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Effects of Calcium Addition during Enzymatic Hydrolysis and Different Pretreatments on Drying Duration of Brown and White Sago Sugars
Nurazureen binti Matnin
(43375)
Bachelor of Science with Honours (Resource Biotechnology)
2016
,... l'usafKlaidnlat Maklumat AK"iifemi}( UNr -~-. K~IDm -~~K~~~A~' ~~~m;K RAWAK
UNl~A5
IIIIIIIIIIIIIII~IIIIIIII 1000272692
Effects of Calcium Addition during Enzymatic Hydrolysis and Different Pre-
treatments on Drying Duration of Brown and White Sago Sugars
Nurazureen binti Matnin (43375)
•
j'
This project is submitted in partial fulfillment of the requirements for the Degree of
Bachelor Science with Honours
(Resource Biotechnology)
Supervisor: Professor Dr Kopli Bujang
I
l
Faculty ofResource Science and Technology Universiti Malaysia Sarawak
2016
ACKNOWLEDGEMENT
First of all, I would like to express my deepest gratitude to God the highest for
granting me the strength to accomplish my final year project. A special thanks and sincere
appreciation dedicated to my supervisor, Professor Dr. Kopli Bujang for his guidance and
support throughout this project. The insight, motivation and experience were certainly
appreciated.
Million thanks also dedicated to all postgraduate students of the Biochemistry
Laboratory, Faculty of Resource Science and Technology especially to my advisor, Miss
Nadia Dayana Sikem for her insight, guidance and willingness to offer suggestions and
commentary for improvements during my thesis topic research. Additionally, I am
indebted to Miss Sharifah binti Mohammad, Mr Muhammad Norhelmi bin Ahmad, Miss
Aimi Nadia binti Saharuddin and Miss Daisy Dinsi for their support and advice throughout
this proj ect.
Next, I am grateful to all the members of my lab and friends for their
companionship and support while perfonning this project. Last but not least, I would like
to thank my family for their understanding and encouragement in many ways throughout
this project.
I
DECLARATION
I hereby declare that no portion of the work has been submitted in the support of an
application for another degree qualification of this or any other university or institution of
higher learning .
........ ... ... ~.... ....... ....... .. . .
(NURAZUREEN BINTI MATNIN)
Resource Biotechnology Programme
Department of Molecular Biology
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
II
flus.t Khidmat MakJumat Akadfmik ~1VERS t,;-\YS A S'.A.:>.AWAK
T ABLE OF CONTENTS
ACKNOWLEDGEMENT I
DECLARATION II
TABLE OF CONTENTS III
LIST OF ABBREVIATIONS VI
LIST OF TABLES VII
LIST OF FIGURES
ABSTRACT 1
VIII
1.0 INTRODUCTION 2
1.1 Overview and Problem Statement 2
1.2 Objectives 4
2.0 LITERATURE REVIEW 5
2.1 Sugar Industry 5
2.1.1 World Market and Demand 5
2.1.2 Sources of Sugar 5
2.1.3 Uses of Sugar 6
2.2 Sago Industry 7
2.2.1 The Sago Palm 7
2.2.2 Production of Sago Starch 8
2.2.3 Utilization and Bioconversion of Sago 8
2.3 Types of Sago Sugars 10
2.3.1 Brown Sago Sugars 11
2.3.2 White Sago Sugars 11
III
,....
123.0 MATERIALS AND METHODS
3.1 Materials
3.1.1
3.1.2
3.1.3
3.1.4
3.2 Methods
3.2.1
3.2.2
3.2.3
3.2.4
Sago Starch 12
Hydrolytic Enzymes 12
Calcium 12
Powdered Activated Charcoal 12
13
Standard Hydrolysis of Sago Starch into Sugar 14
Purification of Sago Sugar 14
Drying and Crystallization of Sago Sugar 14
Analysis of Sago Sugar 15
3.2.4.1 Reducing Sugar using DNS Method 15
3.2.4.2 Total Phenolic Content 16
3.2.4.3 Total Flavonoid Content 17
3.3 Analytical Methods 17
3.3.1 Effects of Calcium during Hydrolysis of Sago Starch 17
3.3.2 Effects of Different Pre-Treatments on Brown Sago Sugars (with
and without addition of Calcium) 18
3.3.2.1 Sedimentation 18
3.3.2.2 Centrifugation 19
I' 3.3.3 Effects of Calcium on White Sago Sugar (with and without
Calcium) 20
3.3.4 Comparison of Drying Time and Crystallization between
Different Pre-Treatments in Brown Sago Sugars (with and
IV
J
20 without Calcium)
,....
