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Alpha-amylase Production by Bacillus licheniformis 1331 using Jackfruit Seeds as

Substrate
Abstract
BY: Rica Mae A. Palongpalong ADVISER: Prof. Percival G. Garcia
Amylases have great significance in biotechnology and are commercially important enzymes in various starch processing
industries. The microbial amylases meet industrial demands and have almost completely replaced chemical hydrolysis of starch in starch
processing industry. This study explored the use of jackfruit seed as carbon source in the production of alpha-amylase. A production
medium composed of ground jackfruit seed and yeast extract powder was mixed with 10% inoculum of Bacillus licheniformis 1331 cultured
in nutrient broth at 30°C for 18 hours. The effects of temperature and carbon to nitrogen ratio were determined. Triplicate analysis was done
at 10%, 15%, 20% carbon source with 20% nitrogen source set at pH 7 and temperature of 30°C and 37°C. It was found out that alpha-
amylase production was optimal at 10% carbon, 20% nitrogen, pH 7, and 37°C. Amylase production was conducted in 1000 ml medium and
incubated with shaking. Samples were taken at regular intervals at 0, 1, 2, 4, 6, 8, 18, 24, 27, and 30 hours of fermentation. Enzyme activity
and protein concentration were determined using Alpha-amylase assay and Bradford assay, respectively. Biomass was determined by dry
cell weight (DCW) analysis. Alpha-amylase production was highest during exponential growth at 4-8 hours of fermentation. Keywords:
Introduction Methodology
alpha amylase, Bacillus licheniformis, jackfruit seed, yeast extract
In the environment abound possible Preparation of jackfruit Production medium Alpha-amylase assay
sources of essential materials or substances seed samples
Biomass
necessary for the development or production of
certain products like enzymes, which will be useful to
humans, plants, and other animals. These substances
can be produced in significantly large amounts with
the advent of biotechnology, but primarily the Preparation of crude
materials and the factors that influence its production Preparation of inoculum enzyme extract
source must be identified. Some wastes from fruits Bradford assay
and vegetables can also be a good substrate for
enzyme production. This particular study aims to
produce an enzyme by a Bacillus sp. using jackfruit
seeds as substrate.
This is an exploratory study to Results 3.5 Conclusion
determine alpha-amylase production by Bacillus 0.18
3
Alpha-
Alpha-amylase activty (U/ml)

0.16

licheniformis using jackfruit seed as substrate. This
0.14
A lp h a -a m y la s e a c t ivit y
0.12
0.1 amylase was successfully
2.5
(U / m l)
study is limited to optimization of cultural and 0.08

P ro t e in c o n c e n t ra t ioproduced using jackfruit
Concentrations (mg/ml)
2 0.06
n
environmental conditions such as temperature and
0.04

1.5 0.02
0
(m g / m l)
seed as solid substrate
carbon/nitrogen ratio. The significance of this study is 1
1 B io m a s s c o n c e n t ra t io n
2
% Carbon, % Nitrogen
(m g / m l)
3

having a starch content
that another substrate will be added to the existing Figure 1. Alpha- of 37.68% in 100 grams
0.5
list of carbon sources for amylase production by amylase activity at seeds. The optimum
0
microorganisms. Many industrial processes use waste
Alpha-amylase activity (U/ml)

pH 7 and 30°C 0 10 20
0.7
30 40 conditions for alpha-
materials from plants or animals. Part of solving the
0.6

ti m e (h )
0.5

0.4
amylase production were
environmental problem of waste management can be 0.3

0.2 pH of 7, temperature of
addressed by the utilization of waste products. Figure 5. Plot of Alpha-
0.1

0
37°C and C/N ratio of 1:2.
Jackfruit seeds as waste from fruit can be a potential
1 2 3

amylase activity, Protein %Carbon, %Nitrogen

Alpha-amylase activity
material for enzyme production. Being a substrate it Figure 2. Alpha- Concentration, and Biomass was highest during
will become a source of income for fruit vendors and amylase activity at pH 7Concentration during course of exponential growth at 4-8