In silico and in vivo analysis of signal peptides effect on recombinant glucose oxidase production in nonconventional yeast Yarrowia lipolytica
Signal peptide (SP) is an important factor and biobrick in the production and secretion of recombinant proteins. The aim of this study was in silico and in vivo analysis of SPs effect on the production of recombinant glucose oxidase (GOX) in Yarrowia lipolytica. Several in silico softwares, namely SignalP4, Signal-CF, Phobius, WolfPsort 0.2, SOLpro and ProtParam, were used to analyse the potential of 15 endogenous and exogenous SPs for the secretion of recombinant GOX in Y. lipolytica. According to in silico results, the SP of GOX was predicted as suitable in terms of high secretory potential and of protein solubility and stability which is chosen for in vivo analysis. The recombinant Y. lipolytica strain produced 280 U/L of extracellular GOX after 7 days in YPD medium. The results show that the SP of GOX can be applied to efficient production of extracellular heterologous proteins and metabolic engineering in Y. lipolytica.
KeywordsYarrowia lipolytica Signal peptide In silico In vivo Glucose oxidase
Compliance with ethical standards
Conflict of interest
The authors declare no conflict of interest.
- Frederick KR, Tung J, Emerick RS, Masiarz FR, Chamberlain SH, Vasavada A, Rosenberg S, Chakraborty S, Schopfer L, Schopter L (1990) Glucose oxidase from Aspergillus niger. cloning, gene sequence, secretion from Saccharomyces cerevisiae and kinetic analysis of a yeast-derived enzyme. J Biol Chem 265:3793–3802PubMedGoogle Scholar
- Humphreys DP, Sehdev M, Chapman AP, Ganesh R, Smith BJ, King LM, Glover DJ, Reeks DG, Stephens PE (2000) High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5′ end of the coding sequence. Protein Expr Purif 20:252–264CrossRefPubMedGoogle Scholar
- Madzak C, Beckerich JM (2013) Heterologous protein expression and secretion in Yarrowia lipolytica. In: Barth G (ed) Yarrowia lipolytica: biotechnological applications. Springer, BerlinGoogle Scholar
- Madzak C, Nicaud JM, Gaillardin C (2005a) Yarrowia lipolytica. Production of recombinant proteins: novel microbial and eukaryotic expression systems. Springer, BerlinGoogle Scholar
- Mirbagheri M, Nahvi I, Emtiazi G, Mafakher L, Darvishi F (2012) Taxonomic characterization and potential biotechnological applications of Yarrowia lipolytica isolated from meat and meat products. Jundishapur J Microbiol 5:346–351Google Scholar
- Sibirny A, Madzak C, Fickers P (2014) Genetic engineering of non-conventional yeast for the production of valuable compounds. In: Darvishi F, Chen H (eds) Microbial biotechnology: progress and trends. CRC Press, Boca RatonGoogle Scholar