3.3.5 Comparison of Drying Time and Crystallization on White Sago
Sugars (with and without Calcium) 20
4.0 RESULTS AND DISCUSSIONS 21
4.1 Effects of Different Pre-Treatments on Brown Sago Sugars (with and
without Calcium) 21
4.2 Effects of Addition of Calcium on White Sago Sugars 24
4.3 Comparison of Drying Time and Crystallization between Different Pre-
Treatments on Brown Sago Sugars (with and without Calcium) 27
4.4 Comparison of Drying Time and Crystallization on White Sago Sugars
(with and without Calcium) 31
5.0 CONCLUSIONS 34
6.0 REFERENCES 35
APPENDICES 38
v
II
ANOVA
AMG
BSS
°C
DE
DNS
DSS
GA
GC
HSS
mL
nm
PAC
QE
rpm
t
TFC
TPC
WSS
LIST OF ABBREVIATIONS
Analysis of Variance
Amyloglucosidase
Brown Sago Sugar
Degree Celsius
Dextrose Equivalent
Dinitrosalicylic Acid
Dried Sago Sugar
Gallic Acid
Glucose Concentration
Hydrolyzed Sago Sugar
millilitre
Nanometre
Powdered Activated Charcoal
Quercetin Equivalent
Revolutions per minute
Ton
Total Flavonoid Content
Total Phenolic Content
White Sago Sugar
VI
II
,.....
Table
Table 1
Table 2
Table 3
Table 4
Table 5
Table 6
Table 7
Table 8
Table 9
Table 10
Table 11
Table 12
Table 13
Table 14
Table 15
•
LIST OF TABLES
Page
Development of food products from sago 9
Development of energy products from sago 10
Reducing sugar, TPC and TFC of different pre-treatments on brown 21
sago sugars
Reducing sugar, TPC and TFC of white sago sugars 24
Drying time between different pre-treatments on brown sago sugars 27
Mass of sugar, glucose recovery and glucose yield between different
pre-treatments on brown sago sugars 28
Drying time of white sago sugars (with and without calcium) 31
Mass of sugar, glucose recovery and glucose yield of white sago
sugars 31
Drying time and production of brown and white sago sugar 38
One-way ANOY A of reducing sugar, TPC and TFC analyses between
different pre-treatments on brown sago sugars (without calcium) 39
One-way ANOY A of reducing sugar, TPC and TFC analyses between
different pre- treatments on brown sago sugars (with calcium) 39
One-way ANOY A of reducing sugar, TPC and TFC analyses of white
sago sugars (with and without calcium) 40
One-way ANOY A of final mass, glucose recovery and glucose yield
between different pre-treatments on brown sago sugars (without 40
calcium)
One-way ANOY A of final mass, glucose recovery and glucose yield
between different pre-treatments on brown sago sugars (with calcium) 41
One-way ANOY A of final mass, glucose recovery and glucose yield
of white sago sugars (with and without calcium) 41
YII
1
LIST OF FIGURES
Figure Page
Figure 1 Flowchart of method involved during production and analysis of brown
and white sago sugars 13
Figure 2 Drying process of; (a) brown sago sugars and (b) white sago sugars. 15
Figure 3 Different pre-treatments used on hydrolyzed brown sago sugars; (a)
refrigerator, and (b) sedimentation in the oven 19
Figure 4 The clear liquid BSS was collected by using centrifugation pre-treatments
Figure 5 Reducing sugar between different pre-treatments on brown sago sugars
Figure 6 Total phenolic and flavonoid content between different pre-treatments on
on brown sago sugars 19
(with and without calcium) 22
brown sago sugars (with and without calcium) 24
Figure 7 Reducing sugar of white sago sugars (with and without calcium) 25
Figure 8 Total phenolic and flavonoid content of white sago sugars (with and
without calcium) 26
Figure 9 Final mass, glucose recovery and glucose yield between different pre
Figure 10 Crystallization of brown sago sugars (without calcium) between different
Figure 11 Crystallization of brown sago sugars (with calcium) between different
Figure 12 Final mass, glucose recovery and glucose yield of white sago sugars (with
Figure 13 Crystallization of white sago sugars; (a) without calcium and (b)with
Figure 14 Graph of concentration and absorbance of stock standard solution of
treatments on brown sago sugars (with and without calcium) 29
pre-treatments 30
pre-treatments 30
and without calcium) 32
addition of calcium 33
glucose concentration 42
Figure 15 Graph of concentration and absorbance of GA stock standard solution 42
Figure 16 Graph of concentration and absorbance of stock standard solution of
Quercetin 43
VIII
,.....
