Talaromyces Thermophilus Research Articles

Characterization of Thermophilic Lignocellulolytic Microorganisms in Composting.

Composting involves the selection of a microbiota capable of resisting the high temperatures generated during the process and degrading the lignocellulose. A deep understanding of the thermophilic microbial community involved in such biotransformation is valuable to improve composting efficiency and to provide thermostable biomass-degrading enzymes for biorefinery. This study investigated the lignocellulose-degrading thermophilic microbial culturome at all the stages of plant waste composting, focusing on the dynamics, enzymes, and thermotolerance of each member of such a community. The results revealed that 58% of holocellulose (cellulose plus hemicellulose) and 7% of lignin were degraded at the end of composting. The whole fungal thermophilic population exhibited lignocellulose-degrading activity, whereas roughly 8–10% of thermophilic bacteria had this trait, although exclusively for hemicellulose degradation (xylan-degrading). Because of the prevalence of both groups, their enzymatic activity, and the wide spectrum of thermotolerance, they play a key role in the breakdown of hemicellulose during the entire process, whereas the degradation of cellulose and lignin is restricted to the activity of a few thermophilic fungi that persists at the end of the process. The xylanolytic bacterial isolates (159 strains) included mostly members of Firmicutes (96%) as well as a few representatives of Actinobacteria (2%) and Proteobacteria (2%). The most prevalent species were Bacillus licheniformis and Aeribacillus pallidus. Thermophilic fungi (27 strains) comprised only four species, namely Thermomyces lanuginosus, Talaromyces thermophilus, Aspergillus fumigatus, and Gibellulopsis nigrescens, of whom A. fumigatus and T. lanuginosus dominated. Several strains of the same species evolved distinctly at the stages of composting showing phenotypes with different thermotolerance and new enzyme expression, even not previously described for the species, as a response to the changing composting environment. Strains of Bacillus thermoamylovorans, Geobacillus thermodenitrificans, T. lanuginosus, and A. fumigatus exhibiting considerable enzyme activities were selected as potential candidates for the production of thermozymes. This study lays a foundation to further investigate the mechanisms of adaptation and acquisition of new traits among thermophilic lignocellulolytic microorganisms during composting as well as their potential utility in biotechnological processing.

An invasive infection caused by the thermophilic mold Talaromyces thermophilus

BackgroundIncreasing incidence of invasive infections caused by rare fungi was observed over the recent years.CaseHere, we describe the first reported case of an infection caused by the thermophilic mold Talaromyces thermophilus. Cultivation and, hence, identification of this fastidious organism is challenging since standard incubation conditions are not sufficient. Retrospective analysis of patient samples and in vitro experiments demonstrated that testing for fungal antigens, i.e., the cell wall components galactomannan and β-1,3-d-glucan, is a promising tool.

Fungal community assembly during a high-temperature composting under different pasteurization regimes used to elaborate the Agaricus bisporus substrate

Agaricus bisporus cultivation is based on a selective substrate prepared by a meticulous composting process where thermophilic and/or thermotolerant fungi might play an important role in straw biomass depolymerization. Since fungi have physiological limitations to survive and grow in high-temperature environments, we set out different pasteurization regimes (57 °C/6 h, 60 °C/2 h, and 68 °C/2 h) to evaluate the impact on the fungal community assembly. The fungal community profile generated by high-throughput sequencing showed shifts in community diversity and composition under different pasteurization regimes. Most of the recovered sequences belong to the Ascomycota phylum. Among 73 species detected, Mycothermus thermophilus, Talaromyces thermophilus, and Thermomyces lanuginosus were the most abundant. In the current study, we outlined that pasteurization regimes can reshape the fungal community in compost which can potentially impact the A. bisporus development.

