Alginate Biosynthesis and Biotechnological Production

2017 ◽  
pp. 1-25 ◽  
Author(s):  
M. Fata Moradali ◽  
Shirin Ghods ◽  
Bernd H. A. Rehm
Keyword(s):  
Biotechnological Production ◽  
Alginate Biosynthesis

scholarly journals Expansion of EasyClone-MarkerFree toolkit for Saccharomyces cerevisiae genome with new integration sites

2021 ◽  
Author(s):  
Mahsa Babaei ◽  
Luisa Sartori ◽  
Alexey Karpukhin ◽  
Dmitrii Abashkin ◽  
Elena Matrosova ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Recombinant Dna ◽  
Green Fluorescence Protein ◽  
Cellular Growth ◽  
Green Fluorescence ◽  
Biotechnological Production ◽  
Yeast Saccharomyces Cerevisiae ◽  
Integration Sites ◽  
Fluorescence Protein ◽  
Integration Efficiency

Abstract Biotechnological production requires genetically stable recombinant strains. To ensure genomic stability, recombinant DNA is commonly integrated into the genome of the host strain. Multiple genetic tools have been developed for genomic integration into baker's yeast Saccharomyces cerevisiae. Previously, we had developed a vector toolkit EasyClone-MarkerFree for stable integration into eleven sites on chromosomes X, XI, and XII of S. cerevisiae. The markerless integration was enabled by CRISPR-Cas9 system. In this study, we have expanded the kit with eight additional intergenic integration sites located on different chromosomes. The integration efficiency into the new sites was above 80%. The expression level of green fluorescence protein (gfp) for all eight sites was similar or above XI-2 site from the original EasyClone-MarkerFree toolkit. The cellular growth was not affected by the integration into any of the new eight locations. The eight-vector expansion kit is available from AddGene.

scholarly journals Biotechnological production of ssDNA with DNA-hydrolyzing deoxyribozymes

2021 ◽  
Vol 2 (2) ◽  
pp. 100531
Author(s):  
Jin Liu ◽  
Hongzhou Gu
Keyword(s):  
Biotechnological Production

scholarly journals Sporulation in solventogenic and acetogenic clostridia

2021 ◽  
Author(s):  
Mamou Diallo ◽  
Servé W. M. Kengen ◽  
Ana M. López-Contreras
Keyword(s):  
Current Knowledge ◽  
Carbon Sources ◽  
Cost Effective ◽  
Biotechnological Production ◽  
Key Points ◽  
Wide Range ◽  
Potential Applications ◽  
A Cell ◽  
Sporulation Initiation ◽  
Solventogenic Clostridia

AbstractThe Clostridium genus harbors compelling organisms for biotechnological production processes; while acetogenic clostridia can fix C1-compounds to produce acetate and ethanol, solventogenic clostridia can utilize a wide range of carbon sources to produce commercially valuable carboxylic acids, alcohols, and ketones by fermentation. Despite their potential, the conversion by these bacteria of carbohydrates or C1 compounds to alcohols is not cost-effective enough to result in economically viable processes. Engineering solventogenic clostridia by impairing sporulation is one of the investigated approaches to improve solvent productivity. Sporulation is a cell differentiation process triggered in bacteria in response to exposure to environmental stressors. The generated spores are metabolically inactive but resistant to harsh conditions (UV, chemicals, heat, oxygen). In Firmicutes, sporulation has been mainly studied in bacilli and pathogenic clostridia, and our knowledge of sporulation in solvent-producing or acetogenic clostridia is limited. Still, sporulation is an integral part of the cellular physiology of clostridia; thus, understanding the regulation of sporulation and its connection to solvent production may give clues to improve the performance of solventogenic clostridia. This review aims to provide an overview of the triggers, characteristics, and regulatory mechanism of sporulation in solventogenic clostridia. Those are further compared to the current knowledge on sporulation in the industrially relevant acetogenic clostridia. Finally, the potential applications of spores for process improvement are discussed.Key Points• The regulatory network governing sporulation initiation varies in solventogenic clostridia.• Media composition and cell density are the main triggers of sporulation.• Spores can be used to improve the fermentation process.