Figure 17 Drying process of brown sago sugar without the addition of calcium
(refrigerator) 44
Figure 18 Drying process of brown sago sugar without the addition of calcium
Figure 19 Drying process of brown sago sugar without the addition of calcium
Figure 20 Drying process of brown sago sugar with the addition of calcium
(oven) 44
(centrifuge) 45
(refrigerator) 45
Figure 21 Drying process of brown sago sugar with the addition of calcium (oven) 46
Figure 22 Drying process of brown sago sugar with the addition of calcium
(centrifuge) 46
Figure 23 Drying process of white sago sugar without the addition of calcium 47
Figure 24 Drying process of white sago sugar with the addition of calcium 47
Figure 25 Crystallization of brown sago sugars (without calcium) between different
pre-treatments; sedimentation in the (a) refrigerator and (b) oven 48
Figure 26 Crystallization of brown sago sugars (with calcium) between different
Figure 27 Crystallization of brown sago sugars (without calcium) between different
Figure 28 Crystallization of brown sago sugars (with calcium) between different
Figure 29 Crystallization of brown sago sugars (without calcium) between different
Figure 30 Crystallization of brown sago sugars (with calcium) between different
pre-treatments; sedimentation in the (a) refrigerator and (b) oven 48
pre-treatments; (a) sedimentation in the refrigerator and (b) centrifuge 49
pre-treatments; (a) sedimentation in the refrigerator and (b) centrifuge 49
pre-treatments; (a) centrifuge and (b) sedimentation in the oven 50
pre-treatments; (a) centrifuge and (b) sedimentation in the oven 50
Figure 31 Crystallization ofwhite sago sugars; (a) (b) (c) without calcium 51
Figure 32 Crystallization of white sago sugars; (a) (b) (c) with calcium 51
IX
:
1
,.....
Effects of Calcium Addition during Enzymatic Hydrolysis and Different PreTreatments on Drying Duration of Brown and White Sago Sugars
Nurazureen binti Matnin
Resource Biotechnology Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
Abstracts
Melroxylon sagu is the starch-producing crop that can produces sago sugar via an enzymatic hydrolysis process. This project has been aiming to identify which pre-treatments (sedimentation and centrifugation) and types of sago sugars can be an alternative to give rise to various qualities of sago sugars. In this project, analyses of white and brown sago sugars were compared by observing the result of drying time, reducing sugar, total phenolic and flavonoid contents, glucose recovery and yield . By the end of this project, sedimentation have been shown to be an excellent pre-treatments of brown sago sugars as it recorded the highest amount of reducing sugar, total phenolic content and total flavonoid content with the shortest drying time (6 days) whereas 221.32 ± 15.54 gIL, 0.099 ± 0.019 mglmL, 0.0097 ± 0.0007 mglmL (without calcium/oven) and 175.44 ± 15.54 giL, 0.091 ± 0.002 mglmL and 0.0119 ± 0.0006 mglmL (with calcium/refrigerator) respectively. Centrifugation is proven to be the most effective pre-treatments as it recorded the highest production of brown sago sugars, which is 298.5 ± 30.41 g (without calcium) and 346.5 ± 6.36 g (with calcium) and yield the finest, powder-like and lighter color of brown sago sugars (with and without calcium). Besides, white sago sugar (without calcium) have the best reducing sugars (175.15 ± 11.l2), TPC (0.00049 ± 0.00046) and TFC (0.00014 ± 0.00012). It is also revealed that white sago sugars (without calcium) have the best results for drying time (6 days), production of white sago sugars (283 ± 10.82 g), glucose recovery (97.30 ± 6.18 %), glucose yield (80.99 ± 5.82 %) and yield the finest, powder-like and whiter color of white sago sugars. However, it does not give different results despite adding calcium in white sago sugars as ANOV A indicated that p = 0.674 (final mass), p = 0.956 (glucose recovery) and p = 0.780 (glucose yield) at (l = 0.05.
