scholarly journals Optimization of a Recombinant Lectin Production in Pichia pastoris Using Crude Glycerol in a Fed-Batch System

Processes ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 876
Author(s):  
Dulce Palmerín-Carreño ◽  
Dania Martínez-Alarcón ◽  
José Luis Dena-Beltrán ◽  
Lineth Juliana Vega-Rojas ◽  
Alejandro Blanco-Labra ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Crude Glycerol ◽  
Reaction Rates ◽  
Biodiesel Production ◽  
Heterologous Proteins ◽  
Fed Batch ◽  
Stirred Tank Bioreactor ◽  
Pure Glycerol ◽  
Area Of Interest ◽  
Batch System

The production of heterologous proteins for medical use is an important area of interest. The optimization of the bioprocesses includes the improvement of time, costs, and unit operations. Our study shows that a lectin fraction from Tepary bean (Phaseolus acutifolius) (TBLF) has cytotoxic effects on colon cancer cells and in vivo antitumorigenic activity. However, the low-yield, time-consuming, and expensive process made us focus on the development of a strategy to obtain a recombinant lectin using engineered Pichia pastoris yeast. Pure glycerol is one of the most expensive supplies; therefore, we worked on process optimization using crude glycerol from biodiesel production. Recombinant lectin (rTBL-1) production and purification were evaluated for the first time by an experimental design where crude glycerol (G65) was used and compared against pure glycerol (G99) in a controlled stirred-tank bioreactor with a fed-batch system. The recombinant lectin was detected and identified by SDS-PAGE, Western blot, and UHPLC–ESI–QTOF/MS analysis. The results show that the recombinant lectin can be produced from G65 with no significant differences with respect to G99: the reaction rates were 2.04 and 1.43 mg L−1 h−1, and the yields were 264.95 and 274.67 mgL−1, respectively. The current low cost of crude glycerol and our results show the possibility of producing heterologous proteins using this substrate with high productivity.

Pichia pastoris fermentation for phytase production using crude glycerol from biodiesel production as the sole carbon source

2009 ◽  
Vol 43 (2) ◽  
pp. 157-162 ◽  
Author(s):  
Shuiquan Tang ◽  
Lindsay Boehme ◽  
Howard Lam ◽  
Zisheng Zhang
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Sole Carbon Source ◽  
Crude Glycerol ◽  
Biodiesel Production ◽  
Phytase Production

Production of recombinant lipase B from Candida antarctica in Pichia pastoris under control of the promoter PGK using crude glycerol from biodiesel production as carbon source

2017 ◽  
Vol 118 ◽  
pp. 123-131 ◽  
Author(s):  
Julia Macedo Robert ◽  
Fernanda Senna Lattari ◽  
Antonio Carlos Machado ◽  
Aline Machado de Castro ◽  
Rodrigo Volcan Almeida ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Carbon Source ◽  
Crude Glycerol ◽  
Candida Antarctica ◽  
Biodiesel Production ◽  
Lipase B ◽  
Recombinant Lipase

scholarly journals Comprehensive clone screening and evaluation of fed-batch strategies in a microbioreactor and lab scale stirred tank bioreactor system: application on Pichia pastoris producing Rhizopus oryzae lipase

2014 ◽  
Vol 13 (1) ◽  
pp. 36 ◽  
Author(s):  
Johannes Hemmerich ◽  
Núria Adelantado ◽  
José Barrigón ◽  
Xavier Ponte ◽  
Astrid Hörmann ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
System Application ◽  
Rhizopus Oryzae ◽  
Stirred Tank ◽  
Fed Batch ◽  
Stirred Tank Bioreactor ◽  
Rhizopus Oryzae Lipase ◽  
Bioreactor System

scholarly journals Influence of trace elements supplementation on the production of recombinant frutalin by Pichia pastoris KM71H in fed-batch process

2013 ◽  
Vol 67 (7) ◽  
Author(s):  
Marcela Wanderley ◽  
Carla Oliveira ◽  
Danyelly Bruneska ◽  
Lucília Domingues ◽  
José Lima Filho ◽  
...  
Keyword(s):  
Trace Elements ◽  
Pichia Pastoris ◽  
Recombinant Protein ◽  
Protein Production ◽  
Batch Process ◽  
Fermentation Medium ◽  
Fed Batch ◽  
Stirred Tank Bioreactor ◽  
Recombinant Pichia Pastoris ◽  
Fed Batch Fermentation

