Professional Documents
Culture Documents
תרכובות חנקן ביין
תרכובות חנקן ביין
בס"ד
Adapted from: Pretorius et al.(2007). Nitrogen management is critical for wine flavour and style. Wine Industry Journal, 2(6), 24-30.
בס"ד
בס"ד
Nitrogen Compounds
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Amines
Amines are a class of compounds derived from ammonia (NH 3) by replacement of one, two, or
three hydrogens to yield primary, secondary, and tertiary amines, respectively.
Additional substitution on the amino nitrogen with a hydrocarbyl group yields a quaternary
ammonium cation whereas protonation of the amino nitrogen yields an aminium cation.
Amines can also be present in molecules containing additional functional groups (e.g. carboxyl,
carbonyl, thiol, etc.), and in grapes, the major amine ‐group ‐containing species are amino acids
and their polymers (e.g., proteins).
תכולה כוללת של חנקן מורכבת מצורה אנאורגנית אחת וצורות אורגניות שונות רבות
מהווה כ 90% -מהחנקן בשיא ההבשלה
ריכוזים משתנים ביחס להבשלת הענבים
החנקן הכולל בזמן ההבשלה הוא כ 140-650 -מ"ג לליטר
הריכוז המינימלי לתסיסה תקינה הוא 50מ"ג לליטר
+NH4 מרכיב חיוני בתסיסה האלכוהולית
ריכוז גבוה יותר ביינות אדומים מאשר בלבנים -בגלל ההשריה ,טמפרטורת תסיסה ואוטוליזה של התאים
חנקן מינרלי
חנקן מינרלי בצורה של מלחי אמוניה נמצא בריכוזים גבוהים מאוד בשלב גידול הגפן -מעל 80%
החנקן המינרלי הופך עם התקדמות ההבשלה לחומצות אמוניות ,פפטידים וכו'
בשיא ההבשלה -מהווה כ 10%בלבד מהחנקן
בס"ד
בס"ד
חומצות אמינו
מורכבות מאטום פחמן מרכזי הקשור קוולנטית לקבוצת אמין ,קבוצת קרבוקסיל ,אטום מימן
ושייר משתנה
שייר הצד של החומצה נקרא גם קבוצת ה R-והוא הגורם המבדיל בין חומצות האמינו השונות.
מהוות את המולקולות העיקריות בחנקן הכולל
חשובות מאד בשל התכונות האנטי-אוקסידנטיות ואנטי מיקרוביאליות שלהן ,תורמות לקיבולת
הבופר של היין ,מהוות אמולסיפריים וכחומרים פעילי שטח
משתתפות במרבית תהליכי התסיסה הכוהלית
”Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine
בס"ד
בס"ד
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Structure: From a structural point of view, the twenty most common amino acids in must
and wine, excepting γ -amino-butyric acid and β-alanine, are α-amino acids, i.e. the
carboxylic acid radical and the amino basic radical are linked to the same carbon atom.
Amino acids are therefore amphoteric molecules. The R radical may be a hydrogen atom
(e.g. glycine or glycocoll).
In other amino acids, the carbon linked to the functional radicals and the R carbohydrate
radical is chiral. This feature gives them the asymmetrical property common to
biologically significant molecules.
Natural amino acids all have a true L configuration. Their optical activity depends on the
type of R radical, but also on the solvent, and even the pH of the solution.
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Other trifunctional amino acids have no marked acid or basic character. They have
hydroxylated (serine, threonine and tyrosine), thiol or sulfide radicals (cysteine and
methionine). Only these trifunctional amino acids are involved in the catalyst property of
enzymes.
In solution, amino acids must be expressed in a double balance state, involving the
ionization of the acid or basic function.
The population of each form is defined by the pH of the medium. At a pH where the
positive and negative charges are in balance, i.e. the isoelectric point, amino acids have
minimum solubility.
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
Chemistry of amines…
בס"ד
The presence of the lone electron pair results in amines and other amine ‐containing compounds
behaving as weak bases.
The nitrogen lone pair of amino acids and many other amines is protonated at wine pH, and thus
many well‐known food chemistry reactions that involve amine groups acting as nucleophiles are
expected to occur very slowly.
Amines with low pKa values will also have minimal volatility and thus no odor in wine.
Amino acids and polypeptides have multiple ionizable groups at wine and juice pH.
• pH < pI – the protein (or amino acid) has a net positive charge.
• pH = pI – the protein has no net charge.
• pH > pI – the protein has a net negative charge.
The importance of pI to wine is that it defines the pH at which a protein will be at minimum
solubility.
Most wine proteins are reported to have pI values just above wine pH, in the range of 4–6, and
will thus have a net positive charge in wine or juice.
As a result, proteins can non‐covalently bond to species that have formal negative charges or are
H‐bond acceptors, such Adapted
as bentonite and etpolyphenols.
from: Waterhouse al., 2016: "Understanding Wine Chemistry"
בס"ד
בס"ד
Profiles showing changes in concentrations of individual amino acids during ripening (grape variety:
Chardonnay) (Millery, 1988)
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Some small peptides also have interesting sweet or bitter flavors that are
useful in winemaking.
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Structure of glutathion and its reaction with quinones produced by the oxidation of phenols
)Proteins( חלבונים
Their structure includes other bonds besides peptide bonds that give the chain
a three-dimensional configuration: spherical, helix, etc.
Grapes and wine contain many proteins with a wide range of molecular weights
(30,000-150,000).