Keywords: Melroxylon sagu, brown sago sugar, white sago sugar, calcium, pre-treatment
Abslrak
Melroxylon sagu adalah hasil kanji yang boleh menghasilkan gula sagu melalui proses hidrolisi enzimalik. Projek ini berlujuan unluk mengkaji kaedah (pemendapan dall pengemparall)dan jenis gula sagll unluk dijadikan a/lernalif yang boleh meningkalkan kualili gula sagu. Dalam projek in i, pembandingan anlara analisis gula sagu perang dan pulih lelah dijalankall dengan membandingkan masa pengeringan, kepekalan glukosa, TPC, TFC, pemulihan dan penghasilan glukosa .. Hasilnya, kaedah pemendapan merupakan kaedah lerbaik dalam gula sagu perang kerana menghasilkan kepekatan gula, TPC dan TFC yang tinggi iaitu 221.32 ± 15.54 giL, 0.099 ± 0.019 mglmL, 0.0097 ± 0.0007 mg/mL (tanpa kalsium/oven) dan 175.44 ± 15.54 giL, 0.091 ± 0.002 mglmL, 0.0119 ± 0.0006 mg/mL (tambahan kalsium/peti sejuk) dengan masa pengerillgan yang singkat (6 hari).Kaedah pengemparan merupakan kaedah yang efektif apabila menghasilkan jumlah gula saguperang yang tinggi iaitu 298.5 ± 30.41 g (tanpa kalsium) dan 346.5 ± 6.36 g (tambahan kalsium) serla menghasilkan gula sagu perang yang paling halus (tanpa dan tambahan kalsium). Selain ilu, gllia sagu pUlih (Ianpa kalsium)mengandungi kepekalan glukosa (175.15 ± 11.12), TPC (0.00049 ± 0.00046) dan TFC (0.00014 ± 0.00012), masa pengeringan (6 hari), berat gula (283 ± 10.82 g), pemulihan glukosa (97.30 ± 6.18 %), penghasilan glukosa (80.99 ± 5.82 %) yang lerbaik serla menghasilkan gula sagu pUlih yang paling halus dan putih. Namun begitu, lambahan kalsium tidak memberi kesan kepada gllia sagu pulili kerana ANOVA menunjukkan p = 0.674 (beral gula), p = 0.956 (pemulihan glukosa) dan p = 0.780 (penghasilan glukosa) pada a = 0.05.
Kata kunci:Melroxylon sagu, gllia sagu perang, gula sagll puti", kalsium, kaedah
1
1.0 INTRODUCTION
1.1 Overview and Problem Statement
Brown and white sago sugars are made from Metroxylon sagu that has been transformed
into sago starch via an extraction process. Metroxylon sagu grows in the hot humid tropics
of South-East Asia and Oceania and contains a valuable starch in its trunk (Flach, 1997).
This pinnate-leaved palm is able to grow naturally in swampy areas or peat soils without
the need of pesticide and also herbicide (Pei-Lang et al., 2006). The sago palm is also
resistance to floods, drought and strong winds because it is an extremely hardy plant.
