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International Journal of Herbal Medicine 2016; 4(6): 34-40
E-ISSN: 2321-2187
P-ISSN: 2394-0514
IJHM 2016; 4(6): 34-40
Received: 07-09-2016
Accepted: 08-10-2016
Jiby John Mathew
Department of Biotechnology,
Mar Augusthinose College,
Ramapuram, Kerala, India
Prem Jose Vazhacharickal
Department of Biotechnology,
Mar Augusthinose College,
Ramapuram, Kerala, India
Sajeshkumar NK
Department of Biotechnology,
Mar Augusthinose College,
Ramapuram, Kerala, India
Anjaly Ashokan
Department of Biotechnology,
Mar Augusthinose College,
Ramapuram, Kerala, India
Correspondence
Prem Jose Vazhacharickal
Department of Biotechnology,
Mar Augusthinose College,
Ramapuram, Kerala, India
Amylase production by Aspergillus niger through
submerged fermentation using starchy food byproducts
as substrate
Jiby John Mathew, Prem Jose Vazhacharickal, Sajeshkumar NK and
Anjaly Ashokan
Abstract
Submerged fermentation holds tremendous fungal potentiality in high biomass yield of alpha-amylase.
Isolation of fungi from bread sample and the rapid screening by plating on starch agar plates led to the
finding of fungal strains capable of producing amylase. These strains were confirmed as Aspergillus
niger by lacto phenol cotton blue staining. The effect of carbon sources of the medium for the activity of
α- amylase from Aspergillus niger utilizing Coconut water, Tapioca water, Rice water and White Yam
water were investigated. The maximum activity of α-amylase was recorded as 0.29 x 10-3 µmoles/sec.
After 7 days of submerged fermentation on white Yam water at pH 7.0 and room temperature 28 °C.
Among the three medium rice water recorded as second (0.09 x 10-3 µmoles/sec) and tapioca water (0.06
x 10-3 µmoles/sec) as third position. The enzyme produced by Aspergillus niger can be used in industrial
process after characterization. The present study suggest that white yam water act as a potent substrate
for industrial production of α-amylase and subjected for further explorations regarding industrial
applications.
Keywords: Submerged fermentation, rice water, Aspergillus niger, tapioca water, amylase production
1. Introduction
Amylases are enzyme that breaks down starch or glycogen. The amylase can be derived from
several sources such as plant, animal and microbes. The major advantage of using
microorganism for production of amylase is in economical bulk production capacity and
microbes are also easy to manipulate to obtain enzymes of desired characteristics [1]. The
microbial amylases meet industrial demands; a large number of them are available
commercially; and, they have almost completely replaced chemical hydrolysis of starch in
starch processing industry [2]. Although many microorganisms produce this enzyme, the most
commonly used for their industrial application are Bacillus licheniformis, Bacillus
amyloliquifaciens and Aspergillus niger. Amylases stand out as a class of enzymes, which are
of useful applications in the food, brewing, textile, detergent and pharmaceutical industries.
They are mainly employed for starch liquefaction to reduce their viscosity, production of
maltose, oligosaccharide mixtures, high fructose syrup and maltotetraose syrup. In detergents
production, they are applied to improve cleaning effect and are also used for starch de-sizing in
textile industry [3, 4].
The use of the submerged fermentation (SmF) is advantageous because of the ease of
sterilization and process control is easier to engineer in these systems. Depending on the strain
and the culture conditions, the enzyme can be constitutive or inducible, showing different
production patterns. Submerged fermentation has been defined as fermentation in the presence
of excess water. Almost all the large-scale enzyme producing facilities are using the proven
technology of SmF due to better monitoring and ease of handling [5]. To meet the growing
demands in the industry it is necessary to improve the performance of the system and thus
increase the conditions, particularly physical and chemical parameters are important in the
development of fermentation processes due to their impact on the economy and practicability
of the process [6]. The growth and enzyme production of the organism are strongly influenced
by medium composition thus optimization of media components and cultural parameters is the
primary task in a biological process [7].
Due to the increasing demand for these enzymes in various industries, there is enormous
interest in developing enzymes with better properties such as raw starch degrading amylases
suitable for industrial applications and their cost effective production techniques [8]. Selection
of appropriate carbon and nitrogen sources or other nutrients is one of the most critical stages
in the development of an efficient and economic process [9].
