Antimicrobial and Biosensors

Food antimicrobials and
biosensors
Dr. Rafat Amin

Topics to be covered
➢ Naturally occurring antimicrobials
➢ Biosensors in food industry
Naturally occurring antimicrobials
An antimicrobial is an agent that kills microorganisms or stops their growth
NEED
➢ Inhibit the growth of microorganisms like bacteria and fungi.
➢ Increase shelf life.
➢ Preserve the appearance of food
➢ Preserve the food characteristics like odor, taste and
➢ food is preserved for a long time
MIC - Minimum Inhibitory Concentration
is the lowest concentration of an antimicrobial agent needed to stop the microbe from
reproducing
MBC - Minimum Bactericidal Concentration
is the lowest concentration of an antimicrobial agent needed to kill the microbe

Classification of naturally occurring antimicrobials
1. Plants with natural antimicrobial activities

2. Antimicrobials from microorganisms
3. Antimicrobials from animals
1. Plants with natural antimicrobial activities
• Basil
• Caraway
• Cardamom • Mint
• Cinnamon • Mustard
• Clove • Nutmeg
• Coriander • Onion
• Cumin • Oregano
• Fennel • Peppers
• Garlic • Rosemary
• Grape • Sage
• Green tea • Thyme
• Marjoram
Onion
Good source of bioactive compounds, such as sulphur containing compounds and

flavonoids
Antimicrobial activity of onion is due to thiosulfinates and other volatile organic

compounds and shown inhibition against gram positive and gram negative bacteria.
Anti-inflammatory, antiallergenic, anticarcinogenic antioxidant capacities, antibacterial,

antifungal and antiviral
Onion antiyeast, anti-fungal activity is due to presence of organosulfur-containing

compounds which inhibit the growth of yeast and fungi
Basil
The oil is extracted by steam distillation from the leaves and the flowering tops.
Basil oil has various chemical compounds that include a pinene, camphene, b-pinene, myrcene,
limonene, cis-ocimene,
camphor, linalool, methyl chavicol, y-terpineol, methyl cinnamate and eugenol.
It shows inhibitory effects against fungi, molds.
Basil oil used in postharvest protection of mango fruits against anthracnose disease.
Post harvest spraying of sweet basil oil on table grapes inhibit fungal decay.
Whole and ground dry basil leaves at 5-10% reduce aflatoxin contamination in grains like
sorghum, ground nuts, maize.
Cardamom (E.Cardamomum)
➢ Contains bioactive components like 1-8 cineole, terpinyl acetate,

phenolics, limonene etc.
➢ Possesses, antimicrobial activities and inhibit the growth of bacteria,

virus, fungi, molds.
➢ Experimental observations undertaken to assure essential oil and

oleoresins of large cardamom as food preservatives on sweet orange
juice.
SEAWEED
➢ Rich in several secondary metabolite, such as polyphenols (phlorotannins,

fucoxanthin, flavonoids) and polysaccharides (fucoidan, laminarans).
➢ methanol:toluene (3∶1) best solvent for extracting the antimicrobial

principles and it was found a lipophilic compound.
➢ Brown seaweed extract reduced maximum specific growth rate to (96.8-

99%) and highly active against Gram negative bacteria
➢ Stocheospermum marginatum seaweed showed activity against

Klebsiella pneumoniae.
2. Animals with natural antimicrobial activities
Lactoperoxidase
➢ Glycoprotein with high isoelectric point and one Fe group, Occurs in milk, saliva and
tears of many mammals.
➢ Function is to oxidize thiocynate and some halides to generate products and affects the
cytoplasmic membrane that kill or inhibit growth microorganisms.
➢ Exert bacteriostatic effect on Listeria, Staphylococcus
➢ Addition of LP to starter culture for yogurt production inhibited the acid production
and new yogurt produced with extended shelf life
• Used in oral care products toothpaste

