Towards closed carbon loop fermentations: co-feeding of Yarrowia lipolytica with glucose and formic acid

A novel fermentation process was developed in which renewable electricity is indirectly used as a fermentation substrate, synergistically decreasing both the consumption of sugar as a first generation carbon source and emission of the greenhouse gas CO2. To achieve this, a glucose-based process is co-fed with formic acid, which can be generated by capturing CO2 from fermentation offgas followed by electrochemical reduction with renewable electricity. This ‘closed carbon loop’ concept is demonstrated by a case study in which co-feeding formic acid is shown to significantly increase the yield of biomass on glucose of the industrially relevant yeast species Yarrowia lipolytica. First, the optimal feed ratio of formic acid to glucose is established using chemostat cultivations. Subsequently, guided by a dynamic fermentation process model, a fed-batch protocol is developed and demonstrated on laboratory scale. Finally, the developed fed-batch process is proven to be scalable to pilot scale. An extension of this proven concept to also recycle the O2 that is co-generated with the formic acid to the fermentation process for intensification purposes, and a potential further application of the concept to anaerobic fermentations are discussed.

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Co-production of polyhydroxybutyrate (PHB) and coenzyme Q10 (CoQ10) via no-sugar fermentation—a case by Methylobacterium sp. XJLW

Annals of Microbiology ◽

10.1186/s13213-021-01632-w ◽

2021 ◽

Vol 71 (1) ◽

Author(s):

Peiwu Cui ◽

Yunhai Shao ◽

Yanxin Wang ◽

Rui Zhao ◽

Huihui Zhan ◽

...

Keyword(s):

Mass Spectrometry ◽

Energy Source ◽

Fermentation Process ◽

Genetically Modify ◽

Tween 80 ◽

Batch Process ◽

Gas Chromatography Mass Spectrometry ◽

Fed Batch ◽

Enhanced Production ◽

Chromatography Mass Spectrometry

Abstract Purpose To explore a competitive PHB-producing fermentation process, this study evaluated the potential for Methylobacterium sp. XJLW to produce simultaneously PHB and coenzyme Q10 (CoQ10) using methanol as sole carbon and energy source. Methods The metabolic pathways of PHB and CoQ10 biosynthesis in Methylobacterium sp. XJLW were first mined based on the genomic and comparative transcriptomics information. Then, real-time fluorescence quantitative PCR (RT-qPCR) was employed for comparing the expression level of important genes involved in PHB and CoQ10 synthesis pathways’ response to methanol and glucose. Transmission electron microscope (TEM), gas chromatography/mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), Fourier transformation infrared spectrum (FT-IR), and liquid chromatography/mass spectrometry (LC-MS) methods were used to elucidate the yield and structure of PHB and CoQ10, respectively. PHB and CoQ10 productivity of Methylobacterium sp. XJLW were evaluated in Erlenmeyer flask for medium optimization, and in a 5-L bioreactor for methanol fed-batch strategy according to dissolved oxygen (DO) and pH control. Results Comparative genomics analysis showed that the PHB and CoQ10 biosynthesis pathways co-exist in Methylobacterium sp. XJLW. Transcriptomics analysis showed that the transcription level of key genes in both pathways responding to methanol was significantly higher than that responding to glucose. Correspondingly, strain Methylobacterium sp. XJLW can produce PHB and CoQ10 simultaneously with higher yield using cheap and abundant methanol than using glucose as sole carbon and energy source. The isolated products showed the structure characteristics same to that of standard PHB and CoQ10. The optimal medium and cultural conditions for PHB and CoQ10 co-production by Methylobacterium sp. XJLW was in M3 medium containing 7.918 g L-1 methanol, 0.5 g L-1 of ammonium sulfate, 0.1% (v/v) of Tween 80, and 1.0 g L-1 of sodium chloride, under 30 °C and pH 7.0. In a 5-L bioreactor coupled with methanol fed-batch process, a maximum DCW value (46.31 g L-1) with the highest yields of PHB and CoQ10, reaching 6.94 g L-1 and 22.28 mg L-1, respectively. Conclusion Methylobacterium sp. XJLW is potential for efficiently co-producing PHB and CoQ10 employing methanol as sole carbon and energy source. However, it is still necessary to further optimize fermentation process, and genetically modify strain pathway, for enhanced production of PHB and CoQ10 simultaneously by Methylobacterium sp. XJLW. It also suggests a potential strategy to develop efficiently co-producing other high-value metabolites using methanol-based bioprocess.

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Co-production of biopolymers and quinone via no-sugar fermentation--a case by Methylobacterium sp. XJLM

10.21203/rs.3.rs-254045/v1 ◽

2021 ◽

Author(s):

Peiwu Cui ◽

Yunhai Shao ◽

Yanxin Wang ◽

Rui Zhao ◽

Huihui Zhan ◽

...

