Trichoderma reesei strains including QM6a (ATCC 13631) and RUT-C30 (ATCC 56765) were maintained on potato dextrose agar plate (PDA) at 28 °C for 7 days for spore collection. Escherichia coli DH5α used as cloning host was culture at 37 °C in Luria–Bertani (LB) medium. Agrobacterium tumefaciens AGL1 was used to transform the gene to T. reesei strains. To induce enzyme production, the conidial suspension (0.5 mL, 1 × 107 conidia/mL) was inoculated into a 50-mL Erlenmeyer flask containing 10 mL of Sabouraud Dextrose Broth (SDB) and incubated for 40 h on an orbital shaker at 200 rpm at 28 °C. The culture was then transferred into a flask containing 10 mL of inducing fermentation medium at 10% inoculum ratio (v/v). The flasks were incubated on an orbital shaker at 200 rpm at 28 °C for 1 week. The wheat bran and Avicel medium were prepared as follows: 0.4% KH2PO4, 0.28% (NH4)2SO4, 0.06% MgSO4·7H2O, 0.05% CaCl2, 0.06% urea, 0.3% tryptone, 0.1% Tween-80, 0.5% CaCO3, 0.001% FeSO4·7H2O, 0.00032% MnSO4·H2O, 0.00028% ZnSO4·7H2O, 0.0004% CoCl2, 2% wheat bran, 3% microcrystalline cellulose. We also used minimal medium (MM) containing 0.5% (NH4)2SO4, 1.5% KH2PO4, 0.06% MgSO4, 0.06% CaCl2, 0.0005% FeSO4·7H2O, 0.00016% MnSO4·H2O, 0.00014% ZnSO4·7H2O, and 0.0002% CoCl2 with 1% lactose or 1% xylan used as carbon source for 3 days of fermentation.
Rut c30
Manufactured by American Type Culture Collection
Sourced in Italy, United States
The RUT-C30 is a laboratory equipment designed for cell culture applications. It provides a controlled environment for the growth and maintenance of cells. The RUT-C30 offers temperature, humidity, and CO2 regulation to support optimal cell culture conditions.
Automatically generated - may contain errors
Lab products found in correlation
Triton x 100, by Merck Group (1 mentions)
Carboxymethylcellulose, by Merck Group (1 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Citric acid, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Potato dextrose agar (pda), by Merck Group (1 mentions)
Malic acid, by Merck Group (1 mentions)
4 nitrophenyl butyrate, by Merck Group (1 mentions)
3 5 dinitrosalicylic acid, by Merck Group (1 mentions)
2 6 dimethoxyphenol, by Merck Group (1 mentions)
Tris hcl buffer, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Na phosphate buffer, by Merck Group (1 mentions)
Qm9414, by American Type Culture Collection (1 mentions)
7 protocols using rut c30
1
Trichoderma reesei Enzyme Production and Screening
Check if the same lab product or an alternative is used in the 5 most similar protocols
Trichoderma reesei strains including QM6a (ATCC 13631) and RUT-C30 (ATCC 56765) were maintained on potato dextrose agar plate (PDA) at 28 °C for 7 days for spore collection. Escherichia coli DH5α used as cloning host was culture at 37 °C in Luria–Bertani (LB) medium. Agrobacterium tumefaciens AGL1 was used to transform the gene to T. reesei strains. To induce enzyme production, the conidial suspension (0.5 mL, 1 × 107 conidia/mL) was inoculated into a 50-mL Erlenmeyer flask containing 10 mL of Sabouraud Dextrose Broth (SDB) and incubated for 40 h on an orbital shaker at 200 rpm at 28 °C. The culture was then transferred into a flask containing 10 mL of inducing fermentation medium at 10% inoculum ratio (v/v). The flasks were incubated on an orbital shaker at 200 rpm at 28 °C for 1 week. The wheat bran and Avicel medium were prepared as follows: 0.4% KH2PO4, 0.28% (NH4)2SO4, 0.06% MgSO4·7H2O, 0.05% CaCl2, 0.06% urea, 0.3% tryptone, 0.1% Tween-80, 0.5% CaCO3, 0.001% FeSO4·7H2O, 0.00032% MnSO4·H2O, 0.00028% ZnSO4·7H2O, 0.0004% CoCl2, 2% wheat bran, 3% microcrystalline cellulose. We also used minimal medium (MM) containing 0.5% (NH4)2SO4, 1.5% KH2PO4, 0.06% MgSO4, 0.06% CaCl2, 0.0005% FeSO4·7H2O, 0.00016% MnSO4·H2O, 0.00014% ZnSO4·7H2O, and 0.0002% CoCl2 with 1% lactose or 1% xylan used as carbon source for 3 days of fermentation.
