Cellobiohydrolase is one of the cellulase enzymes which is involved in degradation of cellulose, the most abundant biopolymer in nature. Trichoderma species are known as a useful source of cellulytic enzymes. In this research cellulase enzymes activity in 30 isolates of Trichoderma sp. were studied by rapid screening on medium containing swollen cellulose and isolate Trichoderma reesei (PTCC5142) was selected on the basis of clearing zone diameter (16 mm). The enzyme activity was measured in the optimum conditions for production of cellobiohydrolase enzyme in Trichoderma. In order to obtain the optimum conditions: the selected isolate was grown in Mandels media with CMC, Avicel and Filter paper as carbon source at pH 4, 5,6, and7. Lactose and cellubiose were added to media as inducer agents. Media were incubated at different temperatures (25, 28 and 320C) for 14 days. Samples were collected in 24h intervals, and enzyme activity was measured. Results showed that the optimum conditions for hyperproduction of cellobiohydrolase were pH=5, CMC as carbon source, temperature 28 0C, lactose as inducer on 7th day. After screening the cellobiohydrolase activity of all 30 isolates in the optimized conditions, results showed that isolate Trichoderma reesei (PTCC5142) with the highest enzyme activity (4.45U/ml) was the best producer of cellobiohydrolase enzyme.

Background & Objective: In the last decades, many successful anticancer fungal metabolites have been obtained, and fungi have shown great potential to produce beneficial anticancer drug compounds. In this article, the effects of the reference strain of Trichoderma reesei ethanolic extract on the Hepa1-6 cell line were studied. Materials & Methods: The fungus was initially cultured in the potato dextrose agar media, and then, it was transferred to yeast peptone dextrose broth. Afterward, the ethanolic and aquatic extracts were prepared, and were analyzed by chromatography and mass spectrometry (GC-MS). Then, the cell lines were treated with different doses of the extracts. The cytotoxic activity of fungal extracts were evaluated by MTT assay, Ferric reducing ability of plasma (FRAP) assay and Ferric Reducing Antioxidant Power (FRAP) Assay. Alkaline phosphatase (ALP) and Lactate dehydrogenase (LDH) measurement tests were done on the culture and cell fractions. Hematoxylin & Eosin staining was performed for microscopic studies, and an oxidant attack-antioxidant defense test was performed on the cell fraction. Results: GC-MS determined compounds such as cineal, cyclohexene, hydroxyethyl benzaldehyde, terpinenyl acetate, dihydro esthophyllene, propionic acid, hexadecanoic acid, and octadecanoic acid were extracted from the fungal extract. The half maximal inhibitory concentration (IC50) was between 7-8%, and the highest cytotoxic effectiveness was 1015%. An increase in the levels of ALP, LDH enzymes, and total protein was observed. Conclusion: Therefore, the findings suggest that the extract of T. reesei has inhibitory effects on Hepa1-6 cancer cell lines.

b-glucosidase, is one of the cellulase enzymes system, hydrolyses cellobiose or cellooligoschrides to glucose.In this research for production of b-glucosidase enzyme, b-glucosidase activity was measured in the optimum conditions in 30 isolates of Trichoderma sp. In order to obtain the optimum conditions: an isolate of Trichoderma (T.reesei PTCC 5142) was grown in Mandels media containing CMC, Avicel and filter paper as carbon source at pH 4,5, 6 and 7. Lactose and cellubiose were added to media as inducer agents. Media were incubated at 25, 29 and 32 oC for 14 days. Samples were collected in 24h intervals, and enzyme activity was measured. Results showed that the optimum conditions for hyperproduction of b-glucosidase were pH=5, CMC as carbon source, lactose as inducer on seventh day at 29 oC. After screening the -glucosidase activity of all 30 isolates in the optimized conditions, it was shown that T.reesei PTCC 5142 had the highest level of -glucosidase specific activity (0.45 U/mg). For characterization and study of b-Glucosidase gene, CTAB method used for genomic DNA extraction.The expected PCR product with 566 bp was amplified with two specific primers (CP11 & CP12). The amplified fragment was confirmed by restriction pattern.

Trichoderma reesei is one of the most important fungi producing cellulose degrading enzymes in nature. In this study, 21 mutated isolate via gamma irradiation of T. reesei were screened for cellulase enzyme production on a colloidal cellulose medium. The concentration of extracellular proteins of wild and mutant isolates were measured using Bradford's method. Avicel, carboxymethyl cellulose and filter paper were used to measure cellulase activity. The purity and composition of enzyme-rich proteins were also evaluated using polyacrylamide gel electrophoresis (SDS-PAGE) test. The highest amount of extracellular protein production was observed in T. r M7 and T. r M17 isolates. Activity of endoglucanase, exoglucanase and total cellulase enzymes in mutated isolate T. r M8 showed the highest levels of enzymatic activity among mutant and wild isolates. The difference in the molecular weight of the enzyme bands showed that the EG 4, Cel 1A, Cel 12A, Cel 45A, Cel 3A, Cel 7A, Cel 6A, Cel 5A and Cel 61A enzymes hydrolyzed the colloidal cellulose, synergictly. These results indicate that, mutation induced by gamma radiation is possible to obtain high producing Trichoderma mutants of cellulase enzymes.

