Beet molasses-based feeding strategy enhances recombinant thermostable glucose isomerase production by Escherichia coli BL21 (DE3)

Yaman, Sena
Çalık, Pınar
The aim of this work was to develop an effective fed-batch feeding strategy to enhance recombinant glucose isomerase (r-GI) production by recombinant Escherichia coli BL21 (DE3) pLysS on an industrially relevant feedstock without the application of an exogenous inducer. Following the batch operation (0 < t < 7H), the effects of pulse and/or continuous feeding of hydrolyzed beet molasses were investigated under five different feeding strategies. The two most promising strategies with respect to r-GI activity were (i) PM-0.05, designed with one pulse feed (t = 7H) followed by a continuous feed and (ii) 2PM(F)-0.05, designed with two consecutive pulse feeds (t = 7 and 10H) followed by a continuous feed. The continuous feeding of molasses for both fermentation strategies employed the same precalculated feeding rate, (o) = 0.05H(-1). The maximum r-GI activities exhibited by PM-0.05 and 2PM(F)-0.05 were 29,050 and 30,642U dm(-3), respectively. On the one hand, compared to PM-0.05 r-GI activity reached its maximum within a shorter cultivation time (t(max) = 2H) at 2PM(F)-0.05, which could be preferable in terms of manufacturing costs and possible risks; on the other hand, PM-0.05 is a simpler fermentation regime compared to 2PM(F)-0.05 with respect to manipulations that should be considered in large-scale production. (C) 2017 International Union of Biochemistry and Molecular Biology, Inc.


Codon optimization of xylA gene for recombinant glucose isomerase production in Pichia pastoris and fed-batch feeding strategies to fine-tune bioreactor performance
Ata, Ozge; Boy, Erdem; Gunes, Hande; Çalık, Pınar (Springer Science and Business Media LLC, 2015-05-01)
The objectives of this work are the optimization of the codons of xylA gene from Thermus thermophilus to enhance the production of recombinant glucose isomerase (rGI) in P. pastoris and to investigate the effects of feeding strategies on rGI production. Codons of xylA gene from T. thermophilus were optimized, ca. 30 % of the codons were replaced with those with higher frequencies according to the codon usage bias of P. pastoris, codon optimization resulted in a 2.4-fold higher rGI activity. To fine-tune bio...
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Angardi, Vahideh; Çalık, Pınar (Wiley, 2013-05-01)
BACKGROUND: The effects of pretreated beet molasses based feeding strategies on thermostable glucose isomerase (GI) production by recombinant Escherichia coli BL21 (DE3) pLysS were investigated. RESULTS: The thermostable GI encoding gene of Thermus thermophilus (xylA) was recombined with pRSETA vector, and the pRSETA::xylA obtained was transferred into E.coli BL21 (DE3) pLysS and used for GI production. The highest soluble GI activity was obtained at t = 30 h, as A = 16 400 U L1 (20.6 U mg1 protein) under m...
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Citation Formats
S. Yaman and P. Çalık, “Beet molasses-based feeding strategy enhances recombinant thermostable glucose isomerase production by Escherichia coli BL21 (DE3),” BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, pp. 944–954, 2017, Accessed: 00, 2020. [Online]. Available: