Microbial Products

A. Primary Metabolites: log phase, use nutrients fast, produce PM

B. Secondary Metabolites: depletion of nutrients, growth retards,
produce SM

Primary Metabolites: Vitamins

Vitamins: cannot be synthesized by higher organisms

But microorganisms are capable of synthesizing (gut)

Studies reveal vitamin deficiencies

Thiamine
Reported beneficial health effects
Riboflavin
Growing vitamin market demand (cost
Pyridoxine
effective)
Folic acid
Genetically engineered MO as alternatives to
Pantothenic acid
chemical synthesis
Biotin
Vitamin B12
Ascorbic acid
b- carotene (provitamin A)
Ergosterol (vitamin D)

Vitamins
Fat soluble
Carotenoids
b-carotene (provitamin A)
Astaxanthin
Poly unsaturated Fatty acids (PUFA; vitamin F)
Docosahexaenoic acid (DHA)
Arachidonic acid (ARA)

Ergosterol (vitamin D)

Water soluble
Riboflavin (vitamin B2)
Cobalamin (vitamin B12)
L-Ascorbic acid (Vitamin C)

R-Pantothenic acid (vitamin B5)

D-Biotin (vitamin H or B7)
Vitamin B1 (Thiamine)
Vitamin B6 (pyridoxol)
Folic acid

Vitamin B12 or Cyanocobalamin

Water soluble vitamin ; complex sructure
Has role in functioning of brain and nervous system, formation of
blood
Contains rare element cobalt

Deficiency causes pernicious anemia which is an causes low Hb, less

RBCs
Pernicious anemia: autoimmune disorder, parietal cells (stomach)
responsible for secreting intrinsic factor are destroyed. Intrinsic
factor is crucial for the normal absorption of B12, so a lack of
intrinsic factor, as seen in pernicious anemia, causes a deficiency of
Vitamin B12
dietary reference intake for an adult ranges from 2 to 3g per day

used in treating cyanide poisoning, prevents brain atrophy in

Alzheimers patients

COMMON INGREDIENT IN ENERGY DRINKS

C63 H88 CoN14 O14P

Pyrrole nitrogen

4 Pyrrole units

cobinamide

Corrin ring
Deep red colour due to corrin
ring
Central Co atom
Coordination state 6
4 of 6 coord sites have pyrrole
ring
5 has dimethylbenzimidazole
group
6 is center of reactivity,
variab;e
CN, OH, Me, 5-deoxyadenosyl
for 4 types of B12

nucleotide

6
2

1
3

5,6-dimethyl
benzimindazole

Commercial production

Chemical syn not feasible

Genera known to produce vit B12

Acetobacterium, Aerobacter, Agrobacterium, Alcaligenes, Azotobacter, Bacillus,
Clostridium, Corynebacterium, Flavobacterium, Micromonospora,
Mycobacterium, Nocardia, Propionibacterium, Protaminobacter, Proteus,
Pseudomonas, Rhizobium, Salmonella, Serratia, Streptomyces,
Streptococcus and Xanthomonas

Most commonly used for industrial production are

20mg/L

Streptomyces griesus
Pseudomonas denitrificans (aerobic)
Salmonella typhimuriu (anaerobic)
Propionibacterium shermanii
GRAS by FDA
(anaerobic)
(Generally Regarded As Safe)

Sanofi-Aventis (FRENCH) use genetically engineered versions to produce vit

B12 under specialized conditions from Propionibacterium since they have no
endotoxins or exotoxins

P. denitrificans also used after strain modification; mutant more efficient

than wild type

Commercial production

Produced in continuous culture with 2 fermenters in series

Addition
of
5,6dimethylbenzimidazol (0.1%)

Glucose
Corn steep
Betaine (5%)
Cobalt (5ppm)
pH 7.5 +

Propionibacterium
freudenreichii

Anaerobic
70h

Cobinamide production
and accumulation

Aerobic
50h

KCN added
CYNACOBALAMIN
80% purity
Used as feed additive
Betaine: sugar beet molasses

Filtrate

Nucleotide synthesized
Combined with cobinamide
To yield 2ppm of cobalamin
Acidification of culture
To 2-3pH/ 100oC
Filter to remove cell debris

Commercial production
ANAEROBIC PHASE

AEROBIC PHASE

2-4 DAYS
5-deoxyadenosylcobinamide produced

5,6-dimethylbenzimidazole
is
added and gets incorporated to
form 5-deoxyadenosylcobalamin