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International Journal of Herbal Medicine
The processing of cassava and white Yam tubers for the
production of nutrient enriched food products is usually
accompanied with the production of stinking wastewater
which usually constitute nuisance to both terrestrial and
aquatic life. After boiling rice also produces a sticking water
with high amount of starch.
The objectives of this work was to study the production of α-
amylase by using Aspergillus niger through submerged
fermentation using four different starchy substrates like Rice
water, White Yam water and Tapioca water as carbon source
and compare the activity of the amylase produced using these
substrates. Enzymes are protein catalysts synthesised by
living system and are important in synthesis as well as
derivative process. Amylase are enzyme that breakdown
starch or glycogen. The amylase can be derived from several
sources such as plant, animal and microbes [10]. Alpha
amylase (endo-1-4 dglucose-Dglucohydrolase 3.2.1.1) belong
to the family of endo amylases. From an industrial point of
view mostly bacterial and fungal source have been used for
the production of alpha amylase. Properties of alpha amylase
such as thermo stability, pH optimum and their other
physicochemical properties are important in the development
of most suitable fermentation process. Alpha amylase can
produce by fungi in large amount but they are not usually heat
stable beyond 40 °C. Bacterial species such as Bacillus
subtilis, B. megaterium, B. amyloliquefaciens and B.
licheniformis produces more heat stable enzymes. Bacterial
species which produce alpha amylase enzyme, if it is often
need to isolation of microorganism that can grow at high
temperature and whose enzyme can function at temperature
up to 95-100 °C.
Kirchhoff was the first scientist to report the discovery of
alpha amylase in 1811.Starch is an abundant source of
carbohydrate. It consists of amylopectin and amylose.
Amylopectin is formed from linked alpha, 1-4 chain of
glucose with linked (α, 1-6) branch points and amylase
consists of chain of glucose α,1-4 linked. α amylase this
enzyme breakdown α-1,4 glycoside linkage of starch and
related products in an endo fashion and produce
oligosaccharides. If the mode of action, properties and
products of hydrolysis is depend upon the source of enzyme
[11].
Microorganism associated with α-amylase production.
Industrial enzymes have been produced from plant, animal
and micro-organism, but the plant and animal source is rather
because of several reasons. If the concentration of enzyme in
plant source is generally low but starch processing in
industrially required as large quantities of enzyme. On the
other hand if the enzyme from animal source origin is from
the byproduct of meat industry and so it is supply is limited.
However the α-amylase from microbial source can be
produced in abundant quantities. Microorganism utilized
different nitrogen, carbon sources for the production of α-
amylase nitrogen source such as yeast extract, peptone,
ammonium sulphate casein, ammonium nitrate, chicken
feathers, carbon source such as corn starch, potato starch,
cane sugar etc.
α-amylase is produced by bacterial species of Bacillus such as
B. subtilis, [12, 13] B. licheniformis etc. Are generally preferred
for the property of thermo stabilities the enzyme α-amylase
utilized in various fermentation processes extreme
thermophillic bacteria such as Rhodothermus marinus and
mesophilic bacteria such as B. megaterium, B. macerans and
B. coagulans are generally utilized while in the case of most
thermo stable α-amylase utilized in industry is produced from
B. licheniformis [14, 15]. And highly thermo stable α- amylase
are also obtained in hyper thermophilic and thermophilic
archea such as Pyrococcus furiosus, Thermococcus
hydrothermalis [16, 17].
Fungi as a source of material for α-amylase production. α-
amylase producing strain of yeast; fungi and actinomycetes
were isolated. Especially aspergillus species are also source of
α-amylase. It has gained more attention because of the easy
availability and high productivity of the fungi, which are also
suitable for genetic manipulation. Different species of
aspergillus such as A. niger, A. oryzae, A. flavous, A. tamarie,
A. fumigatus have frequently used for the production of α-
amylase [18-27]. Pencillium species such P. chrysogenum and
P. camemberti also used for the production of α- amylase and
also in cheese production of α-amylase was obtained from
thermophilic fungi species such as Hemicola insolen, H.
lanaginosa, H. stellata etc. From the industrial point of view
some species of yeast such as Candida tsukubaensis,
Filobasidium capsuligeum, Lipomyas kononenkoae,
Saccharmycopsis capularis, Saccharomyces cerevisiae have
been used for the production of α-amylase [18, 4].
Industrial uses of α-amylase: Bacterial amylase plays an
important role in industrial production process. Many
industrial processes involving manufacturing such as
industrial, environment process and food biotechnology
utilizes the enzyme.
