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Contents
General Main Root Causes ............................................................................................................................................2
1. Organosulfur compounds ................................................................................................................. 2
2. Non-sulfur garlic phytochemicals, .................................................................................................... 3
3. Aminoacid Interactions ..................................................................................................................... 4
4. Storage Temperature Conditions...................................................................................................... 4
5. Environment pH ................................................................................................................................ 5
6. Incidental Additives........................................................................................................................... 5
7. Active Alliinase (4.4.1.4) in Dehydrated Garlic ................................................................................. 6
Mechanism of Garlic Greening from organosulphur Compounds .................................................................................7
Thiosulphinate Base Pigments .................................................................................................................. 8
Reactants – Aminoacid Requirments for Pigmentation ........................................................................... 8
Enzyme - Alliinase (4.4.1.4) - Properties and Stability ............................................................................ 11
Summary
Main root causes of garlic greenings are listed based on the literature review, which involves
enzymatic process of Sulphur-organic compounds break down and their interaction with amino
acids, flavonoids and incidental additives. Based on the explained mechanism of pigmentation
evolvement, it can be lead to conclusion that the most common cause in green color
evolvement in meat product marinades may be failure to denature Alliinase during dehydration
process. In addition, it is worth mentioning that warehousing conditions become a factor too.
While operators prefer to store garlic in lower temperatures to stop germination, studies have
revealed that storing garlic under 20°C will favor greening. Alliinase activity may be added as
part of Quality Assessment for dehydrated granulated garlic or garlic juices used in meat
Marinades.
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GREENING OF GARLIC IN MEAT MARINADES
Based on the literature search the greening of the garlic may happen from the followings:
1. Organosulfur compounds
The discoloration is due to pigments that form between sulfur compounds in garlic and amino acids.
When the garlic tissue is disrupted, as happens in processing, an enzyme is liberated and reacts with it
to form thiosulfinates compounds that then react with the natural amino acids in the garlic to form blue
pigments.
S-allyl-L-cysteine sulfoxide (alliin) accounts for approximately 80% of cysteine sulfoxides in garlic (Figure
1). When raw garlic cloves are crushed, chopped, or chewed, an enzyme known as alliinase is released.
Alliinase catalyzes the formation of sulfenic acids from L-cysteine sulfoxides (Figure 2). Sulfenic acids
spontaneously react with each other to form unstable compounds called thiosulfinates. In the case of
alliin, the resulting sulfenic acids react with each other to form a thiosulfinate known as allicin (half-life
in crushed garlic at 23°C is 2.5 days). The formation of thiosulfinates is very rapid and has been found to
be complete within 10 to 60 seconds of crushing garlic. Color development in these situations was
extensively studied [Kubec, R., Hrbáčová,M., Musah,R and, and Velíšek,J (2004) : Allium Discoloration:
Precursors Involved in Onion Pinking and Garlic Greening, Journal of Agricultural and Food Chemistry, 52
(16), 5089-5094].
The pH optimum for pigment formation has been found to be between 5.0 and 6.0 for all thiosulfinates
tested. [Kubec, R., Velíšek, J. (2007): Allium Discoloration: The Color-Forming Potential of Individual
Thiosulfinates and Amino Acids: Structural Requirements for the Color-Developing Precursors, J. Agric.
Food Chem., 55 (9), pp 3491–
3497]
Crushing garlic does not
change its γ-glutamyl-L-
cysteine peptide content. γ-
Glutamyl-L-cysteine peptides
include an array of water
soluble dipeptides, including
γ-glutamyl-S-allyl-L-cysteine, γ-glutamylmethylcysteine, and γ-glutamylpropylcysteine
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GREENING OF GARLIC IN MEAT MARINADES
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GREENING OF GARLIC IN MEAT MARINADES
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GREENING OF GARLIC IN MEAT MARINADES
Controlled by Temperature, J. Agric. Food Chem., 56 (3), pp 941–945]. Greening in crushed garlic cloves
increases with increasing GGT expression at low temperature, while alliinase expression is not affected.
[Jungeun Cho, Minkyu Park, Doil Choi, Seung Koo Lee (2011): Cloning and expression of γ‐glutamyl
transpeptidase and its relationship to greening in crushed garlic (Allium sativum) cloves, J. Foos Sci and
Agric., (92) 2, pp 253-257].
5. Environment pH
Studies conducted by Chinese for producing of “Laba” Garlic, (Chinese pickled garlic where Garlic Cloves
are expected to be green), have indicated that greening is promoted in acid environment [Bing Bai, Fang
Chen, Zhengfu Wang, Xiaojun Liao, Guanghua Zhao, and Xiaosong Hu (2005): Mechanism of the
Greening Color Formation of “Laba” Garlic, a Traditional Homemade Chinese Food Product, J. Agric.
