Optimization of process parameters for glucoamylase production under solid state fermentation by a newly isolated Aspergillus species Article address: https://app.dimensions.ai/details/publication/pub.1011235545?and_facet_journal=jour.1313739

4. INTRODUCTION

5. GLUCOAMYLASE - AN INTRODUCTION
 Also called
1. Amyloglucosidase
2. Glucogenic enzyme
3. Starch glucogenase
4. Gamma amylase
5. Exo-1,4-a-Dglucan glucanohydrolase
 An exoamylase
 hydrolyses single glucose units from the nonreducing ends of amylose
and amylopectin.
 Sometimes 1,6-a-linkages at the branching points of amylopectin

6. PRODUCTION
 Produced by various microorganisms, including bacteria; fungi and yeasts
 Fungi: Aspergillus oryzae, Aspergillus niger, Aspergillus flavus and Rhizopus
oryzae
 Bacteria: Bacillus species, Pseudomonas and Lactobacilli
 Traditionally, produced by SmF and used in a one-way process in solution. In
recent years, SSF processes have been applied for the production of this
enzyme.
 Currently, industrial demand is met by production in SmF, generally
employing genetically modified strains.

7. APPLICATIONS
 Food industry
 Fermentation industry
 Textile industries
 Paper industries

8. Advantages of SSF over SmF
Higher Productivity
Better-Oxygen Circulation
Low-cost Media
Less effort in Downstream Processing
Scarce-operational Problems
Simple Technology

9. AIM OF THE STUDY:
 To investigate the production of glucoamylase under SSF conditions.
 To report the factors that influence the maximization of glucoamylase
production by Aspergillus sp. A3 through SSF.

10. MATERIALS AND METHODS

11. MATERIALS AND METHODS
Aspergillus
niger,A3
Isolation from
soil
Growth on
Potato dextrose
agar
Transfer to
inoculum
Incubation
(38C,6.5pH) at
rotary
shaker(220rpm)
for 48 hrs
Glucose 20g/l
Potassium nitrate 1g/l
Calcium carbonate 0.1g/l
Salt solution 50ml
Composition of the inoculum medium
K2HPO4 0.5g/l
MgSO4.7H2O 0.5g/l
FeSO4.7H20 0.01g/l
NaCL 0.5g/l
Composition of Salt Solution

13. SUBSTRATE INOCULUM
LEVEL(in %)
TEMPERATURE
(in C)
INCUBATION
TIME(Hours)
INITIAL MOISTURE
CONTENT(in %)
Wheat Bran 5 30 96 50
Wheat Bran 5 30 96 60
Wheat Bran 5 30 96 70
Wheat Bran 5 30 96 80
Wheat Bran 5 30 96 90

14. SUBSTRATE INOCULUM
LEVEL(in %)
TEMPERATURE
(in C)
INCUBATION
TIME(in Hours)
INITIAL MOISTURE
CONTENT(in %)
pH
Wheat Bran 5 30 96 80 3
Wheat Bran 5 30 96 80 4
Wheat Bran 5 30 96 80 5
Wheat Bran 5 30 96 80 6
Wheat Bran 5 30 96 80 7

15. SUBSTRATE INOCULUM
LEVEL (%)
TEMPERATURE
(C)
INCUBATION
TIME (Hours)
INITIAL MOISTURE
CONTENT (%)
pH
Wheat Bran 5 30 24 80 5
Wheat Bran 5 30 48 80 5
Wheat Bran 5 30 72 80 5
Wheat Bran 5 30 96 80 5
Wheat Bran 5 30 120 80 5
Wheat Bran 5 30 144 80 5
Wheat Bran 5 30 168 80 5

16. SUBSTRATE INOCULUM
LEVEL (%)
TEMPERATURE
(C)
INCUBATION
TIME (Hours)
INITIAL MOISTURE
CONTENT (%)
pH SALT LEVEL
(v/w)
Wheat Bran 5 30 120 80 5 1:10
Wheat Bran 5 30 120 80 5 1.5;10
Wheat Bran 5 30 120 80 5 2:10
Wheat Bran 5 30 120 80 5 2.5:10