Glucose And Fructose Industry: Many industry use α-
amylase for the production of glucose. This enzyme
hydrolysis starch and convert it into maltose and glucose.
Alpha amylase is widely used in many starch processing
industries for the production of glucose [38, 19].
In bakery industry the α-amylase play an important role in
improvement of quantity, aroma, taste and porosity of the
product. This enzyme is the major part of bread used in
Russia, USA and the European countries. α-amylase can also
affect anti-salting in baking bread and help to improve the
softness of bread [34]. Enzyme has significant role in the
improvement of detergent quality by affecting bleaching. The
addition of enzyme increases the stability and effectiveness of
the bleach in laundry’s detergent and soap bar composition
[19].
Fermentable sugar is produced by the conversion of starch
with the help of alpha amylase. Starch such as grains, potatoes
etc. are required for the production of ethyl alcohol [33].
Starches increase the stiffness of the finished products after
washing out the cloths. Alpha amylase is used as a resizing
agent. It improves paper quality, protect against mechanical
injury and increase the stiffness and strength in paper. It
readily hydrolysis the starch polymer into fructose and
glucose which increase the digestibility of carbohydrates [39,
40].
Amylase are enzymes that breakdown starch or glycogen. The
amylase can be derived from several sources such as plant,
animal, and microbes. The major advantage of using
microorganism for production of amylase is in economical
bulk production capacity and microbes are also easy to
manipulate to obtain enzymes of desired characteristic [1]. The
microbial amylase meet industrial demands: a large number of
them are available commercially and they have almost
completely replaced chemical hydrolysis of starch processing
industry [2]. They are mainly employed for starch liquefaction
to reduce their viscosity, production of maltose,
oligosaccharide mixture, high fructose syrup etc.
The use of the submerged culture is advantage because of the
ease of sterilization and process control is easier to engineer
in these system depending on the strain and the culture
condition. Most of the enzyme are produced by submerged
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International Journal of Herbal Medicine
fermentation is becoming popular for producing enzyme due
to it is inherent advantage example higher yield, improved
oxygen circulation, less energy requirement, minimum effort
in downstream processing, effect of process variables, namely
incubation period, temperature, initial moisture content, pH of
the medium, supplementary carbon source, supplementary
nitrogen source, and inoculums level on production of α-
amylase have been studied.
The most effective amylase are those that are thermo stable
they are generally preferred as their application minimize
contamination risk and reduce reaction time thus enabling
considerable energy save. Amylase has potential application
in a wide range, number of industrial processes such as food,
fermentation, textile, paper, detergent and pharmaceutical
industries. Starch is an important storage product of many
economically important crops such as wheat, rice, maize,
tapioca, coconut water and potato. Starch converting enzyme
is used in the production of maltodextrin, modified starch or
glucose and fructose syrups. Amylase are a group of
hydrolyses which can specifically cleave glycosidic bond in
starch. There are two important group of amylase which
includes glucoamylase and α-amylase. Microbial amylase has
successfully replaced chemical hydrolysis of starch in starch
processing industries. Amylase has been derived from several
fungi, yeasts, bacteria and actinomycetes, however, enzyme
from fungal and bacterial source have dominated application
in industrial sectors. Major advantage of using fungi for
amylase production is the economical bulk production
capacity.
The amylase derived from several species, source such as
plant, animal and microbes. However the enzyme from fungal
source has dominant application in industrial sectors. The
major advantage of using microorganism has economical bulk
production. The production process of α-amylase mainly they
can be two major methods for large scale production of α-
amylase are solid state fermentation and submerged
fermentation.
Many factors involved in the production and optimization of
α-amylase such as nitrogen and carbon source supplied, metal
ions, pH, and temperature. Nitrogen source used for the
production of α-Amylase: Various nitrogen source including
corn steep liquor, casein, yeast extract, tryptone, ammonium
nitrate, sodium nitrate and ammonium chloride are utilized for
the production of α-amylase in basal medium. Organic
nitrogen source like peptone, yeast extract, usually having
stimulating effect peptone is the best for the production α-
amylase [41-43, 26]. Mainly some organism including such as
bacillus species and aspergillus species such as A. flavus, A.
niger and bacillus species including B. subtilis, B.
licheniformis
Carbon source used for the production of α-Amylase: α-
Amylase is produced from many source of carbon such as
fructose, glucose, maltose, galactose, sucrose, lactose,
dextrose industrial waste like syrup and molasses, agriculture
waste involving sugarcane and rice husk [26]. Microorganism
for the production of α-amylase such as B. subtilis, B.
licheniformis, A. oryzae etc. Metal Ion: Metal ion play an
important role for the production of α- amylase because of
most α-amylase are metallo enzyme.