Statistical optimization of cellulases by Talaromyces thermophilus utilizing Saccharum spontaneum, a novel substrate

Abstract Background At present, cellulases are the most important enzymes worldwide, and their demand has been increasing in the industrial sector owing to their notable hydrolysis capability. Results In the present study, contrary to conventional techniques, three physical parameters were statistically optimized for the production of cellulase by thermophilic fungi by using response surface methodology (RSM). Among all the tested thermophilic strains, the best cellulase producing fungus was identified as Talaromyces thermophilus – both morphologically and molecularly through 5.8S/ITS rDNA sequencing. The central composite design (CCD) was used to evaluate the interactive effect of the significant factors. The CCD was applied by considering incubation period, pH, and temperature as the model factors for the present investigation. A second-order quadratic model and response surface method revealed that the independent variables including pH 6, temperature 50 °C, and incubation period 72 h significantly influenced the production of cellulases. The analysis of variance (ANOVA) indicated that the established model was significant (P ≤ 0.05) and showed the high adequacy of the model. The actual and predicted values of CMCase and FPase activity showed good agreement with each other and also confirmed the validity of the designed model. Conclusions We believe the present findings to be the first report on cellulase production by exploiting Kans grass (Saccharum spontaneum) as a substrate through response surface methodology by using thermophilic fungus, Talaromyces thermophilus. How to cite: Abdullah R, Tahseen M, Nisar K et al. Statistical optimization of cellulases by Talaromyces thermophilus utilizing Saccharum spontaneum, a novel substrate. Electron J Biotechnol 2021;51. https://doi.org/10.1016/j.ejbt.2021.03.007.

High-level expression and characterization of the thermostable leucine aminopeptidase Thelap from the thermophilic fungus Thermomyces lanuginosus in Aspergillus niger and its application in soy protein hydrolysis

Leucine aminopeptidase (LAP), an exopeptidase that releases amino acid residues, especially leucine, from the N-terminus of polypeptides, is often applied to debitter protein hydrolysate in the food industry. However, there are no thermostable and high activity enzymes that can be used in the food industry. In this study, we obtained the highly active and thermostable leucine aminopeptidases screened from the thermophilic fungi Thermomyces lanuginosus, Talaromyces thermophilus, and Malbranchea cinnamomea. The activity of the recombinant leucine aminopeptidase Thelap was significantly increased to 2771.5 U/mL, as mediated by the CRISPR/Cas9 tool. The recombinant Thelap was easily purified from fermentation broth by Ni-affinity chromatography, and the specific activity of the purified Thelap was increased to 7449.6 U/mg. The recombinant Thelap showed optimal activity at pH 8.5 and 75 °C and remained above 70% of the maximum activity over a wide temperature range (30–80 °C). With regard to temperature stability, Thelap retained more than 90% activity when it was incubated at 65–75 °C for 2 h. K+ and Co2+ increased the enzyme activity of the recombinant Thelap, while Ba2+, Mn2+, Ni2+, Ca2+, Mg2+ and SDS inhibited its enzyme activity, and the inhibition capacity of Mg2+ was the weakest. Upon application in soy protein hydrolysis, Thelap could significantly increase the degree of hydrolysis and remove more hydrophobic amino acids from the N-terminal region of the polypeptide to decrease the bitterness.

Agricultural wastes as substrates for β-glucosidase production by Talaromyces thermophilus: Role of these enzymes in enhancing waste paper saccharification

ABSTRACTIn the present study, we investigated a potent extracellular β-glucosidases secreted by the thermophilic fungal strain AX4 of Talaromyces thermophilus, isolated from Tunisian soil samples. This strain was selected referring to the highest thermostability of its β-glucosidases compared to the other fungal isolates. The β-glucosidase production was investigated by submerged fermentation. The optimal temperature and initial pH for maximum β-glucosidase production were 50°C and 7.0, respectively. Several carbon sources were assayed for their effects on β-glucosidase production, significant yields were obtained in media containing lactose 1% (3.0 ± 0.36 U/ml) and wheat bran 2% (4.0 ± 0.4 U/ml). The combination of wheat bran at 2% and lactose at 0.8% as carbon source enhanced β-glucosidase production, which reached 8.5 ± 0.28 U/ml. Furthermore, the β-glucosidase-rich enzymatic juice of T. thermophilus exhibited significant synergism with Trichoderma reesei (Rut C30) cellulases for pretreated waste paper (PWP) hydrolysis. Interestingly, the use of this optimal enzymatic cocktail increased 4.23 fold the glucose yield after saccharification of waste paper. A maximum sugar yield (94%) was reached when using low substrate (2%) and enzyme loading (EC1).

