Intergeneric protoplast fusion between Fusobacterium varium and Enterococcus faecium for enhancing dehydrodivanillin degradation

Chen, Wei; Ohmiya, Kunio; Shimizu, Sakayu

doi:10.1128/aem.53.3.542-548.1987

Cited by 20 publications

(18 citation statements)

References 27 publications

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“…The protoplast fusion technique has been studied (4, 8-11, 16, 20) extensively as a means of genetic transfer or recombination. We have previously reported a successful intergeneric protoplast fusion system between Fusobacterium varium and Enterococcus faecium under anaerobic conditions for enhancing dehydrodivanillin (DDV) degradation (2,3). Our results suggested that genetic transformation and recombination can take place through protoplast fusion between bacterial strains that are distantly related taxonomically.…”

mentioning

confidence: 73%

“…), kanamycin sulfate (Takeda Chemical Industries Co.), restriction enzyme PstI (Nippon Gene Co., Ltd), polyethylene glycol (PEG) 6000 and Congo red (Wako Pure Chemical Co.), API 20A test kit for the identification of anaerobes and API ZYM test kit for the assay of enzymatic activities (API System S.A., La Balme Les Grottes, 38390-Montalieu Vercieu, France). o-Carboxymethyl cellulose (CMC) samples were generously provided by Daiichi Kogyo Seiyaku Co. All the other reagents were the same as those described previously (2,3) and were commercial products of analytical grade.…”

Section: Methodsmentioning

confidence: 99%

“…In this report, we describe an anaerobic mutant strain, FEM29, derived from the anaerobic recombinant FE7 (3).…”

mentioning

confidence: 99%

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Escherichia coli Spheroplast-Mediated Transfer of pBR322 Carrying the Cloned Ruminococcus albus Cellulase Gene into Anaerobic Mutant Strain FEM29 by Protoplast Fusion

Chen

Ohmiya

Shimizu

1988

Appl Environ Microbiol

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Intergeneric protoplast fusion between Escherichia coli HB101 with pBR322 carrying the cloned o-(carboxymethyl)cellulase (CMCase) gene of Ruminococcus albus (Pro-Leu-Apr Kms) and an anaerobic mutant strain, FEM29 (Trp-His-Aps Kmr), with dehydrodivanillin-degrading activity was performed in the presence of 40% polyvinyl alcohol 300 under aerobic and anaerobic conditions to transfer the cloned cellulase gene into the mutant. The mutant FEM29 had a unique property. When it was incubated in liquid medium with 1% glucose and sucrose, protoplasts could be produced autogenously and regenerated on the agar slant. E. coli spheroplasts formed from a plasmid-amplified overnight culture after 10 min of treatment with lysozyme (20 ,Lg/ml) in a hypertonic solution (0.01 M Tris hydrochloride [pH 7.5], 0.4 M mannitol). Protoplast regeneration rates of FEM29 and HB101 were 30 and 83%, respectively, on the agar-yeast extract medium. Apr Kmr fusants were obtained at high frequency: 1.7 x 10-2 anaerobically and 8.2 x 10-3 aerobically. These fusants showed 23 to 57% of CMCase and dehydrodivanillin-degrading activities, respectively, as compared with parental strains. All the fusants isolated were gram-negative rods with main phenotypes such as urease and catalase activities as in HB101 and esterase and chymotrypsin activities as in FEM29. Southern hybridization experiments suggested that pBR322 with the cloned CMCase gene existed autonomously in the fusant cells. This is the first report describing transfer of pBR322 with a cloned cellulase gene into an anaerobic mutant by polyvinyl alcohol-mediated fusion with an E. coli spheroplast.