During the 7-day fermentation run, adenosylcobalamin is predominantly

secreted from the biomass and accumulates in the fermentation broth
in milligram amounts.
The down- stream steps comprise filtration, cyanide treatment,
chromatography, extraction, and crystallization yielding vitamin B12 in
high purity.
If to be used for treatment further purification (95-98% Purity)

Commercial production

Pseudomonas denitrificans: strain improvements resulted in increase in yeild

From 0.6mg/L to 60mg/L
Glucose : common carbon
Alcohols (methanol, ethanol, isopropanol)
Hydrocarbons(alkanes, decane, hexadecane)

With methanol 42mg/L was obtained using Methanosarcina barkeri

Riboflavin (Rf) or Vitamin B2

Water soluble
Essential for growth and reproduction; key role in energy metabolism, ketone bodies,
fats, CHO and protein metabolism
Deficiency leads to cheliosis (fissures around mouth), glossitis (purple tounge) and
dermatitis
Required in coenzymes FAD (flavin adenine dinucleotide) and FMN (flavin
mononucleotide)
Used as an orange-red food colour additive, designated in Europe as E101
7,8-dimethyl-10- (D-19-ribityl) isoalloxazine
Participates in O-R reactions
Flavin is ring moiety with yellow
colour to oxidized form

Isoalloxazine ring

Isoalloxazine ring

Ribitol

FAD
E101

FMN
E101a

genes encoding the riboflavin biosynthetic enzymes are well conserved among bacteria and fungi

INDUSTRIAL USE
Processed food is often fortified by the use of riboflavin as a colorant or vitamin
supplement.
The main application (70%) of commercial riboflavin is in animal feed, since productive
livestock, especially poultry and pigs, show growth retardation and diarrhea in case of
riboflavin deficiency.
According to a report by SRIC, a consulting company in Menlo Park (California), in 2005
the need for industrially produced riboflavin was estimated at 65007000 tons per
year.

Commercial production
Glucose
1/3 production by
direct fermentation
rd

Acetone butanol fermentation

Clostridium acetobutylicum
C. butylicum riboflavin as
by product

50% by biotransformation
using Bacillus pumulis
D-ribose
20% production by Chemical synthesis

Riboflavin

Ashbya gossypii
Candida famata
Bacillus subtillis (genetically modified)

Commercial production
Phase I

use of glucose, accumulation of pyr, pH acidic, growth stops, no Riboflv

Phase II decr pyr, incr in ammonia, alkalinity incr, prod of Riboflv in form of FAD and FMN
Phase III autolysis, cell disruption, release of free FAD, FMN and riboflv

Carbon sources: glucose, acetate, methanol, aliphatic hydrocarbons

Major riboflavin producers are DSM Nutritional Products

(Switzerland) and Hubei Guangji (Hubei Province, China), both using
genetically engineered B. subtilis production strains, and BASF (first in
Germany but now in South Korea), employing genetically engineered A.
gossypii.

Ascorbic acid or Vitamin C

Used in collagen biosynthesis, protects against nitrosamines, free radicals

Deficiency causes scurvy

Precursor for its chemical synthesis can be obtained by biological methods

feed applications of L-ascorbic acid account for only 10%, whereas the main
uses are in the
pharmaceutical industry (50%),
food (25%), and
beverages (15%).
Pharmaceutical applications include stimulation of collagen synthesis (especially
cosmetic products) and high antioxidant capacity, used for the reported
health benefits in the prevention of flu, heart diseases, and cancer, as well as
an antidote for poisoning.
The food and beverage industry predominantly exploits the antioxidant
capacity of L-ascorbic acid to extend durability, prevent discoloration,
and to protect flavor and nutrient contents of their products.

Submerged bioreactor fermentation

Erwinia sp.
Acetobacter sp.
Gluconobacter sp.

2,5-diketogluconic acid

2,5-diketogluconic acid
reductase

D-glucose

(200g)

Glucuronic acid

Corynebacterium sp.

2-keto L-gluconic acid

Bacillus
megaterium

L-ASCORBIC ACID

D-sorbitol
sorbitol dehydrogenase
Acetobacter xylinum,
A,suboxydans

Cloning of gene

L-Sorbose
chemical oxidation

2,5-diketogluconic acid
Reductase of

2 keto L gulonic acid

Corynebacterium into
Erwinia herbicola

Enol form of
2 keto L gulonic acid
Reichstein Grussner synthesis

acid treatment
L-ASCORBIC ACID (100g)

Gluconolactone

L-Gluconolactone
L-Gluconolactone
dehydrogenase

L-ASCORBIC ACID

- carotene or provitamin A
Provitamin A -----> Vitamin A (intestine)

Fat soluble
Deficiency leads to night blindness
Best source is liver and whole milk also coloured fruits and vegetables

Isoprene derivatives
Tetraterpenoids with eight isoprene residues
400 naturally occurring carotenoids: b-carotene, a-carotene, d-carotene, lycopene,
zeaxanthin

Carotenoids Used as food colorants and animal feed supplements for poultry
and aquaculture, carotenoids play an increasing role in cosmetic and
pharmaceutical applications due to their antioxidant properties.
The pigments are often regarded as the driving force of the nutraceutical
boom, since they not only exhibit significant anticarcinogenic activities but also
promote ocular health, can improve immune response, and prevent chronic
degenerative diseases.

Commercial production
Submerged Fermentation process

Microbial fermentation
Blakeslea trispora (high yeild; 7g/L)
Phycomyces blakesleeanus
Choanephora cucurbitarum

Corn starch, soyabean meal, -ionone, antioxidants

stimulators

Trisporic acid: act as microbial sex hormone, improves yield

-Ionone: incr -carotene syn by incr enzyme activity
Purified deodorized kerosene increases solubility of hydrophobic
substrates

Recovery: carotene rich mycelium used as feed additive

Mycelium is dehydrated by methanol, extracted in methylene chloride
and crystallized which is 70-85% pure

DSM Nutritional Products (Switzerland) and BASF (Germany)

dominate the market with their chemical synthesis processes,
but Chinese competitors are catching up.

Halophilic green microalgae Dunaliella salina. It accumulates the pigments in oil

glo- bules in the chloroplast interthylakoid spaces, protecting them against
photoinhibition and photodestruction.
Excessive pigment formation in D. salina is achieved by numerous stress factors
like high temperature, lack of nitrogen and phosphate but excess of carbon, high
light intensity, and high salt concentration, the latter two having the highest
impact.
Dried D. salina biomass for sale contains 1016% carotenoids, mainly -carotene.
In addition crystalline material obtained after extraction with edible oil is also
sold.

Primary Metabolites: Organic Acids

Organic acids are produced by through metabolisms of carbohydrates. They accumulate in
the broth of the fermenter from where they are separated and purified.

Glycolysis
Krebs cycle

I. Terminal end products

(pyruvate, alcohol)

lactic acid
Propionic acid

II. Incomplete oxidation of sugars

(glucose)

citric acid
Itaconic acid
Gluconic acid

III. Dehydrogenation of alcohol with O2

acetic acid

Manufactured on large scale as pure products or as salts

CITRIC ACID: industrial uses

Flavoring agent
In food and beverages
Jams, candies, deserts,
frozen fruits, soft
drinks, wine
Antioxidants
preservative

and

Chemical industry
Antifoam
Treatment of textiles
Metal
industry,
pure
metals +citrate (chelating
agent)

Acidifyer
Flavoring
Chelating agent
Primary metabolite
Present in all organisms

Agent for stabilization of

Fats, oil or ascorbic acid
Stabilizer for cheese
preparation

Pharmaceutical industry
Trisodium citrate (blood
preservative)
Preservation of ointments
and cosmetics
Source of iron

Detergent cleaning industry

Replace polyphosphates

Commercial Production
Strains that can tolerate high sugar and low pH with reduced
synthesis of undesirable by products (oxalic acid, isocitric acid,
gluconic acid)
Glucose
Glucose
Pyruvate

Aspergillus niger
A. clavatus
Pencillium luteum

MEDIUM
CYTOPLASM

Pyruvate
CO2

Pyr carboxylase

OXA
Malate

Pyruvate
Pyr Dehydrogenase

Acetyl CoA

MITOCHONDRIA

Malate
CO2 OXA

Fumarate

Succinyl CoA

Citrate
synthase

citric acid

a-KG

100g sucrose --- 112g any citric acid or 123g citric acid-1hydrate

Factors for regulation

CARBOHYDRATE SOURCE: sugar should be 12-25%
Molasses (sugar cane or sugar beet)
Starch (potato)
Date syrup
Cotton waste
Banana extract
Sweet potato pulp
Brewery waste
Pineapple waste
High sugar conc incr uptake and production of citric acid
TRACE METALS:
Mn2+, Fe3+, Zn2+ incr yield
Mn2+ incr glycolysis
Fe3+ is a cofator for enzymes like aconitase
pH: incr yield when pH below 2.5, production of oxalic acid and gluconic acid is
suppressed and risk of contamination is minimal
DISSOLVED O2: high O2, sparging or incr aeration can affect if interrupted
NITROGEN SOURCE: addition of ammonium stimulates overproduction, molasses is
good source of nitrogen

