Optimized invertase expression and secretion cassette for improving Yarrowia lipolytica growth on sucrose for industrial applications.
Yarrowia lipolytica requires the expression of a heterologous invertase to grow on a sucrose-based substrate. This work reports the construction of an optimized invertase expression cassette composed of Saccharomyces cerevisiae Suc2p secretion signal sequence followed by the SUC2 sequence and under the control of the strong Y. lipolytica pTEF promoter. This new construction allows a fast and optimal cleavage of sucrose into glucose and fructose and allows cells to reach the maximum growth rate. Contrary to pre-existing constructions, the expression of SUC2 is not sensitive to medium composition in this context. The strain JMY2593, expressing this new cassette with an optimized secretion signal sequence and a strong promoter, produces 4,519 U/l of extracellular invertase in bioreactor experiments compared to 597 U/l in a strain expressing the former invertase construction. The expression of this cassette strongly improved production of invertase and is suitable for simultaneously high production level of citric acid from sucrose-based media.
Department of Biotechnology and Food Microbiology, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37/41, 51-630, Wroclaw, Poland.
Zbigniew
Z
Lazar
Tristan
T
Rossignol
Jonathan
J
Verbeke
Anne-Marie
AM
Crutz-Le Coq
Jean-Marc
JM
Nicaud
Małgorzata
M
Robak
D018533:Biomass; D019149:Bioreactors; D001709:Biotechnology; D019343:Citric Acid / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D005632:Fructose / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006868:Hydrolysis; D011401:Promoter Regions, Genetic / Q000235:genetics; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D013395:Sucrose / Q000378:metabolism*; D025062:Yarrowia / Q000187:drug effects / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D043324:beta-Fructofuranosidase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; D029701:Saccharomyces cerevisiae Proteins; D019343:Citric Acid; D005632:Fructose; D013395:Sucrose; C468437:SUC2 protein, S cerevisiae; D043324:beta-Fructofuranosidase; D005947:Glucose