Protoplast formation, L-colony growth, and regeneration ofClostridium beijerinckii NRRL B-592 and B-593 andClostridium acetobutylicum ATCC 10132

Birrer, Gregory A.; Chesbro, William; Zsigray, Robert M.

doi:10.1007/bf01577356

Cited by 6 publications

(10 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As shown in this Table, two wild clostridial strains (i.e., C. acetobutylicum and C. beijerinckii ) were fused with the thermally stable C. thermocellum . Fused strains were prepared following the general protocol proposed by Birrer et al . All fused strains were stored in ultra‐low freezer at ‐80 °C together with the wild strains of Ca and Cb .…”

Section: Methodsmentioning

confidence: 99%

“…The fusion process that was conducted in our laboratory consists of three major steps: Formation of protoplast, fusion and regeneration (of cell wall around the fused protoplast). To prepare for the first step, overnight cultures of the wild strains were harvested by centrifugation at 10,600 xg for 10 min and then were diluted 1:4 with fresh, sterile CBM broth containing 0.4% or 0.8% glycine . After 45–60 min when the cultures become nearly 100% motile, the osmotic strength of each culture was increased by suspending in 5 ml protoplasting medium (PPM).…”

Section: Methodsmentioning

confidence: 99%

“…The protoplast pellets can be directly used if the fusion is to be performed the same day or can be stored by re‐suspending in fresh PPM without lysozyme. The protoplast remains intact in this medium . Protoplast formation was performed for C. acetobutylicum, C. beijerinckii and C. thermocellum individually.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Enhanced biobutanol production using novel clostridial fusants in simultaneous saccharification and fermentation of green renewable agriculture residues

Begum

Dahman

2015

Biofuels Bioprod Bioref

View full text Add to dashboard Cite

Thermostable Clostridia species were developed in the present study through protoplast fusion between mesophilic and thermophilic clostridial species. Two parental strains of Clostridia species, Clostridium acetobutylicum (ATCC 4259) and Clostridium beijerinckii (ATCC BA101), along with three fused strains, Clostridium acetobutylicum Clostridium thermocellum (CaCt), Clostridium beijerinckii Clostridium thermocellum (CbCt) and Clostridium acetobutylicum Clostridium beijerinckii (CaCb), were examined for biobutanol production using wheat straw (WS) as a feedstock in a batch process of simultaneous saccharification and fermentation (SSF). The objective of the study was to use the thermotolerant fused strains to enhance enzymatic activity during the SSF by raising the incubation temperature and thereby eventually increasing biobutanol production. Results from the current study indicated that the fused strains were able to produce the required enzymes at higher temperature essential for the saccharification of WS (i.e., endoglucanase, exoglucanase, and β‐glucosidase). This will have a major impact on eliminating costs associated with adding enzymes to the saccharification process. Fused strain CbCt achieved the highest biobutanol production of up to 14.13 g/L (i.e., a yield of 0.29) at 45 °C with total sugar consumption of 82%. Enzymatic activity of CbCt was found to be 61.67 FPU (filter paper unit) which was the highest in comparison with other fused strains evaluated in this study. These results indicate a breakthrough for the technological and economical obstacles associated with industrialization of the SSF process. © 2015 Society of Chemical Industry and John Wiley & Sons, Ltd

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Enhanced biobutanol production using novel clostridial fusants in simultaneous saccharification and fermentation of green renewable agriculture residues

Begum

Dahman

2015

Biofuels Bioprod Bioref

View full text Add to dashboard Cite

show abstract

“…Yeast extract, 2.5g Casamino acids, 1g L-asparagine. These ingredients were mixed in 930ml H 2 O and the mixture was stirred and brought to boiling before autoclaving at 121 o C for 20 min [94]. Upon cooling, 10ml of stock solution A and 40ml of solution B was added to the basal mixture.…”