• Preservative in ground beef products
Pleurocidin
A 25 amino acid peptide isolated from the skin mucus membrane of the winter flounder
Active against Gram-positive and Gram-negative bacteria.
Effective against foodborne organisms including Vibrioparahe, molyticus, L.

monocytogenes, Saccharomyces cerevisiae, and Penicillium expansum
It was inhibited by magnesium and calcium, which may limit the use of this AMP
Antimicrobial activity was reported at levels well below the legal limit for nisin (10,000
IU/g) without significant effect on human red blood cells
Chitosan
Natural biopolymer obtained from the exoskeletons of crustaceans (crabs and

shrimp) and arthropods
Used as active material for its antifungal activity and antibacterial activity.
E. coli and the strong antibacterial activity of chitosan was also observed
against S. aureus
several bacteria and yeast were exposed to chitosan concentrations varying from 40
to 750 mg/l. Gram-negative bacteria seemed to be very sensitive for the applied
chitosan (MIC< 0.006% (w/v)
The antimicrobial effect of chitosan were observed on strawberries during 12 days.

Defensin
Group of antimicrobial peptides widely found in nature including mammalian epithelial cells
and Immune cells.
Immune response is the first line of defense and occurs soon after pathogen exposure
Function by binding to the microbial cell membrane, and once embedded, forming pore-like
membrane defects that allow efflux of essential ions and nutrients.
Human defensins are active in vitro against Gram-positive and Gram-negative bacteria, fungi,
enveloped viruses, and other pathogens
3. Antimicrobials from microorganisms
Reuterin
Produced by some strains of Lactobacillus reuteri, during the metabolism of glycerol to

1,3-propanediol activity against Gram-negative and Gram-positive bacteria, yeasts,
and filamentous fungi
Lactobacillus reuteri found in the gut of other mammals and birds
Resistant to the action of proteolytic and lipolytic enzymes
Exhibit bacteriostatic activity against Listeria monocytogenes, E. coli , S. choleraesuis ,

Y. enterocolitica,
Pediocin
Produced by strains of Pediococcus acidilactici and P. pentosaceus and is designated GRAS
The bacteriocins produced by P. acidilactici are AcH, PA-1, JD, and 5, and those produced from P.
pentosaceus are A, N5p, ST18, and PD1
Pediocin AcH has proven efficacy against both spoilage and pathogenic organisms, including L.
monocytogenes, S. aureus, and Cl.Perfringens
Control growth of L. monocytogenes in fermented meat product such as sausage and

frankfurters.
Pediocin approved by USDA for use in reduced nitrite bacon to aid in prevention of botulinum
toxin production
Recently, a pediocin by P.acidilactici containing formulation is marketed under the commercial

name Alta 2341
Sakacin
Bacteriocin produced by certain strains of the lactobacillus sakei, 43 amino acid peptide
Inhibit the growth of gram positive bacteria including food pathogen
Target on bacterial cytoplasmic membrane so leakage of cellular component and depletion

of proton motive force.
Sakacin A influence Listeria monocytogens growth, a microorganism able to grow at 4 °C up

to 1600 AU ml-1
Sakacin-A one of the bacteriocins control growth of L. monocytogenes in pasteurized

ground beef
Nisin
Only natural antimicrobial peptide approved by the FDA for use as a food preservative
Obtained from the culturing of L. lactis on natural substrates.
Nisin is soluble in water and 100 & 300 IU/g decrease growth of L.monocytogens, S.aureus
Functions by interacting with the phospholipids in the cytoplasmic membrane of bacteria,

thus disrupting 450 membrane function and preventing outgrowth of spores
Highly active against many of the Gram-positive bacteria, Completely degraded in alimentary
tract so safe to use
Used as a Food preservative in a number of Thermal processed foods, particularly in Dairy

products, Canned foods, Plant protein Foods and Cured Meat products and Marine Products.
Natamycin
➢ Also known as Pimaricin

