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Effect of Nitrogen Sources and Vitamins Addition on Baker's Yeast

Fermentation Activity

Article · June 2017

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 Egyptian Sugar Journal, June 2017 Vol.9 57 -66

Effect of Nitrogen Sources and Vitamins Addition on Baker's

Yeast Fermentation Activity
Zohri1*A. A., Fadel M.2, M. Hmad3 and H. F. El-sharkawey3

1 Botany and Microbiology Department, Faculty of Science, Assiut University Egypt.

2
Microbial Chemistry Department, National Research Center, Cairo, Egypt.
3 Chemical Factories of Hawamdia, Giza, Egypt.

*
Corresponding author: Abdel-Naser A. Zohri email: zohriassiut@yahoo.com Tel:
01006464109
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Abstract

The source and content of nitrogen and vitamins in the fermentation medium are very
important in the development of Baker's yeast production since they both affect the
growth of Saccharomyces cerevisiae. Furthermore, the composition of the growth
medium and the environmental conditions are known to affect the cell membrane and
fatty acid composition.The aim of this work was to study the effect of nitrogen sources
and biotein as well as calciumpanthonate on the biomass yield and fermentative power
of baker,s yeast. Results appeared that urea was the most suitable nitrogen source than
diammonium phosphate, ammonium sulphate and ammonium nitrate. In the presence
of urea as nitrogen source, the yeast biomass yield reached to 152.9 g/L and the
fermentation power was 1900 cm3of CO2 evolved from 10 g yeast sample that
contains 28% dry matter. Markedly enhancing in biomass yield and fermentative
power were obtained when biotin and calciumpantothinate added to growth.

Keywork: Fatty acid; Saccharomyces cerevisiae; Nitrogen; Biotin, Ca-pantothenate.

Introduction
Nitrogenous compounds are important components for yeast production.
Low levels of yeast assimilable nitrogenous compounds have been related to
lower fermentation rates and longer fermentation time (Bely et al.,1990).These
effects also depend on the nitrogen source, sincegrowth on good nitrogen source
seems to yield high growth rates (TerSchureet al.,2000).The chemical
composition of yeast cells depends mainly on the cultivation conditions and
composition of the growth medium. In fact, the synthesis of yeast biomass is
58 Zohri A. A., et. Al. (2017) Egyptian Sugar Journal, Vol.9 : 57 - 66
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clearly dependent on the content of nitrogen in the growth medium, since the
proportion of nitrogenous compounds in yeast cells is about 50% (by weight).
Abromovet al. (1994) reported that cells can synthesize all the required proteins
from inorganic nitrogen provided by ammonium salts and organic carbon. In
yeast cell composition, fatty acids is one of the most variable parameters and it
is clearly influenced by environmental factors (Ratllendge and Evans,1989).

Saccharomyces cerevisiae is known as being a natural rich source of B-

complex vitamins. However, it is very difficult to guarantee constant levels of
B-complex vitamins in brewer’s by-products and even baker’s yeast. Baker’s
yeast production is supplemented with known levels of B-complex vitamins and
therefore we can guarantee that this product contains always an amount of each
vitamin.As the use of nutritional supplements increases, there is a growing
consumer demand for more sophisticated information on the products they are
consuming. Some of the questions on vitamin yeast concern the importance of
B-complex vitamins in the diet, the bioavailability of organic vitamin yeast
versus inorganic sources and toxicity issues. We hope that this information will
be useful to those considering using vitamin supplements. We feel that B-
Complex Vitaminsare anatural, convenient, safe and economical way to
increase β-complex vitamins intake.

Biotin functions as the coenzyme transferring carbon dioxide in four

reactions: pyruvate carboxylase, a key step in gluconeogenesis; acetylCoA
carboxylase, the first step of fatty acid synthesis; and in two other important

carboxylases. Pantothenic acid has a central role in a wide variety of metabolic

pathways. It is required for the synthesis of coenzyme A (CoA), which is
required in the metabolism of carbohydrates, amino acids, and fatty acids.It also
plays an important role in the synthesis of the sterols like cholesterol.
Pantothenic acid is essential for the synthesis of acetylcholine, which is a
 Effect of nitrogen sources and vitamins addition on baker's yeast …..… 59
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chemical mediator of nervous communication. Calcium pantothenate is known
as vitamin B5 or pantothenic acid.

