Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae

Halka, Lisa; Nowacki, Christian; Kleinschmidt, Alica; Koenen, Kevin; Wichmann, R.

doi:10.1186/s13568-018-0662-8

Cited by 5 publications

(5 citation statements)

References 18 publications

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“…According to the diffusional exponent (0.65), the FeedBeads ® release mechanism is due to diffusion and erosion of the matrix ( Padmaa Paarakh et al, 2018 ). The glycerol release kinetic determined for the glycerol FeedBeads ® was in the same order as previously found for the glucose FeedBeads ® , being 1,34 t 0,77 and 1,63 t 0,74 mg of glucose per hour and one FeedBeads ® ( Prielhofer et al, 2015 ; Halka et al, 2018 ).…”

Section: Resultssupporting

confidence: 66%

Enabling growth-decoupled Komagataella phaffii recombinant protein production based on the methanol-free PDH promoter

Bernat-Camps

Ebner²,

Schusterbauer³

et al. 2023

Front. Bioeng. Biotechnol.

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The current transition towards the circular bioeconomy requires a rational development of biorefineries to sustainably fulfill the present demands. The use of Komagataella phaffii (Pichia pastoris) can meet this challenge, since it has the capability to use crude glycerol as a carbon-source, a by-product from the biodiesel industry, while producing high- and low-added value products. Recombinant protein production (RPP) using K. phaffii has often been driven either by the methanol induced AOX1 promoter (PAOX1) and/or the constitutive GAP promoter (PGAP). In the last years, strong efforts have been focused on developing novel expression systems that expand the toolbox variety of K. phaffii to efficiently produce diverse proteins that requires different strategies. In this work, a study was conducted towards the development of methanol-free expression system based on a heat-shock gene promoter (PDH) using glycerol as sole carbon source. Using this promoter, the recombinant expression is strongly induced in carbon-starving conditions. The classical PGAP was used as a benchmark, taking for both strains the lipase B from Candida antarctica (CalB) as model protein. Titer of CalB expressed under PDH outperformed PGAP controlled expression in shake-flask cultivations when using a slow-release continuous feeding technology, confirming that PDH is induced under pseudo-starving conditions. This increase was also confirmed in fed-batch cultivations. Several optimization rounds were carried out for PDH under different feeding and osmolarity conditions. In all of them the PDH controlled process outperformed the PGAP one in regard to CalB titer. The best PDH approach reached 3.6-fold more specific productivity than PGAP fed-batch at low μ. Compared to the optimum approach for PGAP-based process, the best PDH fed-batch strategy resulted in 2.3-fold higher titer, while the specific productivity was very similar. To summarize, PDH is an inducible promoter that exhibited a non-coupled growth regulation showing high performance, which provides a methanol-free additional solution to the usual growth-coupled systems for RPP. Thus, this novel system emerges as a potential alternative for K. phaffii RPP bioprocess and for revaluing crude glycerol, promoting the transition towards a circular economy.

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Section: Resultssupporting

confidence: 66%

Enabling growth-decoupled Komagataella phaffii recombinant protein production based on the methanol-free PDH promoter

Bernat-Camps

Ebner²,

Schusterbauer³

et al. 2023

Front. Bioeng. Biotechnol.

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show abstract

“…This could have increased the availability of aldehyde for conversion to ALKs. Interestingly, the OD 600 of the cultures after 96 h was higher when glucose was provided by batch feeding (OD 600 = 14.5) than single feeding (OD 600 = 10.1), suggesting that the Crabtree effect could have been reduced by batch feeding, which resulted in increased biomass and ALK production ( Halka et al, 2018 ). Although the ALK titer achieved with BYΔ6OYGA/pAT-ALK by continuous glucose feeding was slightly lower (85.8%) than the highest ALK titer produced in BYΔ6OYGA/pGT-ALK under galactose-dependent condition (1,540 μg/L, Figure 5A , Condition II), it may be improved by optimizing the glucose concentration and feeding strategy during the fed-batch cultivation.…”

Section: Resultsmentioning

confidence: 99%

Engineering an Alcohol-Forming Fatty Acyl-CoA Reductase for Aldehyde and Hydrocarbon Biosynthesis in Saccharomyces cerevisiae

Foo

Rasouliha

Susanto

et al. 2020

Front. Bioeng. Biotechnol.