Bujang (2014) pointed out that the sago palm is the only commodity that able to grow in
peat soil which occupies around 75% of coastal plains and lowland river basins in the state
ofSarawak in East Malaysia. This major advantage makes sago palm better than other cash
crops.
Bujang (2010) stated that recently the global consumption of sago starch had accounted for
about 3% of the total world market which initially were dominated by com, potato and
tapioca starches and achieved between 200,000 to 300,000 tons per annum. The
productivity of sago palm was measured to be four times than of paddy rice because it
contains a large amount of starch. Bujang (2010) stated that one ton of sago starch can be
converted into one ton of liquid sugar by using Ishizaki process and then produce 640 litres
of ethanol. Sugar derived from sago starch has the highest concentration at 205 giL with
recovery ofmore than 100% dextrose equivalent (DE) compared to com and tapioca starch
(Bujang, 2011). Besides, sago sugar contains mostly glucose (94%), with maltose and
other impurities, both at 3% each (Bujang, 2011). Nowadays, the consumption of sugar
derived from sugarcane and sugar beet contribute to certain health problem especially
diabetes. Thus, high glucose content in sago sugar may act as alternative for diabetics to
2
control and maintain the sugar level in their body. Furthermore, the presence of flavonoids
has increases the potential of sago sugar to be utilized in pharmaceutical industry (Bujang,
2015).
This study has been almmg to use sago sugar as the ongm to study the effects of
sedimentation, centrifugation and addition of calcium during solid liquid separation on its
quality and drying duration. Drying time of brown and white sago sugars is the interest of
the study because different methods of sugar processing can give rise to various quality of
sago sugar. It could shot up the production of brown and white sago sugars in compromise
with quality performance. The production of white sago sugar does not undergo different
pre-treatments because it will be purified by filtration using powdered activated charcoal
(PAC). Moreover, there are several analyses such as glucose concentration determination;
total phenolic and flavonoid content estimation are conducted on brown and white sago
sugar. Consequently, there are several problems occurred in conversion of sago sugar.
Different pre-treatments (sedimentation and centrifugation) before drying of brown sago
sugars may show different productivity of sugars production. After hydrolysis process,
there is sediment left in the hydrolysate which may not only contain the remaining of
valuable sugar, but also impurities that may contribute to bitter taste in sugar production.
Moreover, sedimentation of brown sago sugar is time consuming meanwhile centrifugation
technique is costly and may lead to fmancial constraint.
3
1.2 Objectives
The objectives of this research are:
I. To identify the effects of calcium addition as a stabilizer on the production and
drying time of brown and white sago sugars.
2. To determine the effects of glucose concentrations, total phenolic compounds and
flavonoid content with different pre-treatments, sedimentation and centrifugation
on brown and white sago sugars.
3. To study the effects of sedimentation and centrifugation on drying time of brown
and white sago sugars.
4
~~I(bidmat MaJdumat Akademik ~ .•~"" ",Ut..4Vs,,. S4IUW,41(
2.0 LITERATURE REVIEW
2.1 Sugar Industry
Today, global production of sugar is increasing from developed to developing countries.
Up to now, sugar consumption and demand have increased rapidly based on the increase of
population indeveloping countries.
2.1.1 World Market and Demand
In crop year 2013114, the scale of global sugar production is about 174.8 million tonnes
and achieves average growth rate of 2 %/year (Pham et al., 2014). According to Pham et
al. (2014), the largest producing countries are Brazil (22% of total production), India
(15%), China (8%) and Thailand (6%) with sugar trading volume on the world market
around 55-60 million tonnes. The largest sugar producer and exporter in the world, Brazil
is also one ofthe largest ethanol manufacturers and consumers (Martinelli et al., 2011). A
huge impact on sugar supply and price in the world will happened due to the decision to
expand ethanol production in this country (Pham et al., 2014).
