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CASE STUDY ON THE ANTIMICROBIAL

PROPERTIES OF NISIN

MINI PROJECT REPORT

(U15BTT403 – Food Process Engineering)

Submitted by

BHALAJI. V. G (15BBT053)

DHIRANESH. N (15BBT056)

ROBIN RICHARD. I (15BBT057)

in partial fulfillment for the award of the degree

of

BACHELOR OF TECHNOLOGY
IN
BIOTECHNOLOGY

KUMARAGURU COLLEGE OF TECHNOLOGY

(An autonomous institution affiliated to Anna University, Chennai)

ANNA UNIVERSITY, CHENNAI

CHENNAI-600025
APRIL 2017

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 BONAFIDE CERTIFICATE

ANNA UNIVERSITY: CHENNAI

Certified that this “CASE STUDY ON THE

project report
ANTIMICROBIAL PROPERTIES OF NISIN” is the bonafide work of

BHALAJI. V. G - Register No. 15BBT053

DHIRANESH. N - Register No. 15BBT056

ROBIN RICHARD. I - Register No. 15BBT057

Who carried out the project work under my supervision?

SIGNATURE SIGNATURE
Dr. A. Thirumurugan Dr.N Saraswathy,
Asssistant Professor Professor & Head

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CONTENTS

ABSTRACT------------------------------------------------------------ 4

INTRODUCTION----------------------------------------------------- 5

MODE OF ACTION-------------------------------------------------- 6

CONCLUSION-------------------------------------------------------- 8

REFERENCES-------------------------------------------------------- 9

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CASE STUDY ON THE ANTIMICROBIAL PROPERTIES OF NISIN

ABSTRACT

Nisin is a commercially added food preservative, commonly tagged as E234

and is used in the processing of many food stuffs like dairy products (milk, cheese
etc.), meats and beverages. Chemically, it is a polycyclic peptide with the formula,
C143H230N42O37S7. It is naturally synthesized by the bacterium Lactococcus lactis
when grown on natural substrates like milk or dextrose.

Nisin is receiving a new focus in recent times for its exceptional anti-
microbial properties, including drug resistant bacteria and tumor cells. Besides the
food industry, Nisin finds applications in the pharmaceutical industry and
cosmetics. Since a lot of antibiotic substances and other anti-microbial products are
being neutralized by many pathogenic bacteria which have become drug-resistant
through mutation and improper administration of the drugs, research in the anti-
microbial properties of Nisin will enable us to tackle the difficulties in the
treatment of drug-resistant cases.

Courtesy: www.21food.com

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 INTRODUCTION

Nisin is chemically a polycyclic peptide of molecular weight 3354.07

g/mol and is composed of some unusual amino acid residues, like lanthionine
(Lan), methyllanthionine (MeLan), didehydroalanine (Dha), and
didehydroaminobutyric acid (Dhb). These are added during the post-
translational modification of the precursor peptide 57-mer to produce Nisin.

Courtesy: Apex Tribune

Nisin belongs to a class of molecules called lantibiotics, which includes

subtilin and epidermin. It appears colorless and powdery, boils at 2,966˚C and has
a density of 1.402 g/mL.

Courtesy: Indiamart

It is heat stable, non-toxic, and sensitive to digestive proteases. It is typically

found to be added in food stuffs in the range of ~1-25ppm, according to the food
type and regulatory approvals. Nisin finds an indispensible use in the food industry
due to its excellent anti-microbial activity which is as follows.

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MODE OF ACTION

Nisin is a broad-spectrum lantibiotic bacteriocin, which is effective against

most gram-positive bacteria especially Lactobacillus lacti, Listeria
monocytogenes, Staphylococcus aureus, Bacillus cereus, Clostridium botulinum,
etc. L.lacti is more commonly associated with spoilage. L.monocytogenes is a
known pathogen causing listerosis. S.aureus and C.botulinum are common causes
of food poisoning. Nisin acts on these through two methods:

 Nisin binds to the static area of bacteria, preventing resistance to the

antibiotic action
 Nisin attacks biofilms, which are essentially bacterial colonies deterring
antibiotic activity.

Nisin also acts upon spores effectively. Gram-negative bacteria are protected
by an extra layer of mucus, but become susceptible to Nisin when they are
subjected to heat shock or coupled with a chelating agent like EDTA.

Recent studies have shown the effectiveness of Nisin against cancer cells in
higher concentrations than used in food preservation. Typical dosage of Nisin in
foods is around 0.25 to 37.5 mg/kg, while a dose of 800 mg/kg is needed to kill
cancer cells. In addition, highly purified Nisin ZP doubled the efficiency during
the studies.

Oral Cancer cells treated with Nisin and undergoing apoptotic cellular changes
(Courtesy: Yvonne Kapila)

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 Recent research by Dr. Yvonne Kapila et al., University of Michigan on Nisin’s
potential as a an anti-microbial and anti-cancerous substance, predicts the future
application of Nisin will be in the treatment of 30 different types of cancer, skin
infections, respiratory and abdomen ailments and oral diseases.

Nisin treated oral biofilm

(Courtesy: Yvonne Kapila)

The advantages of Nisin based on its anti-microbial activity are:

 Good, natural preservative which decreases the required temperature.

 Shorten the time in heat processing of foods
 Replaces chemical preservatives
 Improves the look, texture, flavor and nutritional value of food
 Saves energy and thereby lower production costs
 Does not affect naturally occurring micro biota
 Selectively affects gram-positive and at cases, gram-negative bacteria
which have become resistant to antibiotics.
 Can be manufactured easily from cultures of Lactococcus lactis (In fact,
natural mother’s milk contains good levels of Nisin due to the
presence of L.lactis culture in the milk)