Cloning of Levansucrase from Leuconostoc mesenteroides and Its Expression in Pichia pastoris
A gene (m1ft) encoding levansucrase from Leuconostoc mesenteroides has been previously cloned in Escherichia coli. Presently, m1ft was cloned and secretively expressed in Pichia pastoris. Methanol induction of recombinant M1FT resulted in the production of active levansucrase (PM1FT). PM1FT-5 was expressed as an active form, but the protein accumulated mainly inside the cells, representing about 5% of total cell proteins. M1FT was secreted into the culture supernatant with a maximum yield of 14,400 U/L using fed-batch fermentations. P. pastoris-derived M1FT displayed catalytic activities comparable to those of the E. coli-derived M1FT. PM1FT was glycosylated at its 2 potential N-glycosylation sites.
- Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol. Rev. 24: 45-66 (2000)
- Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr. Opin. Biotech. 13: 329-332 (2002)
- Lee CCW, Tina G, Wong DWS, Robertson GH. An episomal expression vector for screening mutant gene libraries in Pichia pastoris. Plasmid 54: 80-85 (2005)
- Rhee SK, Song KB, Kim CH, Park BS, Jang EK, Jang KH. Levan. Vol. V, pp. 351-377. In: Biopolymers, Polysaccharides I: Polysaccharides from Prokaryotes. De Baets S, Vandamme EJ, Steinbuchel A (eds). Wiley-VCH Verlag GmbH & Co., Weinheim, Germany (2002)
- Scotti PA, Chambert R, Petit-Glatron MF. Extracellular levansucrase of Bacillus subtilis produced in yeast remains in the cell in its precursor form. Yeast 10: 29-38 (1994)
- Trujillo LE, Arrieta JG, Dafhnis F, Garcia J, Valdes J, Tambara Y, Perez M, Hernandez L. Fructo-oligosaccharides production by the Gluconacetobacter diazotrophicus levansucrase expressed in the methylotrophic yeast Pichia pastoris. Enzyme Microb. Tech. 28: 139-144 (2001)
- Kang HK, Seo MY, Seo ES, Kim D, Chung SY, Kimura A, Day DF, Robyt JF. Cloning and expression of levansucrase from Leuconostoc mesenteroides B-512 FMC in Escherichia coli. Biochim. Biophys. Acta 1727: 5-15 (2005)
- Rose MD, Winstern F, Hieter P. Methods in Yeast Genetics: A Laboratory Course Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA. pp. 122-123, 140-141 (1990)
- Xu B, Yang YJ, Huang ZX. Cloning and overexpression of gene encoding the pullulanase from Bacillus maganoensis in Pichia pastoris. J. Microbiol. Biotechn. 16: 1185-1191 (2007)
- Kang HK, Park JY, Ahn JS, Kim SH, Kim D. Cloning of a gene encoding dextranase from Lipomyces starkeyi and its expression in Pichia pastoris. J. Microbiol. Biotechn. 19: 172-177 (2009)
- Invitrogen. Pichia Expression Kit. Protein Expression. A Manual of Methods for Expression of Recombinant Proteins in P. pastoris. Cat. No. K1710-01. Invitrogen Co., Carlsbard, SD, USA (1996)
- Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 (1970)
- Bradford MM. A rapid and sensitive method for the quentitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254 (1976)
- Tanriseven A, Robyt JF. Interpretation of dextransucrase inhibition at high sucrose concentration. Carbohyd. Res. 245: 97-104 (1993)
- Miller AW, Robyt JF. Detection of dextransucrase and levansucrase on polyacrylamide gels by the periodic acid-Schiff stain: Staining artifacts and their prevention. Anal. Biochem. 156: 357-363 (1988)
Kang HK, Oh JS, Kim D. Molecular characterization and expression analysis the glucansucrase DSRWC from Weissella cibaria synthesizing a
$\alpha$(1 $\rightarrow$6) glucan. FEMS Microbiol. Lett. 292: 33-41 (2009)
- Daly R, Hearn MT, Expression of heterologous proteins in Pichia pastoris: A useful experimental tool in protein engineering and production. J. Mol. Recognit. 18: 119-138 (2005)
- Bollok M, Resina D, Valero F, Ferrer P. Recent patenta on the Pichia pastoris expression system: Expanding the toolbox for recombinant protein production. Recent Pat. Biotechnol. 3: 192-201 (2009)
- Scotti PA, Praestegaard M, Chambert R, Petit-Glatron MF. The targeting of Bacillus subtilis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N-terminus mature part. Yeast 12: 953-963 (1996)
- Hestrin S, Avigad G. The mechanism of polysaccharide production from sucrose: Transfer of fructose to C-1 of aldoses by levansucrase. Biochem. J. 69: 388-398 (1958)
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