Antioxidant Activity of ß-glucan Derived

from Yeast and Mold Culture Using Fruits

Waste as Fermentation Substrate

CASEY DIO
BN17280279

UNIVERSITI MALAYSIA SABAH

FAKULTI SAINS MAKANAN DAN PEMAKANAN
FOOD SCIENCE AND BIOPROCESSING
2018
INTRODUCTION
 3
Introduction

Antioxidant is any substance that
delays, prevents or removes
oxidative damage toa target
molecule. (Halliwell,2007)
Beta-Glucans are polysaccharides of
D-glucose monomers linked through
β-glycosidic bond. Beta glucan could
be found in a variety of natural
sources such as yeast, mushrooms,
bacteria, algae, barley and oat. (Zhu,
Du, Bian, & Xu, 2015)
A. Oryzae and S.Cerevisiae in food
fermentation industries
prompted industrial applications
of A. oryzae to be listed as
Generally Recognized as Safe
(GRAS) by the Food and Drug
Administration (FDA) in the USA.
 4
Introduction

Fungal Cell Wall Yeast Cell Wall

(Cheol et al. 2013) (Philipp,2016)

91,87 Million 506,64 Million Kg 0,92 Million Ton

Kg (ATAP,2016) (ATAP,2016)
(Badan Pusat
Statistik, 2016)
 5

Problem statement
1. Extracted glucan from microorganism produce a very few yield and have
a difficult procedure to purify.
2. Research on production of β-glucan from cultures of microorganisms by
utilizing fruit waste as a fermentation medium has never been done. So
the properties may increase or decrease.
3. Lack of study on using β-Glucanase enzyme to determine β-glucan.
 6

Justification of the Study

• Beta glucan extracted from the cell wall of microorganism(1,6-β-glucan)
is known to have a much more biology activity then obtained from plant.
• To Utilizing agricultural byproduct into a valuable product such as beta-
glucan.
• The Research on determining the antioxidant of the β-Glucan
obtained from Saccharomyces cerevisiae and Aspergillus Oryzae by
utilizing fruit waste as a fermentation medium can be used as
reference for literature study.
 7

Objective
1. Determining the best fruit waste used to producting the β-glucan
from S. cerevisiae and A. Oryzae
2. Determining the property of the extracted β-glucan from S. cerevisiae
and A. Oryzae
3. Determining the physiochemical of the extracted β-glucan from S.
cerevisiae and A. Oryzae
 8

MATERIAL AND
METHOD

Bring the attention of your audience over a key

concept using icons or illustrations
RESULTS
 Working culture
Table 2. Inoculum Size
Mikroorganism Initial Cell (cfu/ml)
S. cerevisiae 1,28 x 107
A. Oryzae 4,3 x 102

Picture 1. Picture 2.
Working Culture Working Culture
S. cerevisiae A.Oryzae
Table 1. Crude Fiber of Fruit waste
Waste Crude
Fiber(%)
Pineapple Peel 19,81
Banana Peel 15,65
Papaya Peel 14,75
Picture 3. Papaya, Pineapple and Banana peel in powder from
 Total Plate Count of S.Cerevisiae in Fruit Waste

2%(w/v) of Fruit Waste 5%(w/v) of Fruit Waste

9 9

Viable Cells (Log cfu/ml)

Viable Cells (Log Cfu/ml)

8 8
7 7
6 6
5 5
4 4
3 3
2 2
1 1
0 0
Day 0 Day 1 Day 2 Day 3 Day 4 Day 0 Day 1 Day 2 Day 3 Day 4
Time Time

Control 2% Banana 2% Pineapple 2% papaya Control 5% Banana 5% Pineapple 5% Papaya

Figure 1. Growth of S.Cerevisiae in 2%(w/v) Fruit Waste Figure 2. Growth of S.Cerevisiae in 2%(w/v) Fruit Waste
 10%(w/v) of the fruit waste
10

Viable Cells (Log Cfu/ml)

