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Презентация на тему Cellulase. Introduction (source)

Introduction (source) Cellulase refers to an entourage of enzymes produced chiefly by fungi, bacteria and protozoans that catalyze cellulolysis (i.e. the hydrolysis of cellulose).  However, there are also cellulases produced by a few other types of
Cellulase Introduction  (source)   Cellulase refers to an entourage of enzymes produced CellulaseCellobiohydrolaseswhose major activity involves the cleavage of cellobiose residues consecutively from the Some species of fungi and bacteria are able to exhaustively digest crystalline Other Names  Other names for 'endoglucanases' are: endo-1,4-beta-glucanase, carboxymethyl cellulase (CMCase), endo-1,4-beta-D-glucanase, Types of reactions/ Classification   General types of cellulases based on Choice of host organism Strain engineeringThermostable cellulases productionNowadays, most of the studies about production of thermostable Homologous overexpression in bacteria Some studies report the use of directed evolution Heterologous overexpression The strategies based in heterologous expression are focused in the Future targets for genetic manipulation and optimization will include the use of Schematic representation of the experimental approach and on-site enzyme production in a Cultivation Media Medium 1 ((without carbon source )NaNO3, 2.0, KH2PO4, 1.0, MgSO4⋅7H2O, Harvest and Separation of Enzymes 6 ml6 ml6 ml6 mlThese were then Uses Thank you for attention!!!
Слайды презентации

Слайд 2 Introduction (source)
Cellulase refers to an entourage of

Introduction (source)  Cellulase refers to an entourage of enzymes produced chiefly

enzymes produced chiefly by fungi, bacteria and protozoans that

catalyze cellulolysis (i.e. the hydrolysis of cellulose).
 

However, there are also cellulases produced by a few other types of organisms, such as some termites and the microbial intestinal symbionts of other termites.
 

Several different kinds of cellulases are known, which differ structurally and mechanistically.


Слайд 3 Cellulase


Cellobiohydrolases

whose major activity involves the cleavage of cellobiose

CellulaseCellobiohydrolaseswhose major activity involves the cleavage of cellobiose residues consecutively from

residues consecutively from the ends of the cellulose chains


Endoglucanases

whose major activity involves the cleavage
of β-glycosidic bonds in the cellulose chain

they are necessary for the efficient hydrolysis of cellulose to soluble oligosaccharides


Слайд 4

Some species of fungi and bacteria are able

Some species of fungi and bacteria are able to exhaustively digest

to exhaustively digest crystalline cellulose in pure culture are

said to have complete or true cellulases.

The majority of organisms that produce cellulases can only hydrolyze the cellulose in their diets to certain extent. they are known as incomplete cellulases.

These cellulases unable to digest cellulose exhaustively can still generate sufficient amount of glucose for their producers. Endogenous cellulases of termites belong to this category.






Complete vs. incomplete cellulases


Слайд 5 Other Names
Other names for 'endoglucanases' are: endo-1,4-beta-glucanase,

Other Names Other names for 'endoglucanases' are: endo-1,4-beta-glucanase, carboxymethyl cellulase (CMCase), endo-1,4-beta-D-glucanase,

carboxymethyl cellulase (CMCase), endo-1,4-beta-D-glucanase, beta-1,4-glucanase, beta-1,4-endoglucan hydrolase, and celludextrinase.

The other types of cellulases are called exocellulases.

Слайд 6 Types of reactions/ Classification

General types of

Types of reactions/ Classification  General types of cellulases based on

cellulases based on the type of reaction catalyzed:

Cleaves internal

bonds at Endocellulase (EC 3.2.1.4) randomly amorphous sites that create new chain ends.
Cellobiase (EC 3.2.1.21) or beta-glucosidase hydrolyses the exocellulase product into individual monosaccharides.
Cellulose phosphorylases depolymerize cellulose using phosphates instead of water.


Слайд 7 Choice of host organism

Choice of host organism

Слайд 8 Strain engineering
Thermostable cellulases production
Nowadays, most of the studies

Strain engineeringThermostable cellulases productionNowadays, most of the studies about production of

about production of thermostable cellulases are focused on the utilization

of cellulase-producing thermo/alkalophiles and also, on the improvement of cellulase production by optimizing its nutritional and environmental necessities or by engineering new highproducer recombinants or cellulase-producing transgenic plants, such as transgenic tobacco

Слайд 9 Homologous overexpression in bacteria
Some studies report the

Homologous overexpression in bacteria Some studies report the use of directed

use of directed evolution techniques in combination with a

rational design to overexpress cellulases in their own bacterial source. Genera such as Bacillus (B. subtilis) and Clostridium (C. thermocellum) were used as a homologous cellulases production system, their easy genetic modification and other proper features.
However, the use of these bacteria has disadvantages such as low protein yields, high production costs or need of enriched media

Слайд 10 Heterologous overexpression
The strategies based in heterologous expression

Heterologous overexpression The strategies based in heterologous expression are focused in

are focused in the use of non-cellulolytic micro/organisms that

have high production ratio for expressing microbial cellulases
Bacteria such as E. coli, different species from the genus Bacillus, Pseudomonas fluorescens, Ralstonia eutropha and Zymomonas mobilis;
yeasts such as Saccharomyces cerevisiae and Pichia pastoris and filamentous fungi from genera Aspergillus and Trichoderma genera
are the most used in research and industry, considered as host systems for producing recombinant enzymes. Furthermore, cell cultures of mammals, plants or insects and transgenic plants and/or animals are used for protein expression

Слайд 11
Future targets for genetic manipulation and optimization will

Future targets for genetic manipulation and optimization will include the use

include the use of the cellulolytic system of Clostridium thermocellum

for engineering new strains, depending of the concrete industrial application and the fully characterization of the promising thermophilic bacterium Caldicellulosiruptor bescii.

Слайд 13 Schematic representation of the experimental approach and on-site

Schematic representation of the experimental approach and on-site enzyme production in

enzyme production in a cellulose-to-ethanol process.
Björn Alriksson et al.

Appl. Environ. Microbiol. 2009;75:2366-2374

Слайд 14 Cultivation Media


Medium 1 ((without carbon source )

NaNO3,

Cultivation Media Medium 1 ((without carbon source )NaNO3, 2.0, KH2PO4, 1.0,

2.0, KH2PO4, 1.0, MgSO4⋅7H2O, 0.5, and (mg.L-1) FeSO4, 10.0. The

pH of the medium was adjusted to 6.5.

Medium 2

Peptone, 6.0, KH2PO4, 1.0, MgSO4⋅7H2O, 1.72, KCl, 0.5, and (mg.L-1) FeSO4, 10.0. The pH of the medium was adjusted to 5.5.


Слайд 15 Harvest and Separation of Enzymes








6 ml
6 ml
6

Harvest and Separation of Enzymes 6 ml6 ml6 ml6 mlThese were

ml
6 ml
These were then centrifuged at 5000 rpm for 15

minutes and the supernatant was collected to 10 mL sterile tubes and stored at -20ºC for further use in enzyme assays

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