scholarly journals Production of Recombinant Trichoderma reesei Cellobiohydrolase II in a New Expression System Based on Wickerhamomyces anomalus

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Dennis J. Díaz-Rincón ◽  
Ivonne Duque ◽  
Erika Osorio ◽  
Alexander Rodríguez-López ◽  
Angela Espejo-Mojica ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Recombinant Expression ◽  
Expression System ◽  
Enzymatic Extract ◽  
Native Strain ◽  
Wickerhamomyces Anomalus ◽  
Optimum Ph ◽  
Hydrolysis Of Cellulose ◽  
Cellobiohydrolase Ii ◽  
Hydrolysis Of

Cellulase is a family of at least three groups of enzymes that participate in the sequential hydrolysis of cellulose. Recombinant expression of cellulases might allow reducing their production times and increasing the low proteins concentrations obtained with filamentous fungi. In this study, we describe the production of Trichoderma reesei cellobiohydrolase II (CBHII) in a native strain of Wickerhamomyces anomalus. Recombinant CBHII was expressed in W. anomalus 54-A reaching enzyme activity values of up to 14.5 U L−1. The enzyme extract showed optimum pH and temperature of 5.0–6.0 and 40°C, respectively. Enzyme kinetic parameters (KM of 2.73 mM and Vmax of 23.1 µM min−1) were between the ranges of values reported for other CBHII enzymes. Finally, the results showed that an enzymatic extract of W. anomalus 54-A carrying the recombinant T. reesei CBHII allows production of reducing sugars similar to that of a crude extract from cellulolytic fungi. These results show the first report on the use of W. anomalus as a host to produce recombinant proteins. In addition, recombinant T. reesei CBHII enzyme could potentially be used in the degradation of lignocellulosic residues to produce bioethanol, based on its pH and temperature activity profile.

scholarly journals Hydrolyses of α- and β-cellobiosyl fluorides by cellobiohydrolases of Trichoderma reesei

1993 ◽  
Vol 291 (3) ◽  
pp. 883-888 ◽  
Author(s):  
A K Konstantinidis ◽  
I Marsden ◽  
M L Sinnott
Keyword(s):  
Trichoderma Reesei ◽  
Biological Function ◽  
Fluoride Ion ◽  
Cellobiohydrolase I ◽  
Ion Release ◽  
Cellulase Complex ◽  
Cellobiohydrolase Ii ◽  
Glycoside Hydrolysis ◽  
Kinetics Of ◽  
Hydrolysis Of

Cellobiohydrolase II hydrolyses alpha- and beta-D-cellobiosyl fluorides to alpha-cellobiose at comparable rates, according to Michaelis-Menten kinetics. The stereochemistry, absence of transfer products and strict hyperbolic kinetics of the hydrolysis of alpha-cellobiosyl fluoride suggest that the mechanism for the alpha-fluoride may be the enzymic counterpart of the SNi reaction observed in the trifluoroethanolysis of alpha-glucopyranosyl fluoride [Sinnott and Jencks (1980) J. Am. Chem. Soc. 102, 2026-2032]. The absolute factors by which this enzyme accelerates fluoride ion release are small and greater for the alpha-fluoride than for the beta, suggesting that its biological function may not be just glycoside hydrolysis. Cellobiohydrolase I hydrolyses only beta-cellobiosyl fluoride, which is, however, an approx. 1-3% contaminant in alpha-cellobiosyl fluoride as prepared and purified by conventional methods. Instrumental assays for the various components of the cellulase complex are discussed.

THE SAP OF THE BIRCH TREE, BETULA PAPYRIFERA MARSH.: I. THE AMYLASE SYSTEM

1942 ◽  
Vol 20b (6) ◽  
pp. 114-120 ◽  
Author(s):  
E. Bois ◽  
W. O. Chubb
Keyword(s):  
Cellulose Acetate ◽  
Betula Papyrifera ◽  
Optimum Conditions ◽  
Maximum Production ◽  
Birch Tree ◽  
Optimum Ph ◽  
Hydrolysis Of Cellulose ◽  
Hydrolysis Of

The amylase system of birch sap consists of a cellobiogenic amylase and most probably a glucogenic amylase. The optimum conditions of pH and temperature are pH 5. 5 at 4 °C. to 6.3 at 60 °C. The maximum production of cellobiose occurred at a temperature of 50 °C. at the optimum pH for this temperature. The optimum conditions for the production of glucose were pH 5.5 and a temperature of 50 °C.It is felt that these new facts throw some additional light on the constitution of starch, and that possibly birch sap might serve as a source of cellobiose, which has heretofore only been prepared commercially by hydrolysis of cellulose acetate.

scholarly journals Hydrolyses of α- and β-cellobiosyl fluorides by Cel6A (cellobiohydrolase II) of Trichoderma reesei and Humicola insolens

2000 ◽  
Vol 345 (2) ◽  
pp. 315-319 ◽  
Author(s):  
Dieter BECKER ◽  
Karin S. H. JOHNSON ◽  
Anu KOIVULA ◽  
Martin SCHÜLEIN ◽  
Michael L. SINNOTT
Keyword(s):  
Trichoderma Reesei ◽  
Active Site ◽  
Substrate Concentration ◽  
Initial Rate ◽  
Enzyme Active Site ◽  
Humicola Insolens ◽  
Cellobiohydrolase Ii ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
Identical Crystal