The aim of this study was a strain-improvement program for Trichoderma reesei PTCC 5142 by using a combination of UV light and NTG (N-methyl-N'-nitro-N-nitrosoguanidine) for enhanced cellulase production. Following mutagenesis after several rounds, mutant A6: 2 was selected from a total of 6500 colonies. Results obtained after 4 days were: Enzyme activity 1.26 U/ml and 0.82 U/ml for exoglucanase and endoglucanase, respectively. The comparative results showed increased production exoglucanase and endoglucanase by mutant A6: 2 than Trichoderma reesei PTCC 5142 to amount 130% for exoglucanase and 156% for endoglucanase.

Cellulolytic complex are enzymes capable of hydrolyzing cellulose. Due to rapid growth in population and industrialization, most countries are required to produce more fuel. Production of bioethanol from lignocellulosic biomass is very challenging due to environmental pollution by fossil fuels. Cellulases play a significant role in biotechnological processes. The cost of production of cellulase is very high. Approximately 50% of the cost of producing bioethanol is used to produce cellulose. Gibberellin regulates the growth of fungi and plants. Cellulase production was significantly improved by gibberellins hormone. The cellulase activity was 4. 29 FPU/ml (when we used 50 μ mol/ml of gibberellins in culture medium). This was 12-fold higher (0. 34 FPU/ml in control solution) compare with untreated hormone and was comparable to that achieved with concentrated by n-butanol. In this experiment, the enzyme activity was increased 223-fold higher compare with control solution.

Variation in the composition of the production media were investigated to optimize the excretion of the cellulolytic enzymes by Trichoderma reesei (CBS383.73) and a Botrytis sp. isolated from the microflora of Iran. The culture filtrare of Trichoderma reesei showed a cellulolytic activity of 300 mFPU/ml after 13 days of growth in a medium containing walseth cellulose (0.5%, w/v) at pH 5. Under similar production conditions, except for the initial pH of 7, the culture filtrate of Botrytis sp. showed an activity of 360 mFPU/ml. Replacement of walseth cellulose with h__2O__2 - treated bagasse, Barely hunsks, wheat husks, or sawdust (0.5% w/v) in the presence of the inducer (0.5% w/v, walseth cellulase) enhanced the extent of enzyme production by Trichoderma reesei to 780 mFPU/mL. The Botrytis sp. did not excreate cellulolytic enzyme using (as the main sources of carbon), the H__2O__2 - treated bagasse or sawdust plus 0.5% walseth cellulose. However, the culture filtrate of the Botrutis sp. showed a cellulolytic activity of 679 or 463 mFPU/ml using the H__2O__2 - treated wheat or barely husks (1% w/v) plus 0.5% walseth cellulose, respectively. In addition, the Calcium hydroxide treatment of the agricultural wastes depressed totally the enzyme secretion by the Botrytis sp. except for the treated sawdust shich enhanced the enzyme production almost by a factor of seven in the absence of the inducer.

Cellulose is the most abundant biopolymer in nature which is made of glucose units with  b(1,4) links. It is hydrolysed to glucose through the cinergistic function of three groups of enzymes namely endogluconases, cellobiohydrolases and  betaglucosidases. The most important enzyme in this enzyme complex is b-1,4 endoglucanase (egl) which has industrial applications. In this study genomic DNA from T.reesei PTCC5142 (high EglI enzyme producer, 0.37 U/ml) and specific primers (EB1 and EF1) were used for eglI amplification. The amplified DNA fragment (1524 bp) was cloned into pBluscript SK(+) and sequenced. Also eglI cDNA was synthesized (1380 bp), cloned and sequenced. Comparison of genomic DNA and cDNA sequences showed that eglI gene contains two introns (70 bp and 57 bp) and codes for a polypeptide with a 459 amino acids long. Alignment of eglI sequence revealed three nucleotide differences with eglI gene from T. reesei VTT-D-80133 which altered two amino acids in its catalytic domain. The superposition, using SWISS-MODEL, of all backbone atoms (1480) between the template structure and eglI of T. reesei VTT-D-80133, rendered a Root Mean Squared Deviation (RMSD) of 0.47 A. While superposition over equivalent 370 C a atoms gives a RMSD=0.4 A.