BIOCHEMICAL ENGINEERING

Biochemical engineering is concerned with

conducting biological processes on an industrial
scale, providing the link between biology and
Chemical Engineering. It is concerned with

Study of the numerous fermentation process in

which the production of alcohols, organic acids,
glycerol, acetone and other substance are end
product.

Study of certain aspect of food manufacturing

processes such as the making of cheese, butter
and pickles in which microbic agency takes a
significant part.

Study of food conservation methods.

Study of biological waste disposal.

Common Features of Biological System

Organism share a common physical structure in

that the cell is the fundamental unit structure of
living organism.

They share common chemical composition as

shown by the present of protein, DNA and RNA.

They perform common metabolic activities; while

these metabolic or chemical activities differ in
detail, the resemblance are more than the
differences.

Biochemical Industries:

1.
Food Industries
3.
Brewery And Distilling Industry
2.
Pharmaceutical Industry
4.
Waste Treatment

Products of Microbial Activity Having

Commercial Importance:

Antibiotics Penicillin; Streptomycin,

Tetracyclines
Organic Solvents ethanol, butanol, acetone
Gases CO2, H2, CH4
Beverages wine, beer, distilled spirits

Foods cheese, fermented milk, soy sauce,

vinegar, citric acid, yeast

Flavoring Agents Monosodium Glutamate, soy

sauce, Nucleotides

Organic Acids lactic acid, acetic acid, butyric

acid, propionic acid, citric acid
Glycerol

Amino Acids lysine, alanine

Steroids

Food and Feed Yeast Bakers Yeast, Brewers

Yeast, Fodders Yeast

Legume Inoculant

Bacterial Insecticides
Vitamins Vit. B12, Riboflavin, Vit. A,
gibberilline
Enzymes amylase , protease, pectinase, invertae

Cell and Its Parts

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cytoplasm

mitochondria nucleus

nucleolus

nuclear membrane

cell membrane

Chromosome (chromatin)

Cell membrane composed of proteins and lipids serve as a barrier from the external
environment. It is semipermeable and selectively permit passage of substance into the cell
while allowing exit of waste.
Cytoplasm contain other organelles and the nucleus

Nucleus regulate metabolic and reproductive activity

Mitochondria contain the respiratory system.

Chloroplasts mitochondria of photosynthetic organism.

Golgi bodies transport materials of cellular product.

Cell wall protective substance of the cell.

Characteristics of Biological Material

Microorganism are chemically very similar to higher animal cells and they can perform
many of the same biochemical reactions. Generally, microorganism exists as single cell.
There are four groups of organism which a biochemical engine is interested namely:

Bacteria

Fungi including yeasts and actinomycetes

Protozoa including algae

Viruses including bacteriophage

The characteristics of biological material could be summarized:

Bacteria contain 50-60% protein, yeast 40-50% and mold 20%, most of which in enzymic.
Some specie have a high lipid (or carbohydrate) content but generally, this is less than 10%.
Viruses contain up to 50% nucleic acid, bacteria 20%, yeast 10% and mold 3%.
Viruses are obligate parasite and require a specific host cell for their multiplication, often
killing the lost in the process. Bacteria are largely unicellular and haploid (one set of genes)
with the nuclear material lying free in the cytoplasm. They multiply by simple fission,
through a primitive mating system had been found in the enteric

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bacteria. Fungi are multicellular with many migrating nuclei. They multiply by apical
extension of the hythal filaments may produce both sexual and a sexual spores. Yeast,
however grow by budding from single ovoid cell.

The microbial cell genetic properties are preserved in their DNA. Four nucleotive (NMP,
TMP, CMP, GMP) are polymerized in two strands bonded to each other to form a double
nelix. The base are arranged such that adenine in one strand in complementary to thiamin in
the other, while guanine is complementary to cytosine. When the DNA replicates, the order
of bases in DNA is conserved by the two strands separating, each acting as template, while
nucleotive complementary to those existing strand are polymerized to those existing strand
are polymerized to form two double-stranded molecules identical with the original one.
Errors may occur in the copying process which give rise to mutant.