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confidence: 73%

Section: Methodsmentioning

confidence: 99%

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Escherichia coli Spheroplast-Mediated Transfer of pBR322 Carrying the Cloned Ruminococcus albus Cellulase Gene into Anaerobic Mutant Strain FEM29 by Protoplast Fusion

Chen

Ohmiya

Shimizu

1988

Appl Environ Microbiol

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“…Polyethylene glycol (PEG)-induced cell fusion is one of the means to transfer DNA between bacterial cells, and numerous instances have been reported for intraspecific, interspecific, and intergeneric protoplast fusion (Akamatsu & Sekiguchi, 1983;Chen et al, 1986Chen et al, , 1987Cocconcelli et al, 1986;Baigorí et al, 1988;van der Lelie et al, 1988;van der Vossen et al, 1988;Gokhale & Deobagkar, 1989). The genetic materials used in these studies include either plasmids or chromosomal DNA.…”

Section: Introductionmentioning

confidence: 99%

Effect of Bacillus subtilis BsuM restriction-modification on plasmid transfer by polyethylene glycol-induced protoplast fusion

Maehara

Itaya

Ogura

et al. 2011

FEMS Microbiol Lett

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Polyethylene glycol (PEG)-induced cell fusion is a promising method to transfer larger DNA from one cell to another than conventional genetic DNA transfer systems. The laboratory strain Bacillus subtilis 168 contains a restriction (R) and modification (M) system, BsuM, which recognizes the sequence 5'-CTCGAG-3'. To study whether the BsuM system affects DNA transfer by the PEG-induced cell fusion between R(+)M(+) and R(-)M(-) strains, we examined transfer of plasmids pHV33 and pLS32neo carrying no and eight BsuM sites, respectively. It was shown that although the transfer of pLS32neo but not pHV33 from the R(-)M(-) to R(+)M(+) cells was severely restricted, significant levels of transfer of both plasmids from the R(+)M(+) to R(-)M(-) cells were observed. The latter result shows that the chromosomal DNA in the R(-)M(-) cell used as the recipient partially survived restriction from the donor R(+)M(+) cell, indicating that the BsuM R(-)M(-) strain is useful as a host for accepting DNA from cells carrying a restriction system(s). Two such examples were manifested for plasmid transfer from Bacillus circulans and Bacillus stearothermophilus strains to a BsuM-deficient mutant, B. subtilis RM125.

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“…into Bacillus subtilis were achieved. More recently, the successful use of intergeneric protoplast fusion to transfer genomic traits such as lignin degradation (6) and nitrogen fixation (36) in bacteria has been reported. The expression of cellulases in bacteria could be improved in both the quality and quantity of the desired enzyme as required by introducing various kinds of mutations into the relevant genes.…”

mentioning

confidence: 99%

Differential Expression of Xylanases and Endoglucanases in the Hybrid Derived from Intergeneric Protoplast Fusion between a Cellulomonas sp. and Bacillus subtilis

Gokhale

Deobagkar

1989

Appl Environ Microbiol

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A stable hybrid obtained by protoplast fusion between a Cellulomonas sp. and Bacillus subtilis exhibits an altered pattern of enzyme induction with different cellulosic substrates. Unlike in the Cellulomonas sp., xylanase was induced in the hybrid organism specifically by xylan, and endoglucanase was induced by carboxymethyl cellulose. The amount and specific activity of xylanase produced by the hybrid were more than those produced by the Cellulomonas sp. P-Glucosidase which is cell bound or intracellular in the Cellulomonas sp. was secreted by the hybrid organism, and relative amounts of extracellular B-glucosidase were high.

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Intergeneric protoplast fusion between Fusobacterium varium and Enterococcus faecium for enhancing dehydrodivanillin degradation

Cited by 20 publications

References 27 publications

Escherichia coli Spheroplast-Mediated Transfer of pBR322 Carrying the Cloned Ruminococcus albus Cellulase Gene into Anaerobic Mutant Strain FEM29 by Protoplast Fusion

Escherichia coli Spheroplast-Mediated Transfer of pBR322 Carrying the Cloned Ruminococcus albus Cellulase Gene into Anaerobic Mutant Strain FEM29 by Protoplast Fusion

Effect of Bacillus subtilis BsuM restriction-modification on plasmid transfer by polyethylene glycol-induced protoplast fusion

Differential Expression of Xylanases and Endoglucanases in the Hybrid Derived from Intergeneric Protoplast Fusion between a Cellulomonas sp. and Bacillus subtilis

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