Fed-batch production of a soluble β-1,4-galactosyltransferase with Saccharomyces cerevisiae

• Published in: Enzyme and microbial technology Vol. 25; no. 3; pp. 201 - 207
• Main Authors: Zigova, Jana, Mahle, Matthias, Paschold, Holger, Malissard, Martine, Berger, Eric G, Weuster–Botz, Dirk
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.08.1999; Elsevier Science
• Subjects: Biological and medical sciences; Biotechnology; Chemical variables control; Enzymes; Feedback control; Fermentation; Flow-injection analysis; Fundamental and applied biological sciences. Psychology; Glucose; Glycosyltransferase; Kalman filter; Methods. Procedures. Technologies; Microbial engineering. Fermentation and microbial culture technology; Microorganisms; Proteins; Respiratory quotient (RQ); Saccharomyces cerevisiae; Feedback control; Glycosyltransferase; Saccharomyces cerevisiae; Respiratory quotient (RQ); Kalman filter; Flow-injection analysis; Human; Recombinant microorganism; Yeast; Enzyme; Transferases; Process control; Glycosyltransferases; Fermentation; Pilot plant scale; Fungi; Dimensioning; β-N-Acetylglucosaminylglycopeptide β-1,4-galactosyltransferase; Analysis method; Production; Ascomycetes; Hexosyltransferases; Recombinant protein; Semicontinuous; Thallophyta; Flow injection
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/S0141-0229(99)00027-7

A feedback-controlled fed-batch process for the recombinant production of a soluble human β-1,4-galactosyltransferase (NdrGal-T) with Saccharomyces cerevisiae was developed and scaled up to the pilot scale. A 5000 U NdrGal-T fermentation run was performed on a 300 l scale. Indirect feedback control of the glucose feeding with RQ data at a set-point of RQ = 1.1 resulted in higher NdrGal-T activities (30 U/l) than direct feedback control of glucose with on-line flow-injection analysis (9 U/l). This increase in final activity of NdrGal-T by a factor of 42 compared to published data makes the fed-batch production of glycosyltransferases with a Saccharomyces cerevisiae expression system competitive with cell-culture systems.