➢ Produced by a pure culture of Streptomyces natalensis bacteria through fermentation
process.
➢ Centrifuged, filtered, and washed, to get pure end product.
➢ Targets ergosterol in the cell wall, a building block of yeasts and molds, which is
responsible for intracellular nutrient transport
➢ Effective at very low levels
➢ Natamycin is approved in different applications with different level in the world
➢ In rats, the LD50 is ≥2300 mg/kg, and doses of 500 mg/kg/day over 2 years caused no
detectable differences in survival rate, growth, or incidence of tumors
Legislatory Aspects
Existing food legislation in most countries would not favor the use of natural compound,
unless this compound have genuinely acquired as GRAS
Lengthening procedure for marketing approval and hampering economic implementation

in practice
As they are new, would require non-toxicity record
Only Nisin and Pediocin were approved by U.S.FDA till date

Issues
Mechanism of action of natural preservatives is needed in order to setup an effective
preservation strategy for each product
Financially difficult in developing commercial bacteriocins preparations, because of the

costly production, low production rates, unstable products
Shown strong bactericidal effects against pathogens in laboratory test systems and weak-
moderate effects in real foods so research required
Biosensors in food industry
What are Biosensors?
Classification of Biosensors
Food quality check
Problem with food analysis
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Food antimicrobials and

Dr. Rafat Amin

An antimicrobial is an agent that kills microorganisms or stops their growth

MBC - Minimum Bactericidal Concentration

is the lowest concentration of an antimicrobial agent needed to kill the microbe

1. Plants with natural antimicrobial activities

Good source of bioactive compounds, such as sulphur containing compounds and

Antimicrobial activity of onion is due to thiosulfinates and other volatile organic

Anti-inflammatory, antiallergenic, anticarcinogenic antioxidant capacities, antibacterial,

Onion antiyeast, anti-fungal activity is due to presence of organosulfur-containing

It shows inhibitory effects against fungi, molds.

➢ Contains bioactive components like 1-8 cineole, terpinyl acetate,

➢ Possesses, antimicrobial activities and inhibit the growth of bacteria,

➢ Experimental observations undertaken to assure essential oil and

➢ Rich in several secondary metabolite, such as polyphenols (phlorotannins,

➢ methanol:toluene (3∶1) best solvent for extracting the antimicrobial

➢ Brown seaweed extract reduced maximum specific growth rate to (96.8-

➢ Stocheospermum marginatum seaweed showed activity against

➢ Exert bacteriostatic effect on Listeria, Staphylococcus

• Used in oral care products toothpaste

Active against Gram-positive and Gram-negative bacteria.

Effective against foodborne organisms including Vibrioparahe, molyticus, L.

Natural biopolymer obtained from the exoskeletons of crustaceans (crabs and

The antimicrobial effect of chitosan were observed on strawberries during 12 days.

Produced by some strains of Lactobacillus reuteri, during the metabolism of glycerol to

Lactobacillus reuteri found in the gut of other mammals and birds

Resistant to the action of proteolytic and lipolytic enzymes

Exhibit bacteriostatic activity against Listeria monocytogenes, E. coli , S. choleraesuis ,

Produced by strains of Pediococcus acidilactici and P. pentosaceus and is designated GRAS

Control growth of L. monocytogenes in fermented meat product such as sausage and

Recently, a pediocin by P.acidilactici containing formulation is marketed under the commercial

Target on bacterial cytoplasmic membrane so leakage of cellular component and depletion

Sakacin A influence Listeria monocytogens growth, a microorganism able to grow at 4 °C up

Sakacin-A one of the bacteriocins control growth of L. monocytogenes in pasteurized

Functions by interacting with the phospholipids in the cytoplasmic membrane of bacteria,

Used as a Food preservative in a number of Thermal processed foods, particularly in Dairy

➢ Also known as Pimaricin

Lengthening procedure for marketing approval and hampering economic implementation

As they are new, would require non-toxicity record

Only Nisin and Pediocin were approved by U.S.FDA till date

Financially difficult in developing commercial bacteriocins preparations, because of the

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