Biosynthesis of Plant-derived flavor compounds

By Dudsadee Uttapap

Biosynthesis of plant-derived flavor compounds

References .
. Flavor Chemistry and Technology, H.B. Heath, G. Reineccius, . Flavor Chemistry, D.B. Min, http://class.fst.ohio-state.edu/fst /default.htm

3.

Biosynthesis of plant-derived flavor compounds, The Plant Journal (2008) 54, 712732

4.

Plant Biochemistry http://www.uky.edu/~dhild/biochem/lecture.html

Flavor compounds
Flavor molecules constitute a heterogeneous group of compounds, with straight-chain, branched-chain, aromatic and heteroaromatic backbones bearing diverse chemical groups such as hydroxyl, carbonyl, carboxyl, ester, lactone, amine, and thiol functions. More than 700 flavor chemicals have been identified and catalogued

Chemical synthesis VS Biosynthesis

Most commercial flavorants are nature identical, which means that they are the chemical equivalent of natural flavors but are chemically synthesized, mostly from petroleum-derived precursors

Bioproduction, including the extraction from natural sources, de novo microbial processes (fermentation), and bioconversion of natural precursors using micro-organisms or isolated enzymes

Biological functions of plant volatiles associated with defensive and attractive roles
Compounds emitted by flowers most probably serve to attract and guide pollinators volatiles might also protect the carbohydrate-rich nectar by inhibiting microbial growth. vegetative plant tissue release volatiles following herbivore damage. Some of these substances attract arthropods that prey upon or parasitize the herbivores. Volatiles also act as direct repellents or toxicants for herbivores and pathogens. In fruits, volatile emission and accumulation facilitate seed dispersal by animals and insects.

vegetative tissues often produce and release many of the volatiles after their cells are disrupted. These volatile flavor compounds may exhibit anti-microbial activity.

Aromatic compounds responsible for odor and flavor of fruits comprise;

Alcohols R-OH Carbonyls RR-CO-R CHO

Esters

R-COO-R
R CO

Lactones
O

Acids

R-COOH Phenols

Estimated world consumption of selected aroma chemicals in flavor and fragrance compositions

CHO

OCH3 OH

Calvin cycle

Amino acid synthesis

N enters roots as NO- or NH+. The NH+ is incorporated into amino acids in roots and leaves and the amino acids accumulate in proteins. The main if not sole function of some proteins is to provide a store of amino

Amino acid synthesis

Glycolysis

isoprenoid biosynthesis proceeds either via the "classical" or most well studied, mevalonate pathway (cytosolic) (for the synthesis of sterols, sesquiterpenes, triterpenoids) or via the non-mevalonate (-deoxy-D-xylulose-phosphate, DXP) pathway for plastidic isoprenoids (carotenoids, phytol [side-chain of chlorophylls], plastoquinone, isoprene, monoterpenes and diterpenes).

Biosynthesis of flavors in vegetables and fruits

Fruit flavors
are formed during brief ripening period

Vegetable flavors
develop when tissue damage occurs (Intact vegetable generally contains few volatiles)

BIOGENESIS OF FRUIT AROMA

develops entirely during ripening period of plant

Minute quantities of lipids, CHO, protein (amino

acids) are enzymatically converted to volatile flavors.

FRUIT FLAVOR COMPOUNDS

Apple n-hexanal, ethyl butyrate, 1-propyl propionate, 1-butyl acetate, trans-2-hexenal, ethyl 2methylbutyrate, 2-methylbutyl acetate, 1hexanol, hexen-1-ol, trans-2-hexen-1-ol, hexyl acetate, Esters; alcohols; aldehydes; ketone; acids; including hexanal; ethyl 2-methyl butyrate alcohols; esters, including amyl acetate, isoamyl acetate, butyl butyrate, amyl butyrate Ethyl acetate, dimethyl disulfide, cis- -hexenyl acetate, methyl octanoate, ethyl octanoate, pentyl alpha pyrone, gamma decalactone