Recent trends in the biotechnological production of tocopherols using in vitro cultures

2021 ◽  
Author(s):  
Lorena Almagro ◽  
Ana Belén Sabater-Jara ◽  
Sarai Belchí-Navarro ◽  
María Ángeles Pedreño
Keyword(s):  
In Vitro Cultures ◽  
Biotechnological Production ◽  
Recent Trends

scholarly journals Metabolic Engineering of Corynebacterium glutamicum for Methanol Metabolism

2015 ◽  
Vol 81 (6) ◽  
pp. 2215-2225 ◽  
Author(s):  
Sabrina Witthoff ◽  
Katja Schmitz ◽  
Sebastian Niedenführ ◽  
Katharina Nöh ◽  
Stephan Noack ◽  
...  
Keyword(s):  
Corynebacterium Glutamicum ◽  
Biotechnological Production ◽  
Methanol Metabolism ◽  
Initial Oxidation ◽  
Content Type ◽  
Oxidation Of Methanol ◽  
Bacillus Methanolicus ◽  
Production Processes ◽  
Starting Point ◽  
Limited Application

ABSTRACTMethanol is already an important carbon feedstock in the chemical industry, but it has found only limited application in biotechnological production processes. This can be mostly attributed to the inability of most microbial platform organisms to utilize methanol as a carbon and energy source. With the aim to turn methanol into a suitable feedstock for microbial production processes, we engineered the industrially important but nonmethylotrophic bacteriumCorynebacterium glutamicumtoward the utilization of methanol as an auxiliary carbon source in a sugar-based medium. Initial oxidation of methanol to formaldehyde was achieved by heterologous expression of a methanol dehydrogenase fromBacillus methanolicus, whereas assimilation of formaldehyde was realized by implementing the two key enzymes of the ribulose monophosphate pathway ofBacillus subtilis: 3-hexulose-6-phosphate synthase and 6-phospho-3-hexuloisomerase. The recombinantC. glutamicumstrain showed an average methanol consumption rate of 1.7 ± 0.3 mM/h (mean ± standard deviation) in a glucose-methanol medium, and the culture grew to a higher cell density than in medium without methanol. In addition, [13C]methanol-labeling experiments revealed labeling fractions of 3 to 10% in the m + 1 mass isotopomers of various intracellular metabolites. In the background of aC. glutamicumΔaldΔadhEmutant being strongly impaired in its ability to oxidize formaldehyde to CO2, the m + 1 labeling of these intermediates was increased (8 to 25%), pointing toward higher formaldehyde assimilation capabilities of this strain. The engineeredC. glutamicumstrains represent a promising starting point for the development of sugar-based biotechnological production processes using methanol as an auxiliary substrate.

scholarly journals Mating of 2 Laboratory Saccharomyces cerevisiae Strains Resulted in Enhanced Production of 2-Phenylethanol by Biotransformation of L-Phenylalanine

2017 ◽  
Vol 27 (2) ◽  
pp. 81-90 ◽  
Author(s):  
Jolanta Mierzejewska ◽  
Aleksandra Tymoszewska ◽  
Karolina Chreptowicz ◽  
Kamil Krol
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Batch Culture ◽  
Fold Increase ◽  
Biotechnological Production ◽  
Aromatic Alcohol ◽  
Enhanced Production ◽  
Yeast Strains ◽  
First Time

2-Phenylethanol (2-PE) is an aromatic alcohol with a rosy scent which is widely used in the food, fragrance, and cosmetic industries. Promising sources of natural 2-PE are microorganisms, especially yeasts, which can produce 2-PE by biosynthesis and biotransformation. Thus, the first challenging goal in the development of biotechnological production of 2-PE is searching for highly productive yeast strains. In the present work, 5 laboratory <i>Saccharomyces cerevisiae</i> strains were tested for the production of 2-PE. Thereafter, 2 of them were hybridized by a mating procedure and, as a result, a new diploid, <i>S. cerevisiae</i> AM1-d, was selected. Within the 72-h batch culture in a medium containing 5 g/L of <smlcap>L</smlcap>-phenylalanine, AM1-d produced 3.83 g/L of 2-PE in a shaking flask. In this way, we managed to select the diploid <i>S. cerevisiae</i> AM1-d strain, showing a 3- and 5-fold increase in 2-PE production in comparison to parental strains. Remarkably, the enhanced production of 2-PE by the hybrid of 2 yeast laboratory strains is demonstrated here for the first time.