The aim of this investigation was designed to study the effect of different
four nitrogen sources (urea, diammonium phosphate, ammonium sulphate and
ammonium nitrate) as well as different two concentrations of each of biotein and
calciumpanthonateas vitamins on the biomass yield and fermentative power of
baker,s yeast.

Materials and Methods

Yeast strain
SaccharomycescerevisiaeF-1001 was obtained from Mcrobial Chemistry
Department collection, National Research Center Dokki, Giza, Egypt.
Chemicals and media
Urea 46% nitrogen was obtained from ABU- QIR Fertilizers and
chemical industries Co., Alexandria, Egypt. Other nitrogen sources
(diammonium phosphate; ammonium sulphate and ammonium nitrate were
obtained from ElsafaCo., Egypt; Echem. Technology Co., Ltd and Richin.
International Trad-Dalin- Co., Ltd, respectively). Phosphoric acid was obtained
from Shuangjing china Magnesium sulphate 99% Heptahyrat, china. Antifoam
struktol j673 was obtained from Schill + seilacher, Hamburg Germany. Biotin
and calciumpantothinate were obtained from Natrol. Co., USA and SIGMA-
ALDAICH Co.Llc, respectively.
The yeast growth liquid medium (YMPG) consists of (g/L): 3 yeast
extract, 3 malt extract,5 peptone and 5 glucose per liter of distilled water at pH
5. Sugar cane molasses collected from Upper Egypt sugar factories and used as
fermentation medium for Baker yeast production. .
60 Zohri A. A., et. Al. (2017) Egyptian Sugar Journal, Vol.9 : 57 - 66
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Inoculum preparation and propagation

Saccharomycescerevisiae strain was sub-cultivated on slants of YMPGA

supplemented with chloramphenicol (0.1 g /l).The cultures were incubated at
33°C for 48 hours.The yeast colons were propagated by transferred to YMPG
sterilized broth and incubated at 33°C for 24 hours. As a 10% V/V inoculum
size was used in each case.
Addition of nitrogen sources and vitamins
Different four sources of nitrogen (urea, di ammonium phosphate
(D.A.P), ammonium nitrate (A.N) and ammonium sulfate (A.S) were tested,
individually, in this study at equivalent 8 % nitrogen level. Based on the
molecular weight of urea used 1.5 g / liter,while for di ammonium phosphate,
ammonium nitrate and ammonium sulfate used 3.29, 2and 3.29 g / liter,
respectivally. On the other hand, two different concentrations of each of the two
tested vitaminswere individually examined: 10 & 20 g of biotin per 10000 liter
of fermented medium (0.1 & 0.2 %) and 25 & 50 g of calciumpantothenate per
10000 liter of fermented medium (0.25 & 0.50 %). All experiments were carried
out in 1000 ml conical flask containing 200 ml of basal sterilized medium
(molasses 8 % total sugar and 0.03% magnesium sulfate) at pH 5.0.The flasks
sterilized by autoclaving at 120 °C for15 min., inoculated with 10% of yeast
propagated inoculumand incubated on rotatry incubator at 200 rpm for 24 hours
at 33°C.
Analysis of baker yeast

Determine of cell biomass: at the end of the yeast growth, the culture was
harvested using centrifugation at 4500 rpm for 20 minutes.The supernatant and
pellet was separated and weighted as gram per liter.
Determine of dry matter (D. M): dry matter determined accordingto the
method described in the Association Official of Analytical Chemistry (A.O.A.C)
(1998).
 Effect of nitrogen sources and vitamins addition on baker's yeast …..… 61
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Determine of alcoholic content: alcoholic percentage determined using
Ebulliometer (Fadelet al., 2014).
Determination of pH: pH values were measured by Microprocessor pH-
mv meter pH 526.
Determine of leavening power: leavening power was defined as volume
cm3of CO2 evolved from 10 g yeast sample that contains 28% dry matter and
determined by ferment graph (SJA) in chemical factories of hawamdia, Giza,
Egypt.