2.1.2 Sources of Sugar
Sugar can be processed from sugar cane (75-80% of the global supply), sugar beet and
Metroxylon sagll. Approximately 75% and 25% of the world's sugar production are
produced from sugar cane and sugar beets, respectively (Potts et al., 2014). About 66% of
the sugar consumption in Malaysia is for domestic uses. According to F AO (2009), the
production of sugarcane in Malaysia is concentrated in the northwest of Peninsular
Malaysia in Perlis and Kedah and its cultivation is relatively small. The sugar recovery is
rather low at only 7%/kg fresh weight even though the annual production of sugarcane is
5
relatively high at 1.3 to 1.6 million tonnes (Bujang, 2010). The higher price of this
commodity is caused by the lack of raw materials and then, increase in industrial
application ofsugarcane (Bujang, 2014). Sugar made from sugarcane is profitable and easy
plant to grow compare to Metroxyion sagu that takes up to 10 years for the plant to be
harvestable (Flach. 1997). However, Bujang (2014) stated that sago starch produced the
highest glucose recovery among other starches with (99% DE), followed by corn starch
(84% DE), then tapioca starch (76% DE) and sweet potato (72%). Sago starch has the
potential to replace sugarcane in the sugar industry because it involves enzymatic
hydrolysis of the starch (HSS) process and purified using PAC (Bujang et ai., 2006). On
the other hand, sugar beet has sugar content of 14-18% and grown primarily in temperate
regions (FAO. 2009). Although the harvest time of sugar beet is shorter than of sugarcane
(5 - 6 months), it is more costly and less competitive compare to sago sugar and sugarcane
(FAO, 2009).
2.1.3 Uses of Sugar
Many types of sugars which include glucose, fructose, sucrose and lactose are found in the
diet on a daily basis. Sugar is usually referring to sucrose or known as table sugar. Sugar
cane and sugar beets contain large quantities of sucrose (Potts et ai., 2014), meanwhile
sago starch contain glucose in large quantities (Bujang et ai., 2011). As an example of
carbohydrates, sugar provides our body with the energy that required by organs and
muscles to function. Moreover, sugar is known as food additive used efficiently in drinks
and foodstuffs. They also have important biological, physical and chemical properties as
well as bringing sweetness. For instance, sugar can be used in the canning, cooking and
freezing of foods to improve flavor and texture, and to preserve natural color and shape
(Potts et ai., 2014). During freezing, sugar protects the surfaces of frozen fresh fruit from
6
contact with air that can produce enzymatic browning (due to oxidation). However,
excessive consumption of sugars, combined with insufficient energy expenditure increased
risk ofoverweight, obesity and serious diseases (Potts et al., 2014).
2.2 Sago Industry
Currently, interest in sago industry has increased rapidly. Sago also is one of the important
commodities which contribute to the economic value in the countries. As a carbohydrate,
sago is required to support human life by preventing a human food crisis due to large world
population.
2.2.1 The Sago Palm
Metroxylon sagu is derived from Javanese word and contains starch in its trunk. According
to Ubi and Dransfield (1987), this sago palm is belongs to the family Palmae Jussieu and
genus Metroxylon Rottboell. There are many uses ofsago-palm derived products. The sago
leaves are used for roof thatch and wall sliding and can be woven into bags, baskets, cages
and rope (Adeni et al., 2010). Spoons and food wrappers also can be made from sago
leaves. On the other side, the rachis of fronds is used for walls or house building. The
cortex of trunk is used as flooring and frring in factory. Plank that is used for building or
making things also can be made. Other parts of the sago palm can be used in traditional
medicines and many different things (Adeni et al., 2010). Moreover, ground pith is rasped
and dried as an animal feed for pigs, horses and chicken. In order to extract good quality
and quantity of sago starch, the pith of sago palm that contains starch need to be separated
from the cellulosic materials, then has to undergo several processes (Bujang and Ahmad,
2000). The starch obtained can be converted into many uses, for instance, dextrose
7
glucose, human food products, animal feed, paper industry, textile industry, foundry
molds, single cell protein production and as a biodegradable filler in plastics.