Citric acid production

Surface fermentation

Solid

liquid

submerged fermentation

Stirred
Bioreactor

Airlift
bioreactor

N alkanes (C9-C23) can also be used to produce citric acid; can

result in excess production of isocitric acid

ACETIC ACID: industrial uses

ACETIC ACID

Incomplete oxidation of ethanol

Vinegar is prepared from alcoholic liquids since ceturies

NAD+

NADH +H+

NADP+

NADP +H+

CH3 CH2OH---- CH3CHO-------- CH3CH(OH)2 -------

Ethanol
acetaldehyde
acetaldehyde hydrate
Alcohol
dehydrogenase

CH3COOH
acetic acid

Acetaldehyde dehydrogenase

Gluconobacter, Acetobacter with acid tolerant A. aceti

One molecule of ethanol one molecule of acetic acid is produced
12% acetic acid from 12% alcohol

Clostridium thermoaceticum

It is an obligate anaerobe, Grampositive, spore-forming, rod-shaped,

thermophilic
organism
with
an
optimum growth temperature of 55
60 o C
and optimum pH of 6.6
6.8.

VINEGAR: 4% by volume acetic acid with alcohol, salts, sugars and esters
flauoring agent in sauces and ketchups, preservative also
Wine, malt, whey (surface or submerged fermentation process)
Surface: trickling generator; fermentale material sprayed over surface, trickle thro
shavings contaning acetic acid producing bacteria; 30oC (upper) and 35oC (lower).
Produced in 3 days.
Submerged: stainless steel, aerated using suction pump, production is 10X higher

Clostridium thermoaceticum (from horse manure) is also able to utilize fivecarbon sugars:
2C5H10O5 --- 5CH3COOH
A variety of substrates, including fructose, xylose, lactate, formate, and
pyruvate, have been used as carbon sources in an effort to lower substrate
costs. This factor is also important if cellulosic renewable resources are to be
used as raw materials.
Typical acidogenic bacteria are Clostridium aceticum, C. thermoaceticum,
Clostridium formicoaceticum, and Acetobacterium woodii. Many can also reduce
carbon dioxide and other one-carbon compounds to acetate.

1mol
2moles

1mol

2moles
1mol

CODH

These enzymes are metalloproteins; for example,

CODH contains nickel, iron, and sulfur; FDH
contains iron, selenium, tungsten, and a small
quantity of molybdenum; and the corrinoid enzyme
(vitamin B12 compound) contains cobalt. C.
thermoaceticum does not have any specific amino
acid requirement; nicotinic acid is the sole essential
vitamin

LACTIC ACID: industrial uses

Technical grade
20-50%

>90%
Intestinal treatment
(metal ion lactates)

Food additive
(sour flour and
dough)

Ester manufacture
Textile industry

Glucose
G3P
G3P dehydrogenase

Pharmaceutical grade

Food grade
>80%

NAD+

Lactic acid

NADH +H+
1,3-biphosphoglycerate
Pyruvate

LDH
(Lactate dehydrogenase)

LACTIC ACID
2 isomeric forms L(+) and D(-) and as racemic mixture DL-lactic acid
First isolated from milk
Toady produced microbial

Heterofermentation

Homofermentation

Other than lactate products

only lactate as product

Lactobacillus
L. delbrueckii
L. leichmanni

Mostly one isomer is produced

Glucose

L. bulgaricus
L.helvetii

Whey (lactose)

L.lactis
------L.amylophilus -------L.pentosus
------

Maltose
Starch
Sulfite waste liquor

LACTIC ACID: production process

1mol of glucose gives 2 moles of lactic acid; L lactic acid is predominantly produced