Results and Discussion

Effect of nitrogen source on biomass yield, pH and ethanol production as
well as fermentation power of S. cerevisiae
Saccharomycescerevisiae encounters agreat variety of nitrogen sources in
its natural habitat (Cooper, 1982), but not all nitrogen source support growth
equally (TerSchureet al.,2000). Four sources of nitrogen (urea, di ammonium
phosphate (D.A.P), ammonium nitrate (A.N) and ammonium sulfate (A.S) were
tested in this study at equivalent nitrogen level. Data presented in Table (1 a&b)

reveal that nitrogen source affects biomass yield, pH and ethanol production as
well as fermentation power of S.cerevisiae. Good result was archived when urea
supplemented in growth medium giving highest biomass with highest
fermentation power. The obtained results can be discussed on the light of the
effect of urea on the pH, as the measured pH was 4.46. Also, applying urea was
more suitable than other tested nitrogen sources. The suitable pH in growth
medium was lead to gain biomass yield andlow ethanol level.The chemical
composition of yeast cells depends mainly on the cultivation conditions and
composition of the growth medium. In fact, the synthesis of yeast biomass is
clearly dependent on the content of nitrogen in the growth medium, since the
proportion of nitrogenous compounds in yeast cells is about 50% (Abromovet
al.,1994).
62 Zohri A. A., et. Al. (2017) Egyptian Sugar Journal, Vol.9 : 57 - 66
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Table (1a): Effect of nitrogen source on biomass yield, pH and ethanol
production of S.cerevisiae.

Urea D.A.P A.N A.S

Nitrogen source
Item
Yeast yield g/l 152.9 152.0 142.4 141.9
pH 4.46 4.22 3.76 3.76
Ethanol % ( v/v) 2.1 2.2 2.3 2.4

Table (1b): Effect of nitrogen source on fermentation power (F.P.) in the first
hour (1h) and second hour (2h) of S. cerevisiae.

N Urea D.A.P A.N A.S

F.P.
1.H. 900 850 600 450
2.H. 1900 1800 1160 950

Effect of biotin level on biomass yield, pH and ethanol production as well as

fermentation powerof S.cerevisiae

Data presented in Table (2a&b) reveal that biotin level in fermentation

medium affects yeast yield, pH and ethanol production as well as fermentation
power of S. cerevisiae. Promising data were achieved when Biotin at low
concentration (10 g/10000 liters) was supplemented in growth medium giving
highest biomass with highest fermentation powerand low ethanol yield than the
high biotin level (20 g/10000 liters). Biotin functions as the coenzyme
transferring carbon dioxide in four reactions: pyruvate carboxylase, a key step in
gluconeogenesis; acetylCoA carboxylase, the first step of fatty acid synthesis;
and in two other important carboxylases.It is assumed that B-Complex has a
central role in a wide variety of metabolic pathways. It is required for the
synthesis of coenzyme A (CoA), which is required in the metabolism of
carbohydrates, amino acids, and fatty acids. It also plays an important role in the
synthesis of the sterols like cholesterol(Andrewset al.,1957).
 Effect of nitrogen sources and vitamins addition on baker's yeast …..… 63
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Table (2a): Effect of biotin level on biomass yield, pH and ethanol
production of S.cerevisiae.

Biotin Level Biomass yield pH Ethanol %

(g/L) v/v
0.1% 160.5 4.92 2.0
0.2% 154.3 5.12 2.1
Control 152.9 4.46 2.1

Table (2b): Effect of biotin level on fermentation power (F.P.) in the

first hour (1h) and second hour (2h) of S. cerevisiae.

Time 1h 2h
Biotin level
0.1% Biotin 950 1800
0.2% Biotin 800 1550
Control 900 1600

Effect of calciumpantothenate level on biomass yield, pH and

Ethanol production as well as fermentation power of S.cerevisiae:

Data presented in Table (3a&b) show that Ca-pantothenate concentration

affects yeast yield, pH and ethanol production as well as fermentation power of
S.cerevisiae.The more promising data were achieved when Capantothenate at
low concentration (25 g/10000 liters) was supplemented in growth medium
giving highest biomass with highest fermentation power and low ethanol yield
thanother tested concentration.This result is similar to those recorded by
Williams et al. (936). They found that the addition of a minute amount of
pantothenic acid in production medium increased the fermentation power of
Baker yeast.
64 Zohri A. A., et. Al. (2017) Egyptian Sugar Journal, Vol.9 : 57 - 66
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Table (3a): Effect of Capantothenate level on biomass yield, pH and
Ethanol production of S.cerevisiae.