AbstractFrutalin, a galactose-specific lectin used to detect specific tumour markers, is a protein with low expression level in breadfruit. In the present study, fed-batch fermentation in a stirred tank bioreactor was used as a strategy to enhance protein production by a recombinant Pichia pastoris KM71H. By using this process, the production of recombinant frutalin was 4-fold higher than the value obtained in shaker flasks batch assays. Supplementation of the fermentation medium with trace elements (Pichia trace minerals, PTM) was also evaluated in order to stimulate production of the recombinant protein. The addition of PTM to the minimum medium afforded a recombinant protein production of 13.4 mg L−1, which was 2.5-fold higher than that achieved from the culture medium without PTM supplementation. These results are significant as the development of strategies to improve the production of recombinant frutalin may broaden its application in cancer diagnosis.

scholarly journals Improved production of human type II procollagen in the yeast Pichia pastoris in shake flasks by a wireless-controlled fed-batch system

2008 ◽  
Vol 8 (1) ◽  
pp. 33 ◽  
Author(s):  
Maria Ruottinen ◽  
Monika Bollok ◽  
Martin Kogler ◽  
Antje Neubauer ◽  
Mirja Krause ◽  
...  
Keyword(s):  
Pichia Pastoris ◽  
Type Ii ◽  
Fed Batch ◽  
Yeast Pichia Pastoris ◽  
Human Type ◽  
Batch System ◽  
Shake Flasks

scholarly journals Archaeal Production of Polyhydroxyalkanoate (PHA) Co- and Terpolyesters from Biodiesel Industry-Derived By-Products

Archaea ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Carmen Hermann-Krauss ◽  
Martin Koller ◽  
Alexander Muhr ◽  
Hubert Fasl ◽  
Franz Stelzer ◽  
...  
Keyword(s):  
Crude Glycerol ◽  
Molar Mass ◽  
Molar Fraction ◽  
Volumetric Productivity ◽  
Biodiesel Production ◽  
Haloferax Mediterranei ◽  
Average Molar Mass ◽  
Melting Temperatures ◽  
Pure Glycerol ◽  
By Products

The archaeonHaloferax mediterraneiwas selected for production of PHA co- and terpolyesters using inexpensive crude glycerol phase (CGP) from biodiesel production as carbon source. CGP was assessed by comparison with the application of pure glycerol. Applying pure glycerol, a copolyester with a molar fraction of 3-hydroxybutyrate (3HB) of 0.90 mol/mol and 3-hydroxyvalerate (3HV) of 0.10 mol/mol, was produced at a volumetric productivity of 0.12 g/Lh and an intracellular PHA content of 75.4 wt.-% in the sum of biomass protein plus PHA. Application of CGP resulted in the same polyester composition and volumetric productivity, indicating the feasibility of applying CGP as feedstock. Analysis of molar mass distribution revealed a weight average molar massMwof 150 kDa and polydispersityPiof 2.1 for pure glycerol and 253 kDa and 2.7 for CGP, respectively; melting temperatures ranged between 130 and 140°C in both setups. Supplyingγ-butyrolactone as 4-hydroxybutyrate (4HB) precursor resulted in a poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyvalerate-co-4-hydroxybutyrate] (PHBHV4HB) terpolyester containing 3HV (0.12 mol/mol) and 4HB (0.05 mol/mol) in the poly[(R)-3-hydroxybutyrate] (PHB) matrix; in addition, this process runs without sterilization of the bioreactor. The terpolyester displayed reduced melting (melting endotherms at 122 and 137°C) and glass transition temperature (2.5°C), increased molar mass (391 kDa), and a polydispersity similar to the copolyesters.

1,3-Propanediol Fermentation with the by-Product Glycerol from Biodiesel Production by a Genetic Modified Klebsiella pneumoniae

2012 ◽  
Vol 512-515 ◽  
pp. 323-329 ◽  
Author(s):  
Hong Juan Liu ◽  
Yu Jie Zhou ◽  
Zhong Zhen Cai ◽  
Yan Sun ◽  
Jian An Zhang ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Acid Concentration ◽  
Batch Fermentation ◽  
Crude Glycerol ◽  
Acid Synthesis ◽  
Biodiesel Production ◽  
Klebsiella Pneumonia ◽  
Lactic Acid Concentration ◽  
Fed Batch ◽  
Fed Batch Fermentation

Nowadays, the utilization of the by-product glycerol has become a common problem with the large amount of biodiesel production. This paper studied the conversion of the by-product glycerol of biodiesel production by Klebsiella pneumonia to 1,3-propanediol. The crude glycerol could be used directly without refining by a lactate dehydrogenase deficient K. pneumoniae and the lactic acid concentration was very low. In the fed-batch fermentation of 7L fermenter, 93.3g/l 1,3-propanediol was obtained and lactic acid concentration was just 2g/l. Analysis showed that D-type lactic acid synthesis was reduced obviously. The fermentation also was scaled up on the 42L fermenter. 1,3-propanediol and 2,3-Butanediol concentration reached 81.5g/l and 33.8g/l, respectively. No lactic acid was detected. The demonstration fermentation in 5000L fermenter was also successfully performed. The final 1,3-propanediol concentration and productivity was 87g/l and 1.2g/(lh). This work was considered useful to the integrated production of biodiesel and 1,3-propanediol.

