Mahatma Education Society’s

Pillai College of Arts, Commerce & Science

Autonomous
Affiliated to University of Mumbai
Re-accredited ‘A’ Grade by NAAC (3 cycles)
ISO 9001:2015 certified
Best College Award by University of Mumbai

T.Y.B.Sc. (SEM-VI) Biotechnology

Unit I

STARTER CULTURE

• Meenakshi Johri
• Assistant Professor & M.Sc I Coordinator
• Department of Biotechnology
STARTER CULTURE
• Starter culture is an active microbial preparation intentionally added
during product manufacture to initiate desirable changes.
• These microbial preparations can consist of LAB, Propionibactera,
yeast and molds.
STARTER CULTURE
• Starter cultures have multifunctional role in dairy fermentations
• Produce acid which help in rapid separation of of curd from whey
• Production of low molecular weight compounds
such as diacetyl contributes to flavour and aroma
• Gas production cause eye formation
Role/Functions of Starter Cultures
 Role/Functions
Role/Functions of Starter Cultures of Starter Cultures
GENERAL CHARACTERISTICS OF LAB
• Gram positive, non-sporing, non-pigmented.
• Microaerophile
• Fastidious- requiring specific amino acids, B vitamins and growth
factors.
• Unable to use complex carbohydrates
 GENERAL CHARACTERISTICS OF
LAB
• 11 genera of LAB
• 4 are commonly used as dairy starter cultures
• Lactobacillus
• Streptococcus
• Lactococcus
• Leuconostoc
 GENERAL CHARACTERISTICS OF
LAB
• LAB are generally associated with nutrient rich habitats containing
simple sugars
• Raw milk
• Meat
• Fruits
• Vegetables
• Wine
• Beer
• Bread
Lactococcus
• Mesophilic microbes used for acid production in dairy
fermentations.
• Cocci usually occur in chains
• Homofermentative
• Grow at 10 C but not at 45 C.
• Weak proteolytic and can use milk protein
• Hydrolyse milk protein casein by proteinase PrtP.
• Two imp subsp.- Lc lactis subsp lactis & Lc lactis sub
cremoris
Lactococcus
Characteristics Lc lactis subsp lactis Lc lactis subsp cremoris
Lactose + +
Galactose + +
Maltose + -
Ribose + -
Growth in 4% salt + -
Arginine hydrolysis + -
Streptococcus
• Only species useful in dairy is S. thermophilus
• Thermophilic- ability to grow at 52 C
• Limited proteolytic activity.
Leuconostoc
• Mesophile
• Heterofermentative cocci
• Do not hydrolyse arginine
• Require various B vitamins for growth
• Produce Diacetyl, CO2, acetoin
• 2 imp. sub species- L. meseteroides, L. lactis
Propionibacteria
• Gram positive rods
• Non spore former
• Produce large amount of propionic acid, acetic acid and carbon
dioxide from sugars.
• Uses both inorganic and organic nitrogen source
• Most strains require biotin
• Used for eye production in cheese
Propionibacteria
Brevibacterium
• Aerobic, Gram positive , pleomorphic rods
• Found on the surface of surface ripened cheese.
• Most often isolated is B.linens.
• B.linens produce yellow orange carotenoid pigment that colours the
surface of the cheese
• Does not use lactose but can grow on the lactate produced during
cheese manufacture.
Brevibacterium
• Grows best at neutral pH.
• Does not grow well on the cheese surface until lactic acid is
neutralized or metabolised.
• Highly proteolytic.
• Degrade amino acids to ammonia and methionine to methanethiol-
strong flavour and odour.
• Other flavouring compounds- butyric acid, caproic acid, etc.
Lactobacillus
• Gram positive rods
• Homofermentative –
• ferment hexose to lactic acid using EMP pathway.
• Do not ferment pentose sugars or gluconate
• Ex. L. delbrueckii subsp.bulgaricus, L. delbrueckii subsp.lactis, L. helveticus
• Thermoduric- 45 C
Lactobacillus
• Heterofermentative
• Use hexose to lactic acid, acetic acid and CO2
• Pentose are also used
• Ex. L.casei
Penicillium
• Two gp used in cheese manufacture is P.camberti and P. roqueforti

• P. camemberti and P. roqueforti are lipolytic and proteolytic. Both

produce methyl ketones and free fatty acids, but the much higher
levels produced by P. roqueforti give blue cheeses their distinctive
flavor and aroma.
Penicillium
• P. camemberti contributes to the flavor of Camembert and Brie
cheeses by producing a complex mixture of compounds, the major
ones being 2-heptanone, 2-heptanol, 8-nonen-2-one, 1-octen-3-ol, 2-
nonanol, phenol, butanoic acid, and methyl cinnamate
USE OF GENETICALLY MODIFIED STARTER
CULTURE
• A. Metabolic Engineering for Diacetyl Production
• Diacetyl is an industrially important buttery flavor and aromatic compound
that is derived from citrate metabolism by LAB.
• Inactivation of the gene encoding α-acetolactate decarboxylase, enzyme
which converts α-acetolactate to acetoin.
• α-acetolactate is precurssor for Diacetyl
A. Metabolic Engineering for Diacetyl Production

• Inactivation of aldB – replacement recombination

• Or natural mutant for aldB can be selected by growth selection in a
medium that contains leucine but not valine
• Leucine is an allosteric activator of α-acetolactate decarboxylase
• Wild type can not grow in medium as leucine stimulate conversion of
α-acetolactate to acetoin and valine is synthesized.
A. Metabolic Engineering for Diacetyl Production

• Most strains can not synthesize branched amino acid.

• Transformstion of cells with recombinant plasmid encoding enzymes
for branched chain amino acid
ENHANCED PHAGE RESISTANCE
• Bacteriophage infection of starter culture can result in failure of
fermentation or loss of product.
• Bacteriophage resistance is one of imp industrial trait that may be
encoded by plasmid DNA.
• Transfer of plasmid DNA by conjugation
• pTRK2030 encodes restriction/modification and abortive infection
defense mechanism