Capantothenate Biomass PH Ethanol % v/v

Level yield (g/L)
0.25 % 159.5 5.0 1.8
0.50 % 153.6 5.1 1.9
Control 152.9 4.46 2.1
Table (3b): Effect of Ca- pantothenate level on fermentation power
(F.P.) in the first hour (1h) and second hour (2h) of S.cerevisiae.

Time 1h 2h
Ca-pantothenate level
0.25 % 1000 2000
0.50 % 700 1300
Control 900 1600

References
Abromov, S.H.A., Efendieva,D.A. andkotecko, S.TS. (1994): Effect of
growth medium on protein content of yeast Saccharomyces cerevisiae. Appl.
Biochem. Microbiol. 30 (2): 225-227.

Andrews, E. A., Chrestman, J. F. and Williams, V. R. (1957): Studies on a

biotin-hexokinase interrelationship. Arch. Biochem. Biophys. 66 (1): 234 –
240.

Bely, M., Sablay, J.M.and Barre, P. (1990):Automatic detection of

assimilable nitrogen deficiencies during alcoholic fermentation inenological
condition J.Ferm.Bioeng. 70 :246-252.

Cooper, T.G. (1982): Nitrogen metabolism in Saccharomyces cerevisiae.In:

The Molecular Biology of the Yeast Saccharomyces (Strathern, J.N., Jones,
E.W. and Broach, J.R., Eds.), pp. 39 - 99. Cold Spring Harbor Laboratory,
Cold Spring Harbor, NY.
 Effect of nitrogen sources and vitamins addition on baker's yeast …..… 65
‫ـــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــــ‬
Fadel, M., Abdel Naser, A. Zohri. Makawy, M., Hsona. M.S. and Abdel-
Aziz A M (2014): Recycling of vinasse in ethanol fermentation and
application in Egyptain Distillery Factory. African Journal of
Biotechnology13 (47):4390 – 4398.

Ratlendge,C. and Evans, C. T. (1989): Lipids and their metabolism in: Rose,
A.H., Harrison, J.S. (eds.): The yeast, vol .3, 2 nd edition Academic Press,
London , pp.367

Steels, E.L, Learmonth, R.P. and Watson, K. (1994): Stress tolerance and
membrane lipid unsaturation in Saccharomyces cerevisiae grown aerobically
or anerobically. Microbiology: 140, 569 – 576.

Ter Schure, E.G., van Rial, N.A.W and Verrips, C.T, (2000): The
role of ammonia metabolism in nitrogen catabolite repression in
Saccharomyces cerevisiae. FEMS 24 (1): 67 – 83.

William, R. J., Mosher, W. A. and Rohrman, E, (1936): The importance of

'pantothenicacid' in fermentation, respiration and glycogen storage. Biochem.
J. 30 (11): 2036–2039.
 ‫‪66‬‬ ‫‪Zohri A. A., et. Al. (2017) Egyptian Sugar Journal, Vol.9 : 57 - 66‬‬
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‫^‪fà≈ÿ^=ïÄŸª‬‬