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Aldehydes are a class of highly versatile chemicals that can undergo a wide range of chemical reactions and are in high demand as starting materials for chemical manufacturing. Biologically, fatty aldehydes can be produced from fatty acyl-CoA by the action of fatty acyl-CoA reductases. The aldehydes produced can be further converted enzymatically to other valuable derivatives. Thus, metabolic engineering of microorganisms for biosynthesizing aldehydes and their derivatives could provide an economical and sustainable platform for key aldehyde precursor production and subsequent conversion to various value-added chemicals. Saccharomyces cerevisiae is an excellent host for this purpose because it is a robust organism that has been used extensively for industrial biochemical production. However, fatty acyl-CoA-dependent aldehyde-forming enzymes expressed in S. cerevisiae thus far have extremely low activities, hence limiting direct utilization of fatty acyl-CoA as substrate for aldehyde biosynthesis. Toward overcoming this challenge, we successfully engineered an alcohol-forming fatty acyl-CoA reductase for aldehyde production through rational design. We further improved aldehyde production through strain engineering by deleting competing pathways and increasing substrate availability. Subsequently, we demonstrated alkane and alkene production as one of the many possible applications of the aldehyde-producing strain. Overall, by protein engineering of a fatty acyl-CoA reductase to alter its activity and metabolic engineering of S. cerevisiae , we generated strains with the highest reported cytosolic aliphatic aldehyde and alkane/alkene production to date in S. cerevisiae from fatty acyl-CoA.

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“…Various microorganisms are employed in fermentation processes to convert sugars to glucose. Yeasts, such as Saccharomyces cerevisiae, are commonly used for ethanol production (Halka et al, 2018). They possess the ability to metabolize glucose through the glycolysis pathway, producing ethanol as the primary end product.…”

Section: Fermentation Of Sugars For Glucose Productionmentioning

confidence: 99%

“…The determination of reaction rates is crucial for understanding the kinetics of glucose production. Experimental techniques such as batch assays, continuous flow reactors, or spectroscopic methods can be employed to measure the rates of glucose release or consumption (Halka et al, 2018). By varying process parameters such as enzyme or acid concentration, temperature, or reaction time, the rates can be determined and used to estimate reaction rate constants and activation energies.…”

Section: Kinetics Of Glucose Productionmentioning

confidence: 99%

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Renewable Carbohydrates: Advancements in Sustainable Glucose Production and Optimization

Mperiju,

Silas,

Aji

et al. 2023

GSSR

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This study explores and optimizes glucose production through various biochemical processes and assesses the potential of diverse feedstock sources to meet the growing demand for renewable carbohydrates. It focuses on glucose production's significance in biological systems and industrial applications, analyzing pathways like enzymatic hydrolysis of polysaccharides and acid hydrolysis of biomass. The kinetics of glucose production are examined, encompassing kinetic models for enzymatic hydrolysis, acid hydrolysis, and fermentation processes. Factors influencing reaction kinetics are explored, and experimental techniques for kinetic parameter estimation are discussed. To address sustainability and resource utilization challenges, the study investigates locally sourced materials like agricultural residues, forest biomass, algal biomass, and food waste as renewable feedstock sources. Optimization strategies for glucose production are presented, using statistical design of experiments and response surface methodology. Techno-economic analysis and life cycle assessments provide a holistic evaluation of environmental and economic aspects associated with glucose production processes. The study's comprehensive approach to glucose production, encompassing both technological advancements and sustainability considerations, offers insights into enzymatic, acid hydrolysis, and fermentation processes, as well as comparing diverse feedstock sources. This knowledge can foster further advancements in the field, benefit industries, and encourage policymakers to promote the integration of renewable carbohydrates in the broader bioeconomy. The research contributes to the global shift towards a greener and more sustainable future, where glucose production plays a key role in building a resilient and eco-conscious society.

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Glucose limited feed strategy leads to increased production of fusicocca-2,10(14)-diene by Saccharomyces cerevisiae

Cited by 5 publications

References 18 publications

Enabling growth-decoupled Komagataella phaffii recombinant protein production based on the methanol-free PDH promoter

Enabling growth-decoupled Komagataella phaffii recombinant protein production based on the methanol-free PDH promoter

Engineering an Alcohol-Forming Fatty Acyl-CoA Reductase for Aldehyde and Hydrocarbon Biosynthesis in Saccharomyces cerevisiae

Renewable Carbohydrates: Advancements in Sustainable Glucose Production and Optimization

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