When protein is required by a cell, the information for its synthesis is transcribed from one
strand of the DNA into m-RNA; the order of bases in DNA code for the order of bases in
m-RNA. The m-RNA associated with ribosomes (non specific, relatively stable organelles)
where the protein in synthesized. The amino acid are activated and associate with a specie
of t-RNA. The t-RNA has a triplet base (anticodon) which is complementary to the triplet of
bases (codon) in m-RNA specifying the particular amino acid.

Each amino acid is specified by at least one codon but may have as many as six at the
ribosome, amino acids are polymerized into protein in the order specified by m-RNA; the
growing polypeptide is attached to the ribosome until the last amino acid is added, when the
ribosome is free to combine with new m-RNA. Free of ribosome, m-RNA is hydrolyzed
and its component nucleotive enters the nucleotive pool for resynthesis.

CELL HIERARCHY Order of molecular organization of cell

Ribase

Nucleic

Carbamyl

Mono-nucleotides
phosphate

Acids
CO2
alpha-keto

Simple

H2O

Proteins

Enzymes

Nucleus
acids

sugars

Complexes

Mitochondria
CELL

N2
Phospho-

Ribosomes
Chloroplasts

Simple

Polysaccharides

Contracile

Pyruvato

sugars

systems

Malate
Acetate

Fatty

Acid
Lipids

Malonate

Glycerol
Precursors

Intermediates

Building Blocks

Macromole-Cells
(MW = 18-44)

(MW = 50-250)

(MW = 100-350)

3 9
(MW = 10 -10 )

Supramolecular
Cell Organwelles Assemblies

6 9
(Particle WT. = 10 -10 )

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HAECKEL THEORY division in the living world

Plant kingdom Flora

Animal kingdom Fauna

Microorganism Protist

Higher Protist or Eucaryotes

Lower Protist or Procaryoutes

Cellular Organisms (3 types)

Unicellular

Multicellular

Coenocytic

Chemical Make-Up of Bacterial Cells:

Proteins made up of amino acids joined by peptide linkage

Lipids or Fats made up of fatty acids; water insoluble components of the cells that can be
extracted by nonpolar solvents.

Carbohydrates polyhydroxy aldehayde or ketone

Nucleotides monomeric units of nucleic acid macromolecules third OH of one

nucleotide and fifth OH of another nucleotide

Cultivation:

Chemical conditions
Temperature

Gas requirement

pH

Other factors

Isolation:

Pour Plate suspension of cell is mixed with the melted agar at 45C and poured into a
petri dish. When agar solidifies, cells are immobilized in the agar and grow into colonies.

Streak Place the sterile, melted and cooled medium is first poured into a sterile petri dish
and allowed to harden thoroughly; then the surface of the hardened agar is inoculated by
streaking the needle of swab across it.
Bacterial Culture System

Batch Culture growth curve characterized by the lag phase, exponential phase, stationary
phase and decline phase.

Lag Phase physiological adjustment of the organism to the environment following

inoculation.

Logarithmic or Exponential Phase exhibits a constant rate of growth where the rate of
increase of cells can be expressed as:

dx
Ux

dt

U = growth

x
=
microbial concentration

t
=
time

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Stationary Phase growth ceases completely due to the exhaustion of nutrients,
accumulation of waste products, change in pH.

Decline Phase irreversible loss of ability to regenerate or reproduce.

Continuous Culture utilizes the bacterial kinetics of the exponential phase. This demands
a continuous flow-in of the nutrients of growth and the corresponding outflow of the
products of the culture.

Sterilization:

Chemical

Mechanical

Thermal
Radiation

5.

Energy Yielding Metabolism : Metabolism of carbohydrates, e.g. glucose.

Anaerobic Fermentation

Glycolysis or EMP Pathway

Alcoholic Fermentation

Aerobic Respiration TCA (Tricarboxylic Acid Cycle) or Krebs Cycle

ENZYMES

Sources plant, animal organs, microbial cells

Types:
Endo enzyme or Intracellular enzyme

Exo enzyme or Extracellular enzyme

Enzyme Theories:

Lock and key theory (Emil Fischer) illustrates the precise fit between enzyme and
substrate.

