Studies on cellulase production by Clostridium thermocellum

Martínez, Duniesky; Shinmyo, Atsuhiko; Madia, Ashwin; Deman, A L

doi:10.1007/bf00504485

Cited by 140 publications

(71 citation statements)

References 8 publications

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“…Maximum CMCase activity was found at pH 7.5 in Aspergillus niger (Z10 wild type strain) as reported by GoKhan et al [15] and at 6-7 in A. niger [16]. Similar findings are reported by Garcia-Martinez et al [17] and Prasetson and Doelle [18] for maximum enzyme production by Clostridium thermocellum and Cellulomonas. Our findings are in accordance with that of theirs.…”

Section: Enzyme Assaysupporting

confidence: 87%

Polyphasic Characterization of a Potential Novel Cellulolytic Bacterium Brevibacillus Brevis Strain St-2

Prasad¹,

Singh

Bedi

2014

JBT

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The tremendous commercial potential of cellulases in a variety of applications remains the driving force for research in this area. The present study was aimed at isolation and screening of promising cellulolytic strains from locally collected soil samples. A promising cellulose degrading bacterial strain designated as St-2 was isolated from agricultural field. Optimization of fermentation media ingredients and environmental factors were done for optimizing growth of the strain which facilitates effective cellulase production. St-2 showed luxuriant growth on sucrose, lactose, mannitol and inositol whereas the growth was moderate on dextrose and fructose. The strain was able to grow in a wide range of pH (5-11) and temperature (4-45 ˚C ) and was tolerant to up to 6% (w/v) of NaCl concentration in the medium. The results indicate wide spectrum adaptability of the strain to variable pH, temperatures and saline concentrations that makes it an advantageous organism to survive in the fluctuating environmental conditions. The optimum pH and temperature for enzyme production were 7 (with 2.11 U/ml CMCase activities and 2.22 U/ml FPase activities) and 37˚C (with 1.73 U/ml CMCase activities and 1.92 U/ml FPase activities), respectively. The morphological and biochemical characteristics of the strain were consistent with those of the genus Brevibacillus. Analysis of the 16S rRNA gene sequence of strain St-2 showed a similarity score of 99% with Brevibacillus brevis (GenBank Accession No. AB271756) having BLAST score ranging between 821 to 827 bits. Brevibacillus brevis Strain St-2 has been deposited in the GenBank database with the Accession No. KF306223.

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Section: Enzyme Assaysupporting

confidence: 87%

Polyphasic Characterization of a Potential Novel Cellulolytic Bacterium Brevibacillus Brevis Strain St-2

Prasad¹,

Singh

Bedi

2014

JBT

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“…Growth on xylose by strain ATCC 27405 (the designation given by the American Type Culture Collection to the 651 strain) was not confirmed by us (304). We also did not observe growth on xylan, a component of hemicellulose (93). Patni and Alexander (264,265) found C. thermocellum 651 to grow on mannitol, glucose, xylose, and fructose.…”

Section: Carbon Source Nutritionmentioning

confidence: 46%

“…However, upon increasing yeast extract to 0.6%, the organism assimilated glucose (304). We developed a chemically defined medium in which the yeast extract was replaced by para-aminobenzoic acid, vitamin B 12 , pyridoxamine, and biotin (140) and found that C. thermocellum could use ammonium, nitrate, urea, or certain amino acids as major nitrogen sources (93). The organism was found to break down xylan but could not use the resulting xylose or xylose oligomers.…”

Section: Strains and Mediamentioning

confidence: 99%

Cellulase, Clostridia, and Ethanol

Demain

Newcomb

2005

Microbiol Mol Biol Rev

810

579

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SUMMARY Biomass conversion to ethanol as a liquid fuel by the thermophilic and anaerobic clostridia offers a potential partial solution to the problem of the world's dependence on petroleum for energy. Coculture of a cellulolytic strain and a saccharolytic strain of Clostridium on agricultural resources, as well as on urban and industrial cellulosic wastes, is a promising approach to an alternate energy source from an economic viewpoint. This review discusses the need for such a process, the cellulases of clostridia, their presence in extracellular complexes or organelles (the cellulosomes), the binding of the cellulosomes to cellulose and to the cell surface, cellulase genetics, regulation of their synthesis, cocultures, ethanol tolerance, and metabolic pathway engineering for maximizing ethanol yield.

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“…Strains of E. coli were maintained on Luria-Bertani medium (Sambrook et al, 1989). C. cellulolyticum ATCC 35319 was routinely grown on GS medium, modified from that described by Garcia-Martinez et al (1980), which contains: KH # PO % , 3n7 mM; K # HPO % , 3n5 mM; urea, 33 mM; MgCl # , 2n5 mM; CaCl # , 0n3 mM; FeSO % , 0n004 mM; sodium β-glycerophosphate, 28 mM; trisodium citrate, 10 mM; MOPS, 48 mM; -cellobiose, 15 mM; cysteine;HCl, 6n3 mM; yeast extract, 0n5% (w\v); pH 7n2. VM medium was derived from GS medium by replacing yeast extract with: -biotin, 0n08 µM; pyridoxamine, 0n02 µM; cyanocobalamin, 0n001 µM; p-aminobenzoic acid, 0n15 µM; thiamin, 0n9 µM; -alanine, 0n22 µM.…”

Section: Methodsmentioning

confidence: 99%

Gene transfer to Clostridium cellulolyticum ATCC 35319

et al. 2000

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Although much is known about the bacterial cellulosome and its various protein components, their contributions to bacterial growth on cellulose and the process of cellulolysis in vivo cannot currently be assessed. To remedy this, the authors have developed gene transfer techniques for Clostridium cellulolyticum ATCC 35319. Firstly, transfer of Tn1545 has been obtained using an Enterococcus faecalis donor. Secondly, IncP-mediated conjugative mobilization of plasmids from Escherichia coli donors has also been achieved. The yield of transconjugants in both cases was low and was probably limited by the suboptimal growth conditions that must of necessity be employed for the co-culture of oligotrophic C. cellulolyticum with copiotrophic donors. A restriction endonuclease was detected in crude extracts of C. cellulolyticum. This enzyme, named CceI, is an isoschizomer of MspI (HpaII). Electrotransformation was employed to establish plasmids containing the replication functions of pAMβ1 (En. faecalis), pIM13 (Bacillus subtilis), pCB102 (Clostridium butyricum), pIP404 (Clostridium perfringens) and pWV01 (Lactococcus lactis subsp. cremoris) in C. cellulolyticum. Transformants were only obtained if the DNA was appropriately methylated on the external C of the sequence 5'-CCGG-3' using either BsuFI methylase in vivo or MspI methylase in vitro. Plasmids based on the pAMβ1 and pIM13 replicons were more stably maintained than one based on the pCB102 replicon. Selection of transformants on solid medium led to low apparent transformation efficiencies (approx. 10 2 transformants per µg DNA) which might, in part, reflect the low plating efficiency of the organism. Selection of transformants in liquid medium led to a higher apparent yield of transformants (between 10 5 and 10 7 transformants per µg DNA). The methods developed here will pave the way for functional analysis of the various cellulosome components in vivo.

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Studies on cellulase production by Clostridium thermocellum

Cited by 140 publications

References 8 publications

Polyphasic Characterization of a Potential Novel Cellulolytic Bacterium Brevibacillus Brevis Strain St-2

Polyphasic Characterization of a Potential Novel Cellulolytic Bacterium Brevibacillus Brevis Strain St-2

Cellulase, Clostridia, and Ethanol

Gene transfer to Clostridium cellulolyticum ATCC 35319

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