Fermentation broth (12-15% glucose, N2, PO4, salts micronutrients)

pH 5.5-6.5/temp 45-50oC/75h
Heat to dissolve Ca lactate
Addition of H2SO4
(removal of Ca SO4)
Filter and concentrate
Addtion of Hexacyanoferrant
(removes heavy metal)
Purification (Ion exchange)
Concentration
Lactic acid

GLUCONIC ACID: Applications

1. Used in stainless steel manufacturing, leather (can remove rust and
calcareous deposits)
2.
3.
4.
5.
6.

Food additive for breverages

Used in Ca and Fe therapy
Na gluconate used in sequestering agent in detergets
Desizing polyester or polyamide fabric
Manufacture of frost and cracking resistant concrete

Bacteria: Gluconobacter, Acetobacter, Pseudomonas, Vibrio

Fungi: Aspergillus, Penicillium, Gliocladium

intracellular

PQQ

Glucose
dehydrogenase

PQQH2

D-gluconolactone

D-Glucose
Extracellular
Inducible

Glucose
oxidase

FAD
H2O2

extracellular

Bacteria
H2O
Lactonase

Gluconic Acid

fungi
FADH2 Fungi

Catalase

O2

High conc of glucose and pH above 4

H2O2 antagonist for other micro-organisms

Submerged fermentation process

Use glucose from corn
H 4.5-6.5
28-30oC for 24h
Incr supply of O2 enhances yield

PQQ: pyrroliquinoline quinone

coenzyme

ITACNIC ACID: Applications

Aspergillus itoconicus and A.terreus

1. Used in plastic industry, paper industry

2. Manufacturing of adhesives

Cis-aconitic acid undergoes decarboxylation

Itaconic acid Oxidase

Itaconic acid

Itatartaric acid
(-) By Ca to incr yield

SECONDARY METABOLITES
ANTIBIOTICS

BROAD SPECTRUM
Control growth of
wide
range
of
unrelated organisms
Tet, Cm

NARROW SPECTRUM
Control growth
selected number
organisms
Pen, Str

Streptomyces,eg. Tetracyclin, actinomycin D,

of
of

ANTIBIOTICS: applications

1. Antimicrobial agents for chemotherapy

2. Antitumour antibiotics eg. Actinomycin D and mitomycin D
3. Food preservative antibiotics eg in canning (chlortetracycline) or fish or meat
preservation (pimarcin, nisin)
4. Antibiotics in animal feed and veterinary medicine eg enduracidin, tylosin and
hygromycin B, theostrepton, salinomycin
5. Control of plant diseases eg blasticidin, teranactin, polyoxin
6. Molecular biology

MODE OF ACTION OF ANTIBIOTICS

DNA GYRASE

RNA ELONGATION

CELL WALL SYNTHESIS

DNA DIRECTED RNA POLYMERASE

DNA

THF
RIBOSOMES
DHF

RNA

PROTEIN SYNTHESIS
(50S INHIBITORS)
PROTEIN SYNTHESIS
(30S INHIBITORS)

PROTEIN SYNTHESIS
(tRNA)
CYTOPLASMIC
MEMBRANE STRUCTURE
AND FUNCTION
PABA

LIPID BIOSYNTHESIS

SYTHETIC ANTIBIOTICS
Selective toxicity: concept, Paul Ehrlich

1. GROWTH FACTOR ANALOGS:

structurally similar to a growth factor required in a micro-organism; small
differences of analogs in authentic growth factor prevent analog to function
in the cell.
A.

SULFA DRUGS: specifically inhibit bacteria (streptococcal infections)

eg. SULFANILAMIDE: is an analog of PABA (p-aminobenzoic acid) which is
part of folic acid and nucleic acid precursor. Combination:
sulfamethoxazole and trimethoprim; disadvantages and advantages

B. ISONIAZID: important growth factor with narrow spectrum only against

Mycobacterium. It interferes with synthesis of mycolic acids, a cell wall
component. It is an analog of nicotinamide (vitamin). Single most effective
drug against tuberculosis.

2. NUCLEIC ACID BASE ANALOGS

URACIL
PHENYLALANINE
THYMINE

5-FLOUROURACIL (Uracil analog)

p-FLOUROPHENYLALANINE
5-BROMOURACIL (thymine analog)

Addition of F or Br does not alter the shape but changes chemical properties
such that the compound does not function in the cell metabolism, thereby
blocking the nucleic acid synthesis.
These analogs are used in treatment of viral and fungal infections and many of
these occur as mutagens.
3. QUINOLONES:
Antibacterial compounds interfere with bacterial DNA gyrase, prevent
supercoiling (packaging of DNA) eg Flouroquinolones like ciprofloxin (UTI,
anthrax). B. anthracis maybe resistant to pencillin. These are effective in both
G+ve and G-ve bacteria since DNA gyrase is present in all.
Also used in beef and poultry for prevention and treatment of respiratory
diseases.