Physicochemical properties of thermotolerant extracellular β-glucosidase from Talaromyces thermophilus and enzymatic synthesis of cello-oligosaccharides

A thermophilic fungus, Talaromyces thermophilus that produces a novel thermotolerant extra-cellular β-glucosidase (Bgl.tls), was isolated from Tunisian soil samples. The enzyme was purified from the culture filtrates of T. thermophilus grown on lactose using gel filtration, ion exchange chromatography and FPLC. The monomeric enzyme had a molecular mass of 116.0 kDa and a high specific activity of 1429 UI/mg. Bgl.tls exhibited optimal activity at pH 5.0 and 65 °C. It was also stable over a wide range of pH (4.0–10.0) and stable at 50 °C for 34 h. Bgl.tls retained about 80% of its initial activity after 1.0 hours of preincubation at 60 °C. The Km and Vmax values recorded for pNPG were 0.25 mM and 228.7 µmol min−1, respectively. Bgl.tls was activated by Mn2+, Mg2+, Ca2+ and Co2+ but obviously inhibited by Fe2+ and Cu2+. It was able to hydrolyze a variety of aryl / alkyl -β-glucosides and disaccharides as well as (1→6) and (1→4)-β-glucosidic linkages and α-glycosidic substrates, thus providing evidence for its broad substrate specificity. The enzyme also displayed high hydrolytic and transglycosylation activities. Overall, this study is the first report on the purification and physicochemical properties of a β-glucosidase secreted by T. thermophilus. The cello-oligosaccharides synthesized by this enzyme within 2 h were mainly cellotriose, cellotetraose and cellopentaose identified by HPLC and ESI-MS techniques.

Expression of a thermo-alkaline lipase gene from Talaromyces thermophilus in recombinant Pichia pastoris

Expression of a thermo-alkaline lipase gene from Talaromyces thermophilus in Pichia pastoris was researched to enhance its production. The lipase gene (TTL) was genetically optimized and inserted into the downstream of AOX1 promoter and α-factor to construct recombinant plasmid pPIC9K-TTL, which was then transformed into P. pastoris via electroporation, producing 220 positive transformants. Stable integration of lipase gene into chromosomal DNA of transformants was confirmed through PCR analysis. Lipase production was performed at lab scale, and an obvious protein band about 39kDa was detected using SDS-PAGE, which suggested lipase gene in recombinant P. pastoris was successfully extracellularly expressed. This lipase worked efficiently at pH range from 8.9 to 10.5, and showed the maximum activity at pH 9.5. After treating at pH 11 for one hour, 75% of its activity could be remained. It was active above 40°C up to 70°C, and the optimal temperature for reaction was 60°C. High thermostability was observed, and more than 70% activity could be kept after one hour treatment at temperatures up to 80°C. Lipase activity was promoted by Ca2+ and inhibited by Zn2+ and Cu2+. This research showed a bright prospect for industrial application of thermo- and alkaline-stable lipase.

Heterologous Expression of an Alkali and Thermotolerant Lipase from Talaromyces thermophilus in Trichoderma reesei.

To heterologously express a Talaromyces thermophilus lipase gene in Trichoderma reesei, an efficient binary vector pChph-pCBH1sigpro-ttl which includes a newly designed cbh1 promoter and hygromycin-resistant marker was constructed. This plasmid was then transformed into T. reesei via improved Agrobacterium EHA 105-mediated transformation. After modification of co-culture conditions and enzymolysis treatment of conidia, 258 transformants were produced. A two-step screening method based on antibiotic resistance and capacity to utilize lactose and tributyrin was introduced to further select promising candidates, which would be additionally verified by PCR analysis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and lipase activity assay. Lipase production was carried out in shaking flasks, and the activity reached 241 IU/mL (7415.4 IU/mg) after 84-h fermentation. It was found that this lipase performed high alkali and thermostable tolerance with the optimal pH 9.5 and temperature 60 °C, and it could retain more than 70 % activity after being disposed in pH 11 or 70 °C for 1 h. This study herein would benefit the genetic engineering of T. reesei and the industrial application of this important fungal lipase.

Characterization of Thermolide Biosynthetic Genes and a New Thermolide from Sister Thermophilic Fungi

Prior chemical analysis of obligate thermophilic fungus Talaromyces thermophilus led to the discovery of thermolides A-F, six previously undescribed members of the lactam-bearing macrolactone class. A combination of chemical screening, genome analyses, and genetic manipulation led to the identification of the thermolide biosynthetic genes from sister thermophilic fungi T. thermophilus and Thermomyces lanuginosus and a new thermolide. The biosynthetic locus for the thermolides' mixed polyketide/amino acid structure encodes a hybrid polyketide synthase-nonribosomal peptide synthetase (PKS-NRPS). Our results reveal the first fungal hybrid iterative PKS-NRPS genes involved in the biosynthesis of bacterial-like hybrid macrolactones instead of typical fungal tetramic acids-containing metabolites. The finding provides an insight into the convergent biosynthetic end products that bridge the gap between the modular and iterative PKS-NRPS hybrids.

