Yarrowia lipolytica

Saccharomyces cerevisiae is well characterized biochemically and genetically and was the organism of choice for most of the eady experiments. However, heterologous expression seems to be better in some of the industrial strains of yeasts such as Pichiapastoris Hansenulapolymorpha Kluyveromyces lactis and Yarrowia lipolytica (25—28). 

An alternative approach to the microbial deracemization of secondary alcohols is to use two different microorganisms with complementary stereoselectivity. Fantin et al. studied the stereoinversion of several secondary alcohols using the culture supernatants of two microorganisms, namely Bacillus stearothermophilus and Yarrowia lipolytica (Figure 5.18) [31]. The authors tested three main systems for deracemization. First, they used the supernatant from cultures of B. stearothermophilus, to which they added Y. lipolytica cells and the racemic alcohols. Secondly, they used the culture supernatant of Y. lipolytica and added B. stearothermophilus cells and the racemic alcohols. Finally, they resuspended the cells of both organisms in phosphate buffer and added the racemic alcohols. The best results were obtained in the first system with 6-penten-2-ol (26) (100% ee and 100% yield). The phosphate buffer system gave... 

Traditional and well-established yeast species are Saccharomyces cerevisiae, Hansenula polymorpha, Klyveromyces lactis, Pichia pastoris and Schizosaccharomyces pombe. With every year that passes they are increasingly being used in industrial and pharmaceutical enzyme production on a large scale. Many further yeasts present interesting features (e.g. Arxula adeninivorans and Yarrowia lipolytica), but are not that widely used. 

Gellissen, G., Kunze, G., Gaillardin, C. et al. (2005) New yeast expression platforms based on methylotrophic Hansenula polymorpha and Pichia pastoris and on dimorphic Arxula adeninivorans and Yarrowia lipolytica - a comparison. FEMS Yeast Research, 5 (11), 1079-1096. 

Madzak, C., Gaillardin, C. and Beckerich, J.M. (2004) Heterologous protein expression and secretion in the non-conventional yeast Yarrowia lipolytica. a review. Journal of Biotechnology, 109 (1—2), 63—81. 

Figure 7.1 The reductions of a-chloroketones catalyzed by Yarrowia lipolytica CECT1240 and Pichia mexicana CECT11015...

In addition to bacterial conversion of L-methionine to cheese aroma compounds, certain cheese-ripening yeasts have been implicated. They include De-baromyces hansenii, Geotrichum candidum, and Yarrowia lipolytica, in addition to Kluyveromyces lactis and Saccharomyces cerevisiae (previously noted). Of these yeasts, Geotrichum candidum was most effective at producing sulfur compounds with the major product being S-methyl thioacetate, with smaller amounts of MT, DMS, DMDS, and DMTS. Kluyveromyces lactis had a similar profile, but produced a much smaller amount of S-methyl thioacetate than did G. candidum. S-Methyl thioacetate is formed by a reaction of MT and acetyl-CoA (Equation 7) ... 

Manufacturing processes for I -decalactone have been T developed by a number of flavour companies using yeast such as Yarrowia lipolytica and S. cerevisiae selected... 

Decanolide (y-decalactone) Ricinoleic acid Yarrowia lipolytica llgL, 55 h, several tons per year Final acidification and temperature increase effect cyclisation of all 4-hydroxydecanoic acid to the corresponding lactone [222, 224, 228]... 

However, expression in a microbial cell is not always straightforward. For example, recombinant enzyme activity maybe different from that of the native enzyme. When incubated in a mixture of hydroperoxides, a HPL from green bell pepper (Capsicum annuum L.) that was expressed in Yarrowia lipolytica favours the production of hexanal although the native enzyme produces the unsaturated aldehyde ds-3-hexenal, both within the green bell pepper itself and when expressed in E. coli [22]. 

Finally, the yeast Yarrowia lipolytica is able to transform ricinoleic acid (12-hydroxy oleic acid) into y-decalactone, a desirable fruity and creamy aroma compound however, the biotransformation pathway involves fi-oxidation and requires the lactonisation at the CIO level. The first step of fi-oxidation in Y. lipolytica is catalysed by five acyl-CoA oxidases (Aox), some of which are long-chain-specific, whereas the short-chain-specific enzymes are also involved in the degradation of the lactone. Genetic constructions have been made to remove these lactone-degrading activities from the yeast strain [49, 50]. A strain displaying only Aox2p activity produced 10 times more lactone than the wild type in 48 h but still showed the same growth behaviour as the wild type. 

Acetobacter aceti Clostridium thermoaceticum Propionibacterium shermanii Aspergillus niger Yarrowia lipolytica... 

FIG. 20 Flow sheet of a novel integrated membrane process for citric acid production from glucose syrups by Yarrowia lipolytica, as proposed by Moresi (1995). 

Bubbico, R., Lo Presti, S., and Moresi, M. 1997. Repeated batch citrate production by Yarrowia lipolytica in a membrane recycle bioreactor. In Engineering Food at ICEF7. Part I (R. Jowitt, ed.), pp. B21-B24. Sheffield Academic Press, Sheffield, UK. 

Papanikolaou, S., Muniglia, L., Chevalot, I., Aggelis, G. and Marc, I. 2002. Yarrowia Lipolytica as a Potential Producer of Citric Acid from Raw Glycerol. J. Appl. Microbiol., 92, 737-744. Papanikolaou, S., Ruiz-Sanchez, P., Pariset, B., Blanchard, F. and Fick, M. 2000. High Production of 1,3-Propanediol from Industrial Glycerol by a Newly Isolated Clostridium Butyricum Strain. J. Biotechnol., 77, 191-208. 

Park, C., Chang, C., and Ryu, D. 2000. Expression and high-level secretion of Trichoderma reesei endoglucanase I in Yarrowia lipolytica. Appl. Biochem. Biotechnol., 87,1-15. 

Fig. 10. Sequence of the Cu regulatory domain (CuRD) of Ace 1 from Saccharomyces cerevisiae. Eight cysteinyl residues form the CuRD. A consensus sequence is derived from orthologues from Candida glabrata (Amt 1) and Yarrowia lipolytica (Crfl). The presence of a large variable segment in the middle of the CuRD suggests that the CuRD consists of two lobes with four Cys residues each. The tetracopper cluster is likely buried between these two-lobe polypeptides.

Rodrigues, G. and Pais, C. 2000. The influence of acetic and other weak carboxylic acids on growth and cellular death of the yeast Yarrowia lipolytica. Food Technology and Biotechnology 38 27-32. 

Finogenova, T.V., Morgunov, I.G., Kamsolova, S.V., and Chernyavskaya, O.G. 2005. Organic acid production by the yeast Yarrowia lipolytica A review of prospects. Applied Biochemistry and Microbiology 41 418-425. 

Shah, N.D., Chattoo, B.B., Baroda, R.M., and Patiala, V.M. 1993. Starch hydrolysate, an optimal and economical source of carbon for the secretion of citric acid by Yarrowia lipolytica. Starch 45 104-109. 

Interest in the yeast Yarrowia lipolytica has also developed over the past few years, due to potentially numerous biotechnological applications. Although this organism is not considered to be a dangerous food spoilage yeast, various cases have been reported from food environments, some associated with severe damage (Rodrigues and Pais, 2000). 

Farbood and Willis(68) in a recent patent application disclosed a process for production of optically active alpha-hydroxy decanoic acid (gamma-decalactone) by growing Yarrowia lipolytica on castor oil as a sole source of carbon. This is a good example of a commercial application of a volatile chemical produced by a microorganism. Yields of up to 6 grams per liter culture media were obtained making this a promising industrial fermentation. 

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