Effect of temperature on α-Amylase activity: Action of
enzymes is time dependent process. Increase in temperature
will lead to an increase in activity of kinetic reaction. High
temperature can affect enzyme activity because enzyme
proteinaceous molecule. Thermo stable α-amylases have been
isolated from organisms such as, B. amyloliquefaciens B.
licheniformis, B. subtilis and A. niger. The effect of
temperature on α-amylase action has been reported previously
in such studies. Optimum temperature was noted to be 65 °c
at low substrate concentration and 75 °C at high substrate
concentration. If the temperature range of Bacillus species up
to about 30-70 °C and Aspergillus species about 30-40 °C [18,
30, 31].
Effect of pH on α-Amylase activity: The pH on enzyme
stability and activity is also depending on time and
temperature. In general, enzymes are less stable at high
temperature over time at pH value near the limit of the
optimum. α-Amylase are stable a pH range of 4-11 [15].
Rice water, tapioca water and white yam water has been used
as a potent substrate for the production of amylase by A. niger
in submerged fermentation. The cheap starchy agro-by
products have been reported to be a good substrate for the
cost effective production of alpha amylase. The synthetic
media are used for the production of amylase fungal species
have been studied a lot for the production of alpha amylase
[16].
Coconut water (Thenga vellam in Malayalam) is the
suspension of starch, sugars and minerals obtained by
draining ripe coconut (Cocos nucifera) for the extraction of
oil.
Rice water (Kanji vellam in Malayalam) is the suspension of
starch obtained by draining boiled rice (Oryza sative) or by
boiling rice. Rice water is also a milky liquid which contain
vitamin B, E and mineral. Rice water is relatively containing
good source of carbohydrate, calcium, iron, vitamin. The
protein content is about 36-58% and presence fat is 16-25%.
Tapioca water (Kappa vellam in Malayalam) (Manihot
esculenta) is a suspension of starch is obtained by boiling
pieces of tapioca with water. The use of cassava in submerged
fermentation such as the microorganism have a very fast
growth rate, they can be easily modified genetically for
growth on a particular substrate under particular cultural
condition, the protein content is quiet high varying from 35-
60%.
White Yam water (Kachil vellam in malayalam) (Dioscorea
rotundata) have an average crude protein content is 4-7% and
the starch content about 75.6-84.5% and which contain high
carbohydrate content more than 85% and fat contain 0.17g.
Given lacking qualitative and quantitative data on various
starchy substrates in Kerala, objective of this study were to
screens a variety of easily available and inexpensive starchy
plant materials as substrate for the production of α-amylase
using Aspergillus niger through sumerged fermentation.
2. Materials and Methods
2.1 Sample collection/Substrate collection and sterilization
Rice water, Tapioca water and White Yam water were
obtained by draining boiled small pieces of fresh tuber or by
boiling small pieces fresh tuber until it completely dissolves
into the water. They are stored in sterile bottles under aseptic
condition until use.
2.2 Isolation of Aspergillus niger
A piece of bread was kept in a moist condition at room
temperature in dark for 2 days. The bread sample was serially
diluted and different dilutions were inoculated on potato
dextrose agar (PDA) medium. The slants were incubated at 30
°C 4 days. Fungal cultures were observed on PDA medium.
The fungal strain was subjected to lactophenol cotton blue
staining for studying the morphology. The fungal culture was
confirmed as Aspergillus niger by studying the morphology
and the spore colour.
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International Journal of Herbal Medicine
2.3 Lacto phenol cotton blue staining
Place a drop of Lacto phenol Cotton Blue Solution on a slide.
Using an inoculating needle carefully spread the fungal
culture into a thin preparation. Place a cover slip edge on the
drop and slowly lower it. Observe under low to high power
objectives of microscope. Lactic acid acts as a preservative
for fungi. The phenol portion kills the fungi. The cotton blue
stains the fungal elements. Fungal elements are stained a deep
blue; background is pale blue.