Banana

Peach

Lipids Polysaccharide Proteins/Enzymes

Malonyl CoA Fatty acid Acetyl-CoA Carbohydrate Pyruvate Acetyl CoA Terpene Mevalonyl CoA Amino acid
Shikimic acid

Lignins

Cinnamic acid

Aliphatic
Acids Alcohols Esters Carbonyls lactones

Terpenes
Sesquiterpenes Hydrocarbons Alcohols Carbonyls monoterpenes

Methyl-Branched Alcohols Acids Esters carbonyls

Aromatic
Alcohols Acids Esters carbonyls

Biosynthesis of fruit volatiles

Flavorants from carbohydrate metabolism

Only a limited number of natural volatiles originate directly from carbohydrates without prior degradation of the carbon skeleton.

Furanones and pyrones

fruit constituents

Furanones and pyrones

Carbohydrate-derived flavor molecules, including -hydroxy-,-dimethyl-(H)furanone (furaneol), ,-dimethyl--methoxy-(H)-furanone (methoxyfuraneol), hydroxy--methyl-(H)-furanone (norfuraneol), -ethyl--hydroxy--methyl-(H)furanone (homofuraneol), -hydroxy--methylene--methyl-(H)- furanone (HMMF) and -hydroxy--methyl-H-pyran--on (maltol).

Flavorants from carbohydrate metabolism

Glucose (C)

Pyruvate (C)
CO -O +O -O

Glycolysis
Lactate

Ethanol TCA Cycle

Pyruvic acid

CH3COCOOH CH3COOH CH3COSCoA

Acetic acid

Acetyl CoA + CO2 Malonyl CoA

HOOCCH2COSCoA HOOCCH2COOH

Malonic Acid

Flavorants from carbohydrate metabolism

the most interesting is terpene biosynthesis
Terpenoids are enzymatically synthesized from acetyl CoA and pyruvate provided by the carbohydrate pools in plastids and the cytoplasm. Terpenoids constitute one of the most diverse families of natural products, with over 40 000 different structures of terpenoids Many of the terpenoids produced are non-volatile and are involved in important plant processes such as membrane structure (sterols), photosynthesis (chlorophyll side chains, carotenoids), redox chemistry (quinones) and growth regulation (gibberellins, abscisic acid, brassinosteroids)

Important plant-derived volatile terpenoids.

Biosynthesis of Terpenes
isoprene is derived from acetyl-CoA

Classification of Terpenes

Apocarotenoid formation

Carotenoid substrates are oxidatively cleaved to yield the apocarotenoid derivatives (right).

Some of the volatile organic compounds in wine come from the grape's skin, or exocarp, while others come from the grape's flesh, or mesocarp. Organic acids give wine its tartness, and sugars give it sweetness. Terpenes provide floral or fruity flavors. Norisoprenoids impart a honeylike character. Thiols are the sulfur-based compounds behind complex wine aromas such as guava, passionfruit or grapefruit but when thiols go wrong, they can make a wine taste "funky."

Lipids
metabolic pathway for lipid biosynthesis plays a significant role in flavor formation.

Oxidation via lipoxygenase Alpha-, Beta-oxidation

products; acids, alcohols, diketones, ketones, esters of these compounds.

Oxidation via Lipoxygenase

Lipoxygenase activity is believed to be the major source of volatiles in plants.
Lipoxygenase enzymes (dioxygenase) catalyze reactions between O2 and polyunsaturated fatty acids

Substrate: unsaturated fatty acid (linoleic and linolenic acids). Major products: volatile C and C aldehydes and alcohols

Linolenic acid-derived flavor molecules.

AAT, alcohol acyl CoA transferase; ADH, alcohol dehydrogenase; AER, alkenal oxidoreductase; AOC, allene oxide cyclase; AOS, allene oxide synthase; HPL, hydroperoxide lyase; JMT, jasmonate methyltransferase; LOX, lipoxygenase; OPR, oxo-phytodienoic acid reductase; Z,E-EI, Z,E-enal isomerase.