2.2.2 Production of Sago Starch
Several processes have to be taken for production of sago starch. According to Bujang and
Yusop (2006), steps taken in sago starch production include debarking, pulping, extraction,
dewatering, drying and packing. Sago starch was extracted from the pith after removing
the cortex, rachis and leaflets from the pith (Singhal et al., 2008). The liquefied starch was
poured and precipitated in a settling container. Then, the starch settled on the bottom of the
container was washed several times until white starch was obtained (Kamal et al., 2007).
Sago is grown commercially for the production of sago starch and conversion to animal
food or to ethanol in Malaysia, Indonesia and Papua New Guinea (Singhal et al., 2008).
One of the main potential of sago starch is that it has multifunctional uses and can be
utilized for energy and food products.
2.2.3 Utilization and Bioconversion of Sago
Sago can be utilized and converted into many uses. Nowadays, the products made from
sago are marketable and provide a regular cash flow to the sago farmers. Thus, the
utilization and bioconversion of sago has been developed. Table 1 shows the development
of food products by using several enzymes and substrate. Sugars, single cell protein, lactic
acid, kojic acid and cyclodextrin are some of food products that can be produced from
sago.
8
Table 1: Development offood products from sago
DescriptionFood Product Enzyme! References
Substrate
The starch slurries were enzymatically (Bujang,Sugars • Termamyl-120L
hydrolysed for 4-6 hours at the optimum 2014)
concentration and then, purified using powdered
activated charcoal (PAC) by filtration . Sugar
also can be obtained from sago hampas because
hampas is the fibrous wastes that are discarded
with sago effluent into the river.
• Amyloglucosidase
Spirulina platensis Spirulina platensis has very high protein (Bujang,
content. The cyanobaterium cultivated in sago 2014)
Single
Cell Protein effluent is treated for production of single cell
protein.
L-Iactic acid, the only form of lactate that is (Bujang,
absorbed entirely by the human body is 2014)
Lactic Acid Lactococcus
lactis 10-1 produced from fermentation of starch by
Lactococcus lactis 10-1. It is importance in the
food and cosmetic industries.
I In order to produce high yields of kojic acid by (Ros farizan I Aspergillus flavus, gelatinized sago starch has
Kojic Acid Aspergillus sp.
et al., 2002)been chosen as a carbon source in different
fermentation modes In an 8-L stirred tank (Satoko
fermentor. It is importance In the field of et ai., 2006)agriculture, food science, medicine and
agriculture and recently, there are reports on the
genotoxicity of kojic acid.
Cyclodextrin Cyclodextrin Cyclodextrin glycosyltransferase produced by (Charoenlap
(CD) from glysyltranferase various microorganisms, such as Bacillus sp. et aI., 2004)
sago starch produced synthesizes the CD enzymatically by converting
particularlyby Bacillus starch into CD. CD IS useful in food ,
sp. pharmaceutical, cosmetic and agricultural
applications.
9
On the other side Table 2 shows the development of energy products by using several
enzymes and substrate. Starch to ethanol, fiber to ethanol and effluent to biodiesel are
some ofenergy products that can be produced from sago.
Table 2: Development of energy products from sago
Energy
Product
Enzyme/Substrate Description References
Starch • Nitrogen sources
(mielU or com steep
The sago starch is used as substrate together
with nitrogen sources (mieki or com steep
(Bujang,
2014) to Ethanol
liquor)
• Saccharomyces
cereviceae CSI-l (for
continuous ethanol
fermentation)
liquor) as the alternative to yeast extract for
production of ethanol for biofuel.
Fiber
to Ethanol
• Cellulose
• p-gLucosidase
--Enzymatic hydrolysis of steam-treated sago
fiber production of sugars from cellulose can
be used as substrate for production ofbiofuel.
(Bujang,
2014)
Eftluent Scenedesmus dimorphus Scenedesmus dimorphus is cultured in l6L of
modified sago effluent with NaHC03.
(Bujang,
2014) to
Scenedesmus is the preferred species for oil Biodiesel
yield in the production ofbiodiesel. A process
used in conversion lipid to biodiesel, trans
esterification is the most economical with 98%
conversion yield.
