Enhancing Production of Amino Acids from Bacillus spp. Using Batch and Fed-batch Fermentation Strategies
Abou-Taleb, Khadiga;
Abstract
Aims: Production of amino acids from black strap sugar cane molasses by Bacillus sp. R22EG1
strain and Bacillus sp. R20EG2 using batch and fed-batch (pulsed and continuous feeding) cultures
was investigated to achieve the maximum concentration of free amino acids.
Place and Duration of Study: Department of Agricultural Microbiology, Faculty of Agriculture, Ain
Shams University, Cairo, Egypt, between December 2011 and March 2012.
Methodology: Two Bacillus strains namely R22EG1 and R20EG2, were used as amino acid
produces. The amino acid producing bacteria were grown in the bioreactor with batch and fedbatch
cultivation. The fed-batch fermentations were performed in two strategies using pulsed and
continuous feeding of black strap sugar cane molasses. In the first strategy (fed-batch by pulsed
feeding), the amount of black strap sugar cane molasses (50 ml.L-1) was added to the fermentation
vessel. Two, three and four additions of this amount of black strap sugar cane molasses were also
added during the first 12 and 48 h of cultivation periods. In the second strategy, the black strap
sugar cane molasses was fed continuously during the first 12, 18 and 24 h of cultivation periods at
rates of 4.17, 2.78 and 2.08 ml.L-1.h-1, respectively (fed-batch by continuous feeding). The cell dry
weight, amino acid concentration and residual sugar were determined as well as the growth and
production parameters were calculated.
Results: The biological activity of Bacillus sp. R22EG1 and Bacillus sp. R20EG2 strains during production of amino acids on 5% black strap sugar cane molasses medium for 72 h at 30°C was
investigated in bioreactor as a batch and fed-batch cultures. In batch culture, the highest figures of
amino acid concentration (2.30 and 2.83 g.L-1), yield (9.52 and 11.71%), and conversion coefficient
(11.09 and 13.44%) were recorded after 72 h and 60 h of fermentation periods by Bacillus sp.
R22EG1 and R20EG2 strains, respectively, whereas the maximum productivity, approximately,
0.044 and 0.059 g.L-1.h-1 were observed after 12 h and 24 -36 h for Bacillus sp. R22EG1 a nd
R20EG2 strains, respectively. Two feeding strategies (pulsed and continuous) were studied during
production of amino acids using fed-batch culture. The highest cell dry weight, amino acid
concentration and yield were recorded after three pulsed molasses addition during the first 12 h of
fermentation periods with specific addition rate of 0.204 ml.L-1.h-1 (0.099 g.L-1.h-1 sugar) for the
tested strains. The continuous feeding rate of 4.17 ml.L-1.h-1 (2.01 g.L-1.h-1 sugar) was more
favorable than pulsed feeding during 12 h for amino acid production in fed-batch culture, as it
increased the amino acid concentration by Bacillus sp. R22EG1 and R20EG2 strains,
approximately, (1.53 & 1.42) fold than pulsed feeding and about (1.64 & 1.59) fold than that
produced in batch bioreactor technique, after a 48 h fermentation period. The highest content of
free amino acid species in culture supernatants was glutamic acid produced by both strains.
Conclusion: The maximum production of amino acids from continuous fed–batch by Bacillus sp.
R22EG1 strain and Bacillus sp. R20EG2 was 3.76 and 4.49 g.L-1 with continuous feeding at 4.17
ml.L-1.h-1 (2.01 g.L-1.h-1 sugar) at 48 h, respectively. These results were 1.53 & 1.42 fold higher than
pulsed feeding and 1.64 & 1.59 fold higher than batch fermentation by Bacillus sp. R22EG1 and
R20EG2 strains, respectively. The highest content species of free amino acids was glutamic acid
using a Bichrom 30 amino acid analyzer.
strain and Bacillus sp. R20EG2 using batch and fed-batch (pulsed and continuous feeding) cultures
was investigated to achieve the maximum concentration of free amino acids.