17. SUBSTRATE INOCULUM
LEVEL(%)
TEMPERATUR
E (C)
INCUBATION
TIME (Hrs)
INITIAL
MOISTURE
CONTENT (%)
pH SALT
LEVEL
Wheat Bran 5 20 120 80 5 2:10
Wheat Bran 5 25 120 80 5 2:10
Wheat Bran 5 30 120 80 5 2:10
Wheat Bran 5 35 120 80 5 2:10
Wheat Bran 5 40 120 80 5 2:10
Wheat Bran 5 45 120 80 5 2:10

18. SUBSTRATE INOCULUM
LEVEL (%)
TEMPERATURE
(C)
TIME(Hrs) MOISTURE
CONTENT (%)
pH SALT LEVEL
Wheat Bran 5 30 120 80 5 2:10
Wheat Bran 10 30 120 80 5 2:10
Wheat Bran 15 30 120 80 5 2:10
Wheat Bran 20 30 120 80 5 2:10

19. SUBSTRAT
E
AMOUNT OF
SUBSTRATE
(g/250ml)
INOCULUM
LEVEL(%)
TEMPERATUR
E
(C)
INCUBATION
TIME (Hr)
MOISTUR
E
CONTENT
(%)
pH
SALT LEVEL
Wheat
2 10 30 120 80 5 2:10
Wheat
10 10 30 120 80 5 2:10
Wheat
15 10 30 120 80 5 2:10
Wheat
20 10 30 120 80 5 2:10

20. Carbon Sources
Glucose
Starch
Lactose
Fructose
Sucrose
Nitrogen Sources
Casein
Corn Steep Liquor
Yeast Extract
Soya bean Meal
Peptone
Urea

21. RESULTS

29. DISCUSSION

30. SUBSTRATE SELECTION:
 In SSF, the selection of a suitable solid substrate is a critical factor
 The order of substrate suitability was
wheat bran > green gram bran > corn flour > black gram bran > jowar
flour > maize bran > barley > wheat rawa > rice bran
 Different solid substrates were found to effect the production of
enzymes[15]
 wheat bran was found to be the best substrate[16]
 Wheat bran is suitable for necessary manipulation[17]

31. MOISTURE CONTENT
 High enzyme titre (92.0 U/g) was attained when the initial moisture level was
80% (Fig.1)
Increase in
Moisure content
Reduced porosity
of Wheat bran
Limited Oxygen
Transfer
Low Moisture
Content
Reduced Solubility
of nutrients
Low degree of
swelling

32. pH
 The maximum glucoamylase production (109.0 U/g) was obtained at pH 5.0.
 Metabolic activities are sensitive to changes in Ph
 Glucoamylase production by Aspergillus sp. A3 is affected if pH level is higher
or lower compared to the optimum value

33. INCUBATION TIME,SALT CONCENTRATION, TEMPERATURE,
INOCULUM LEVEL & SUBSTRATE VOLUME
The strain used produced
 High titres of enzyme (125.0 U/g) at 120 h incubation
 High enzyme titre(141.0 U/g) at a salt solution concentration to wheat bran weight
ratio of 2:10
 High enzyme titres (142.0 U/g) at 30C
 High enzyme titres (153.0 U/g) at 10% (v/w) inoculum level
 High enzyme titres (169.0 U/g) when the substrate weight-to-flask volume was 1:50
ratio

34. SUPPLEMENTS
 Addition of sugars as additives resulted in better glucoamylase production with
fructose (213.0 U/g)
 Among nitrogen sources,urea exhibited better glucoamylase production (247.0
U/g) followed by corn steep liquor and peptone.

35. CONCLUSION:
Maximum productivity of glucoamylase (247.0 U/ g)
 Fructose as additive 1% w/w
 Urea as additive 1% w/w
 Incubation time of 120 h
 Incubation temperature at 30 8C
 2:10 (v/ w) ratio of salt solution to weight of wheat bran
 Inoculum level 10% v/v
 Moisture content of solid substrate 80%
 1:50 ratio of substrate weight to flask volume
 pH 5.0

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