Biotechnology In Tea

Professor: Ph.D. Akhavan Sepahi

Suppliers: Abolfazl Ehsani
Alireza Mehrdadfar
Introduction
• Tea refers to:
– the plant Camellia sinensis (Thea sinensis)
– the dried, processed leaf manufactured from it
– extracts derived from the leaf, and the beverages prepared from
the leaf or extract of the species
Originated in Southeast Asia in an area that includes China and India,
and probably Myanmar, Laos and Vietnam
Tea varieties
• Two major varieties of Camellia sinensis are
recognized:

Chinese variety (sinensis) a smaller-leaved (5-12 cm) plant

Assam variety (assamica) a large-leaved (15-20 cm) plant

Introduction
• The first harvest is obtained after 4-5 years

• The shrub can be used for 60-70 years

• Harvesting season depends upon the region and climate and

lasts for 8-9 months per year or leaves can be plucked at
intervals of 6-9 days all year round

• The younger the plucked leaves, the better the tea quality

• The white-haired bud and the two adjacent youngest leaves are
plucked (famous “two-leaves and bud” formula)
 Types of Tea
• The enzymatic oxidation of tea
leaves is referred to as Black Tea
fermentation
Green Tea
• If the enzymes are allowed to act,
they turn green leaf black (black
tea) Yellow Tea

• If the enzymes are inactivated by

Red Tea
heat, as in blanching, then the leaf (Oolong)
remains green (green tea)
 Black
Constituent Fresh Tea
Composition of Tea Tea
(%, dry weight basis) Phenolic compounds 30 5
Oxidized phenolic 0 25
compounds
Protein 15 15
Amino acids 4 4
Caffeine 4 4
Crude fiber 26 26
Other carbohydrates 7 7
Lipids 7 7
Pigments 2 2
(chlorophyll and
caroteniods)
Volatile compounds 0.1 0.1
Minerals 5 5
Phenolic Compounds
• Phenolic compounds make up 25–35% of the dry matter
content of young, fresh tea leaves.
• Flavanol compounds are 80% of the phenols
• During fermentation the flavanols are oxidized
enzymatically to compounds which are responsible for the
color and flavour of black tea.
• The reddish-yellow color of black tea extract is largely due to
theaflavins and thearubigins
Flavonoid
• Flavonoids (or bioflavonoids) - are a class of plant secondary metabolites.
• They can be classified into:

• flavonoids, derived from 2-phenylchromen-4-one (2-phenyl-1,4-benzopyrone)

structure (examples: quercetin, rutin).
• isoflavonoids, derived from 3-phenylchromen-4-one (3-phenyl-1,4-benzopyrone)
structure
• neoflavonoids, derived from 4-phenylcoumarine (4-phenyl-1,2-benzopyrone)
structure.

• The three flavonoid classes above are all ketone-containing compounds, and as
such, are flavonoids and flavonols. The terms flavonoid and bioflavonoid have
also been more loosely used to describe non-ketone polyhydroxy polyphenol
compounds which are more specifically termed flavanoids, flavan-3-ols (or
catechins).
Flavanol
• Flavan-3-ols (flavanols) are a class of flavonoids – term is
mainly used for non-ketone polyhydroxy polyphenols –
Flavanoids
• These compounds include the catechins and the catechin
gallates.

Epicatechin (EC)

Chemical structure of Flavan-3-ol

Epigallocatechin (EGC)
Black Tea
• During fermentation the flavanols are oxidized
enzymatically to compounds which are responsible for the
color and flavor of black tea.

• The reddish-yellow color of black tea extract is largely due to

theaflavins and thearubigins.

• The astringent taste is caused primarily by flavonol-3-

glycosides.

• The catechins are turned from the monomer structure to

become the dimers that are the theaflavins and the
oligomers that are thearubigins
Enzymes
1) Polyphenol oxidases- located within cell of leaf epidermis &
activity rises during withering & rolling
2) Shikimate dehydrogenase- reversibly interconverts
dehydroshikimase & shikimate via phenylalanine pathway
3) Phenylalanine ammonia lyase- catalyse cleavage of phnylalanine
into ammonia & cinnamate
4) Proteinases – cause protein hydrolysis during withering resulting
in rise of peptides & free amino acids
5) Chlorophyllases participate in the degradation of chlorophyll
and transaminases in the production of precursors for aroma
constituents.
Amino acids
• Constitute about 1% of dry matter of tea leaves
• Of this 50% is thiamine & rest consists of protein forming amino
acids
• Β- alanine is also present
• Green tea contains more thiamine than black tea

5-N-ethyl-glutamine
Pigments
 Chlorophyll is degraded during tea processing.

 Chlorophyllides and pheophorbides (brownish in colour) are

present in fermented leaves, both being converted to
pheophytines (black) during the firing step.

 Fourteen carotenoids have been identified in tea leaves. The

main carotenoids are xanthophylls, neoxanthin, violaxanthin and
β-carotene

 The content decreases during the processing of black tea.

Manufacture of black tea

Withering
• Reduces moisture from about
75-80% to 55-65%
• Moisture reduction converts
the turgid leaf to a flaccid
material that is easily handled
Withering
• Enzymatic protein hydrolysis yields amino acids of which a part
is transaminated to the corresponding keto acids.
• Both types of acids provide a precursor pool for aroma
substances
• Chlorophyll degradation has significance for the appearance of
the end-product.
• Conversion of chlorophyll into chlorophyllide, a reaction
catalyzed by the enzyme chlorophyllase – more extensive is
undesirable as give rise to pheophorbides (brown) and not the
desired olive black pheophytins.
• Increased cell permeability during withering favors the
fermentation procedure.
Manufacture of black tea

Rolling
• It establishes proper
conditions for enzymatic
oxidation of the flavanols by
atmospheric oxygen
Rolling
• Tea leaf is macerated and the substrate and enzymes are brought
together; - The subsequent enzymatic oxidative reactions are
designated as a prerequisite for fermentation
• In this processing step, the pigments are formed primarily as a result
of phenolic oxidation by the PPO. In addition, oxidation of amino
acids, carotenoids and unsaturated fatty acids, preferentially by
oxidized phenols, is of importance for the formation of odorants
• The enzymatic oxidation of flavanols via the corresponding
oquinones gives theaflavins - bright red color, good solubility
• A second, heterogeneous group of compounds, found in tea after the
enzymatic oxidation of flavanols, are the thearubigins - a group of
compounds responsible for the characteristic reddish-yellow color of
black tea extracts
Manufacture of black tea

Fermentation
• Conversion of colourless
catechins to a complex
mixture of yellow-orange to
red-brown substances
Fermentation
• Aroma development during fermentation is accompanied
by an increase in the volatile compounds typical of black
tea.
• They are produced by Strecker degradation reactions of
amino acids with oxidized flavanols.
Manufacture of black tea

Firing
Ends fermentation process
and reduce moisture
content to 3%
Firing
• During this step there is an initial rise in enzyme activity
then all the enzymes are inactivated.
• Conversion of chlorophyll into pheophytin is involved in
reactions leading to the black colour of tea.
• A prerequisite for these reactions is high temperature and
an acidic environment. The undesired brown colour is
obtained at higher pH’s.
• The astringent character of teas is decreased by the
formation of complexes between phenolic compounds and
proteins
Thank you for your attention