9
8
7
6
5
4
3
2
1
0
Day 0 Day 1 Day 2 Day 3 Day 4
Time

Control 10% Banana 10% Pineapple 10% Papaya

Figure 3. Growth of S.Cerevisiae in 10%(w/v) Fruit Waste

 Production of β-glucan in Broth without waste (Control)
β-glucan extraction based on Noppawat et al. (2017)
Table 4. Crude dried β-glucan yield
Mikroorganism Yield (%)
Saccharomyces
0,1592
Cerevisiae
Aspergillus Oryzae
13,2032

Picture 4.
Picture 5. Dry Crude
Dry Basis of Crude
Beta glucan from A.Oryzae
Beta glucan from S. Cerevisiae
Production of β-glucan in Broth without waste (Control)

Table 5. pH from the medium during fermentation with S.Cerevisiae and A.Oryzae

Microorganism Day 0 Day 1 Day 2 Day 3 Day 4 Day 5

Saccharomyces
Cerevisiae
6,7 6,16 6,2 6,04 6,7 6,6
Aspergillus
Oryzae 6,89 6,84 6,3 6,1 6,23 6,64
 Glucose Standard Curve
3

2.5 y = 0.017x + 0.8697

R² = 0.9984
Abosrbance (490 nm)
2

1.5
Glucose
Linear (Glucose)

0.5

0
0 10 20 30 40 50 60 70 80 90 100
Concentration (ppm)

Graphic 1. Glucose Standard Curve

 β-glucanase Enzyme

Picture 6. Media production of Enzyme Beta Glucanase Picture 7. Supernatan from the media Production
 Beta-glucanase Enzyme Activity from S.Cerevisiae
and A.Oryzae
12

10

0
day 1 day 3 day 5
S. cerevisiae 9.810958235 10.81291902 3.827428824
A. oryzae 0.005827255 3.680958235 6.152330784

S. cerevisiae A. oryzae

Graphic 2. β-glucanase enzyme activity from the supernatant of the media broth
Further Study Needed
• Determining the glucose of each waste
• Total Plate Count From A.Oryzae
• Beta glucan extraction with the media of best concentration of the fruit
waste with A.Oryzae and S. Cerevisiae
• Analyzing of the property and physiochemical of the beta glucan
Conclusion
• 5% Concentration of each fruit waste have higher population than 2% and
10% on S.Cerevisiae
• 5% Pineapple peel have the highest population of S.Cerevisiae than 5%
banana peel,5% papaya peel. 5% Pineapple peel will produce more higher
yield
• A.Oryzae have higher yield than S. Cerevisiae in media without waste
(control)
• Beta Glucanase Enzyme produced from S.Cerevisiae have higher activity
than A.Oryzae
References
• Badan Pusat Statistik.2016. Konsumsi Buah dan Sayur Susenas Edisi Maret 2016.
• Cheol M, Lee et al. 2013. Raman Spectroscopy and Density Functional Theory Calculations of
Beta Glucan and Chitins in Fungal Cell Walls.
• Kementerian Pertanian. 2016. Komoditas Pertanian Sub Sektor Hortikultura. Nenas. ISSN :
1907-1507
• Kementerian Pertanian. 2016. Komoditas Pertanian Sub Sektor Hortikultura. Pisang. ISSN :
1907-1507
• Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 4. Oxford: Clarendon; 2007.
• Philipp H. Fesel and Alga Z. 2016. Beta-Glucan : Crucial Component of the fungal cell wall and
elusive MAMP in Plants.
• Zhu, F. M., Du, B., Bian, Z. X., & Xu, B. J. (2015). Beta-glucans from edible and medicinal
mushrooms: characteristics, physicochemical and biological activities. Journal of Food
Composition and Analysis, 41, 165e173.
• Noppawat et al. 2017. Extraction of β-glucan from Saccharomyces cerevisiae : comparison of
different extraction methods and in vivo eassesment of immunomodulator effect in mice.
Thank You !