Enzymes​- biological catalysts that are protein S-->P

molecules in nature. *Product concentration increase with time

-produced by living cells *Substrate concentration decrease with time
-catalyze the making and breaking
of chemical bonds Lock and Key Theory​- on of the original
Simple Enzyme Kinetics​- rate of enzyme theories for enzyme-substrate complex
reaction and how it is affected by chemical - there is a topographical,
and physical structural compatibility
conditions between enzyme and
Kinetic studies of enzymatic reactions​- substrate
provide info about basic mechanism of the
enzyme reaction Assumptions used to derive reaction rate
Rate equations​- applied in calculating time, equation
yields, etc. 1. ​T​otal enzyme concentration is constant
during the reaction.
2. ​A​mount of enzyme is very small compared
to amount of substrate.
3. ​P​roduct concentration is very low.

Approaches to derive rate equations

Michaelis-Menten Approach
Briggs-Haldane Approach
Numerical Solution
Assumed that the product releasing step is much slower than the
reversible reaction (heterogeneous catalytic reactions)
*Product formation and substrate consumption is proportional to
the enzyme-substrate concentration
*Equation analogous to Langmuir isotherm equation
Assume dC​ES​/dt = 0 (pseudo- or quasi-steady state) (homogeneous
catalytic reactions);
Same with M-M except for K​M
For simultaneous differential equations

Plots
Langmuir Plot Most satisfactory
Lineweaver-Burk Plot
Eadie-Hofstee Plot Gives slightly better weighing than L-B Plot
Disadvantage: Rate of reaction (r) appears on both coordinates.

Bioreactor​- device within which biochemical transformations are caused

- called fermenter (living cells), enzyme reactor (enzyme); transformation by living cells or
in vivo cellular components (enzymes)
Fermentation​- both aerobic, anaerobic metabolism (modern); anaerobic metabolism (origina​l)
In vivo​- ​in life; biological reaction in a living cell or organism

Batch or Steady-State - simplest reactor configuration

Plug-Flow Reactor - equipped with agitator (to mix); pH controller/buffer solution
-ideal batch reactor is well-mixed (homogeneous content)
Continuous - ideal reactor based on the assumption that reactor contents are well-mixed
Stirred-Tank Reactor -concentrations of outlet stream and reactor are the same
(CSTR) -(steady-state) substrate conc. Of reactor is constant (dCs/dt=0)
Dilution rate (D)​- equal to reciprocal of the residence time (T)

Inhibition of Enzyme Reactions

Modulator (or effector)​- substance that can combine with enzymes to alter catalytic activities
Inhibitor​- modulator which decreases enzyme activity and rate of reaction (noncompetitively,
competitively, or partially competitively)

Types of Inhibition
Competitive Inhibitor has strong resemblance to the substrate;
 Inhibitor and substrate compete for the active site of an enzyme
Noncompetitive Inhibitors interact with enzyme in different ways;
Can bind to enzyme ir- or reversibly (active site or some other region);
Resultant complex is inactive

Influences on Enzyme Activity:

1. Concentration of Components
2. pH
3. Temperature
4. Shear

*Immobilization of enzymes may introduce ​mass-transfer resistance​ (absent in free soluble enzymes)
1. Large particle size of immobilized enzyme
2. |Inclusion of enzyme in polymeric matrix
Steps of Hypothetical Path (see photo)
1. ​T​ransfer from bulk liquid to relatively unmixed liquid layer (external mass-transfer resistance)
2. ​D​iffusion through relatively unmixed layer (external MTR)
3. ​D​iffusion from the surface of the particle to the active site of the enzyme (intraparticle MTR)

External MTR​- if an enzyme is immobilized on the surface of an insoluble particle

- rate of mass transfer is proportional to the driving force
-rate of substrate transfer is equal to that of substrate consumption
Enzyme Kinetics​- branch of enzymology that deals with the factors affecting the rates of
enzyme-catalyzed reactions
Important factors:
1. Enzyme concentration
2. Ligand concentrations
3. pH
4. Ionic strength
5. Temperature
Uni Un​i-commonly used Cheland nomenclature
- simplest enzyme-catalyzed reaction involves a single substrate going to a single product
Rapid equilibrium conditions​- E, S, ES equilibrate very rapidly compared to the rate at which ES
breaks down to E+P (see photo)
Henri-Michaelis-Menten equation​-gives the instantaneous or initial velocity relative to Vmax at a
given substrate concentration
-no more than 5% of substrate be utilized over the assay period
Equilibrium Constant
*​Many reactions in nature are ​REVERSIBLE​ and do not need to proceed to completion.
*At equilibrium, net velocity is​ ZERO ​(velocity forward = velocity backward)
K​eq​ ​-equilibrium constant / position of equilibrium

*Enzyme reaction is influenced by pH (both in vivo and in vitro)

*Chemical reaction depends on the temperature acc to ​Arrhenius equation​ (see photo).
*Increase in temperature increases the rate of reaction (atoms have greater energies and greater
tendencies to move)
*When temperature increases, ​denaturation​ (begins at 45-50​o​C) destroys activity of enzyme
molecules due to unfolding of protein chain after the breakage of weak bonds.
*Mechanical force that an enzyme solution usu encounters is ​fluid shear​ (flowing fluid, shaking of
vessel, stirring).
*Mechanical forces disturb the elaborate shape of an enzyme molecule

Conjugate Acid-Base Pair (Bronsted)

Acid- donates protons
Base- accepts protons
Ionization of Water:
Considered in two ways: Described as:
1. Simple dissociation to yield H​+​ and OH​-​ ions 1. Dissociation constant (K​d​)
2. In terms of conjugate acid-base pairs 2. Ionization constant (K​I​)
3. Special constant of water (K​W​)

*Water is ​amphotheric​ (can both be acid and base).