‫ﺗﺄﺛﲑ ﺇﺿـﺎﻓﺔ ﻣﺼـﺎﺩﺭ ﺍﻟﻨﻴﱰﻭﺟﲔ ﻭﺍﻟﻔﻴﺘﺎﻣﻴﻨﺎﺕ ﻋﻠﻰ ﻛﻔﺎءﺓ ﺍﻟﺘﺨﻤﲑ ﻟﻠﺨﻤﲑﺓ ﺍﳉﺎﻓﺔ‬

‫‪‬‬
‫‪‬‬
‫‪‬‬
‫‪ ‬‬
‫‪‬‬
‫‪‬‬

‫ﻤﺼدر وكﻤ�ﺔ اﻟﻨﯿﺘروﺠﯿن واﻟﻔﯿﺘﺎﻤﯿﻨﺎت ﻓﻰ وﺴـط اﻟﺘﺨﻤﯿر ﻫﺎم ﺠداً ﻓﻰ إﻨﺘﺎج ﺨﻤﯿرة اﻟﺨ�ﺎز‬
‫ﺤﯿث ان كﻼﻫﻤﺎ ﯿؤﺜر ﻓﻰ ﻨﻤـو اﻟﺨﻤﯿرة ‪.‬‬
‫اﻛﺜر ﻤن ذﻟك ﻓﺈن ﺘركﯿب وﺴـط اﻟﻨﻤو واﻟظروف اﻟﺒﯿﺌ�ﺔ ﻤﻌروف إﻨﻬﺎ ﻤؤﺜرة ﻋﻠﻰ اﻟﻐﺸﺎء اﻟ�ﻼ زﻤﻰ‬
‫ﻟﺨﻠ�ﺔ اﻟﺨﻤﯿرة وﺘركﯿب اﻻﺤﻤﺎض اﻟدﻫﻨ�ﺔ‪.‬‬
‫ﻫدف ﻫذا اﻟ�ﺤث ﻫو دراﺴـﺔ إﻀﺎﻓﺔ اﻟﻤﺼﺎدر اﻟﻨﯿﺘروﺠﯿﻨ�ﺔ وﻓﯿﺘﺎﻤﯿن اﻟﺒﯿوﺘﯿن وﻓﯿﺘﺎﻤﯿن �ﺎﻨﺜوﺜﯿﻨﺎت‬
‫اﻟﻛﺎﻟﺴﯿوم ﻋﻠﻰ اﻟﻛﺘﻠﺔ اﻟﺤﯿو�ﺔ وﻗوة اﻟﺘﺨﻤﯿر ﻟﺨﻤﯿرة اﻟﺨ�ﺎز‪.‬‬
‫اﻟﻨﺘﺎﺌﺞ اظﻬرت ان اﻟﯿور�ﺎ ﻫﻰ اﻓﻀـﻞ ﻤﺼدر ﻨﯿﺘروﺠﯿﻨﻰ �ﺎﻟﻤﻘﺎرﻨﺔ ﺒ�ﺎﻗﻲ اﻟﻤﺼﺎدراﻟﻤﺨﺘﺒرة ‪ ،‬وﺠود‬
‫اﻟﯿور�ﺎ كﻤﺼدر ﻨﯿﺘروﺠﯿﻨﻰ زاد اﻟﻨﺘﺎﺌﺞ ﻤن اﻟﻛﺘﻠﺔ اﻟﺤﯿو�ﺔ ﻟﻠﺨﻤﯿرة اﻟﻰ ‪ ١٥٢٬٩‬ﺠم‪/‬ﻟﺘر وﻗوة‬
‫اﻟﺘﺨﻤﯿر اﻟﻰ ‪ ٢٩٠٠‬ﺴم‪ ٣‬ﻤن ﺜﺎﻨﻰ اﻛﺴـﯿد اﻟﻛر�ون اﻟﻨﺎﺘﺞ ﻤن ‪ ١٠‬ﺠم ﺨﻤﯿرة ﺘﺤﺘوى ﻋﻠﻰ‬
‫‪ %٢٨‬ﻤﺎدة ﺠﺎﻓﺔ ‪.‬‬
‫ﺤدﺜت ز�ﺎدة ﻤﻠﺤوظﺔ ﻓﻰ اﻟﻛﺘﻠﺔ اﻟﺤﯿو�ﺔ وﻗوة اﻟﺘﺨﻤﯿر ﻟﺨﻤﯿرة اﻟﺨ�ﺎز ﻓﻰ وﺠود كﻞ ﻤن ﻓﯿﺘﺎﻤﯿن‬
‫اﻟﺒﯿوﺘﯿن واﻟ�ﺎﻨﺜوﺜﯿﻨﺎت اﻟﻛﺎﻟﺴـﯿوم‪.‬‬

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