24.

1 INTRODUCTION TO ENZYMES AND THEIR MODES OF ACTION

Enzymes are exceptionally particular, proteinaceous, macromolecular biocatalysts that trigger the rate
and improve the specificity of metabolic chemical reactions. Emil Fischer proposed the ‘‘lock and key’’
model (Fig. 24.1) for the stereochemical relationship between catalysts and their substrates (Fischer,
1894). The actuated fit model of Koshland (Koshland Jr, 1958) illuminated the idea of substrate
specificity in light of the requirement for a transition state adjustment by the catalyst dynamic site.
According to him, catalysts are adaptable structures; the dynamic site is ceaselessly reshaped by
interactions with the substrate as the substrate connects with the compound (Koshland Jr, 1958). The
Henri–Michaelis–Menten mathematical statement portrays the chemical energy with the assistance of
the rate equation (Michaelis and Menten, 1913).

Ε + S Ε + P ΕSk f kcat kr

In the above equation, E denotes the enzyme, S denotes the substrate, ES denotes the enzyme–
substrate complex, P denotes the product, and kf, kr, and kcat denote the rate constants. Monod,
Wyman, and Changeux (Changeux and Edelstein, 2005) added to the hypothesis of allosteric regulations.
Allosteric sites are binding sites on catalysts, which shape feeble noncovalent bonds with the coupling
substrates. Allosteric collaborations can both restrain and enact catalysts along these lines, controlling
the chemical movement (Changeux and Edelstein, 2005). Enzymes are mainly proteins with essential,
auxiliary, tertiary, and quaternary structures. Essential structures are made out of amino acid sequences
with C-terminal and N-terminal areas.

Optional structures incorporate alpha helices and beta sheets. There are super optional structures,
which are a mix of the alpha helices and beta sheets (Fig. 24.2). Tertiary structures are three-
dimensional structures of the optional structure and amino acids. The accompanying outline (Fig. 24.3)
demonstrates the folding of a polypeptide chain, illustrating the progressive system of the protein
structure from the essential structure or amino acid sequence through the auxiliary structure and the
tertiary structure (Dill et al., 1995). For a protein to be organic dynamic, two or more polypeptide chains
are obliged, and these polypeptides are called subunits. A combination of these subunits adds to the
quaternary structure. A few catalysts require extra nonprotein chemical groups called cofactors or
coenzymes for productively catalyzing the responses. Samples for cofactors incorporate heme, flavin,
nicotinamide, ubiquinone, etc. Table 24.1 shows different cofactors and theirs catalysts. The following
section further discusses the significance of proteins in industries and how they have effectively
supplanted the chemical catalysts. Significance is given to imperative compounds used in biorefineries.

24.2 CHEMICAL CATALYSIS VERSUS BIOCATALYSIS

The previous section gave some thought regarding the general system of protein action. This section will
focus on why enzyme-based catalysis is better over the ordinary chemical catalysis, particularly in
biorefineries. Industries that incorporate chemicals depend on nonrenewable vitality and assets. The
effluents from such industries are harmful to the ecosystem, as they are generally not recyclable or
degradable. On the other hand, enzymes are naturally occurring biocatalysts and can catalyze complex
reactions in an extremely proficient way. There are about 3000 enzymes of which just 150–170 are
utilized commercially (Fischer, 1894). Biocatalysis works at lower temperatures and produces less toxic
waste, outflows, and byproducts in contrast to the conventional chemical processes. New biocatalysts
with enhanced selectivity and better catalytic activity are utilized as a part of different assembling and
waste-degrading processes (Koshland Jr, 1958; Michaelis and Menten, 1913; Changeux and

Edelstein, 2005; Dill et al., 1995). Biological generation frameworks are intrinsically appealing

in light of the fact that they utilize the essential renewable assets like sunlight, water, and carbon
dioxide to yield high-quality products that generally have no or very low toxicity, thereby not
compromising the safety of the environment (Fischer, 1894). A report, “New Biocatalysts: Essential Tools
for a Sustainable 21st Century Chemical Industry”, throws light on the crucial role of biocatalysts in
establishing a sustainable chemical industry. Table 24.2 presents the differences in crucial parameters
between the chemical and bioprocess (Gavrilescu, 2004).

24.3 ADVANTAGES OF BIOCATALYST-BASED PROCESSES

1. Biocatalysts are faster in action and more adaptable than identical chemical catalysts. 2. Biocatalysts
can catalyze an expanded scope of responses, stable at high temperatures and highly soluble. 3. Novel
equipment has made the designing of enzymatic catalysts easier so that enzymes that are more
appropriate to catalyze a reaction can be designed (Gavrilescu, 2004). In contrast to conventional
chemical catalysis, enzyme catalysis is highly specific (Dill et al., 1995; Gavrilescu and Nicu, 2004) and
works under ambient temperatures, pressures, and pHs (Michaelis and Menten, 1913; Van Berkel,
2000). Unlike numerous procedures of conventional synthetic chemistry, enzymes require nontoxic and
noncorrosive conditions. Comparisons of the costs, environmental impacts, and reaction conditions of
chemical- and bio-based synthesis are given in Table 24.3.

24.4 IMPORTANCE OF BIOCATALYSTS OVER CHEMICAL CATALYSTS

1. They have high chemo-, regio-, and stereo-selectivities.

2. They need gentle response conditions, particularly when the substrate or product is chemically or
thermally labile

3. Biocatalysis can ordinarily be performed in fluid environments (Fischer, 1894).

24.5 ENZYMES IN BIOREFINERY

It is understood that metabolic building, or atomic level control of metabolic pathways in full or part,
provides transgenic organisms with new and improved abilities for delivering chemicals. A future
bioethanol-based chemical industry, for instance, will depend on biotechnology for the following:

1. generation of high-return, transgenic corn-mixed bags having starch that is promptly opened for
enzymatic hydrolysis to glucose;

2. production of designed compounds for a significantly enhanced bioconversion of starch to sugars;

3. genetically improved ethanol tolerant microorganisms that can quickly mature sugars to ethanol; and
4. ability to recuperate ethanol utilizing high-proficiency, low-cost biohandling (Koshland Jr, 1958).
Whereas the above features of an enzyme-based process are advantageous for biofuel

generation, the following section will focus on enzyme applications in biorefineries