Thermolides, Potent Nematocidal PKS-NRPS Hybrid Metabolites from Thermophilic Fungus Talaromyces thermophilus

Macrocyclic PKS-NRPS hybrid metabolites represent a unique family of natural products mainly from bacteria with broad and outstanding biological activities. However, their distribution in fungi has rarely been reported, and little has been reported regarding their nematocidal activity. Here we describe an unprecedented class of PKS-NRPS hybrid metabolites possessing a 13-membered lactam-bearing macrolactone, thermolides A-F (1-6) from a thermophilic fungus Talaromyces thermophilus. We showed that 1 and 2 displayed potent inhibitory activity against three notorious nematodes with LC(50) values of 0.5-1 μg/mL, as active as commercial avermectins. This work provided a new class of promising lead compounds for nematocide discovery.

The effect of Talaromyces thermophilus cellulase-free xylanase and commercial laccase on lignocellulosic components during the bleaching of kraft pulp

Biobleaching studies of kraft pulp with cellulase-free xylanase produced by Talaromyces thermophilus and commercial laccase were done in order to investigate their potential applications in various industries. The efficacy of xylanase in reducing the amount of bleaching agent needed was demonstrated by using 20 IU xylanase per gram of oven-dried pulp at 3% concentration prior to the bleaching at pH 7.0 and 70 °C for 120 min of treatment. This amount of xylanase acts on hemicelluloses; it releases chromophores and reduced sugars, and shows the further, decrease in kappa number of pulp without much change in viscosity. Also, a commercial laccase used at 40 IU per gram of oven-dry pulp at 3% concentration exhibited the optimum bleach boosting of the pulp at pH 7.0 and 50 °C after 180 min of treatment. Enzymatic treatment of pulp with the two enzymes (xylanase and laccase) required a smaller amount of chlorine to obtain the same results when pulp was treated with chlorine only. Enzymatic pretreatment of pulp followed by a chemical treatment with 3% NaOCl makes it possible to obtain the same results with chemicals at 7% NaOCl, resulting in 42% reduction in chlorine consumption.

Purification, physico-chemical and kinetic properties of the deglycosylated Talaromyces thermophilus lipase

The Talaromyces thermophilus strain produces only one form of lipase called TTLI. When the culture medium was concentrated and stored at 4°C during a few days, we noticed the appearance of a second short form of lipase named TTLII. This second form was purified to homogeneity using gel filtration and FPLC-Anion exchange chromatography. The NH2-terminal 24 amino acid residues were found to be identical to those of TTLI. The treatment of the TTLI with endoglycosidase H decreased its apparent molecular weight from 39 to 30kDa which corresponds to the molecular weight of TTLII. This difference was mostly attributed to the N-glycosylation of the enzyme. In fact, the glycan chain content and concavaline A–Sepharose affinity column confirmed that the TTLII was completely deglycosylated. Compared to TTLI, the TTLII activity was completely decreased over a broad range of temperature and pH. Furthermore, the deglycosylation of the enzyme reduced its specific activity by 50% toward different substrates; strongly suggest that the N-glycans are determinants for optimal catalytic activity and thermal stability of this enzyme. Covalent immobilization of the enzymes on supports suggests the involvement of the glycan moiety in enzyme–polymer interactions. In the case of TTLI the glycan moiety can constitute an extra site for the covalent linkage of the enzyme on the carrier.