2.4 Determination of Amylase activity
The Aspergillus niger isolate was tested for amylase
production by starch hydrolysis. When starch agar medium
was inoculated with the organism and subsequently flooded
with iodine solution, the zone of clearance around the
microbial growth indicated the production of amylase and the
fungal isolate was taken for amylase production.
2.5 Enzyme production by Solid State Fermentation
The Aspergillus niger was subjected to solid state
fermentation in different substrates like rice water, tapioca
water and white yam water, replicated four times each; which
was used as liquid substrates for submerged fermentation.
Each substrate was taken in about half in all the bottles 1% of
inoculums was inoculated after sterilization and incubated at
room temperature for six days.
2.6 Enzyme extraction
25 ml of 0.1M phosphate buffer saline (pH 7) was added to
each of the inoculated substrate beds and was vigorously
shaken in rotary shaker for 15 min at 120 rpm. The mixture
was filtered through cheese cloth and centrifuged at 8000 rpm
at 4 °C for 15 min. The supernatant was filtered through
cheesecloth and the filtrate was used as the crude enzyme
preparation. Enzyme amylase was assayed by
Dinitrosalicyclic acid method.
2.7 Determination of Amylase activity
Enzyme assay was carried out by DNS method in which
0.5ml enzyme was reacted with substrate (1% starch in 100
mM Tris buffer) under standard reaction conditions and the
reaction was stopped by adding DNS reagent, amount of
maltose released was determined by comparing the
absorbance reading of the test enzyme at 540 nm with the
standard graph plotted by reacting the known concentration of
maltose ranging from 0.05 mg/ml to 0.5 mg/ml. One unit
amylase activity was defined as amount of enzyme that
releases 1 micromoles of maltose per minute under standard
reaction conditions.
The culture supernatants were collected separately. 4 test
tubes were taken and marked sample, pure blank (PB),
substrate blank (SB) and enzyme blank (EB). With the help of
a pipette, 2 ml of phosphate buffer was transferred to all the
tubes. 1ml of starch was added to all tubes except PB & SB.
1% Sodium Chloride was added to all the test tubes. 1ml of
distilled water was added to PB & SB. The contents of the test
tubes were mixed well and then incubated for 5mins at 37 °C.
After incubation crude enzyme was added to all the test tubes
except PB & EB, and distilled water is added to PB & EB.
The contents of the test tubes were mixed well and incubated
for 10 mins at 37 °C. After incubation 1ml of 2N NaOH were
added to all the test tubes. The reducing sugars liberated were
assayed calorimetrically by the addition of 1ml
Dinitrosalicyclic acid (DNS) reagent. The contents of the test
tubes were mixed well and incubated in boiling water bath for
10 mins. Intensity of the colour developed was read at 520 nm
using a calorimeter. A standard graph was plotted and the
enzyme activity was calculated. One unit of enzyme activity
was defined as the amount of enzyme required to liberate
1μmol of sugar per minute under the standard assay
conditions and enzyme activity is expressed in terms of IU per
gram fermented substrates.
2.8 Statistical analysis
The survey results were analyzed and descriptive statistics
were done using SPSS 12.0 (SPSS Inc., an IBM Company,
Chicago, USA) and graphs were generated using Sigma Plot 7
(Systat Software Inc., Chicago, USA).
Table 1: Details of substrates used in submerged fermentation
Substrate Common Name Source Plant
Tapioca water Kappa vellam Manihot esculenta
Rice water Kanji vellam Oryza sative
White Yam water Kachill vellam Dioscorea rotundata
Coconut water Thenga vellam Cocos nucifera
Fig 1: Production of amylase using submerged fermentation using
various substrates, coconut water (top left); white yam water (top
right); rice water (bottom left); tapioca water (bottom right).
Fig 2: Amylase produced by submerged fermentation using various
substrates and their enzymatic activity, coconut water (top left);
white yam water (top right); rice water (middle left); tapioca water
(middle right).
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International Journal of Herbal Medicine
3. Results and discussion
3.1. Isolation of Fungi
Four different fungal isolates differentiated on the basis of
colony morphology were obtained after spreading. All the
four isolates were subcultures by point inoculation and used
for further studies.
3.2. Screening of Fungal Isolates for Alpha Amylase
Production
All the four fungal isolates were subjected to screening
procedure and after completion of incubation period plates
were flooded with iodine solution and observed for zone of
hydrolysis.
3.3. Identification of the Isolate Showing Maximum
Hydrolysis
Based on morphological studies, and lactophenol cotton blue
staining characteristics the isolate was identified as
Aspergillus niger.