Some unstable proteins are responsible for protein casse in white wines.
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Amide nitrogen
פירוט
as well as ethyl carbamate which is very strictly controlled for health ...בהמשך
Reasons:
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Nucleic nitrogen
Pyrazines
Pyrazines are heterocycles with six links containing two nitrogen atoms
and four carbon atoms bearing radicals, which largely determine their
olfactory impact.
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
Structure of Pyrazines
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
)Urea( אוראה
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
Urea cycle and its relationship with the Krebs cycle. Comparative catabolism of arginine בס"ד
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
חומר מסרטן
פעם שימש כחומר משמר והיום אסור לשימוש
הריכוז הממוצע ביין הוא כ 8 -מיקרוגרם לליטר
ריכוז מקסימאלי לפי החוק האמריקאי הוא 15מיקרוגרם לליטר
תנאים המגבירים את ריכוזו:
. 1זן הענבים והוספת מרכיבים חנקניים לקרקע
.2תסיסה בטמפרטורות גבוהות וארוכות
.3תסיסה אלכוהולית עם שזרות
.4טמפרטורת אחסון של יין מבוקבק
”Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine
בס"ד
בס"ד
”Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine
בס"ד
בס"ד
Adapted from: Ribéreau-Gayon, et al. (2007): “Handbook of Enology: The Chemistry of Wine”
בס"ד
בס"ד
Nitrogen is present in grape must in different forms: inorganic (ammonium) and organic
(amino acids, peptides and proteins).
Nitrogen that can be used by yeast during alcoholic fermentation is called “assimilable
nitrogen.” The needs for assimilable nitrogen are different according to the wine yeast
used.
Yeast cells also contain nitrogen in the form of proteins, peptides particularly tripeptides
and amino acids.
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
YAN (yeast assimilable nitrogen) includes free α-amino acids (AA), ammonium and some
peptides.
Inactivated yeast-based nutrients can also be used by wine yeast to efficiently carry out
alcoholic fermentation.
Proline is the only free α-amino acid (AA) not assimilated by yeasts during alcoholic
fermentation, even if it is one of the AAs most abundant in grape must.
The main free AAs (in quantity) in grapes include proline, arginine, glutamate.
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
• It represents an important nutritional factor for yeasts during alcoholic fermentation due
to its function in protein synthesis and sugar transport.
• It is essential for the biosynthesis of wine quality markers like higher alcohols, thiols
and esters by wine yeast. YAN concentrations in natural grape musts range from about 60
mg/L to 500 mg/L, depending on grape variety, vintage and microclimate.
A low initial YAN concentration has been shown to cause slow and sluggish
fermentations.
YAN measurement is a good way for a winery to assess the general state of grapes in
terms of nitrogen quantity, but not necessarily nitrogen quality, which is another key
parameter from the point of view of fermentation and the sensory quality of the final wine.
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Organic nitrogen has been shown to be the most efficient and complete nutrient to
complete fermentation and achieve the full aromatic potential of the grape.
In must, one-third of the nitrogen is found in ammonium form and two-thirds in amino
acids, but musts are often deficient in YAN (less than 150 mg/L).
Nitrogen from external sources can be added to the must to assist with fermentation.
2. Organic nitrogen: in the form of proteins, peptides, tripeptides and free amino acids that
are part of the cells from yeast.
When yeast cells are inactivated through various production processes (inactivated
yeast and yeast autolysate), the nitrogen found in the yeast proteins, peptides,
tripeptides and free amino acids is available for the fermenting yeast cells to use during
alcoholic fermentation
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
As soon as must has a nitrogen deficiency at the end of the growth phase, there is a
decrease in protein synthesis and sugar transport activity.
YAN addition to nitrogen deficient must leads to a significant decrease in fermentation length
by reactivating protein synthesis and increasing sugar transport speed, which results in an
increase in the fermentation rate.
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
As a result, the nitrogen composition of the must can modulate the aroma profile of a wine.
Organic nutrients have also been shown to positively influence the formation of aroma
compounds when added during alcoholic fermentation.
Thiols: During fermentation it has been shown that excessive inorganic ammonium or
addition at the beginning of alcoholic fermentation limits the release of varietal thiols by the
yeast. Catabolic repression by the inorganic nitrogen in the form of ammonium prevents the
synthesis of amino acid transporters in fermenting yeast. It also limits the entry of thiol
precursors of the cysteinylated form into the cell and, consequently, their intracellular
conversion into volatile thiols.
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Adapted from: Ortiz-Julien, et al., 2016. Nitrogen Plays Many Roles During Fermentation, WINEMAKING PRACTICAL WINERY & VINEYARD
בס"ד
בס"ד
Adapted from: Pretorius et al.(2007). Nitrogen management is critical for wine flavour and style. Wine Industry Journal, 2(6), 24-30.
בס"ד
בס"ד
Adapted from: Pretorius et al.(2007). Nitrogen management is critical for wine flavour and style. Wine Industry Journal, 2(6), 24-30.
בס"ד
בס"ד
Adapted from: Pretorius et al.(2007). Nitrogen management is critical for wine flavour and style. Wine Industry Journal, 2(6), 24-30.
בס"ד
בס"ד
א יור סכמאטי של יצירת חומרים נדיפים במהלך התסיסה האלכוהולית בשמרי יין ((Swiegers, Bartowsky, Henschke & Pretorius, 2005
…To be continued