A two-stage temperature control strategy enhances extracellular secretion of recombinant α-cyclodextrin glucosyltransferase in Escherichia coli

Abstract

The effects of temperature on extracellular secretion of the α-cyclodextrin glucosyltransferase (α-CGTase) from Paenibacillus macerans JFB05-01 by Escherichia coli were investigated. When protein expression was induced at constant temperature, the greatest amount of extracellular recombinant α-CGTase was produced at 25 °C. Higher or lower induction temperatures were not conducive to extracellular secretion of recombinant α-CGTase. To enhance extracellular secretion of α-CGTase by E. coli, a two-stage temperature control strategy was adopted. When expression was induced at 25 °C for 32 h, and then the temperature was shifted to 30 °C, the extracellular α-CGTase activity at 90 h was 45% higher than that observed when induction was performed at a constant temperature of 25 °C. Further experiments suggested that raising the induction temperature can benefit the transport of recombinant enzyme and compensate for the decreased rate of recombinant enzyme synthesis during the later stage of expression. This report provides a new method of optimizing the secretory expression of recombinant enzymes by E. coli.

Enhanced AmB Production in Streptomyces nodosus by Fermentation Regulation and Rational Combined Feeding Strategy

“…As shown in Table 2 , the maximum biomass at different temperatures of 24, 26, 28, 30, and 32°C was 28, 32, 30, 34, and 35%, respectively, which indicated that high temperature (30°C) with the highest AmB titer (11.01 g/L) could accelerate cell growth. In particular, we postulated that some enzymes associated with AmB biosynthesis, such as ketoreductase, cytochrome P450 enzymes, etc., may have higher optimum temperatures and promoted product synthesis by affecting the activity of related enzymes, which was consistent with other studies (Peng et al, 2010 ; Li et al, 2017 ). Furthermore, when the temperature exceeded 30°C, the cell growth was continuous, but the biosynthesis of AmB was essentially inhibited ( Table 2 ).…”

Efficient production of heat-stable antifungal factor through integrating statistical optimization with a two-stage temperature control strategy in Lysobacter enzymogenes OH11

“…However, the temperatures required for microbial growth and product formation differ in most cases [ 14 ]. A phased temperature control strategy can provide ideal temperatures for both cell growth and product synthesis and is widely used in generating fermentation products such as α-cyclodextrin glucosyltransferase [ 27 ], 1,3-propanediol [ 14 ], and glutathione [ 28 ].…”