Genomics Characterization of an Engineered Corynebacterium glutamicum in Bioreactor Cultivation Under Ionic Liquid Stress

Corynebacterium glutamicum is an ideal microbial chassis for production of valuable bioproducts including amino acids and next generation biofuels. Here we resequence engineered isopentenol (IP) producing C. glutamicum BRC-JBEI 1.1.2 strain and assess differential transcriptional profiles using RNA sequencing under industrially relevant conditions including scale transition and compare the presence vs absence of an ionic liquid, cholinium lysinate ([Ch][Lys]). Analysis of the scale transition from shake flask to bioreactor with transcriptomics identified a distinct pattern of metabolic and regulatory responses needed for growth in this industrial format. These differential changes in gene expression corroborate altered accumulation of organic acids and bioproducts, including succinate, acetate, and acetoin that occur when cells are grown in the presence of 50 mM [Ch][Lys] in the stirred-tank reactor. This new genome assembly and differential expression analysis of cells grown in a stirred tank bioreactor clarify the cell response of an C. glutamicum strain engineered to produce IP.

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Genomics Characterization of an engineered Corynebacterium glutamicum in Bioreactor Cultivation under Ionic Liquid Stress

10.1101/2021.09.29.462453 ◽

2021 ◽

Author(s):

Deepanwita Banerjee ◽

Thomas Eng ◽

Yusuke Sasaki ◽

Aparajitha Srinivasan ◽

Asun Oka ◽

...

Keyword(s):

Ionic Liquid ◽

Corynebacterium Glutamicum ◽

Shake Flask ◽

Differential Expression Analysis ◽

Distinct Pattern ◽

Stirred Tank ◽

Bioreactor Cultivation ◽

Stirred Tank Bioreactor ◽

Scale Transition

AbstractCorynebacterium glutamicum is an ideal microbial chassis for the production of valuable bioproducts including amino acids and next-generation biofuels. Here we resequence engineered isopentenol (IP) producing C. glutamicum BRC-JBEI 1.1.2 strain and assess differential transcriptional profiles using RNA sequencing under industrially relevant conditions including scale transition and compare the presence vs. absence of an ionic liquid, cholinium lysinate ([Ch][Lys]). Analysis of the scale transition from shake flask to bioreactor with transcriptomics identified a distinct pattern of metabolic and regulatory responses needed for growth in this industrial format. These differential changes in gene expression corroborate altered accumulation of organic acids and bioproducts, including succinate, acetate, and acetoin that occur when cells are grown in the presence of 50mM [Ch][Lys] in the stirred-tank reactor. This new genome assembly and differential expression analysis of cells grown in a stirred tank bioreactor clarify the cell response of a C. glutamicum strain engineered to produce IP.

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Shake-flask and bench-scale stirred tank bioreactor production optimization of a thermoalkaline protease fromBacillus cereusSIU1 using one-factor-at-a-time and response surface (statistical) methodologies

Biocatalysis and Biotransformation ◽

10.3109/10242422.2014.913582 ◽

2014 ◽

Vol 32 (3) ◽

pp. 173-187 ◽

Cited By ~ 4

Author(s):

Sanjay Kumar Singh ◽

Satyendra Kumar Garg

Keyword(s):

Response Surface ◽

Shake Flask ◽

Production Optimization ◽

Stirred Tank ◽

Stirred Tank Bioreactor ◽

Bench Scale

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Kinetics and modelling of batch fermentation for the production of organic solvent tolerant and thermostable lipase by recombinant E. coli / Organik çözücü toleranslı ve ısıya dayanıklı rekombinan E. coli lipaz üretiminin kinetiği ve grup fermentasyonu modellemesi

Turkish Journal of Biochemistry ◽

10.1515/tjb-2015-0017 ◽

2015 ◽

Vol 40 (4) ◽

Author(s):

Arbakariya Bin Ariff ◽

Rubina Nelofer ◽

Raja Nor Zaliha Raja Abdul Rahman ◽

Mahiran Basri

Keyword(s):

Cell Growth ◽

Shake Flask ◽

Lipase Production ◽

Stirred Tank ◽

Thermostable Lipase ◽

Stirred Tank Bioreactor ◽

Monod Equation ◽

E Coli ◽

Organic Solvent Tolerant ◽

Solvent Tolerant

AbstractObjective: Kinetics of organic solvent tolerant and thermostable lipase production by recombinant E. coli in shake flask level and 2 L stirred tank bioreactor level was studied to observe the variations of important kinetic parameters at two different levels of bioprocess.Methods: Unstructured models based on Monod equation for growth and Luedeking-Piret equation for lipase production and glucose consumption were used to predict cell growth, lipase production and glucose utilization. The shake flask fermentation experiments were carried out at different initial glucose and yeast extract concentrations using recombinant bacterial strain E. coli BL21. Lipase production was also carried out using 2L stirred tank bioreactor for comparison.Results: In all cases, the data fitted well to the proposed models. The highest growth and lipase activity were obtained at 25 g/L glucose and 25 g/L yeast extract. Cell growth (6.42 g/L) and lipase production (65.32 IU/mL) in 2 L stirred tank bioreactor was comparable to those obtained in shake flask fermentations. The calculated value of growth associated constant (9.874 IU/g/h) was much higher than that of non-growth associated constant (0.022 IU/g/h) in bioreactor as well as in shake flasks. The values of maximum specific growth rate (μm) and glucose saturation constant (KS) for shake flask fermentations, calculated from Monod equation, were 0.476 h-1 and 5.237 g/L respectively.Conclusion: From the modelling exercise, it was concluded that the lipase production is dominantly growth associated process. The kinetic parameter values for fermentations in shake flask and 2L stirred tank bioreactor were comparable, indicating that the bioprocess could be transferred into larger scale.

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