Simultaneous Improvement of Limonene Production and Tolerance in Yarrowia lipolytica through Tolerance Engineering and Evolutionary Engineering

2021 ◽  
Author(s):  
Jian Li ◽  
Kun Zhu ◽  
Lin Miao ◽  
Lanxin Rong ◽  
Yu Zhao ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Evolutionary Engineering

scholarly journals Evolutionary engineering improved D-glucose/xylose co-fermentation of Yarrowia lipolytica

2019 ◽  
Author(s):  
Linlin Zhou ◽  
Zhiqiang Wen ◽  
Zedi Wang ◽  
Yuwei Zhang ◽  
Rodrigo Ledesma-Amaro ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Glucose Transporter ◽  
Transcriptional Profiling ◽  
Evolutionary Engineering ◽  
Xylose Utilization ◽  
Missense Mutations ◽  
Microbial Lipids ◽  
Xylose Metabolism ◽  
Adaptive Laboratory Evolution ◽  
Xylose Uptake

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.

scholarly journals Does co-expression of Yarrowia lipolytica genes encoding Yas1p, Yas2p and Yas3p make a potential alkane-responsive biosensor in Saccharomyces cerevisiae?

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0239882
Author(s):  
Yasaman Dabirian ◽  
Christos Skrekas ◽  
Florian David ◽  
Verena Siewers
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yarrowia Lipolytica ◽  
High Throughput Screening ◽  
Screening Tools ◽  
Evolutionary Engineering ◽  
Transportation Fuels ◽  
Sensing System ◽  
Commercial Scale ◽  
Genes Encoding ◽  
Native Host

Alkane-based biofuels are desirable to produce at a commercial scale as these have properties similar to current petroleum-derived transportation fuels. Rationally engineering microorganisms to produce a desirable compound, such as alkanes, is, however, challenging. Metabolic engineers are therefore increasingly implementing evolutionary engineering approaches combined with high-throughput screening tools, including metabolite biosensors, to identify productive cells. Engineering Saccharomyces cerevisiae to produce alkanes could be facilitated by using an alkane-responsive biosensor, which can potentially be developed from the native alkane-sensing system in Yarrowia lipolytica, a well-known alkane-assimilating yeast. This putative alkane-sensing system is, at least, based on three different transcription factors (TFs) named Yas1p, Yas2p and Yas3p. Although this system is not fully elucidated in Y. lipolytica, we were interested in evaluating the possibility of translating this system into an alkane-responsive biosensor in S. cerevisiae. We evaluated the alkane-sensing system in S. cerevisiae by developing one sensor based on the native Y. lipolytica ALK1 promoter and one sensor based on the native S. cerevisiae CYC1 promoter. In both systems, we found that the TFs Yas1p, Yas2p and Yas3p do not seem to act in the same way as these have been reported to do in their native host. Additional analysis of the TFs suggests that more knowledge regarding their mechanism is needed before a potential alkane-responsive sensor based on the Y. lipolytica system can be established in S. cerevisiae.

scholarly journals Does co-expression of Yarrowia lipolytica genes encoding Yas1p, Yas2p and Yas3p make a potential alkane-responsive biosensor in Saccharomyces cerevisiae?

2020 ◽  
Author(s):  
Yasaman Dabirian ◽  
Christos Skrekas ◽  
Florian David ◽  
Verena Siewers
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Yarrowia Lipolytica ◽  
High Throughput Screening ◽  
Screening Tools ◽  
Evolutionary Engineering ◽  
Transportation Fuels ◽  
Sensing System ◽  
Commercial Scale ◽  
Genes Encoding ◽  
Native Host

ABSTRACTAlkane-based biofuels are desirable to produce at a commercial scale as these have properties similar to our current petroleum-derived transportation fuels. Rationally engineering microorganisms to produce a desirable compound, such as alkanes, is, however, challenging. Metabolic engineers are therefore increasingly implementing evolutionary engineering approaches combined with high-throughput screening tools, including metabolite biosensors, to identify productive targets. Engineering Saccharomyces cerevisiae to produce alkanes can be facilitated by using an alkane-responsive biosensor, which can potentially be developed from the native alkane-sensing system in Yarrowia lipolytica, a well-known alkane-assimilating yeast. This putative alkane-sensing system is, at least, based on three different transcription factors (TFs) named Yas1p, Yas2p and Yas3p. Although this system is not fully elucidated in Y. lipolytica, we were interested in evaluating the possibility of translating this system into an alkane-responsive biosensor in S. cerevisiae. We evaluated the alkane-sensing system in S. cerevisiae by developing one sensor based on the native Y. lipolytica ALK1 promoter and one sensor based on the native S. cerevisiae CYC1 promoter. In both systems, we found that the TFs Yas1p, Yas2p and Yas3p do not seem to act in the same way as these have been reported to do in their native host. Additional analysis of the TFs suggests that more knowledge regarding their mechanism is needed before a potential alkane-responsive sensor based on the Y. lipolytica system can be established in S. cerevisiae.

scholarly journals Evolutionary Engineering Improved d-Glucose/Xylose Cofermentation of Yarrowia lipolytica

2020 ◽  
Vol 59 (39) ◽  
pp. 17113-17123 ◽  
Author(s):  
Linlin Zhou ◽  
Zhiqiang Wen ◽  
Zedi Wang ◽  
Yuwei Zhang ◽  
Rodrigo Ledesma-Amaro ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Evolutionary Engineering

Effect of essential oil of Citrus cinensis cv new hall – Citrus aurantium (indigenous in Greece) upon growth of Yarrowia lipolytica

Planta Medica ◽  
2006 ◽  
Vol 72 (11) ◽  
Author(s):  
O Gortzi ◽  
S Papanikolaou ◽  
S Lalas ◽  
M Galiotou-Panayotou ◽  
P Mitliaga
Keyword(s):  
Essential Oil ◽  
Yarrowia Lipolytica ◽  
Citrus Aurantium

Effect of Yarrowia lipolytica RO25 cricket-based hydrolysates on sourdough quality parameters

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

scholarly journals Aeration and Stirring in Yarrowia lipolytica Lipase Biosynthesis during Batch Cultures with Waste Fish Oil as a Carbon Source

Fermentation ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 88
Author(s):  
Paulina Snopek ◽  
Dorota Nowak ◽  
Bartłomiej Zieniuk ◽  
Agata Fabiszewska
Keyword(s):  
Carbon Source ◽  
Fish Oil ◽  
Yarrowia Lipolytica ◽  
Lipolytic Activity ◽  
Culture Conditions ◽  
Agitation Speed ◽  
Oxygen Deficit ◽  
Shear Stresses ◽  
Process Selection ◽  
Batch Cultures

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.

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