BAG and pretreated RH, respectively. The mechanism of cellulase production explored through SEM analysis and the location of cellulase enzymes of the isolate was also presented. Agro-waste cocktail supplementation provides an alternative method for an efficient production of cellulase.

Matched MeSH terms: Cellulose/metabolism
Fulltext Hemocyanin facilitates lignocellulose digestion by wood-boring marine crustaceans

Besser K, Malyon GP, Eborall WS, Paro da Cunha G, Filgueiras JG, Dowle A, et al.

Nat Commun, 2018 12 03;9(1):5125.
PMID: 30510200 DOI: 10.1038/s41467-018-07575-2

Woody (lignocellulosic) plant biomass is an abundant renewable feedstock, rich in polysaccharides that are bound into an insoluble fiber composite with lignin. Marine crustacean woodborers of the genus Limnoria are among the few animals that can survive on a diet of this recalcitrant material without relying on gut resident microbiota. Analysis of fecal pellets revealed that Limnoria targets hexose-containing polysaccharides (mainly cellulose, and also glucomannans), corresponding with the abundance of cellulases in their digestive system, but xylans and lignin are largely unconsumed. We show that the limnoriid respiratory protein, hemocyanin, is abundant in the hindgut where wood is digested, that incubation of wood with hemocyanin markedly enhances its digestibility by cellulases, and that it modifies lignin. We propose that this activity of hemocyanins is instrumental to the ability of Limnoria to feed on wood in the absence of gut symbionts. These findings may hold potential for innovations in lignocellulose biorefining.

Matched MeSH terms: Cellulose/metabolism
Comprehensive studies on optimization of cellulase and xylanase production by a local indigenous fungus strain via solid state fermentation using oil palm frond as substrate

Tai WY, Tan JS, Lim V, Lee CK

Biotechnol Prog, 2019 05;35(3):e2781.
PMID: 30701709 DOI: 10.1002/btpr.2781

The high cost of cellulases remains the most significant barrier to the economical production of bio-ethanol from lignocellulosic biomass. The goal of this study was to optimize cellulases and xylanase production by a local indigenous fungus strain (Aspergillus niger DWA8) using agricultural waste (oil palm frond [OPF]) as substrate. The enzyme production profile before optimization indicated that the highest carboxymethyl cellulose (CMCase), filter paper (FPase), and xylanase activities of 1.06 U/g, 2.55 U/g, and 2.93 U/g were obtained on day 5, day 4, and day 5 of fermentation, respectively. Response surface methodology was used to study the effects of several key process parameters in order to optimize cellulase production. Of the five physical and two chemical factors tested, only moisture content of 75% (w/w) and substrate amount of 2.5 g had statistically significant effect on enzymes production. Under optimized conditions of 2.5 g of substrate, 75% (w/w) moisture content, initial medium of pH 4.5, 1 × 106 spores/mL of inoculum, and incubation at ambient temperature (±30°C) without additional carbon and nitrogen, the highest CMCase, FPase, and xylanase activities obtained were 2.38 U/g, 2.47 U/g, and 5.23 U/g, respectively. Thus, the optimization process increased CMCase and xylanase production by 124.5 and 78.5%, respectively. Moreover, A. niger DWA8 produced reasonably good cellulase and xylanase titers using OPF as the substrate when compared with previous researcher finding. The enzymes produced by this process could be further use to hydrolyze biomass to generate reducing sugars, which are the feedstock for bioethanol production.

Matched MeSH terms: Cellulose/metabolism

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