scholarly journals Engineering xylose utilization in Yarrowia lipolytica by understanding its cryptic xylose pathway

2016 ◽  
Vol 9 (1) ◽  
Author(s):  
Gabriel M. Rodriguez ◽  
Murtaza Shabbir Hussain ◽  
Lauren Gambill ◽  
Difeng Gao ◽  
Allison Yaguchi ◽  
...  
Author(s):  
Linlin Zhou ◽  
Zhiqiang Wen ◽  
Zedi Wang ◽  
Yuwei Zhang ◽  
Rodrigo Ledesma-Amaro ◽  
...  

Abstract Background: Yarrowia lipolytica is considered as a promising biorefinery chassis for production of microbial lipids, the important precursors of advanced biofuels. Unfortunately, wild Yarrowia lipolytica is unable to consume xylose, the major pentose in lignocellulosic hydrolysates. A recombinant strain Yarrowia lipolytica yl-XYL+ can utilize xylose to produce microbial lipids efficiently, but its xylose uptake is severely delayed in the presentence of D-glucose. Therefore, it is critical to develop co-fermenting D-glucose and xylose strains and study the underlying mechanisms.Results: In this study, an adaptive laboratory evolution (ALE) is performed to engineering the strains in the medium containing xylose and D-glucose analog 2-deoxyglucose (dG). After four stages of evolution over a total of 64 days, we obtained for the first time a strain of Y. lipolytica (yl-XYL+*04*10) with derepressed xylose metabolism. Xylose uptake kinetics showed that it could efficiently utilize xylose in the presence of 10 g/L dG or D-glucose. Transcriptional profiling analysis revealed that relative expression level of YALI0_C04730g and YALI0_D00363g (both encoding xylose-specific transporter) was significantly up-regulated. Besides, we found that missense mutations N373T and G270A in YALI0_E23287g (encoding a D-glucose transporter) and YALI0_E15488g (encoding a hexokinase) respectively.Conclusions: These results indicate that these are important gene targets responsible for improved xylose utilization in the evolved Yarrowia lipolytica. Our work provides a new approach for breeding Yarrowia lipolytica and paved the way for future pentose metabolic engineering.


2021 ◽  
Author(s):  
Caleb Walker ◽  
Bruce Dien ◽  
Richard J Giannone ◽  
Patricia Slininger ◽  
Stephanie R Thompson ◽  
...  

Yarrowia lipolytica is an oleaginous yeast exhibiting robust phenotypes beneficial for industrial biotechnology. The phenotypic diversity found within the undomesticated Y. lipolytica clade from various origins illuminates desirable phenotypic traits not found in the conventional laboratory strain CBS7504, which include xylose utilization, lipid accumulation, and growth on undetoxified biomass hydrolysates. Currently, the related phenotypes of lipid accumulation and degradation when metabolizing non-preferred sugars (e.g., xylose) associated with biomass hydrolysates  are poorly understood, making it difficult to control and engineer in Y. lipolytica To fill this knowledge gap, we analyzed the genetic diversity of five undomesticated Y. lipolytica strains and identified singleton genes and genes exclusively shared by strains exhibiting desirable phenotypes. Strain characterizations from controlled bioreactor cultures revealed that the undomesticated strain YB420 used xylose to support cell growth and maintained high lipid levels while the conventional strain CBS7504 degraded cell biomass and lipids when xylose was the sole remaining carbon source. From proteomic analysis, we identified carbohydrate transporters, xylose metabolic enzymes and pentose phosphate pathway proteins stimulated during the xylose uptake stage for both strains. Furthermore, we distinguished proteins in lipid metabolism (e.g., lipase, NADPH generation, lipid regulators, β-oxidation) activated by YB420 (lipid maintenance phenotype) or CBS7504 (lipid degradation phenotype) when xylose was the sole remaining carbon source. Overall, the results relate genetic diversity of undomesticated Y. lipolytica strains to complex phenotypes of superior growth, sugar utilization, lipid accumulation and degradation in biomass hydrolysates.


Planta Medica ◽  
2006 ◽  
Vol 72 (11) ◽  
Author(s):  
O Gortzi ◽  
S Papanikolaou ◽  
S Lalas ◽  
M Galiotou-Panayotou ◽  
P Mitliaga

LWT ◽  
2021 ◽  
pp. 111760
Author(s):  
Samantha Rossi ◽  
Luigi Parrotta ◽  
Stefano Del Duca ◽  
Marco Dalla Rosa ◽  
Francesca Patrignani ◽  
...  

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 88
Author(s):  
Paulina Snopek ◽  
Dorota Nowak ◽  
Bartłomiej Zieniuk ◽  
Agata Fabiszewska

Yarrowia lipolytica is one of the most studied non-conventional forms of yeast, exhibiting a high secretory capacity and producing many industrially important and valuable metabolites. The yeast conceals a great biotechnological potential to synthesize organic acids, sweeteners, microbial oil, or fragrances. The vast majority of bioprocesses are carried out in bioreactors, where suitable culture conditions are provided. In the current study, the effect of agitation speed (200–600 rpm) and air flow rate (0.0375–2.0 dm3/(dm3 × min)) on the biomass yield and lipase activity of Y. lipolytica KKP 379 is analyzed in a growth medium containing waste fish oil. The increase of aeration intensity limited the period of oxygen deficit in the medium. Simultaneously, an increase in lipolytic activity was observed from 2.09 U/cm3 to 14.21 U/cm3; however, an excessive agitation speed likely caused oxidative or shear stresses, and a reduction in lipolytic activity was observed. Moreover, it is confirmed that the synthesis of lipases is related to oxygen consumption, pH, and the yeast growth phase, and appropriate process selection may provide two advantages, namely, the maximum use of the waste carbon source and the production of lipolytic enzymes that are valuable in many industries.


2021 ◽  
pp. 124659
Author(s):  
Satish Kommoji ◽  
M. Gopinath ◽  
Polinati Satya Sagar ◽  
D. Yuvaraj ◽  
J. Iyyappan ◽  
...  

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