Section: Medium Preparation For Fusionmentioning

confidence: 99%

“…Formation of protoplast, fusion and regeneration (of cell wall around the fused protoplast). To prepare for the first step, overnight cultures of the wild strains were harvested by centrifugation at 10,600 x g for 10 min and then were diluted 1:4 with fresh, sterile CBM broth containing 0.4% or 0.8% glycine [94]. After 45-60 min when the cultures become nearly 100% motile, the osmotic strength of each culture was increased by suspending in 5ml Protoplasting medium (PPM).…”

Section: Protoplast Fusion Formationmentioning

confidence: 99%

Enhanced Biobutanol Production in Batch Simultaneous Saccharification and Fermentation From Agriculture Residues Using Novel Clostridial Fusants

Begum¹

2021

Preprint

View full text Add to dashboard Cite

A novel approach was introduced for the enhancement of biobutanol production in ABE fermenetation. Thermostable Clostridia species were developed through protoplast fusion between mesophilic and thermophilic Clostridial species. Two parental strains of Clostridia species, Clostridium acetobutylicum (ATCC 4259) and Clostridium beijerinckii (ATCC BA101), along with three fused strains, Clostridium acetobutylicum Clostridium thermocellum (CaCt), Clostridium beijerinckii Clostridium thermocellum (CbCt) and Clostridium acetobutylicum Clostridium beijerinckii (CaCb), were examined for biobutanol production using wheat straw as a feed stock in a batch process of simultaneous saccharification and fermentation (SSF). The objective of the study was to use the thermotolerant fused strains to enhance enzymatic activity during the SSF by raising the incubation temperature and thereby eventually increasing biobutanol production. Economic and residence time analysis of SSF found that addition of cellulase increases the cost of fermentation and prolongs biobutanol production cycle. However, results from the current study indicated that the fused strains, at the higher temperature, were able to produce the required enzymes for the saccharification of wheat straw (i.e., endoglucanase, exoglucanase, and b- glucosidase). This will have a major impact on eliminating costs associated with adding enzymes as raw material to the saccharification process. Fused strain CbCt achieved the highest biobutanol production of up to 14.13 g/L (i.e., a yield of 0.29) was reached at 45°C with total sugar consumption of 82%. Enzymatic activity of CbCt was found to be 61.67 FPU (Filter Paper Unit) which was the highest in comparison with other fused strains evaluated in this study. These results indicate a breakthrough for the technological and economical obstacles associated with industrialization of the SSF process.

show abstract

Electro-transformation of Clostridium beijerinckii NRRL B-592 with shuttle plasmid pHR106 and recombinant derivatives

Birrer

Chesbro

Zsigray

1994

Appl Microbiol Biotechnol

View full text Add to dashboard Cite

Conditions for transformation of the solventogenic anaerobe Clostridium beijerinckii NRRL B-592 with plasmid DNA via electroporation are described. Shuttle plasmid pHR106 and two derivatives constructed in this study were transferred and were expressed in this organism. One recombinant derivative of pHR106 was constructed by separately subcloning the clostridial tetracycline (tetP) resistance genes into pHR106. The second vector conferring erythromycin resistance was obtained via in-vivo recombination. The new constructs, termed pRZL and pRZE respectively, were then transferred to C. beijerinckii in order to evaluate their potential as shuttle vectors. The recombinant plasmids were shown to transfer to C. beijerinckii and were expressed as autonomously replicating vectors. The use of these plasmids as cloning and shuttle vectors for C. beijerinckii is discussed.

show abstract

Protoplast formation, L-colony growth, and regeneration ofClostridium beijerinckii NRRL B-592 and B-593 andClostridium acetobutylicum ATCC 10132

Cited by 6 publications

References 11 publications

Enhanced biobutanol production using novel clostridial fusants in simultaneous saccharification and fermentation of green renewable agriculture residues

Enhanced biobutanol production using novel clostridial fusants in simultaneous saccharification and fermentation of green renewable agriculture residues

Enhanced Biobutanol Production in Batch Simultaneous Saccharification and Fermentation From Agriculture Residues Using Novel Clostridial Fusants

Electro-transformation of Clostridium beijerinckii NRRL B-592 with shuttle plasmid pHR106 and recombinant derivatives

Contact Info

Product

Resources

About