scholarly journals Production of 1,3-Propanediol from Pure and Crude Glycerol Using Immobilized Clostridium butyricum

Catalysts ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 317 ◽  
Author(s):  
Igor Dolejš ◽  
Monika Líšková ◽  
Vladimír Krasňan ◽  
Kristína Markošová ◽  
Michal Rosenberg ◽  
...  
Keyword(s):  
Crude Glycerol ◽  
Value Added ◽  
Final Concentration ◽  
Clostridium Butyricum ◽  
Biodiesel Production ◽  
Glycerol Concentration ◽  
Pure Glycerol ◽  
Average Productivity ◽  
Used Cooking Oil ◽  
Repeated Batch

The present study describes the production of the value-added chemical 1,3-propanediol (1,3-PD) from crude glycerol, a waste by-product formed during biodiesel production. The efficiency, robustness, and stability of the process were improved by immobilization of the anaerobic bacterium Clostridium butyricum into a polyvinyl alcohol (PVA) hydrogel. The highest average productivity, 6.8 ± 0.2 g/(L·h), was achieved in 10 consecutive, repeated batch fermentations, with an initial concentration of pure glycerol 45.5 ± 0.7 g/L, after 2.5 hours. The highest final concentration and yield of 1,3-PD, 28.3 ± 0.6 g/L, and 0.42 ± 0.01 g/g, respectively, were achieved in eleven repeated batch fermentations, after increasing the initial pure glycerol concentration to 70.4 ± 1.9 g/L. Two different types of crude glycerol, produced from used cooking oil (UCO) and rapeseed oil (RO), were tested in repeated batch fermentations, with an average productivity achieved of 2.3 ± 0.1 and 3.5 ± 0.3 g/(L·h), respectively. The highest final concentration and yield of 1,3-PD, 12.6 ± 0.9 g/L, and 0.35 ± 0.02 g/g, respectively, were observed in fifteen repeated batch fermentations with RO crude glycerol. An excellent stability of the immobilized anaerobic bacteria and increase of productivity in fermentation of crude glycerol was demonstrated.

scholarly journals Biodiesel-Derived Glycerol Obtained from Renewable Biomass—A Suitable Substrate for the Growth of Candida zeylanoides Yeast Strain ATCC 20367

2019 ◽  
Vol 7 (8) ◽  
pp. 265 ◽  
Author(s):  
Laura Mitrea ◽  
Floricuța Ranga ◽  
Florinela Fetea ◽  
Francisc Vasile Dulf ◽  
Alexandru Rusu ◽  
...  
Keyword(s):  
Crude Glycerol ◽  
Carbon Sources ◽  
Biodiesel Production ◽  
Bioreactor Cultivation ◽  
Renewable Biomass ◽  
Pure Glycerol ◽  
Cultivation Process ◽  
By Products ◽  
Candida Zeylanoides ◽  
Main Carbon

Used kitchen oil represents a feasible and renewable biomass to produce green biofuels such as biodiesel. Biodiesel production generates large amounts of by-products such as the crude glycerol fraction, which can be further used biotechnologically as a valuable nutrient for many microorganisms. In this study, we transesterified used kitchen oil with methanol and sodium hydroxide in order to obtain biodiesel and crude glycerol fractions. The crude glycerol fraction consisting of 30% glycerol was integrated into a bioreactor cultivation process as a nutrient source for the growth of Candida zeylanoides ATCC 20367. Cell viability and biomass production were similar to those obtained with batch cultivations on pure glycerol or glucose as the main nutrient substrates. However, the biosynthesis of organic acids (e.g., citric and succinic) was significantly different compared to pure glycerol and glucose used as main carbon sources.

Technologies for Processing of Crude Glycerol from Biodiesel Production: Synthesis of Solketal and Its Hydrolysis to Obtain Pure Glycerol

2018 ◽  
Vol 91 (9) ◽  
pp. 1478-1485 ◽  
Author(s):  
G. S. Dmitriev ◽  
L. N. Zanaveskin ◽  
A. V. Terekhov ◽  
V. O. Samoilov ◽  
I. A. Kozlovskii ◽  
...  
Keyword(s):  
Crude Glycerol ◽  
Biodiesel Production ◽  
Pure Glycerol
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