Induced Fit Theory (Koshland) Enzyme undergoes structural or conformational changes

brought about or induced by the substrate.

Michaelis- Menten Theory Enzyme activity depends on substrate concentration.

V
Equation: Vm x S
 V = Velocity of enzymatic reaction
Km S
Vm = limiting (maximum) velocity S = substance
concentration

Km = Michaelis Menten constant

EMP Pathway

Glucose
ATP

ADP

Glucose-6-Phosphate

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Fructose-6-Phosphate
ATP

ADP

Fructose-1,6-Diphosphate

Clyceraldehyde-3-Phosphate Dihydroxyacetone Phosphate

1, 3-Diphosphoglyceric Acid

2 ADP

2 ATP

3-Phosphoglyceric Acid

2-Phosphoglyceric Acid

Phospoenolpyruvic Acid 2 ADP 2 ATP

Pyruvic Acid

2 NADH

2 NAD Lactic Acid

TCA or Krebs Cycle

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Proteins
Carbohydrate

Lipid

Amino Acids
Pyruvate

Fatty acids

CO2
2H
Acetyl Co-A
Oxaloacetate

Citrate
CO2
Malate

Cis-Atconitate
Fumarate

Isocitrate
Succinate

Ketoglutarate
2H

2H

2H
2H
Electron Transport and Oxidative Phosphorylation

ATP

General Properties of Enzymes:

Enzymes are proteins

Enzymes have electrical charges which depends on the pH and two pK values

Enzymes have definite three dimensional structures

primary structure amino acid sequence of the polypeptide chain (most important and
specific structure; determines secondary and tertiary structures)

secondary structure spatial arrangement of polypeptide chains into helices, pleated sheet
and random coil structures.

tertiary structure three dimensional arrangement of helices, pleated sheet

and random coil structures in enzyme.

quaternary structure aggregation of enzyme submits or molecules into multi submit or

multi enzyme complexes.

Non-covalent bonds that contribute to the three dimensional structure of enzymes:

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Hydrogen bond Interpeptide Hydrogen Bond; Side Group Hydrogen Bond.
Ionic Bond (electrostatic attraction)

Non-polar Side Chain Interaction (Hydrophobic Bond)

Polar Side Chain Interaction

Enzymes are efficient catalysts and catalyze a wide range of chemical reactions.

Six General Classes of Enzymes and Functions:

Oxido-reductases oxidize or reduce substrates by transferring hydrogen or electrons

Transferases remove groups (excluding hydrogen) and transfer them to acceptor

molecules (excluding water); transfer of groups.

Hydrolases for hydrolytic reactions. It catalyze the splitting of a covalent bond of the
substrate and that of a water molecule with the subsequent addition of the hydrogen and
hydroxide to the two fragments of the substrate molecule.

Lyases remove groups from the substrate after than by hydrolysis to form a double bond
or conversely, add groups to the double bonds.

Isomerases catalytic isomerizations. It causes isomerization to the substrate.

Ligases or Synthetases cause condensation of two molecules by splitting a phosphate

bond.

Enzyme Nomenclature:

Based on their source

Papais papaya
Bromelin pineapple

Pepsin stomach

Based on the substrate used Urease urea Protease protein Cellulase cellulose Sucrase
sucrose

Based on their function Lysozyme to loosen Catalase catalyst Oxidase oxidation

Systematic method

Substrate nature of reaction

glucose oxidase

glycerol ester hydrolase

Definition of Terms:

Aerobe An organism that grow in the presence of air or O2.

Amino Acid A sub-unit of the protein structure.

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Anaerobe An organism that grow in the absence of oxygen.

Autolysis dissolution of the cells by self-produced lytic enzyme.

Autotrophs organism that grow or uses CO2 as its principal carbon source.

Auxotrophs nutritional mutant which require one or more nutrients in addition to those
required by its wild type.

Bacteriacidal an agent that kills bacteria.