3.4. Evaluation of starchy food as Substrates for SSF
Enzyme activity in the extracted enzymes from different
substrates was determined by DNS assay. The average
activity of enzyme produced by Aspergillus niger from
tapioca water as substrate was 0.06 x 10-3 µmoles/sec. An
average activity of 0.09 x 10-3 µmoles/sec obtained for
enzyme produced from rice water. The average activity of
enzyme produced from white yam water were 0.29 x 10-3
µmoles/sec. From the above observations, among the three
starchy substrates used white yam water was found be more
efficient for the production of amylase enzyme.
This indicates that even the percentage of starch is higher in
tapioca (29%) water there is a reduction in the activity of
amylase than other substrates used, white Yam (21%) and rice
water (22%). Higher activity was recorded in the cause of
white Yam water and then rice water. It can be seen that
maximum amylase activity was seen when Dioscorea alata
water was used as substrate followed by rice water, tapioca
water and coconut water. The enzyme activity was maximum
in the box containing Dioscorea alata water as substrate and
it was found to be (0.29 x 10-3 µmols/s) followed by rice soup
(0.09 x 10-3 µmols/s), Tapioca water (0.06 x 10-3 µmols/s) and
coconut water (0.02 x 10-3 µmols/s). Dioscorea alata is the
most efficient substrate which produced amylase under the
culture condition.
The production process of α-amylase, there are two major
method for large scale production of α-amylase are solid state
fermentation and submerged fermentation [28, 29]. Plant
products has been reported to be good substrate for the cost
effective production of alpha amylase. Many factors are
involved in the production and optimization of α- amylase
such as nitrogen and carbon source. If the metal ions play an
important role for the production of α-amylase are
metaloenzyme. Ca²+ and CaCl2 ions are significantly
important for the production of this enzyme.
Effect of temperature and pH on α- amylase activity. Increase
in temperature will lead to an increase in activity reaction of
kinetics, but also accelerate the denaturation induced by
higher physiological temperature. If soluble enzyme is used in
manufacturing process, it is beneficial to operate at the
maximum temperature. The effect of temperature on α-
amylase action has been studied if optimum temperature was
noted to be 65 °C at low substrate concentration [18, 30-32].
The effect of pH on the enzyme activity is depending on the
time and temperature. In general enzymes are less stable at
high temperature over time at pH value near the limit of the
optimum. The optimum pH should be determined to be under
certain conditions. In such case it is important to choose an
enzyme with a pH range from 4 to 11 [15, 33-37]. If the Bacillus
species have pH range between 7-8 and the Aspergillus
species is about the pH range between 3-5.
Among four substrates screened White Yam water gave
highest enzyme production 0.29 x10-3 µmols/s.), which was
almost two times higher than that produced by other
substrates. Dioscorea alata has been a highly reported
substrate producing promising results, among the various
agriculture byproducts substrates used. Widespread suitability
of Dioscorea alata may be due to the presence of sufficient
nutrients. Dioscorea alata especially Dioscorea alata water
are rich source of starch. Dioscorea alta are rich in fiber,
protein and energy contents. These agriculture byproducts
residues are cheap raw materials for amylase production
Table 2: Activity of enzyme produced by Aspergillus niger from
various substrates; tapioca water, rice water and white yam water.
Substrate Trial 1 Trial 2 Trial 3 Trial 4 Average
Tapioca
water
0.05
x10-3
0.05
x10-3
0.60
x10-3
0.70
x10-3
0.06 x10-
3
Rice water 0.07
x10-3
0.09
x10-3
0.09
x10-3
0.12
x10-3
0.09 x10-
3
White yam
water
0.27
x10-3
0.27
x10-3
0.30
x10-3
0.30
x10-3
0.29 x10-
3
Coconut
water
0.02
x10-3
0.02
x10-3
0.03
x10-3
0.03
x10-3
0.02 x10-
3
4. Conclusions
The results obtained in the present study suggest that the
white yam water may act as a potent substrate for industrial
production of α-amylase and subjected for further
explorations regarding industrial applications.
5. Acknowledgements
The authors are grateful for the cooperation of the
management of Mar Augsthinose college for necessary
support. Technical assistance from Binoy A Mulanthra is also
acknowledged. We also thank an anonymous farmer for
proving majority of samples used in the study.
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International Journal of Herbal Medicine
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