Combinatorial approach for improved production of whole-cell 3-aminopropionic acid in recombinant Bacillus megaterium: codon optimization, gene duplication and process optimization

3 Biotech ◽  
2021 ◽  
Vol 11 (7) ◽  
Author(s):  
Subbi Rami Reddy Tadi ◽  
Ganesh Nehru ◽  
Senthilkumar Sivaprakasam
Keyword(s):  
Gene Duplication ◽  
Process Optimization ◽  
Bacillus Megaterium ◽  
Codon Optimization ◽  
Combinatorial Approach ◽  
Whole Cell

GM3 trisaccharide biosynthesis and process optimization using engineered E. coli lysate and whole-cell catalysis

2020 ◽  
Vol 39 (5-6) ◽  
pp. 217-231
Author(s):  
Lipeng Feng ◽  
Jie Shi ◽  
Haofei Hong ◽  
Zhifang Zhou ◽  
Zhimeng Wu
Keyword(s):  
Process Optimization ◽  
Whole Cell ◽  
E Coli ◽  
Whole Cell Catalysis

Detection of mRNA of nprM in Bacillus megaterium ATCC 14581 grown in soil by whole-cell hybridization

1995 ◽  
Vol 163 (4) ◽  
pp. 235-241 ◽  
Author(s):  
Wolfgang H�nerlage ◽  
Dittmar Hahn ◽  
Josef Zeyer
Keyword(s):  
Bacillus Megaterium ◽  
Cell Hybridization ◽  
Whole Cell

Effect of passage through the gut of the earthworm Lumbricus terrestris L. on Bacillus megaterium studied by whole cell hydridization

1997 ◽  
Vol 29 (7) ◽  
pp. 1149-1152 ◽  
Author(s):  
Kathrin Fischer ◽  
Dittmar Hahn ◽  
Wolfgang Hönerlage ◽  
Josef Zeyer
Keyword(s):  
Bacillus Megaterium ◽  
Lumbricus Terrestris ◽  
Whole Cell ◽  
Earthworm Lumbricus

scholarly journals Process optimization of antibody fragment-secreting Bacillus megaterium cultivations and its effect on the cell heterogeneity

2009 ◽  
Vol 25 ◽  
pp. S217
Author(s):  
F. David ◽  
C. Korneli ◽  
M. Schön ◽  
R. Westphal ◽  
E. Franco-Lara
Keyword(s):  
Process Optimization ◽  
Bacillus Megaterium ◽  
Antibody Fragment ◽  
Cell Heterogeneity

In-Cell Crosslinked Enzymes technique: Improving Bacillus megaterium whole-cell biocatalyst stability for the decarboxylation of ferulic acid

2021 ◽  
Author(s):  
Victor Contreras-Jácquez ◽  
Daniel A. Grajales-Hernández ◽  
Mariana Armendáriz-Ruiz ◽  
Jorge Rodríguez-González ◽  
Elisa M. Valenzuela-Soto ◽  
...  
Keyword(s):  
Ferulic Acid ◽  
Bacillus Megaterium ◽  
Whole Cell ◽  
Whole Cell Biocatalyst

CYP105A1 mediated 3-hydroxylation of glimepiride and glibenclamide using a recombinant Bacillus megaterium whole-cell catalyst

2012 ◽  
Vol 157 (3) ◽  
pp. 405-412 ◽  
Author(s):  
Michael Kleser ◽  
Frank Hannemann ◽  
Michael Hutter ◽  
Josef Zapp ◽  
Rita Bernhardt
Keyword(s):  
Bacillus Megaterium ◽  
Whole Cell

scholarly journals An effective combination of codon optimization, gene dosage, and process optimization for high-level production of fibrinolytic enzyme in Komagataella phaffii (Pichia pastoris)

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhiqun Che ◽  
Xiaoyan Cao ◽  
Guiguang Chen ◽  
Zhiqun Liang
Keyword(s):  
Process Optimization ◽  
Copy Number ◽  
Codon Optimization ◽  
Gene Dosage ◽  
Fibrinolytic Enzyme ◽  
Efficient Production ◽  
Combination Strategy ◽  
Thrombolytic Agents ◽  
Komagataella Phaffii

Abstract Background As a main drug for diseased thrombus, some clinically used thrombolytic agents have various disadvantages, safer novel thrombolytic agents are of great demand. This study aimed to achieve high and efficient production of a fibrinolytic enzyme with superior enzymatic properties, by a combination strategy of codon optimization, gene dosage and process optimization in Komagataella phaffii (K. phaffii). Results After codon optimization, the fibase from a marine Bacillus subtilis was expressed and secreted in K. phaffii GS115. Recombinant strains harboring different copies of the fib gene (fib-nc) were successfully obtained via Geneticin (0.25–4 mg/ml) screening on minimal dextrose selection plates and assessment via real-time quantitative PCR. The respective levels of fibase produced by strains expressing fib-5.4c, fib-6c, fib-8c, fib-9c, and fib-12c were 4428, 5781, 7323, 7930, and 2472 U/ml. Levels increased as the copy number increased from 4 to 9, but decreased dramatically at copy number 12. After high cell density fermentation optimization, the highest fibase activity of the strain expressing fib-9c was 7930 U/ml in a shake flask and increased to 12,690 U/ml after 3 days of continuous culture in a 5-L fermenter, which is one of the highest levels of production reported. The recombinant fibase was maximally active at pH 9.0 and 45 °C, and was remarkably stable at pH levels ranging from 5 to 10 and temperatures up to 50 °C. As a metal-dependent serine protease, fibase did not cause hemolysis in vitro and preferentially degraded fibrin directly. Conclusions The combination of codon optimization, gene dosage, and process optimization described herein could be used for the expression of other therapeutic proteins difficult to express. The characteristics of the recombinant fibase suggest that it has potential applications for thrombosis prevention and therapy.

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