Fatty acid precursors (Tomato)

- and -oxidation of fatty acids

the specific pathways in plants are not well understood

Palmitoyl-CoA (:)

+ Acetyl-CoA

Myristoyl-CoA (:)

Formation of pear flavors via beta-oxidation

Lactones

Amino Acid Metabolism

Amino acid metabolism yields short chain aliphatic and aromatic alcohols, acids, carbonyls and esters

They are the primary source of branched chain aliphatic flavor compounds

their pathways have been barely analyzed in plants.

amino acid precursors (Tomato)

Biosynthesis of amino acid-derived flavor compounds

(a) Catabolism of branched-chain amino acids leading to methyl branched flavor compounds, and (b) postulated biosynthesis of sotolon. Formation of aldehyde (a) from amino acids requires the removal of both carboxyl and amino groups. The sequence of these removals is not fully known and could be the opposite to that shown or aldehyde could be formed in one step by aldehyde synthase

Starting amino acids: Tyrosine and phenylalanine products: phenolic/spicy in character

Shikimic acid formation

Vegetable Flavors

Vegetable flavors
flavor again arises from major metabolic processes e.g. Lipids, CHO & amino acids.

the precursors, enzymes and end flavors are quite different from fruits.

The role or importance of S compounds to vegetable flavor is quite significant.

Carbohydrate Fatty acid Amino acid

Nonvolatile Precursors
Linoleic, Linolenic acid Thioglucosinolates Cysteine-sulfoxides Methyl-methionine

Precursor-splitting Enzymes
Lypoxygenase Thioglucosidases C-S-lyases None (Heating)

Carbonyls Alcohols Oxo-acids

Isothiocyanates Nitriles S C O Thiocyanates

Polysulides Alkylthosulfinates

CH3-S-CH3

Formation of flavor in vegetables

Vegetable Flavor Categories

Genus Allium Enzymes produce volatiles from derivatives of cysteine (sulfoxides)

Genus Brassica Enzymes produce volatiles from glucosinolates

Alliaceous vegetables
garlic (Allium sativum L.) onion (Allium cepa L.) chive (Allium schoenoprasum L.) leek (Allium porrum L.)

Characteristic flavors
not exist in the bulb before processing

are produced when the cellular tissues are ruptured by cutting or chewing

flavor is produced very rapidly by the action of an enzyme on the odorless precursors which coexist in the cells

Onion and Gar lic Flavor

Enzymatic reaction of cysteine derivative

GLUCOSINOLATES

Glucosinolate precursors are important to the flavor of both the Brassica and Cruciferae family Cruciferae family includes radish, horseradish, mustard.

Hydrolysis of the glucosinolate

glucosinolate

thioglucosidase

thiocyanate, nitrile, or isothiocyanate & glucose

Natural carbon pools for the production of flavor compounds, and the pathways

Mevalonic acid

Acetate

Mevalonic acid

Isoprene

Shikimic acid

Pyruvate + Erythrose phosphate

Flavorants from carbohydrate metabolism

the most interesting is terpene biosynthesis
most of essential oils get flavor from terpenoids ( carbon)

Limonene - a monoterpene hydrocarbon - is the major terpene in many or most citrus products. Orange > % of the essential oil is limonene, lemon ~ % limonene, yet is of little flavor significance.

Citral - oxygenated monoterpene - seldom comprises > % of the essential oil of lemon - largely carries the lemon flavor.

Methane Ethane Propane Butane Pentane Hexane Heptane Octane Nonane Decane

Undecane Dodecane Tridecane Tetradecane Pentadecane Hexadecane Heptadecane Octadecane Nonadecane Icosane

Henicosane Docosane Tricosane Tetracosane Pentacosane Hexacosane Heptacosane Octacosane Nonacosane Triacontane

Hentriacontane Dotriacontane Tritriacontane Tetracontane Pentacontane Hexacontane Heptacontane Octacontane Nonacontane Hectane

Isoamyl acetate, a strong fruity odor described as similar to banana or pear -Methyl-butyl acetate has a strong apple scent