Keyword(s):

Mass Spectrometry ◽

Energy Source ◽

Fermentation Process ◽

Genetically Modify ◽

Tween 80 ◽

Batch Process ◽

Gas Chromatography Mass Spectrometry ◽

Fed Batch ◽

Enhanced Production ◽

Chromatography Mass Spectrometry

Abstract Purpose To explore a competitive PHB producing fermentation process, this study evaluated the potential for Methylobacterium sp. XJLW to produce simultaneously PHB and coenzyme Q 10 (CoQ 10 ) using cheap and abundant methanol as sole carbon and energy source. Methods The metabolic pathways of PHB and CoQ 10 biosynthesis in XJLW strain were first mined based on the genomic and comparative transcriptomics information. Then, Real-time fluorescence quantitative PCR (RT-qPCR) was employed for comparing the expression level of important genes involved in PHB and CoQ10 synthesis pathways response to methanol and glucose. Transmission electron microscope (TEM), gas chromatography/mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), Fourier transformation infrared spectrum (FT-IR), and liquid chromatography/mass spectrometry (LC-MS) methods, were used to elucidate the yield and structure of PHB and CoQ 10 , respectively. PHB and CoQ 10 productivity of XJLW strain were evaluated in flasks for medium optimization, and in a 5-L bioreactor for methanol fed-batch strategy according to dissolved oxygen (DO) and pH control. Results Comparative genomics and transcriptomics analysis showed that the PHB and CoQ 10 biosynthesis pathways coexist in XJLW strain, and the transcription level of key genes in both pathways response to methanol was significantly higher than that response to glucose. Correspondingly, strain XJLW can produce PHB and CoQ 10 simultaneously with higher yield using cheap and abundant methanol than using glucose as sole carbon and energy source. The isolated products showed the structure characteristics same to that of standard PHB and CoQ 10 . The optimal medium and cultural conditions for PHB and CoQ 10 co-production by XJLW strain was in M3 medium containing 1% (v/v) of methanol, 0.5 g/L of ammonium sulfate, 0.1% (v/v) of Tween 80, and 1.0 g/L of sodium chloride, under 30°C and pH 7.0. In a 5-L bioreactor coupled with methanol fed-batch process, a maximum DCW value (46.31 g/L) with the highest yields of PHB and CoQ 10 , reaching 6.94 g/L and 22.28 mg/L, respectively. Conclusion Methylobacterium sp. XJLW is potential for efficiently co-producing PHB and CoQ 10 employing methanol as sole carbon and energy source. However, it is still necessary to further optimize fermentation process, and genetically modify strain pathway, for enhanced production of PHB and CoQ 10 simultaneously by XJLW. It also suggests a potential strategy to develop efficiently co-producing other high value metabolites using methanol-based bio-process.

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Development of a Fed-Batch Process for the Production of a Recombinant Protein X in CHO-GS System – Case Study from the Cell to Reactor Process Ready for Pilot Scale Cultivation

Cells and Culture ◽

10.1007/978-90-481-3419-9_125 ◽

2010 ◽

pp. 723-725

Author(s):

Véronique Chotteau ◽

Yun Jiang ◽

Jeannette Westin ◽

Kerstin Dahlenborg ◽

Anna Sjöblom-Hallén ◽

...

Keyword(s):

Recombinant Protein ◽

Batch Process ◽

Pilot Scale ◽

Fed Batch ◽

Protein X

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Integrated L-phenylalanine separation in an E. coli fed-batch process: from laboratory to pilot scale

Bioprocess and Biosystems Engineering ◽

10.1007/s00449-002-0279-8 ◽

2002 ◽

Vol 25 (2) ◽

pp. 85-96 ◽

Cited By ~ 13

Author(s):

Maass D. ◽

Gerigk M. ◽

Kreutzer A. ◽

Weuster-Botz D. ◽

Wubbolts M. ◽

...

Keyword(s):

Batch Process ◽

Pilot Scale ◽

Fed Batch ◽

E Coli

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Implementation and process analysis of pilot scale multi-phase anaerobic fermentation and digestion of faecal sludge in Ghana

Gates Open Research ◽

10.12688/gatesopenres.12754.1 ◽

2017 ◽

Vol 1 ◽

pp. 10

Author(s):

Justin Shih ◽

Ato Fanyin-Martin ◽

Edris Taher ◽

Kartik Chandran

Keyword(s):