2.3 Types of Sago Sugars
There are two types of sago sugars studied; hydrolyzed sago sugars (brown sago sugars)
and purified liquid sago sugars (white sago sugars).
10
2.3.1 Brown Sago Sugars
Consumption of brown sago sugar is beneficial to health due to the presence of
antioxidant, which analyzed based on total phenolic content (TPC) and total flavonoid
content (fFC). Flavonoids are based on a C6-C3-C6 backbone and belong to polyphenol
family. As the secondary metabolite, it plays vital function in plant such as growth
regulators, gene expression modulators, intracellular signaling and also stress responses
(Kamtekar et al., 2014). The most important is that they can acts as anticancer, antioxidant
and anti-inflammatory properties on activated cell lines of animal and possible role in
several chronic diseases prevention involving oxidative stress that acts as their protective
effect against low-density lipoprotein (LDL) oxidation (Sudha et at. , 2011). According to
Bujang (2015), brown sago sugar has the highest total phenolic and total flavonoid
contents at 20% (w/v). Furthermore, types of flavonoid that has been discovered in brown
sago sugar are Gallic acid, Quercetin and Kaempferol (Bujang, 2015).
2.3.2 White Sago Sugars
White sago sugar contains mostly glucose (94%), maltose (3%) and other impurities (3%)
(Bujang et al., 2011). Several types of mono- or oligo-saccharides as impurities (3%) are
produced when the hydrolytic enzymes used contain a-amylase and attacks gelatinized
starch randomly (Bujang, 2015). PAC is used to purify brown sago sugars into white sago
sugars (Bujang et al., 2012). This process is known as purification of BSS. Different
amount of PAC gives different recovery of sugars (Bujang et al., 2012). Based on the
research, the yield of white sago sugars was lower when higher amount of PAC was used
during purification (Bujang, 2011). According to Ang et al. (2006), the absorption between
PAC is higher towards protein and color but lower towards glucose and lactate.
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3.0 MATERIALS AND METHODS
3.1 Materials
3.1.1 Sago Starch
Sago starch was obtained from Herdsen Sago Industries in Pusa, Sarawak.
3.1.2 Hydrolytic Enzymes
Termamyl-SC (alpha-amylase from Bacillus licheniform is 120KNU-S/g)
Amyloglucosidase (AMG) from Aspergillus niger, ~ 300 U/mL purchased
Novozyme Biomass Kitt were used for sago starch hydrolysis (Bujang, 2012).
and
from
3.1.3 Calcium
Calcium powder obtamed from calcium granule was added into another set of enzymatic
hydrolysis of sago sugar in both brown and white sago sugars.
3.1.4 Powdered Activated Charcoal (PAC)
AC was used in purification ofHSS (with and without calcium) aided by a vacuum pump
produce Purified Sago Sugar (PSS). Sterilized PAC which was washed with 0.2 M HCI
deionized water was used for all samples (Bujang et al., 2012). Then, cellulose acetate
filter paper was used to filter hydrolyzed sago sugar.
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3.2 Method
Flowchart presented in Figure 1 shows the summary of experimental method involved
during production and analysis of brown and white sago sugars. Detail methods for each
step are explained in the next proceeding pages.
Standard Enzymatic Hydrolysis of Sago Starch into Sugar (With and Without Calcium)
- Enzymatic hydrolysis:
- Liquefaction (Tennamyl-SC)
- Saccharification (Amyloglucosidase)
- Effect ofdifferent pre-treatments on brown sago sugar:
- Sedimentation in the Refrigerator
- Sedimentation in the Oven
- Centrifugation
Purification of Sago Sugar
Drying and Crystallization of Sago Sugar
Analysis of Sago Sugar:
- Reducing Sugar using DNS method
- Total Phenolic Contents using Follin-Ciocalteu method
- Total Flavonoid Contents using colorimetric method
Statistical Analysis
Jlpre 1: Flowchart ofmethod involved during production and analysis of brown and wh ite sago sugars
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