Repeated-batch production of glucoamylase using recombinant Saccharomyces cerevisiae immobilized in a fibrous bed bioreactor.
The recombinant Saccharomyces cerevisiae strain C468/pGAC9 has an unstable hybrid plasmid pGAC9, which directs production of glucoamylase. A fibrous cotton material with a good adsorption capability for recombinant S. cerevisiae cells was used as the immobilization matrix in an internal loop airlift-driven fibrous bed bioreactor (ILALFBB) system. With batch cultures in the ILALFBB, the fraction of plasmid-carrying cells was 72% after more than 2 days cultivation, which was two times higher than that in the conventional free-cell culture. Correspondingly, a high activity of glucoamylase (GA; 113 U/l) was achieved with a high productivity of 43 U/l/h. The ILALFBB system also maintained a high fraction of viable plasmid-carrying of 74% for glucoamylase production during repeated-batch cultures, achieving a high glucoamylase activity of 140 U/l with a productivity of 19-130 U/l/h in all 14 batches studied during 19.8 days. The stable and long-term glucoamylase production from the ILALFBB was attributed to the effect of cell immobilization on plasmid stability. Plasmid-carrying cells were preferentially retained in the fibrous matrix because of their ability to adhere to the fiber surface and to form cell aggregates higher than those of plasmid-free cells. The repeated batch using immobilized cell of recombinant S. cerevisiae in the ALALFBB system thus provides a feasible method for stable, long-term and high-level production of glucoamylase.
Kilonzo
Peter M
PM
Department of Chemical and Biochemical Engineering, University of Western Ontario, London, ON N6A5B9, Canada.
Margaritis
Argyrios
A
Bergougnou
Maurice A
MA
D019149:Bioreactors / Q000382:microbiology*; D018914:Cells, Immobilized / Q000201:enzymology*; D005087:Glucan 1,4-alpha-Glucosidase / Q000096:biosynthesis*; D003368:Gossypium; D050296:Microbial Viability; D010957:Plasmids / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis*
D029701:Saccharomyces cerevisiae Proteins; D005087:Glucan 1,4-alpha-Glucosidase
Bioresour Technol. 2009 Jul;100(13):3403-9
19297150
Appl Microbiol Biotechnol. 2009 Feb;82(2):241-8
19015846
Cytotechnology. 2004 Jan;44(1-2):1-14
19003225
Appl Biochem Biotechnol. 2008 Mar;145(1-3):59-67
18425612
Bioresour Technol. 2001 Jan;76(1):1-8
11315804
Biotechnol Prog. 1996 Jul-Aug;12(4):449-56
8987473
Biotechnol Bioeng. 1998 Aug 20;59(4):407-18
10099354
J Biotechnol. 1997 May 23;55(1):31-41
9226960
Enzyme Microb Technol. 2001 Jan 2;28(1):89-99
11118602
J Ind Microbiol Biotechnol. 2009 Sep;36(9):1157-69
19504139
J Biotechnol. 2003 Feb 27;101(1):81-7
12523972
Appl Microbiol Biotechnol. 2000 May;53(5):568-74
10855717
Bioresour Technol. 2009 Jan;100(1):374-9
18684618
Appl Environ Microbiol. 1996 Jun;62(6):1951-7
8787393
Appl Microbiol Biotechnol. 2009 Aug;84(2):261-9
19319524
Biotechnol Bioeng. 2001 Sep;76(2):164-74
11505386
Appl Microbiol Biotechnol. 1997 Jan;47(1):1-11
9035405