Food Chem., 2005, 53 (18), pp 7103–7107]. Soaking aged garlic in 5% (v/v, pH 2.33) acetic acid solution,
after 2 days, the garlic cloves turned green. On day 4, pigment(s) diffused from garlic cloves to the
pickling solution. The solution exhibits two maximal absorbances at ∼440 and ∼590 nm, corresponding
to yellow and blue species, respectively, the combination of which creates the green coloration. With
increasing time from 4 to 25 days, the concentration of both yellow and blue species increases at nearly
the same rate, while after 25 days, the concentration of the yellow species increases faster than that of
the blue species. They found that, most thiosulfinates (∼85%) in garlic cloves were converted within 4
days, suggesting that thiosulfinate conversion is proportional to the formation of the pigments. UV−vis
spectral measurements and pH results suggest that the color formation occurs by two kinds of
processes: one enzymatic and the other nonenzymic. Low pH (2.0−3.0) favors nonenzymic reactions,
while high pH (6.0 or above) is conducive to enzymatic reactions. Thus, the ideal pH for the entire
process of garlic greening is between 4.0 and 5.0.
6. Incidental Additives
Copper from water source, or other can create green colored chelated complexes. The other sources of
copper might be the butter or lemon juice.
In addition, there is reported that Iodine from Iodized salts may turn garlic blue.
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GREENING OF GARLIC IN MEAT MARINADES
[Kubec, R., Hrbáčová,M., Musah,R and, and Velíšek,J (2004) : Allium Discoloration: Precursors Involved in Onion Pinking and Garlic
Greening, Journal of Agricultural and Food Chemistry, 52 (16), 5089-5094]
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GREENING OF GARLIC IN MEAT MARINADES
In garlic, abundant green-blue compounds formed from 19/20 and 23, which are the key color-
developing compounds occurring in crushed garlic.
It has been found that an unsubstituted primary amino group and a free carboxyl group are essential
structural features required for amino compounds to be able to generate the pigments.
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GREENING OF GARLIC IN MEAT MARINADES
Out of the thiosulfinate analogs tested, only those containing at least a three-carbon chain with the ß-
carbon bearing a hydrogen atom yielded the pigments. Thiosulfonates, sulfoxides, sulfides, and di
Sulfides did not form any colored products.
The pH optimum for pigment formation has been found to be between 5.0 and 6.0 for all thiosulfinates
tested (Kubec R, Velisek J (2007) Allium discoloration: The color-forming potential of individual
thiosulfinates and amino acids: Structural requirements for the color-developing precursors. Journal of
Agricultural and Food Chemistry 55, 3491-3497).
Investigation of the variation in thiosulfinates and related compounds between fresh bulbs and cloves,
and commercial garlic products, indicated that thiosulfinates were released only from garlic
cloves and garlic powder products. (Lawson LD, Wang Z-Y, Hughes BG (1991): Identification and HPLC
quantitation of the sulfides and dialk(en)yl thiosulfinates in commercial garlic products. Planta Medica
57, 363-370).
The analysis indicated that thiosulfinates were released only from garlic cloves and garlic powder
products.
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GREENING OF GARLIC IN MEAT MARINADES
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GREENING OF GARLIC IN MEAT MARINADES
Alliinase in garlic has been studied extensively and its properties are summarized below.
[http://lem.ch.unito.it/didattica/infochimica/2006_aglio/pagine/when.html#terzo]
It has been suggested that there are two different alliinase activities for garlic, one being specific for
substrate (+)-S-2-propenyl-L-cysteine sulphoxide and (+)-S-1-propenyl-L-cysteine sulphoxide at an
optimum pH of 4.5 and the other specific for substrate (+)-S-methyl-L-cysteine sulphoxide at an
optimum pH of 6.5. Both of these alliinases are irreversibly deactivated at pH 1.5-3 and >9.
Although alliinase can act on both (+) and (-) sulphoxides, it shows a preference for the (+) isomers. 3
The transformation rate of thiosulphinates from their relative precursors is also different while
maximum formation of diallyl thiosulphinate (allicin) can reach 100% within 1 minute, it takes more than
5 minutes for allyl methyl thiosulphinate to reach 100%.
The activity of alliinase is also pH dependent. Below pH 4, alliinase is partially deactivated whilst at pH
4.5-7, alliinase has very high activity. The effects of pH adjustment during the blending of garlic cloves on
the formation of flavor compounds were
studied by Yu and Wu 1 who showed that
maximum allicin formation occurred around
pH 6.5.
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Thermal destruction of Alliinase follows the first order kinetics with Z-value=9.4 °C. The effectiveness of
thermal treatment to eliminate pungent flavor and inhibit green discoloration increased with increasing
temperatures. Optimum blanch treatment was reached at 80 °C for 1.1 min. [Rejano, L., de Castro, A.,
Sánchez, A., Casado, J., Montaño, A. (2004): Thermal kinetics of pungency loss in relation to the quality
of pickled garlic, International Journal of Food Science & Technology 39(3):311 – 317]
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