+ Expand
Trichoderma reesei strains including QM6a (ATCC 13631) and RUT-C30 (ATCC 56765) were maintained on potato dextrose agar plate (PDA) at 28 °C for 7 days for spore collection. Escherichia coli DH5α used as cloning host was culture at 37 °C in Luria–Bertani (LB) medium. Agrobacterium tumefaciens AGL1 was used to transform the gene to T. reesei strains. To induce enzyme production, the conidial suspension (0.5 mL, 1 × 107 conidia/mL) was inoculated into a 50-mL Erlenmeyer flask containing 10 mL of Sabouraud Dextrose Broth (SDB) and incubated for 40 h on an orbital shaker at 200 rpm at 28 °C. The culture was then transferred into a flask containing 10 mL of inducing fermentation medium at 10% inoculum ratio (v/v). The flasks were incubated on an orbital shaker at 200 rpm at 28 °C for 1 week. The wheat bran and Avicel medium were prepared as follows: 0.4% KH2PO4, 0.28% (NH4)2SO4, 0.06% MgSO4·7H2O, 0.05% CaCl2, 0.06% urea, 0.3% tryptone, 0.1% Tween-80, 0.5% CaCO3, 0.001% FeSO4·7H2O, 0.00032% MnSO4·H2O, 0.00028% ZnSO4·7H2O, 0.0004% CoCl2, 2% wheat bran, 3% microcrystalline cellulose. We also used minimal medium (MM) containing 0.5% (NH4)2SO4, 1.5% KH2PO4, 0.06% MgSO4, 0.06% CaCl2, 0.0005% FeSO4·7H2O, 0.00016% MnSO4·H2O, 0.00014% ZnSO4·7H2O, and 0.0002% CoCl2 with 1% lactose or 1% xylan used as carbon source for 3 days of fermentation.
2
Characterizing T. reesei Strains
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following T. reesei strains were used throughout this study: the wild-type strain QM6a (ATCC 13631) and the carbon catabolite derepressed strain Rut-C30 (ATCC 56765). Both strains were maintained on malt extract agar.
3
Fungal Strain Isolation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T. reesei RUT-C30 (ATCC 56765) was purchased from ATCC (Manassas, VA) and T. atroviride Ta13 (MUT 6701) was isolated from
wheat seeds (Algeria) and deposited at the Mycotheca Universitatis
Taurinensis (MUT, Turin, Italy).
2,6-Dimethoxyphenol, 3,5-dinitrosalicylic
acid, 4-nitrophenyl butyrate, acetonitrile, bovine serum albumin,
carboxymethyl cellulose, citrate solution, citric acid, formic acid,
malic acid, methanol, Na acetate buffer, Na phosphate buffer, Na phosphate–citrate,
potato dextrose agar, trichloroacetic acid, Tris-HCl buffer, and Triton
X-100 were purchased from Merck (Darmstadt, Germany).
wheat seeds (Algeria) and deposited at the Mycotheca Universitatis
Taurinensis (MUT, Turin, Italy).
2,6-Dimethoxyphenol, 3,5-dinitrosalicylic
acid, 4-nitrophenyl butyrate, acetonitrile, bovine serum albumin,
carboxymethyl cellulose, citrate solution, citric acid, formic acid,
malic acid, methanol, Na acetate buffer, Na phosphate buffer, Na phosphate–citrate,
potato dextrose agar, trichloroacetic acid, Tris-HCl buffer, and Triton
X-100 were purchased from Merck (Darmstadt, Germany).
