Enhanced production of insect raw-starch-digesting alpha-amylase accompanied by high erythritol synthesis in recombinant Yarrowia lipolytica fed-batch cultures at high-cell-densities

Feeding strategy impacts heterologous protein production in Yarrowia lipolytica fed‐batch cultures—Insight into the role of osmolarity

“…Consistently, our previous studies have shown that such a mode of cultivation had beneficial effect on the production of heterologous alpha‐amylase by Y . lipolytica , as the titer of the extracellular protein was increased by 160 fold (max activity 22,979.2 AU L −1 AUl −1 h −1 AUl −1 h −1 ; Celińska, Borkowska, & Białas, ), when compared with previously obtained result in batch cultivation (142.84 AU L −1 AUl −1 h −1 AUl −1 h −1 ; Celińska, Borkowska, & Białas, ). Another interesting example of fed‐batch cultures optimization was presented in a series of papers by (Chang, Ryu, Park, & Kim, ; Chang, Ryu, Park, Kim, & Ogrydziak, ; Kim et al, ), which concerned production of rice alpha‐amylase in recombinant Y .…”

“…Fed‐batch mode of culturing coupled with high‐cell density was frequently reported to be a strategy of choice for intensification of heterologous protein production in Y . lipolytica (Celińska, Borkowska, & Białas, ; Kim et al, ; Nicaud et al, ; Park, Chang, & Ryu, ). As shown by (Nicaud et al, ), application of fed‐batch high‐cell density culture resulted in 90‐fold higher production of extracellular lipase and aminopeptidase by a recombinant, multicopy strain.…”

“…Fed‐batch strategy facilitates development of more robust processes due to overcoming several limitations of batch cultures, like substrate exhaustion or lack of control of microbial growth and respiration, which contribute to underperformance of such cultures (Lee, Lee, Park, & Middelberg, ; Turki et al, ; Van Oort, ). It has been stated that fed‐batch cultivation is the generally preferred strategy for large‐scale production of heterologous proteins by yeasts (Hensing, Rouwenhorst, Heijnen, van Dijken, & Pronk, ), especially, when coupled with high‐cell‐density culturing (Celińska, Borkowska, & Białas, ; Kim, Park, Ryu, & Kim, ; Tapia, Vázquez‐Ramírez, Genzel, & Reichl, ).…”

Filamentous fungi-like secretory pathway strayed in a yeast system: peculiarities of Yarrowia lipolytica secretory pathway underlying its extraordinary performance

“…Enhanced production of the targeted, secretory protein may be achieved by several different strategies, including (i) engineering the DNA coding sequence itself, (ii) modification of a genetic construct structure, (iii) optimization of culturing conditions, and (iv) engineering molecular mechanisms of protein synthesis and secretion pathway (Graf et al 2009; Liu et al 2012). Within each of the proposed strategies, various solutions have been proposed, and some of them were conducted in Y. lipolytica , like (i) optimization of codon usage within the coding sequence (Celińska et al 2015; Celińska 2015; Dulermo et al 2017), (ii) amplification of the gene copy number, manipulation with regulatory elements contained in the genetic construction, mainly promoter (Le Dall et al 1994; Gasmi et al 2011b; Gasmi et al 2011a; Gasmi et al 2012; Dulermo et al 2017; Larroude et al 2018), (iii) bioprocess engineering, optimization of cultivation conditions (Chang et al 1998; Kim et al 2000; Nicaud et al 2002; Celińska et al 2017a), and (iv) co-synthesis of chaperones, manipulation with protein folding, and maturation mechanisms, engineering secretory tags (De Pourcq et al 2012; Celińska et al 2018). Nevertheless, the vast majority of studies on enhanced production of secretory proteins in Y. lipolytica was focused on strategies from (i) to (iii), while the approaches directing secretory pathway were largely neglected.…”

Erythritol production by yeasts: a snapshot of current knowledge

“…When cultivated in a glycerol medium, this strain also exhibited an important erythritol production. Although amylase activity was the primary objective and erythritol was a side product, its concentration and productivity (84 g/L and 0.7 g/Lh respectively) were on a par with other erythritol production processes, and could contribute to the profitability of this process (Celińska, Borkowska, & Białas, ). A similar co‐production of erythritol along with an enzyme was reported in Y. lipolytica .…”