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- Title
Efficient secretion of inulin fructotransferase in Pichia pastoris using the formaldehyde dehydrogenase 1 promoter.
- Authors
Zhan, Rongrong; Mu, Wanmeng; Jiang, Bo; Zhou, Liuming; Zhang, Tao
- Abstract
Inulin fructotransferase (IFTase) has received considerable attention due to its ability to catalyse inulin hydrolysis to difructose anhydride (DFA III), a natural low-calorie functional sweetener. In the present study, for the first time, we describe the expression of IFTase in Pichia pastoris under the control of the formaldehyde dehydrogenase 1 promoter ( PFLD1). Using this system, we achieved efficient secretion with four substrate fed-batch strategies in a 3-L fermenter. The co-feeding induction strategy with methylamine hydrochloride and methanol achieved the maximum extracellular IFTase activity of 62.72 U mL, which was 3.2-fold higher than that obtained with the wild-type strain. In addition to methanol, carbon sources such as glucose and glycerol could also be utilised by PFLD1-controlled P. pastoris for IFTase production using methylamine hydrochloride induction. However, we found that glycerol and glucose should be strictly controlled at low concentrations of 0.5-1.5 % (v/v) and 1-1.5 % (w/v), respectively. The use of glycerol and glucose demonstrated that P. pastoris was also attractive for IFTase production via methanol-free cultivation strategies. This study may provide the basis for the industrial use of this recombinant IFTase for the production of DFA III.
- Subjects
PICHIA pastoris; FORMALDEHYDE dehydrogenase; FERMENTATION; BIOCHEMICAL substrates; METHYLAMINES; METHANOL
- Publication
Journal of Industrial Microbiology & Biotechnology, 2014, Vol 41, Issue 12, p1783
- ISSN
1367-5435
- Publication type
Article
- DOI
10.1007/s10295-014-1516-2