Prévalence et mécanismes moléculaires de la résistance aux antifongiques azoles d’Aspergillus fumigatus au sein d’une cohorte de patients atteints de mucoviscidose

Taxonomy is a dynamic discipline and name changes of fungi with biotechnological, industrial, or medical importance are often difficult to understand for researchers in the applied field. Species belonging to the genera Aspergillus and Penicillium are commonly used or isolated, and inadequate taxonomy or uncertain nomenclature of these genera can therefore lead to tremendous confusion. Misidentification of strains used in biotechnology can be traced back to (1) recent changes in nomenclature, (2) new taxonomic insights, including description of new species, and/or (3) incorrect identifications. Changes in the recent published International Code of Nomenclature for Algae, Fungi and Plants will lead to numerous name changes of existing Aspergillus and Penicillium species and an overview of the current names of biotechnological important species is given. Furthermore, in (biotechnological) literature old and invalid names are still used, such as Aspergillus awamori, A. foetidus, A. kawachii, Talaromyces emersonii, Acremonium cellulolyticus, and Penicillium funiculosum. An overview of these and other species with their correct names is presented. Furthermore, the biotechnologically important species Talaromyces thermophilus is here combined in Thermomyces as Th. dupontii. The importance of Aspergillus, Penicillium, and related genera is also illustrated by the high number of undertaken genome sequencing projects. A number of these strains are incorrectly identified or atypical strains are selected for these projects. Recommendations for correct strain selection are given here. Phylogenetic analysis shows a close relationship between the genome-sequenced strains of Aspergillus, Penicillium, and Monascus. Talaromyces stipitatus and T. marneffei (syn. Penicillium marneffei) are closely related to Thermomyces lanuginosus and Th. dupontii (syn. Talaromyces thermophilus), and these species appear to be distantly related to Aspergillus and Penicillium. In the last part of this review, an overview of heterothallic reproduction in Aspergillus and Penicillium is given. The new insights in the taxonomy of Aspergillus, Penicillium, and related genera will help to interpret the results generated with comparative genomics studies or other studies dealing with evolution of, for example, enzymes, mating-type loci, virulence genes, and secondary metabolite biosynthetic gene clusters.

Production, Partial Purification and Some Properties of α-L-Arabinofuranosidase from Chaetomium thermophile and Talaromyces thermophilus

Production, Partial Purification and Some Properties of α-L-Arabinofuranosidase from Chaetomium thermophile and Talaromyces thermophilus

Catalytic properties of Talaromyces thermophilus α-l-arabinofuranosidase and its synergistic action with immobilized endo-β-1,4-xylanase

Abstract When grown on wheat bran, Talaromyces thermophilus produces a wide spectrum of hemicellulases, mainly endo-β-1,4-xylanase, α- l -arabinofuranosidase and β-xylosidase. The extracellular α- l -arabinofuranosidase was purified to homogeneity by sequential operation of ammonium sulfate precipitation, Q-sepharose column chromatography, gel filtration on Sephacryl S-200 and MonoQ column. The pure α- l -arabinofuranosidase had a specific activity of 49 U/mg of protein and was purified 26.7-fold. The molecular mass of the enzyme was estimated to be 35 kDa, determined by SDS-PAGE and by gel filtration. The α- l -arabinofuranosidase exhibited maximal activity at pH 6.0–7.0 and an optimal temperature at 55 °C. The half-life of the α- l -arabinofuranosidase at 60 °C was approximately 2 h and it was very stable over a wide pH range for 24 h at 4 °C. The apparent Michaelis constant K m value of the α- l -arabinofuranosidase was 0.77 mM for p -nitropeny l -α- l -arabinofuranoside. The turnover number ( K cat ) and catalytic efficiency ( K cat / K m ) were found to be 14.3 s −1 and 1.8 104 M −1 s −1 , respectively. Metal ions such as Hg 2+ and Cu 2+ inhibited enzyme activity, whereas it was strongly activated by Mn 2+ . The α- l -arabinofuranosidase was specific for the α-linked arabinoside in the furanoside configuration and can also retain 52% of its activity in the presence of p -nitrophenyl-β- d -xylopyranoside as substrate. α- l -arabinofuranosidase acted synergistically with the immobilized endo-β-1,4-xylanase for the breakdown of alkali-extracted arabinoxylan and in the improvement of xylobiose and monosaccharide production.

A novel thermoactive and alkaline lipase from Talaromyces thermophilus fungus for use in laundry detergents

The newly isolated fungus Talaromyces thermophilus was found to exhibit an extracellular lipase activity depending on the growth conditions. A 39 kDa monomeric lipase (TTL) was purified from the culture supernatant using ammonium sulphate precipitation, gel filtration and anion exchange chromatography. TTL N-terminal amino acid sequence showed a high similarity with Thermomyces lanuginosa lipase, a well-known lipase with multiple applications. The specific activity of about 7300 ± 122 and 9868 ± 139 U mg −1 was obtained using tributyrin and olive oil emulsion as substrate, respectively, at pH 9.5 and 50 °C. TTL maximum specific activities were found to be 24,110 ± 390 U mg −1 on trioctanoin. TTL was found to be fairly stable and active on long chain triglycerides at pH 9.5. Interestingly, TTL was found to be resistant to interfacial denaturation since it did not require any detergent to show its maximum activity on pure triglycerides. These findings are particularly important for lipase applications, in particular when variable temperatures and high pH values can be encountered, as well as its ability to show high levels activity in the presence of various surfactants and compatibility with some commercial wash agents and bleach agents.