Bacteriostatic an agent that inhibits the bacterial growth by its presence without causing
death.

Binary fission cell division where the mother cell split into daughter cells.

Biomass cellular mass e.g. bacterial cell or fungal mass.

Biosynthesis a change occurring within the bacterial cell such as the building up of
protein, carbohydrates from simple cell.

Broth a liquid medium wherein an active growth of microorganism is in progress;

fermentation liquor wherein the process of fermentation is in progress.

Beer fermentation liquor wherein fermentation process had just terminated.

Enzyme an organic catalyst produced by the cell. Constitutive always present in the
cell; substrate independent; induced present in the cell only when needed, substrate
dependent; extracellular (exo enzyme) activity is outside the cell; digestive or hydrolytic
in its activity. Intracellular (endoenzyme) activity is inside the cell usually involved in the
bio-synthesis of macromolecules and energy-yielding reactions.

Facultative having the characteristics that permit alternate responses under different
conditions; e.g. facultative anaerobe can grow aerobically.

Fermentation process in which chemical changes are brought about in an organic

substance through the action of enzyme elaborated by microorganism.

Generation time the time required for a mother cell to divide into two daughter cells.

Inoculant starting microbial culture in fermentation process.

Heterotroph an organism that uses predominantly organic carbon for energy and
photoplasmic building block.

Lag phase early period of growth where the organism adjust to its new environment.

Logarithmic growth phase in the microbial life cycle when the cells are dividing
exponentially.

Mesophile an organism that grow optimally at 20C to 37C

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Microbial something of the organism; from the word microbe which means
microorganism.

Microaerophile an organism that prefers to grow under reduced oxygen tension but
sometimes increased carbon dioxide tension.

Mutation a chemical change in the genetic code of the cell

Mycelium a mass of hyphae of a mold or actinomycetes colony.

Obligate strict or restricted; e.g. obligate aerobe can grow only in the presence of oxygen;
obligate thermophile can grow only at elevated temperatures in relation to others.

Phage a bacterial virus

Photoplast that part of t he cell which includes the cell membrane and its content.

Phototroph an organism exhibiting wild type nutrition.

Putrefaction decomposition of protein under anaerobic conditions.

Respiration oxidation of the chemical compound by the organism coupled with the
released of energy; Aerobic-final oxygen acceptor is an inorganic substance like SO3, NO3
other than molecular oxygen.

Strain a classification of specie of an organism identifying is a part from the rest by virtue
of specificity of source.

Substrate nutrient for cellular growth.

Thermophile an organism that grow optimally above 45C.

Viable state of the living cell where it is still capable of division.

D N A deoxyribonucleic acid holder of the genetic information of the cell.

R N A ribonucleic acid responsible for the coding and the transfer of genetic
information.

SCP single cell protein protein obtained for the growth or culture of unicellular
microorganism.

Cultivation process of propagating organism by providing the proper environmental

condition such as nutrient, temperature and PA.

Isolation separation of a particular microorganism from the mixed population that exist in
nature.

Psychrophile organism whose optimum temperature for growth is 10C.

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Inoculum serum containing microorganism.

Some useful utilization of microorganisms are the following:

Food Processing:

soy sauce aspergillus sojae vinegar acetobacter

nata acetobacter xylinum pickles lactic acid bacteria

milk products like cheese and sour cream tempe soybean rhizopus
oligosporus ontjom peanut cake neurospora sitophilia

Alcoholic Beverages

wine, beer, sake, tapuy, basi, whisky, rum, lambanog

Fuel

Alcohol saccharomyces cerevisial xymomones mobilis biogas mixture of

microorganisms
methane producing bacteria

Production of:

antibiotics

penicillin penicillium chrysogenum streptomycin streptomyces griseus

tetracyclines streptomyces aureofaciens polymixins bacillus polymyxa

amino acids
glutamic acid corynabacterium glutamicum

vitamins

B-carotene choanspore conjuncta Blakaslea trispora

riboflavin ashbye gossypti

Enemothecium ashbili
Vitamin B12
hormones
Gibberallins Gibberella fugikunol