New generation
Flouroquinolnes

Ouinolones

NATURALLY OCCURING ANTIBIOTICS

FROM BACTERIA, FUNGI
LESS THAN 1% OF 1000S OF ANTIBIOTICS ARE USEFUL BECAUSE OF TOXICITY
OR LACK OF UPTAKE BY HOST CELLS
Natural antibiotics can be artificially modified to enhance their efficacy then they are
semi-synthetic antibiotics

Broad spectrum antibiotics: effective against both gram +ve and gram-ve
Narrow may also be beneficial to target specific group of bacteria eg. Vancomycin:
narrow spectrum effective for gram positive pencillin resistant Staphylococcus,
Bacillus, Clostridium
Targets for antibiotics maybe
ribosomes (Cm and Str for Bacteria and Cyclohexamide for eukarya), Cell
wall, cytoplasmic membrane, lipid biosynthesis, enzymes, DNA replication and
transcription elements
Protein synthesis, Transcription (RNA poly, RNA elongation etc)

Produced By Fungi
B-LACTAMS (b-lactam ring)
Penicillin
Cephalosporins

Produced by Prokaryotes
AMINOGLYCOSIDES (amino sugars with glycosidic linkage)
MACROLIDES (lactone ring bonded to sugars)
TETRACYLINES (Streptomyces)
PEPTIDE ANTIBIOTICS (Daptomycin, (Streptomyces)
PLATENSIMYSIN (Streptomyces)

Beta Lactam Antibiotics

1.
2.
3.
4.

PENICILLINS,
CEPHALOSPORINS,
MONOBACTAMS AND
CARBAPENEMS

PENCILLIN---------LACTAM ANTIBIOTIC

Alexander Fleming

Pencillin G and V (natural)

Penicillium chrysogenum

Pencillin G first clinically useful antibiotic

For Gram positive bacteria

Used for
Pneumococcal
Streptococcal infections

6-AMINOPENICILLIANIC ACID

Ampicillin, carbencillin
Slight modification in N-acyl groups results in semi synthetic penicillin which is able to
act on gram negative bacteria (goes past outer membrane) to act on cell wall
MANY BACTERIA HAVE BETA LACTAMASE HENCE THOSE BACTERIA ARE
PENCILLIN RESISTANT
EG. Oxacillin and Methicillin beta lactamase resistant semi synthetic antibiotics
MECHANISM OF ACTION

Pencillins block cell wall synthesis: transpeptidation (cross linking 2 glycan peptide
chains)
Transpeptidases bind to pencillin hence they are called PENCILLIN BINDING
PROTEINS (PBP)
Newly synthesized bacterial wall is no longer cross linked and has poor strength
PBP also stimulates release of AUTOLYSINS (ENZYMES TO DIGEST CELL WALL)
Osmotic pressure differences cause lysis
VANCOMYCIN: does not bind PBPs but D-alanyl- Dalanine peptide to block
transpeptidation
BECAUSE OF SELECTIVE PROCESS B-LACTAMS DO NOT AFFTECT HOST CELLS
AND MECHANISM IS UNIQUE TO BACTERIA

MECHANISM OF ACTION
Natural penicillin: i.e. V and G are effective against several gram positive bacteria
They are effective against b-lactamase producing MO (enz which can hydrolyze penicillins)
Eg. Staphylococcus aureus
Production of penicillin is used: 45% (human), 15% (animal health) and
45% for production of semi synthetic penicillin

P. notatum, P.chrysogenum and its mutant strain which is a high yeilding strain (Q176)
Genetically engineered strains for improved pencillin production are being used now

UDP deriv of NAM and NAG are

synthesized
Sequentially aa are added to UDPNAM to form NAM -pentapeptide
ATP is used, no tRNA or ribosomes
involved in peptide bond formation