Biotechnological Production of Taxol and Related Taxoids: Current State and Prospects

2009 ◽  
Vol 9 (1) ◽  
pp. 109-121 ◽  
Author(s):  
O. Exposito ◽  
M. Bonfill ◽  
E. Moyano ◽  
M. Onrubia ◽  
M. Mirjalili ◽  
...  
Keyword(s):  
Biotechnological Production ◽  
Current State

Expanding lysine industry: Biotechnological production of l-lysine and its derivatives

2021 ◽  
Author(s):  
Jiaping Wang ◽  
Cong Gao ◽  
Xiulai Chen ◽  
Liming Liu
Keyword(s):  
Biotechnological Production

scholarly journals The algT Gene of Pseudomonas syringae pv. glycinea andNew Insights into the Transcriptional Organization of thealgT-muc Gene Cluster

2006 ◽  
Vol 188 (23) ◽  
pp. 8013-8021 ◽  
Author(s):  
Alexander Schenk ◽  
Michael Berger ◽  
Lisa M. Keith ◽  
Carol L. Bender ◽  
Georgi Muskhelishvili ◽  
...  
Keyword(s):  
Gene Cluster ◽  
Pseudomonas Syringae ◽  
Azotobacter Vinelandii ◽  
Regulatory Gene ◽  
In Planta ◽  
Phytopathogenic Bacterium ◽  
Reverse Transcription Pcr ◽  
Alginate Production ◽  
Alginate Biosynthesis

ABSTRACT The phytopathogenic bacterium Pseudomonas syringae pv. glycinea infects soybean plants and causes bacterial blight. In addition to P. syringae, the human pathogen Pseudomonas aeruginosa and the soil bacterium Azotobacter vinelandii produce the exopolysaccharide alginate, a copolymer of d-mannuronic and l-guluronic acids. Alginate production in P. syringae has been associated with increased fitness and virulence in planta. Alginate biosynthesis is tightly controlled by proteins encoded by the algT-muc regulatory gene cluster in P. aeruginosa and A. vinelandii. These genes encode the alternative sigma factor AlgT (σ22), its anti-sigma factors MucA and MucB, MucC, a protein with a controversial function that is absent in P. syringae, and MucD, a periplasmic serine protease and homolog of HtrA in Escherichia coli. We compared an alginate-deficient algT mutant of P. syringae pv. glycinea with an alginate-producing derivative in which algT is intact. The alginate-producing derivative grew significantly slower in vitro growth but showed increased epiphytic fitness and better symptom development in planta. Evaluation of expression levels for algT, mucA, mucB, mucD, and algD, which encodes an alginate biosynthesis gene, showed that mucD transcription is not dependent on AlgT in P. syringae in vitro. Promoter mapping using primer extension experiments confirmed this finding. Results of reverse transcription-PCR demonstrated that algT, mucA, and mucB are cotranscribed as an operon in P. syringae. Northern blot analysis revealed that mucD was expressed as a 1.75-kb monocistronic mRNA in P. syringae.

scholarly journals Microbial Production of Glyceric Acid, an Organic Acid That Can Be Mass Produced from Glycerol

2009 ◽  
Vol 75 (24) ◽  
pp. 7760-7766 ◽  
Author(s):  
Hiroshi Habe ◽  
Yuko Shimada ◽  
Toshiharu Yakushi ◽  
Hiromi Hattori ◽  
Yoshitaka Ano ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Growth Parameters ◽  
Operating Time ◽  
Enantiomeric Excess ◽  
Gluconobacter Oxydans ◽  
Acetic Acid Bacteria ◽  
Culture Broth ◽  
Glyceric Acid ◽  
Bacterial Strains ◽  
Biotechnological Production

ABSTRACT Glyceric acid (GA), an unfamiliar biotechnological product, is currently produced as a small by-product of dihydroxyacetone production from glycerol by Gluconobacter oxydans. We developed a method for the efficient biotechnological production of GA as a target compound for new surplus glycerol applications in the biodiesel and oleochemical industries. We investigated the ability of 162 acetic acid bacterial strains to produce GA from glycerol and found that the patterns of productivity and enantiomeric GA compositions obtained from several strains differed significantly. The growth parameters of two different strain types, Gluconobacter frateurii NBRC103465 and Acetobacter tropicalis NBRC16470, were optimized using a jar fermentor. G. frateurii accumulated 136.5 g/liter of GA with a 72% d-GA enantiomeric excess (ee) in the culture broth, whereas A. tropicalis produced 101.8 g/liter of d-GA with a 99% ee. The 136.5 g/liter of glycerate in the culture broth was concentrated to 236.5 g/liter by desalting electrodialysis during the 140-min operating time, and then, from 50 ml of the concentrated solution, 9.35 g of GA calcium salt was obtained by crystallization. Gene disruption analysis using G. oxydans IFO12528 revealed that the membrane-bound alcohol dehydrogenase (mADH)-encoding gene (adhA) is required for GA production, and purified mADH from G. oxydans IFO12528 catalyzed the oxidation of glycerol. These results strongly suggest that mADH is involved in GA production by acetic acid bacteria. We propose that GA is potentially mass producible from glycerol feedstock by a biotechnological process.

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