Secretory production of Zymomonas mobilis levansucrase by the methylotrophic yeast Hansenula polymorpha

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Title
Secretory production of Zymomonas mobilis levansucrase by the methylotrophic yeast Hansenula polymorpha
Author(s)
Buem Seek Park; A Vladimir; Chul Ho Kim; Sang Ki Rhee; Hyun Ah Kang
Bibliographic Citation
Enzyme and Microbial Technology, vol. 34, no. 2, pp. 132-138
Publication Year
2004
Abstract
To direct the secretory expression of Zymomonas mobilis levansucrase (LevU) in the methylotrophic yeast Hansenula polymorpha, the Z. mobilis levU gene was fused to the inulinase signal sequence under the control of the methanol oxidase promoter (PMOX). The vector containing the P MOX-levU expression cassette was introduced into H. polymorpha and multiply integrated into the host chromosomes. Recombinant LevU was expressed as an active and soluble form in H. polymorpha transformants, but the recombinant protein accumulated mainly inside the cells, representing about 5% of total cell proteins in shake-flask cultivations. Interestingly, a significant fraction of the recombinant LevU was secreted into the culture supernatant with a maximum yield of 12,200U/l using fed-batch fermentations with methanol feeding. H. polymorpha-derived recombinant LevU displayed catalytic activities comparable to those of the Escherichia coli-derived recombinant LevU, producing levans with high molecular weights.
Keyword
hansenula polymorpharecombinant levansucrasesecretory expressionzymomonas mobilis
ISSN
0141-0229
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.enzmictec.2003.09.005
Type
Article
Appears in Collections:
Jeonbuk Branch Institute > Microbial Biotechnology Research Center > 1. Journal Articles
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