Process Model ◽

Fermentation Process ◽

Volatile Fatty Acids ◽

Anaerobic Fermentation ◽

Management Practice ◽

Process Analysis ◽

Pilot Scale ◽

Model Framework ◽

Capital Costs ◽

Faecal Sludge

Background. In Ghana, faecal sludge (FS) from on-site sanitation facilities is often discharged untreated into the environment, leading to significant insults to environmental and human health. Anaerobic digestion offers an attractive pathway for FS treatment with the concomitant production of energy in the form of methane. Another innovative option includes separating digestion into acidogenesis (production of volatile fatty acids (VFA)) and methanogenesis (production of methane), which could ultimately facilitate the production of an array of biofuels and biochemicals from the VFA. This work describes the development, implementation and modeling based analysis of a novel multiphase anaerobic fermentation-digestion process aimed at FS treatment in Kumasi, Ghana. Methods. A pilot-scale anaerobic fermentation process was implemented at the Kumasi Metropolitan Assembly’s Oti Sanitary Landfill Site at Adanse Dompoase. The process consisted of six 10 m3 reactors in series, which were inoculated with bovine rumen and fed with fecal sludge obtained from public toilets. The performance of the fermentation process was characterized in terms of both aqueous and gaseous variables representing the conversion of influent organic carbon to VFA as well as CH4. Using the operating data, the first-ever process model for FS fermentation and digestion was developed and calibrated, based on the activated sludge model framework. Results and Conclusions. This work represents one of the first systematic efforts at integrated FS characterization and process modeling to enable anaerobic fermentation and digestion of FS. It is shown that owing to pre-fermentation of FS in public septage holding tanks, one could employ significantly smaller digesters (lower capital costs) or increased loading capabilities for FS conversion to biogas or VFA. Further, using the first-ever calibrated process model for FS fermentation and digestion presented herein, we expect improved and more mechanistically informed development and application of different process designs and configurations for global FS management practice.

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Effect of addition of anaerobic fermented OFMSW (organic fraction of municipal solid waste) on biological nutrient removal (BNR) process: preliminary results

Water Science & Technology ◽

10.2166/wst.1998.0078 ◽

1998 ◽

Vol 38 (1) ◽

pp. 327-334 ◽

Cited By ~ 7

Author(s):

P. Pavan ◽

P. Battistoni ◽

P. Traverso ◽

A. Musacco ◽

F. Cecchi

Keyword(s):

Nutrient Removal ◽

Fermentation Process ◽

Organic Waste ◽

Anaerobic Fermentation ◽

Pilot Scale ◽

Biological Nutrient Removal ◽

Organic Fraction ◽

P Release ◽

Pure Methanol ◽

P Removal

The paper presents results coming from experiments on pilot scale plants about the possibility to integrate the organic waste and wastewater treatment cycles, using the light organic fraction produced via anaerobic fermentation of OFMSW as RBCOD source for BNR processes. The effluent from the anaerobic fermentation process, with an average content of 20 g/l of VFA+ lactic acid was added to wastewater to be treated in order to increase RBCOD content of about 60-70 mg/l. The results obtained in the BNR process through the addition of the effluent from the fermentation unit are presented. Significant increase of denitrification rate was obtained: 0.06 KgN-NO3/KgVSS d were denitrified in the best operative conditions studied. -Vmax shows values close to those typical of the pure methanol addition (about 0.3 KgN-NO3/KgVSS d). A considerable P release (35%) was observed in the anaerobic step of the BNR process, even if not yet a completely developed P removal process.

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Production of Rhamnolipid Biosurfactant from Fed Batch Culture by Pseudomonas aeruginosa using Multiple Substrates

Current Nutrition & Food Science ◽

10.2174/1573401314666181107100127 ◽

2020 ◽

Vol 16 (6) ◽

pp. 928-933

Author(s):

Jujjavarapu S. Eswari

Keyword(s):

Pseudomonas Aeruginosa ◽

Batch Process ◽

Time Interval ◽

Fed Batch ◽

Multiple Substrates ◽

Surface Active Agents ◽

Limiting Nutrients ◽

Limiting Nutrient ◽

Surface Active ◽

Rhamnolipid Biosurfactant

Objective: Biosurfactants are the surface active agents which are used for the reduction of surface and interfacial tensions of liquids. Rhamnolipids are the surfactants produced by Pseudomonas aeruginosa. It requires minimum nutrition for its growth as it can also grow in distilled water. The rhamnolipids produced by Pseudomonas aeruginosa are extra-cellular glycolipids consisting of L-rhamnose and 3-hydroxyalkanoic acid. Methods: The fed-batch method for the rhamnolipid production is considered in this study to know the influence of the carbon, nitrogen, phosphorous substrates as growth-limiting nutrients. Pulse feeding is employed for limiting nutrient addition at particular time interval to obtain maximum rhamnolipid formation from Pseudomonas aeruginosa compared with the batch process. Results: Out of 3 fed batch strategies constant glucose fed batch strategy shows best and gave maximum rhamnolipid concentration of 0.134 g/l.

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