4
Fungal Transformation and Cellulase Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Escherichia coli DH5α was used as the cloning host for plasmid construction. Agrobacterium tumefaciens AGL-1 was used as a T-DNA donor for fungal transformation. T. reesei RUT-C30 (CICC 13052, ATCC 56765) was purchased from China Center of Industrial Culture Collection. T. reesei KU70, where ku70 was deleted in RUT-C30 [57 (link)], was provided friendly by Professor Wei Wang from East China University of Science and Technology. E. coli DH5α and A. tumefaciens AGL-1 were cultivated in Luria–Bertani (LB) with 220 rpm at 37 °C and 28 °C, respectively. T. reesei were grown on potato dextrose agar (PDA) plates for conidia production and in TMM [58 (link)] with 2% (w/t) cellulose for cellulase production at 28 °C with 220 rpm. The TMM medium was as followed: Tryptone, 0.75 g/L; Yeast extract, 0.25 g/L; Urea, 1.00 g/L; (NH4)2SO4, 4.00 g/L; KH2PO4, 6.59 g/L; Maleic acid, 11.6 g/L; FeSO4 * 7H2O, 0.005 g/L; MnSO4 * H2O, 0.0016 g/L; ZnSO4 * 7H2O, 0.0014 g/L; CoCl2 * 6H2O, 0.002 g/L; MgSO4, 0.60 g/L; CaCl2, 0.60 g/L; Tween 80, 0.186 ml/L [58 (link)]. When glutamine was the sole nitrogen source, different concentrations of glutamine were used to replace all the nitrogen sources of TMM. All chemicals used in this research were ordered from Sigma-Aldrich, USA.
5
Cultivation and Maintenance of Trichoderma reesei
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Escherichia coli DH5α was used for plasmid amplification. T. reesei Rut-C30 (ATCC 56765), NG14 (ATCC 56767), and PC-3-7 (ATCC 66589) were purchased from ATCC (American type culture collection). T. reesei strain QM6a (ATCC 13631), QM9414 (ATCC 26921), and RL-P37 (NRRL 15709) were respectively purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen), Institute of Microbiology-Chinese Academy of Sciences, and NRRL (Agriculture Research Service Culture Collection). E. coli was cultured in Luria broth (LB) medium. All the strains of T. reesei were cultivated at 28 °C (200 rpm) in 2× Mandels’ medium (1.0 g/L yeast extract, 3 g/L peptone, 0.6 g/L urea, 2.8 g/L (NH4)2SO4, 4.0 g/L KH2PO4, 0.5 g/L CaCl2, 0.6 g/L MgSO4∙7H2O, 5 mg/L FeSO4∙7H2O, 1.6 mg/L MnSO4∙4H2O, 1.4 mg/L ZnSO4∙7H2O, and 20 mg/L CoCl2∙6H2O) in which 2% glucose, 2% lactose, or 1% (w/v) Avicel was used as carbon source [31 ]. In addition, all these strains were maintained on potato dextrose agar (PDA) plates at 28 °C for the generation of conidia.
6
Cryogenic Storage of T. reesei Spores
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
T. reesei RUT-C30 (ATCC 56765) was received from ATTC on October 2013, spread on PDA plates and incubated until sporulation. Spores were harvested with 50 % glycerol solution then stored at −80∘C. Spore solution concentration was 6e9 mL −1. Culture media are prepared according to [53 (link)] (case with 25 m M dipotassium phthalate) and supplemented with 12.5 g L −1 glucose. Feeding solutions (stoichiometric mix of carbon and nitrogen sources) were prepared according to [52 (link)].
7
Fungal Decay and Enzyme Production
Check if the same lab product or an alternative is used in the 5 most similar protocols
Trichoderma reesei RUT-C30 (ATCC 56765), Trametes versicolor A1-ATF (Forest Pathology Culture Collection, University of Minnesota, USA), and Rhodonia placenta MAD 698-R (ATCC 44394) were maintained on potato dextrose agar (PDA) plates for routine subculturing. For R. placenta and T. versicolor, aspen wood wafer cultures were set up as described previously (8 (link)). After 2 to 4 weeks of fungal colonization, wood wafers were harvested. Sections corresponding to early wood decay (0 to 5 mm, with 0 being the hyphal front) were sliced as previously described (12 (link)). Trichoderma reesei , which does not effectively colonize and degrade fresh wood (41 (link), 42 (link)), was used here as a reference GH producer that is widely known for its commercial applications. For this purpose, T. reesei was cultured at 25°C at 150 rpm in liquid medium (2% malt extract, 0.2% yeast extract), inoculated with five edge-transferred disks (3-mm diameter) of actively growing mycelia on PDA. At day 7, the supernatant was collected by centrifugation at 4,000 × g at 4°C for 10 min.
+ Expand
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!