We have measured the hydrolyses of α- and β-cellobiosyl fluorides by the Cel6A [cellobiohydrolase II (CBHII)] enzymes of Humicola insolens and Trichoderma reesei, which have essentially identical crystal structures [Varrot, Hastrup, Schülein and Davies (1999) Biochem. J. 337, 297-304]. The β-fluoride is hydrolysed according to Michaelis-Menten kinetics by both enzymes. When the ~ 2.0% of β-fluoride which is an inevitable contaminant in all preparations of the α-fluoride is hydrolysed by Cel7A (CBHI) of T. reesei before initial-rate measurements are made, both Cel6A enzymes show a sigmoidal dependence of rate on substrate concentration, as well as activation by cellobiose. These kinetics are consistent with the classic Hehre resynthesis-hydrolysis mechanism for glycosidase-catalysed hydrolysis of the ‘wrong’ glycosyl fluoride for both enzymes. The Michaelis-Menten kinetics of α-cellobiosyl fluoride hydrolysis by the T. reesei enzyme, and its inhibition by cellobiose, previously reported [Konstantinidis, Marsden and Sinnott (1993) Biochem. J. 291, 883-888] are withdrawn. 1H NMR monitoring of the hydrolysis of α-cellobiosyl fluoride by both enzymes reveals that in neither case is α-cellobiosyl fluoride released into solution in detectable quantities, but instead it appears to be hydrolysed in the enzyme active site as soon as it is formed.

scholarly journals Hydrolysis of Cellulose from Oil Palm Empty Fruit Bunch using Aspergillus niger

2021 ◽  
Vol 226 ◽  
pp. 00042
Author(s):  
Sri Sugiwati ◽  
Suaidah Suaidah ◽  
Eka Triwahyuni ◽  
Muryanto Muryanto ◽  
Yosie Andriani ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Oil Palm ◽  
Hydrolytic Activity ◽  
Reducing Sugar ◽  
Empty Fruit Bunch ◽  
Hydrolysis Time ◽  
Glucose Assay ◽  
Optimum Ph ◽  
Hydrolysis Of Cellulose ◽  
Hydrolysis Of

Oil palm empty fruit bunch (OPEFB) constitutes a great source of lignocellulosic biomass, mainly comprising of 66.97 % of holocellulose (cellulose and hemicellulose) and 24.45 % of lignin. This present work aimed to hydrolyze cellulose present in OPEFB to form glucose with the aid of Aspergillus niger. A. niger is a type of filamentous fungi able to produce cellulase, a multi-enzyme complex consisting of an endoglucanase, exoglucanase, and β-glucosidase, able to converting cellulose into glucose. The glucose produced is then fermented to produce bioethanol. The present study compared hydrolytic activity of cellulose between OPEFB with pretreatment using NaOH 10 % and OPEFB without pretreatment, concerning temperature, pH, and hydrolysis time. The concentration of reducing sugar derived from cellulosic hydrolysis was determined by using a glucose assay of 3.5-dinitrosalicylic acid. The results showed that the optimum temperature for hydrolysis of cellulose OPEFB (pretreated and untreated) was at 40 °C and the optimum pH was 5.0 for OPEFB-untreated and 5.5 for OPEFB-pretreated. Hydrolysis of cellulose at 40 °C and 3 d yielded reducing sugar 13.01 mg mL−1 and 1.16 mg mL−1 for OPEFB-untreated and OPEFB-pretreated, respectively.

scholarly journals Cellobiohydrolase from Trichoderma reesei

1983 ◽  
Vol 215 (3) ◽  
pp. 677-683 ◽  
Author(s):  
M Nummi ◽  
M L Niku-Paavola ◽  
A Lappalainen ◽  
T M Enari ◽  
V Raunio
Keyword(s):  
Trichoderma Reesei ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Culture Liquid ◽  
Protein Band ◽  
Immunoaffinity Chromatography ◽  
Amorphous Cellulose ◽  
Single Protein ◽  
Hydrolysis Of Cellulose ◽  
Hydrolysis Of

A 1,4-beta-D-glucan cellobiohydrolase (EC 3.2.1.91) was purified from the culture liquid of Trichoderma reesei by using biospecific sorption on amorphous cellulose and immunoaffinity chromatography. A single protein band in polyacrylamide-gel electrophoresis and one arc in immunoelectrophoresis corresponded to the enzyme activity. The Mr was 65 000. The pI was 4.2-3.6. The purified enzyme contained about 10% hexose. The enzyme differs from previously described cellobiohydrolases in being more effective in the hydrolysis of cellulose.

Hydrolysis of barley (1→3), (1→4)-β-d-glucan by a cellobiohydrolase II preparation from Trichoderma reesei

1995 ◽  
Vol 26 (2) ◽  
pp. 109-119 ◽  
Author(s):  
Kirsti Henriksson ◽  
Anita Teleman ◽  
Tapani Suortti ◽  
Tapani Reinikainen ◽  
Johanna Jaskari ◽  
...  
Keyword(s):  
Trichoderma Reesei ◽  
Cellobiohydrolase Ii ◽  
Hydrolysis Of
Sign in / Sign up

Export Citation Format

Share Document