UDP tansfers NAG to bactoprenolNAM peptapeptide. For

pentaglycine use special glycyltRNA moc but not ribosomes

Transport of completed NAM-NAGpepntapeptide across membrane

Transfer of UDP-NAMpentapeptideto bactoprenol PO4

LIPID I

Bactoprenol carrier moves back

across membrane by losing one PO4
for a new cycle

LIPID II

Attached to growing end of PG

chain and incr by one repeat unit

Bactoprenol is a 55 carbon alcohol and linked to NAM by pyrophosphate

In S. aureus pepntapeptide has L-lys and in

E. coli DAP

UDP glucose

Final step is TRANSPEPTIDATION which creates peptide cross links between PG

chains. The enzyme removes terminal D-alanine as cross link is formed

The b-lactam group of antibiotics includes an enormous diversity of natural and

semi-synthetic compounds that inhibit several enzymes associated with the final
step of peptidoglycan synthesis.
All of this enormous family are derived from a b-lactam structure: a four-membered
ring in which the b-lactam bond resembles a peptide bond. The multitude of chemical
modifications based on this four-membered ring permits the astonishing array of
antibacterial and pharmacological properties within this valuable family of
antibiotics.
Clinically useful families of b-lactam compounds include the penicillins,
cephalosporins, monobactams and carbapenems. Many new variants on the b-lactam
theme are currently being explored. Certain b-lactams have limited use directly as
therapeutic agents, but may be used in combination with other b-lactams to act as blactamase inhibitors.

Co-amoxyclav, for example is a combination of amoxycillin and the blactamase inhibitor clavulanic acid. During cross-linking of the peptidoglycan
polymer, one D-alanine residue is cleaved from the peptidoglycan precursor and
this reaction is prevented by b-lactam drugs.
More recent studies have shown that the activity of this class of drugs is more
complicated and involves other processes as well as preventing cross-linking of
peptidoglycan.

B-lactamase
An increasing number of bacteria are penicillin resistant. Penicillinase-resistant
penicillins such as methicillin, nafcillin, and oxacillin are frequently employed against
these bacterial pathogens.
Although penicillins are the least toxic of the antibiotics, about 1 to 5% of the
adults in the United States are allergic to them. Occasionally a person will die of a
violent allergic re- sponse; therefore patients should be questioned about penicillin
allergies before treatment is begun.

MRSA
VRSA

CEPHALOSPORINS

Cephalosporium: Cephalosporin C
B-lactam ring

Dihydrothiazine ring (6 member)

cefatrioxone

Same mode of action with broader spectrum than penicillins

Resistant to b-lactamases
Hence used to treat infections which are penicillin resistant
Used to treat Nesseria gonorrhea (STD)

Most cephalosporins (including cephalothin, cefoxitin, ceftri- axone,

and cefoperazone) are administered parenterally.
Cefoperazone is resistant to destruction by b-lactamases and
effective against many gram-negative bacteria, including Pseudomonas
aeruginosa.
Cephalexine and cefixime are given orally rather than by injection.

7-ACA: 7- aminocephalosporanic acid nucleus structure in all cephalosporins

G+ = G-

G+ > G-

R1

R2
G+ < G-

TETRACYCLINES

Broad spectrum
Effective for G+ and G- (mycoplasmas, rickettesia, chlamydia)
Used for combatting stomach ulcer (Helicobacter pylori)
Inhibit protein synthesis by blocking binding of amino acyl tRNA to ribosome (A site)

BASIC STRUCTURE

Napthacene ring
Chlortetracycline and oxytetracycline are most commonly used in human and veterinary
diseases and for preservation of meat, fish and poultry

Three members of the tetracycline family.

Tetracycline lacks both of the groups that are
shaded. Chlortetracycline (aureomycin) differs
from tetracycline in having a chlorine atom
(blue); doxycycline consists of tetracycline
with an extra hydroxyl (purple).

TETRACYCLINES
Str. aureus. S.flavus
S. rimosus, S. antibioticus

Streptomyces aureofaciens
20 diff species producing mix of tet
Genetic modification
Polyketide synthesis

Antibiotics synthesized by successive condensation of small carboxylic acids

Like acetate, butyrate, propionate, malonate

High doses of tetracycline may result in nausea, diarrhea, yellowing of teeth in

children, and damage to the liver and kidneys.

AMINOGLYCOSIDES

Oligosaccharide antibiotics

Structurally all contain a cyclohexane ring and amino sugars bound by glycosidic
linkages
Bind to the 30S small ribosomal subunit and interfere with protein synthesis in at least
two ways. They directly inhibit protein synthesis and also cause misreading of the
genetic message carried by mRNAprolonged use can cause kidney damage and hearing
loss

Streptomycin, kanamycin, neomycin, and tobramycin are synthesized by

Streptomyces, whereas gentamicin comes from a related bacterium,
Micromonospora purpurea.

