Module 2.

Classification of starters

Lesson 2

STARTERS CULTURES AND THEIR CLASSIFICATION

2.1 Introduction

A starter culture is a microbial culture which actually performs fermentation. Starter preparations
assist the beginning of the fermentation process in preparation of various foods and fermented
drinks. A number of bacterial and other microbial strains have been used either in single or in
combination for producing the desired effect in the finished product.

2.2 Definitions

� Starter cultures are bacterial or fungal strains either pure or mixed, used to initiate a
fermentation process.
� Starter culture means selected strains of food-grade microorganisms of known and
stable metabolic activities and that is used to produce fermented foods of desirable
appearance, body, texture and flavour
� Starter culture means the microorganisms that are selected based on their ability to
produce lactic acid for curd production and a low pH to prevent spoilage, produce
metabolites that give desirable flavours: or produce enzymes that ripen the dairy
product.

2.3 Role/Functions of Starter Cultures

The primary function of lactic starters is the production of lactic acid from lactose. In addition to
lactic acid production the starter cultures are also useful in different ways as stated below.

Table 2.1 Functions of starter cultures

Function Result
Acid production Gel formation

Expulsion (syneresis) of whey for texturing

Preservation of milk
 Helps in the development of flavour
Flavour Formation of flavour compounds like diacetyl and
acetaldehyde
Preservation Lowering of pH and redox potential

Production of lactic acid

Production of antibiotics

Production of H2O2

Production of acetate
Gas formation Eye formation in certain cheeses
Production of open texture Ex. blue veined cheese
Stabilizer formation Development of body and viscosity
Ex. Polysaccharide materials
Lactose utilization Reduces the development of gas and off flavours

Suitable for lactose intolerant people

Lowering of redox potential Helps in preservation

Helps in development of flavour

Proteolysis and lipolysis Helpful in the ripening/maturation of cheeses
Miscellaneous compounds Production of alcohol in kefir and kumis

2.4 Classification /Taxonomic Groups as Per Bergey's ManualIntroduction

Starter cultures are generally classified based on their ability to utilize the lactose as shown in
Fig. 2.
 Fig. 2.1 Classification of starter cultures

2.4.1 Bacteria

2.4.1.1 Genus lactococcus

Bergey�s Manual of Systematic Bacteriology (1986), combined all the mesophilic lactic acid
bacteria (LAB) with Lactococcus lactis to form a single species as they possess

1. Identical isoprenoid quinines and the enzyme β-phosphotase

2. Indistinguishable lactic dehydrogenase
3. Identical percentage of guanine and cytosine.
4. High DNA homology

The only properties that distinguish them are plasmid controlled.

Lactococcus lactis subsp lactis Acid producer but non-

flavour producer
Lactococcus lactis subsp cremoris Acid producer but non-
flavour producer
Lactococcus lactis subsp lactis biovar Both acid & flavour
 diacetylactis producer

All the above organisms are mesophilic in nature and their optimum growth temperature is
between 25-30�C. All are homofermentative organisms

2.4.1.2 Genus streptococcus

The members of the Streptococcus are Gram-positive organisms that usually form pairs or
chains. In 1937, Sherman separated the genus according to physiological and growth
characteristics, especially with regards to temperature limitations on growth. Four general
groups designated by Sherman are (1) pyogenic, (2) viridans, (3) enterococcus, and (4) lactic.
This categorization has become somewhat obsolete as relationships between species have been
shown to overlap.

The only species used as starter culture is Streptococcus salivarius subsp thermophilus. This is a
yoghurt culture, which is thermophilic in nature with optimum growth temperature of 38-42�C.
All are homofermentative organisms.

2.4.1.3 Genus leuconostoc

All are heterofermentative organisms capable of producing lactic acid, CO 2 and aromatic
compounds (ethanol and acetic acid) from glucose. These organisms are normally used along
with lactic acid bacteria (LAB) in multiple or mixed strain cheese starter cultures, which
produces flavour compounds.
Leuconostoc cremoris
Leuconostoc citrovorum
Leuconostoc dextranicum

2.4.1.4 Genus lactobacillus

Lactobacillus delbruekii subsp bulgaricus is used for the preparation of yoghurt along with
Streptococcus salivarius subsp thermophilus. These two organisms exhibit a symbiotic
relationship.
Lactobacillus acidophilus is a probiotic culture, used for preparation of acidophilus milk and
other probiotic milk products like Bifighurt, Bioghurt, etc. The members of lactobacillus are
classified based on fermentation of glucose into 3 groups as shown in Figure 2.2.

Fig. 2.2 Classification of lactobacillus based on glucose fermentation

2.4.1.5 Genus bifid bacterium

Found in the gut of infants, intestines of man, various animals and honeybees. These organisms
are generally used in preparation of therapeutic fermented milk products in combination with
yoghurt, acidophilus milk or yakult starter cultures.

Eg: Bioghurt, Biograde, Bifighurt, Cultura �AB, Yakult, Miru-Miru.

Bifid bacterium bifidum, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium
breve, etc.