Diversity of thermophilic fungi in Tengchong Rehai national park revealed by ITS nucleotide sequence analyses

The geothermal sites near neutral and alkalescent thermal springs in Tengchong Rehai National Park were examined through cultivation-dependent approach to determine the diversity of thermophilic fungi in these environments. Here, we collected soils samples in this area, plated on agar media conducive for fungal growth, obtained pure cultures, and then employed the method of internal transcribed spacer (ITS) sequencing combined with morphological analysis for identification of thermophilic fungi to the species level. In total, 102 strains were isolated and identified as Rhizomucor miehei, Chaetomium sp., Talaromyces thermophilus, Talaromyces byssochlamydoides, Thermoascus aurantiacus Miehe var. levisporus, Thermomyces lanuginosus, Scytalidium thermophilum, Malbranchea flava, Myceliophthora sp. 1, Myceliophthora sp. 2, Myceliophthora sp. 3, and Coprinopsis sp. Two species, T. lanuginosus and S. thermophilum were the dominant species, representing 34.78% and 28.26% of the sample, respectively. Our results indicated a greater diversity of thermophilic fungi in neutral and alkaline geothermal sites than acidic sites around hot springs reported in previous studies. Most of our strains thrived at alkaline growth conditions.

Improvement of Highly Thermostable Xylanases Production by Talaromyces thermophilus for the Agro-industrials Residue Hydrolysis

A newly isolated thermophilic fungal strain from Tunisian soil samples was identified as Talaromyces thermophilus and was selected for its ability to produce extracellular hemicellulases when grown on various lignocellulosic substrates. Following the optimization of carbon source, nitrogen source, and initial pH of the growth medium in submerged liquid cultures, yields as high as 10.00 +/- 0.15 and 0.21 +/- 0.02 U/ml were obtained for xylanase and beta-xylosidase, respectively. In fact, wheat bran was found to be a good inducer of hemicellulase enzymes, mainly beta-xylosidase. The optimal temperature and pH of the xylanase activity were 75 degrees C and 8.0, respectively. This enzyme exhibited a remarkable stability and retained 100% of its original activity at 50 degrees C for 7 days at pH 7.0-8.0. The half-lives of the enzyme were 4 h at 80 degrees C, 2 h at 90 degrees C, and 1 h at 100 degrees C. T. thermophilus could therefore be considered as a satisfactory and promising producer of thermostable xylanases. Crude enzyme of T. thermophilus rich in xylanase and beta-xylosidase was established for the hydrolysis of lignocellulosic materials as wheat bran.

Improvement of α-l-arabinofuranosidase production by Talaromyces thermophilus and agro-industrial residues saccharification

This study is an application of an experimental design methodology for the optimization of the culture conditions of alpha-L: -arabinofuranosidase production by Talaromyces thermophilus. Wheat bran and yeast extract were first selected as the best carbon and nitrogen sources, respectively, for enzyme production. A Plackett-Burman design was then used to evaluate the effects of eight variables. Statistical analyses showed that while pH had a negative effect on alpha-L: -arabinofuranosidase production, wheat bran and MgSO(4) had a significantly positive effect. The values of the latter three parameters were further optimised using a central composite design and a response surface methodology. The experimental results were fitted to a second-order polynomial model that yielded a determination coefficient of R(2) = 0.91. The statistical output showed that the linear and quadric terms of the three variables had significant effects. Using optimal conditions, the experimental value of alpha-L: -arabinofuranosidase activity produced was very close to the model-predicted value. The optimal temperature and pH of enzyme activity were 55 degrees C and 7.0, respectively. This enzyme was very stable over a considerable pH range from 4 to 9. The crude enzyme of T. thermophilus rich in alpha-L: -arabinofuranosidase was also used for saccharification of lignocellulosic materials and arabinose production.