Known as reserve antibiotics as they develop resistance quickly

AMINOGLYCOSIDES producing organisms

Streptomycin

Streptomyces griesus

Neomycin B and C

S.fradiae

Kanamycin A, B and C

S.kanamyceticus

Hygromycin B

S.hygroscopicus

Gentamycin

Micromonospora purpurea

Sisimicin

M.inyoensis

MACROLIDES
Antibiotics with a large lactone ring (macrocyclic lactone ring)
Which consists of 12-, 14- and 16-membered lactone rings with 1-3 sugars linked
by glycosidic bond
Effective agaist penicillin resistant MO, G+ org, inhibitb y binding to 50S
ribosome
Clarithromycin (Erythromycin derv)
Used to treat stomach ulcers

Erythromycin : Streptomyces erythreus

14-membred connected to 2 sugars
Genetic modifications by polyketide synthesis

MACROLIDES
Polyene macrolides: lactone rings in range of 26-28
Eg. Nystatin, amphotericin
Actinomycetes are most common organisms which produce them

Erythromycin is a relatively broad-spectrum antibiotic effective against grampositive bacteria, mycoplasmas, and a few gram-negative bacteria. It is used
with patients allergic to penicillins and in the treatment of whooping cough,
diphtheria, diarrhea caused by Campylobacter, and pneumonia from Legionella
or Mycoplasma infections.
Newer macrolides are now in use.
Clindamycin is effective against a variety of bacteria including staphylococci
and anaerobes such as Bacteroides.
Azithromycin is particularly effective against Chlamydia trachomatis.

AROMATIC ANTIBIOTICS

Aromatic rings in structure

Chloroamphenicol, griesofluvin, novobiocin

CHLORAMPHENICOL

Broad spectrum antibiotic against G+ and G- bacteria, rickettesia, chlamydia,

actinomycetes
chloramphenicol binds to 23S rRNA on the 50S ribosomal subunit. It inhibits
the peptidyl transferase and is bacteriostatic.

Streptomyces venezuelae and S.omiyanesis

This antibiotic has a very broad spectrum of activity but unfortunately is quite toxic. One may see allergic responses or
neurotoxic reactions. The most common side effect is a temporary or permanent depression of bone marrow function, leading
to aplastic anemia and a decreased number of blood leukocytes. Chloramphenicol is used only in life-threatening situations
when no other drug is adequate.

GRIESOFULVIN

Penicillium patulum

Maybe attacks chitin biosynthesis hence acts as anti fungal antibiotic

Following a 40-year hiatus in discovering new classes of antibacterial compounds,

three new classes of antibacterial antibiotics have been brought into clinical use:
Cyclic lipopeptides (Daptomycin), Glycylcyclines (tigecycline) and Oxazolidinones
(Linezolid)
PEPTIDE ANTIBIOTICS
Daptomycin : Streptomyces roseosporus used to treat MDR infections
Tigecycline: Tygacil marketed by Wyeth used to treat MDR strains of
Staphylococcus aureus and Acineotobacter baumanii. Mechanism similar to
tetracycline.
Also shows suceptibility to NDML (New Delhi metallo-b-lactamase multidrug
resistant Enterobacteriaceae)
NDML is an enzyme which makes bacteria resistant to broad range of b-lactam antibiotics.
This includes antibiotics of carbapenems for treatment of antibiotics resistant infections.
Termed as SUPERBUGS Such bacteria susceptible to polymixins and tigecyclines

MECHANISM OF DRUG RESISTANCE

Plasmids
R-Plasmids
Superinfection: Clostridium difficile, Candida albicans
Transformation, conjugation, transduction, ABC transporters
Phage therapy

There has been some recent progress in developing new antibiotics that are
effective against drug-resistant pathogens.
Two new drugs are fairly effective against vancomycin-resistant enterococci.
Synercid is a mixture of the streptogramin antibiotics quinupristin and dalfopristin
that inhibits protein synthesis.
A second drug, linezolid (Zyvox), is the first drug in a new family of antibiotics, the
oxazolidinones. It inhibits protein synthesis and is active against both vancomycinresistant enterococci and methicillin-resistant Staphylococcus aureus.