The optimum growth temperature is 37�C- 41�C. Anaerobic conditions are essential for
optimum growth. Milk fermented with bifidobacteria has a distinctive vinegar taste due to the
production of acetate plus lactate from the metabolism of carbohydrates.

2.4.1.6 Genus propionibacterium

Propionibacterium freudenreichii and Propionibacterium shermanii are used in swiss cheese. It

has the ability to produce large gas holes in the cheese during ripening / maturation period. P.
jensenii, P. thoenii and P. acidipropionici are other organisms present in these genera.

2.4.1.7 Genus brevebacterium

Brevebacterium linens is used as starter culture in preparation of bacterial surface ripened

cheeses. It imparts distinctive, reddish orange colour to the rind of (or formation of smear on)
Brick and Limburger cheese or Camembert cheese.

2.4.2 Molds

Moulds are used for the manufacture of some semi soft cheese varieties and in some fermented
milk products. Moulds enhance the flavour and modify slightly the body and texture of curd.

2.4.2.1 White mold

It is used in manufacture of surface mould ripened cheeses like Camembert and Brie cheeses.
Eg: Penicillium camemberti, Penicillium caseicolum, Penicillium candidum

2.4.2.2 Blue mold

It is used in manufacture of internal mould ripened cheeses like Roquefort, Blue Stilton, Danish
blue, Gorgonzola and mycella cheeses.

Eg: Penicillium roquefortii

2.4.2.3 Other molds

Mucor rasmusen � used in Norway for the manufacture of ripened skim milk cheese.

Asperigillus oryzae � used in Japan for the manufacture of Soya milk cheese.

Geotricum candidum� used in the manufacture of Villi a cultured product of Finland. The
mould grows on the surface of the milk to form the white velvety layer.

2.4.3 Yeasts

Yeasts are used in the manufacture of Kefir and Kumiss

2.4.3.1 Kefir grains

Kefir grains consist of a mixture of different microorganisms such as Candida kefir,

Kluyeromyces marxianus, Saccharomyces kefir, Torulopsis kefir.

2.4.3.2 Kumiss

The important starter microflora of kumiss include Torulopsis spp.

Kluyeromyces marxianus var lactis, Saccharomyces cervisiae

2.5 Types of Starters

Starters are grouped under different categories based on composition of microflora, growth
temperature, type of products, flavour production and type of fermentation into the following
categories

2.5.1 Based on the composition of micro flora/ organisms

a. Single: Always used as a single organism in the preparation of dahi or cheese. The
only problem is there will be sudden failure of starter due to bacteriophage attack
which leads to heavy loss to the industry.
b. Paired compatible strain: Two strains of cultures having complementary activities in
know proportion are used. This will reduce chances of culture failures. . In case of
bacteriophage attack, only one type of organism will be affected and the other
organism will carry out the fermentation without any problem.
 c. Mixed Strain: More than two organisms which may have different characteristics like,
acid production, flavour production, slime production etc. in unknown proportion are
used.
d. Multiple mixedstrain: More than two strains in known proportion are used. The
quality and behaviour of these strains is predictable.

2.5.2 Based on the growth temperature

Based on the growth temperature organisms can be divided into mesophilic and thermophilic.

Mesophilic starter cultures: The optimum growth temperature of these cultures is 30�C and
they have a growth temperature range of 22- 40�C. The mesophilic starter cultures generally
contain the organisms of Lactococci.
Ex. Dahi cultures : Lactococcus spp.
Cheddar cheese: Lactococcus lactis subsp lactis, Lactococcus lactis subsp cremoris
Lactococcus lactis subsp lactis biovar diacetylactis, Leuconostoc mesenteroides subsp
cremoris

Thermophilic starter cultures: The optimum growth temperature of these cultures is 40*C and
they have a growth temperature range of 32- 45�C.

: Streptococcus thermophilus

��������������Lactobacillus delbrueckii subsp bulgaricus

��������������Lb. delbrueckii subsp lactis

��������������Lb. Casei

��������������Lb. Helveticus

��������������Lb. plantarum

2.5.3 Product for which used

��� Yoghurt: Streptococcus thermophilus

 ����������������������� Lactobacillus delbrueckii
subsp bulgaricus

�� Swiss cheese: Streptococcus salivarius subsp thermophilus

Lactobacillus delbrueckii subsp bulgaricus

Lactobacillus helveticus
Lactobacillus casei

2.5.4 Based on the flavour production

The starters are grouped into B, D, BD and N type based on their ability of flavour production

B (L) type: Leuconostocs as flavour producer (old name is Betacocccus)

D type: L. lactis subsp lactis biovar diacetylactis

BD (LD) type: Mixer of both of the above cultures

N or O type: Absence of flavour producing organism

2.5.5 Based on the type of fermentation

The starters are classified as homo or hetero fermenter based on end products resulting from
glucose metabolism.

Homo fermentative cultures: eg. Lactococcus lactis subsp lactis

Hetero fermentative cultures eg. Leuconostoc dextranicum