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Enhanced α‐amylase production in recombinant Bacillus brevis by fed‐batch culture with amino acid control

Enhanced α‐amylase production in recombinant Bacillus brevis by fed‐batch culture with amino acid... Fed‐batch culture with controlled L‐amino acid composition was performed to improve production of a recombinant gene product in Bacillus brevis. The maximum recombinant protein (α‐amylase) level and specific activity increased from 5.14 kU/mL and 0.77 kU/mg dry cell in conventional fed‐batch culture to 12.01 kU/mL and 2.64 kU/mg dry cell, respectively, when L‐amino acid concentration was controlled at 5 mM using an asparagine (Asn)‐ and isoleucine (Ile)‐enriched nitrogen source. The L‐amino acid concentration in the culture was monitored by an automatic biotech analyzer and controlled at 2–20 mM using a mixture of polypeptone and yeast extract. Although L‐amino acid concentrations were controlled at low levels, the α‐amylase activity increased only 1.3 times compared to an uncontrolled batch culture; accumulation of ammonium ion was not reduced. When L‐amino acid was controlled at the high level, more cell mass and less recombinant gene product were produced than in those with low control level. To overcome ammonium ion inhibition, the specific amino acids Asn and Ile were substituted to improve the production of gene product. Addition of these amino acids to a flask culture led to an improvement in the enzyme production level and specific activity to 2.9 and 5.1 times, respectively, as high as that without them. Both the control of amino acids at low concentrations and the enrichment of Asn and Ile were effective for the improvement of recombinant protein production from recombinant B. brevis cells. © 1996 John Wiley & Sons, Inc. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology and Bioengineering Wiley

Enhanced α‐amylase production in recombinant Bacillus brevis by fed‐batch culture with amino acid control

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References (41)

Publisher
Wiley
Copyright
Copyright © 1996 Wiley Subscription Services
ISSN
0006-3592
eISSN
1097-0290
DOI
10.1002/(SICI)1097-0290(19960105)49:1<36::AID-BIT5>3.0.CO;2-U
pmid
18623551
Publisher site
See Article on Publisher Site

Abstract

Fed‐batch culture with controlled L‐amino acid composition was performed to improve production of a recombinant gene product in Bacillus brevis. The maximum recombinant protein (α‐amylase) level and specific activity increased from 5.14 kU/mL and 0.77 kU/mg dry cell in conventional fed‐batch culture to 12.01 kU/mL and 2.64 kU/mg dry cell, respectively, when L‐amino acid concentration was controlled at 5 mM using an asparagine (Asn)‐ and isoleucine (Ile)‐enriched nitrogen source. The L‐amino acid concentration in the culture was monitored by an automatic biotech analyzer and controlled at 2–20 mM using a mixture of polypeptone and yeast extract. Although L‐amino acid concentrations were controlled at low levels, the α‐amylase activity increased only 1.3 times compared to an uncontrolled batch culture; accumulation of ammonium ion was not reduced. When L‐amino acid was controlled at the high level, more cell mass and less recombinant gene product were produced than in those with low control level. To overcome ammonium ion inhibition, the specific amino acids Asn and Ile were substituted to improve the production of gene product. Addition of these amino acids to a flask culture led to an improvement in the enzyme production level and specific activity to 2.9 and 5.1 times, respectively, as high as that without them. Both the control of amino acids at low concentrations and the enrichment of Asn and Ile were effective for the improvement of recombinant protein production from recombinant B. brevis cells. © 1996 John Wiley & Sons, Inc.

Journal

Biotechnology and BioengineeringWiley

Published: Jan 5, 1996

Keywords: ; ; ;

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