Place and Duration of Study: Department of Agricultural Microbiology, Faculty of Agriculture, Ain
Shams University, Cairo, Egypt, between December 2011 and March 2012.
Methodology: Two Bacillus strains namely R22EG1 and R20EG2, were used as amino acid
produces. The amino acid producing bacteria were grown in the bioreactor with batch and fedbatch
cultivation. The fed-batch fermentations were performed in two strategies using pulsed and
continuous feeding of black strap sugar cane molasses. In the first strategy (fed-batch by pulsed
feeding), the amount of black strap sugar cane molasses (50 ml.L-1) was added to the fermentation
vessel. Two, three and four additions of this amount of black strap sugar cane molasses were also
added during the first 12 and 48 h of cultivation periods. In the second strategy, the black strap
sugar cane molasses was fed continuously during the first 12, 18 and 24 h of cultivation periods at
rates of 4.17, 2.78 and 2.08 ml.L-1.h-1, respectively (fed-batch by continuous feeding). The cell dry
weight, amino acid concentration and residual sugar were determined as well as the growth and
production parameters were calculated.
Results: The biological activity of Bacillus sp. R22EG1 and Bacillus sp. R20EG2 strains during production of amino acids on 5% black strap sugar cane molasses medium for 72 h at 30°C was
investigated in bioreactor as a batch and fed-batch cultures. In batch culture, the highest figures of
amino acid concentration (2.30 and 2.83 g.L-1), yield (9.52 and 11.71%), and conversion coefficient
(11.09 and 13.44%) were recorded after 72 h and 60 h of fermentation periods by Bacillus sp.
R22EG1 and R20EG2 strains, respectively, whereas the maximum productivity, approximately,
0.044 and 0.059 g.L-1.h-1 were observed after 12 h and 24 -36 h for Bacillus sp. R22EG1 a nd
R20EG2 strains, respectively. Two feeding strategies (pulsed and continuous) were studied during
production of amino acids using fed-batch culture. The highest cell dry weight, amino acid
concentration and yield were recorded after three pulsed molasses addition during the first 12 h of
fermentation periods with specific addition rate of 0.204 ml.L-1.h-1 (0.099 g.L-1.h-1 sugar) for the
tested strains. The continuous feeding rate of 4.17 ml.L-1.h-1 (2.01 g.L-1.h-1 sugar) was more
favorable than pulsed feeding during 12 h for amino acid production in fed-batch culture, as it
increased the amino acid concentration by Bacillus sp. R22EG1 and R20EG2 strains,
approximately, (1.53 & 1.42) fold than pulsed feeding and about (1.64 & 1.59) fold than that
produced in batch bioreactor technique, after a 48 h fermentation period. The highest content of
free amino acid species in culture supernatants was glutamic acid produced by both strains.
Conclusion: The maximum production of amino acids from continuous fed–batch by Bacillus sp.
R22EG1 strain and Bacillus sp. R20EG2 was 3.76 and 4.49 g.L-1 with continuous feeding at 4.17
ml.L-1.h-1 (2.01 g.L-1.h-1 sugar) at 48 h, respectively. These results were 1.53 & 1.42 fold higher than
pulsed feeding and 1.64 & 1.59 fold higher than batch fermentation by Bacillus sp. R22EG1 and
R20EG2 strains, respectively. The highest content species of free amino acids was glutamic acid
using a Bichrom 30 amino acid analyzer.
Other data
Title | Enhancing Production of Amino Acids from Bacillus spp. Using Batch and Fed-batch Fermentation Strategies | Authors | Abou-Taleb, Khadiga | Keywords | Amino acids; bacteria; pulsed fed–batch; continuous fed–batch; bioreactor | Issue Date | 2015 | Publisher | SCIENCEDOMAIN international | Source | Abou-taleb, Kh. A. (2015). Enhancing Production of Amino Acids from Bacillus spp. Using Batch and Fed-Batch Fermentation Strategies. British Microbiology Research Journal, 5 (3): 257-272. | Journal | British Microbiology Research Journal |
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