8/6/13 DT-1: Lesson 8.

LACTOPEROXIDASE-THIOCYANATE (LP) SYSTEM

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B. Tech. (Dairy Technology) ► DT-1 ► Resources ► Lesson 8. LACTOPEROXIDASE-

THIOCYANATE (LP) SYSTEM

Module 4. Procurement of milk

Lesson 8
LACTOPEROXIDASE-THIOCYANATE (LP) SYSTEM

8.1 Introduction
8.2 Effect of LP System against Bacteria
8.3 Method of Adoption
8.4 Practical Application
8.5 Monitoring the Practice
8.6 Effect on Milk Quality

8.1 Introduction

The perishable nature of milk makes it necessary to exercise extra care to preserve it. The
tropical (hot and humid) climate of India further aggravates the problem. Milk meant for
long distance transportation is always refrigerated to maintain its quality. However,
refrigeration may not always be possible or available due to economic and/or technical
reasons. Therefore the need to have a suitable alternative to refrigeration has been felt
since long. An FAO/WHO Expert Panel on Milk Quality suggested addition of hydrogen
peroxide (H2O2) as an alternative. This was not popular for several reasons, mostly related
to the proper control of its usage.

Milk of poor quality could be disguised and passed of as good quality milk

Higher concentration of H2O2 is toxic

Research conducted in the 1960s on colostrum in Sweden led to the discovery of naturally
occurring enzymes and their positive effect on preservation systems. One such enzyme
was lactoperoxidase (LP).

It is an oxido-reductase enzyme

It occurs in milk, saliva, tears, cervical mucus

It is a single polypeptide chain with a molecular weight of 77,000 – 100,000.

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8/6/13 DT-1: Lesson 8. LACTOPEROXIDASE-THIOCYANATE (LP) SYSTEM

Cow milk has 1.4 units/ml of LP, whereas buffalo milk has 0.9 units/ml. The amount of
LP required for preservation is 0.5-1 mg/l, much lower than its concentration in cow milk
(30 mg/l). It is a relatively heat resistant enzyme which retains sufficient activity even after
pasteurization. Thiocyanate is widely distributed in animal tissues and secretions. Its level
is dependent on dietary habits and lifestyle (for example, smokers have a higher level of
thiocyanate in their system). Thiocyanate is the product of detoxification of thioaminoacids
and cyanides and is secreted in urine with a half life of elimination of 2.5 days under
normal renal function. Bovine milk has 2-15 ppm thiocyanate, though higher values have
also been reported.

Lactoperodixase can oxidise thiocyanate ions in the presence of H2O2, which is not
normally present in milk. Even if present, it is in minute quantities, as a result of bacterial
activity.

The LP-thiocyanate system can be explained with the following chemical reactions:

Hypothiocyanous acid dissociates into hypothiocyanite at the pH of milk

There are other end products besides these, such as sulphate, ammonia and carbon
dioxide. The intermediary products of the LP-thiocyanate reaction are implicated in
bacteriostatic and bactericidal activities.

The presence of large quantities of sulphydral groups (-SH) such as those occurring in
boiled milk would reduce/eliminate the antibacterial effect because the OSCN- ions would
then attach to these freely available groups. Raw milk has very few SH- groups and
therefore, the LP-system is most effective in raw milk.

8.2 Effect of LP System against Bacteria

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8/6/13 DT-1: Lesson 8. LACTOPEROXIDASE-THIOCYANATE (LP) SYSTEM

The bacteriostatic/ bacterioacidal effect of the LP system is species- and strain- specific. In
a mixed flora dominated by mesophilic bacteria, the effect is largely bacteriostatic. The
system exhibits bacteriocidal effect on some Gram negative organisms such as
Pseudomonas sp. and E. coli. In case exogenous H2O2 is present, the effect may also be
bacteriostatic. The LP system has an impact on the glycolytic enzymes and hence, the
metabolic activity is adversely affected. Lysis occurs due to leaking of bacterial contents.

Owing to the bacteriostatic effect, it is not possible to hide poor quality of milk. The
high bacterial counts will be revealed during plating, MBRT etc.

The antibacterial oxidation products are not heat stable. The stability is temperature
dependent, leading to complete destruction by pasteurization.

The oxidation products are not stable at neutral pH. Any surplus will decompose to
thiocyanate.

The preservation effect is directly proportional to the thiocyanate concentration,

provided equimolar concentration of H2O2 is present. In practice small quantities of
thiocyanate is added to make up the concentration

The quantity of H2O2 to be added must be monitored. If excess quantity is added

(for example, 300-800 ppm), lactoperoxidase is inactivated and preservation effect is
then largely due to H2O2.

8.3 Method of Adoption

The LP system may be used when technical, economic and/or practical reasons do not
permit chilling. It is not recommended for the use of individual farmers. It should be
practiced only at collection centers with proper cleaning and sanitizing facilities. The
manpower deputed to handle the system should be given adequate training. The dairy that
receives the milk should monitor the system. Appropriate detection methods should be set
up to avoid foul practices. The LP system does not eliminate the need for the normal clean
milk production methods that are adopted at farm levels. It does not exclude the need for
pasteurization before consumption

The preservation effect is temperature-dependent. The preservation effect under

laboratory and field experimental conditions for stored milk which had been produced
hygienically are listed in Table 8.1.

Table 8.1 Preservation effect of LP system at different temperatures

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8.4 Practical Application

At the village collection centre, each clean and washed milk can should be given a 3
min chlorine rinse (400 ppm) immediately before milk is poured into it.

Sodium thiocyanate (NaSCN) should be added @ 14 mg/l milk and mixed well for a
minute with a plunger.

Sodium percarbonate (precursor of H2O2) should be added @ 30 mg/l milk and

stirred well for 2-3 min

This sequence of addition should be followed because the enzyme action starts
when H2O2 added and is completed within 5 min of addition of H2O2.

This rate of addition will provide 75 ppm of NaSCN and 50 ppm of H2O2 as
percarbonate.

The LP activation must be done within 2-3 hours of milking

After 10 hours of milking (or 7 hours of first treatment), a booster dose of 35 ppm
H2O2 (~ 20 mg sodium percarbonate per litre milk) should be mixed into milk.

To make the practice easy, the quantities of NaSCN and sodium percarbonate for a
certain volume of milk (say 40 l) should be distributed to collection points in
prepacked quantities, say for a few weeks at a time.

8.5 Monitoring the Practice

The usage of the system can be monitored through acidity, MBRT, Resazurin reduction
test, total viable count and analysis of thiocyanate concentrations.

8.6 Effect on Milk Quality

Field trials conducted in Kenya, Sudan, Mexico, Sri Lanka, Pakistan and India report
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8/6/13 DT-1: Lesson 8. LACTOPEROXIDASE-THIOCYANATE (LP) SYSTEM

that untreated milk spoiled within two hours whereas treated milk was preserved for
different durations depending on the temperature of milk.

There was no difference on the preservation effect of cow and buffalo milk.

The quality of initial milk was, in all cases, proportional to the time taken for
spoilage.

A strong bacteriostatic effect was reported on mesophilic and thermophilic bacteria

and also spores.

Total and proteolytic psychrotrophs were also affected.

Selected Reading

FAO. 1999. Manual on the use of the LP-system in milk handling and preservation. Global
Lactoperoxidase Programme, Animal Production Service, FAO Animal Production and
Health Division. Food and Agriculture Organization of the United Nations. Rome.

Last modified: Thursday, 27 December 2012, 03:50 PM

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