ENZYME MIKROBIA

MATA KULIAH : ENZYMOLOGI

DEPARTEMEN BIOLOGI
FSM UNDIP
 SOURCES OF ENZYMES
Biologically active enzymes may be extracted from any
living organism:
Of the hundred enzymes being used industrially,
- over a half are from fungi
- over a third are from bacteria with the
remainder divided between animal (8%) and
plant (4%) sources .
 SOURCES ENZYMES
Microbes are preferred to plants and animals as
sources of enzymes because:

- They are generally cheaper to produce.

- Their enzyme contents are more predictable

and controllable.

- Plant and animal tissues contain more

potentially harmful materials than microbes,
including phenolic compounds (from plants).
 Fungal Enzymes
Enzyme EC Sources Application

a-Amylase 3.2.1.1 Aspergillus E Baking

Catalase 1.11.1.6 Aspergillus I Food
Cellulase 3.2.1.4 Trichoderma E Waste
Dextranase 3.2.1.11 Penicillium E Food
Glucose oxidase 1.1.3.4 Aspergillus I Food
Lactase 3.2.1.23 Aspergillus E Dairy
Lipase 3.1.1.3 Rhizopus E Food
Mucor
Rennet 3.4.23.6 E Cheese
miehei
Pectinase 3.2.1.15 Aspergillus E Drinks
Protease 3.4.23.6 Aspergillus E Baking

E: extracellular enzyme; I: intracellular enzyme

 Bacterial Enzymes

Enzyme Sources Application

a-Amylase 3.2.1.1 Bacillus E Starch

b-Amylase 3.2.1.2 Bacillus E Starch
Escherichia
Asparaginase 3.5.1.1 I Health
coli
Glucose Fructose
5.3.1.5 Bacillus I
isomerase syrup
Penicillin Pharmace
3.5.1.11 Bacillus I
amidase utical
Protease 3.4.21.14 Bacillus E Detergent
 HISTORY OF ENZYME PRODUCTION

Cristian Hanse (Denmark) : rennet from

cow.
Takamine (Jepang) : amilase from
Aspergillus oryzae with rice as a
substrate. Takadiastase, the first enzyme
production in industrial scale.
Boidin and Effront : amilase high
temperature resisten from Baccilus
subtilis.
 Growing Enzymes

(1) Cultivate the organisms producing the

desired enzymes.
Production can be regulated
Fermentation conditions can be optimized for
overproduction.
 Growing Enzymes
(2) Cell separated from the media usually by filtration
or something by centrifugation.
Depending on intra/extracellular nature of the
enzyme, the cell or fermentation broth is further
processed.
Recovery of intracellular enzymes is more
complicated and involves the disruption of cells
and removal of debris and nucleic acids.
Increasing permeability of cell membrane
(CaCl2 (salt) or dimethylsulfoxide (DMSO) or
change in pH
Last resort is cell disruption.
Enzyme Production
o Cell Culturing optimum condition
o Fermentation aerobic, in stirred-
tank reactor
o Enzyme purification
o Extracellular enzyme separate cells
o Intracellular enzyme break cell
o Concentrate
o Chromatography
o Crystallization
o Enzyme product: as concentrate,
powder or formulations (mixed
xylanase-pectinase for animal feeds
Enzyme Production
EKSTRAKSI ENZYME
 ENZYME DEVELOPMENT
Strain improvement
Genetic manipulation and classical selection
o Chemical /physical mutagenesis
Recombinant DNA technology
Optimization of cell culturing condition
Enzyme improvement
Chemical modification to change conformation
Protein engineering: change specific amino acid
by site-directed mutagenesis
Change the catalytic condition
Organic solvent
Metal ion
Industrial applications of enzymes
Laundry detergents-addition of lipase, protease, and amylase to
detergents allowed the use of lower temperatures, and the better
removal of specific stains. There has been continual development of
enzyme detergents with increased activity at low and high
temperatures and at alkaline pH values.
Textile industry has used alkaline proteases for degumming silk,
removing a protein from the outside of the silk fibres (Gupta et al.,
2002). Pectinases used to remove cell-wall components from cotton
fibres and glucose oxidase used to bleach the fibres. Cellulases have
been used to treat denim garments as an alternative to
stonewashing in a process known as biostoning (Belghith et al.,
2001).
Leather Replacement of the chemical processes by enzymes should
have considerable advantages. Proteases have been used to de-hair
hides and lipase to remove fat.
Paper Cellulases have been used to assist pulping, and to de-ink
paper. Laccase enzymes that attack lignin have been used to bleach
pulp as an alternative to chlorine bleach (Kirk et al., 2002).
Industrial applications of
enzymes
 Enzymes from Extremophiles
(Eichler, 2001; Haki and Rakshit, 2003; Huber
and Stetter, 1998).
 Extremophiles
The microorganisms found in extreme conditions of
temperature, salt, and pressure belong to the
domains of Bacteria and Archaea, but the majority-
are the Archaea.
Clearly these types of micro-organism will contain
enzymes that can function under extreme
conditions. e.g Thermus aquaticus (Taq polymerase
enzyme for PCR)
 Enzymes and Sources
Proteases
Overproducing strains of Bacillus, Aspergillus,
Rhizopus, and Mucor.
Pectinases
Aspergillus niger.
Lactases
Yeast and Aspergillus.
Lipases
Certain strains of yeast and fungi.
Glucose isomerase
Flavobecterium arborescens or Bacillus coagulans