G. P. Hazlewood

Growth and Survival of Genetically Manipulated Lactobacillus plantarum in Silage

Sharp, R., O'Donnell, A. G., Gilbert, H. G., Hazlewood, G. P.

The growth and persistence of two genetically manipulated forms of Lactobacillus plantarum NCDO (National Collection of Dairy Organisms) 1193 have been monitored in grass silage. Both recombinants...

Expression of a Clostridium thermocellum endoglucanase gene in Lactobacillus plantarum.

Bates, E E, Gilbert, H J, Hazlewood, G P, Huckle, J, Laurie, J I, Mann, S P

Recombinant plasmid pM25 containing the celE gene of Clostridium thermocellum, which codes for an enzymatically active endoglucanase, was transformed into Lactobacillus plantarum by electroporation....

Isolation of proteolytic rumen bacteria by use of selective medium containing leaf fraction 1 protein (ribulosebisphosphate carboxylase).

Hazlewood, G P, Orpin, C G, Greenwood, Y, Black, M E

The principle proteolytic bacteria isolated from bovine rumen contents by virtue of the ability to obtain nitrogen from the proteolysis of leaf fraction 1 protein (ribulosebisphosphate carboxylase,...

Spatial separation of protein domains is not necessary for catalytic activity or substrate binding in a xylanase.

Ferreira, L M, Durrant, A J, Hall, J, Hazlewood, G P, Gilbert, H J

Xylanase A (XYLA) from Pseudomonas fluorescens subspecies cellulosa shows sequence conservation with two endoglucanases from the same organism. The conserved sequence in XYLA, consisting of the...

Characterization of the gene celD and its encoded product 1,4-beta-D-glucan glucohydrolase D from Pseudomonas fluorescens subsp. cellulosa.

Rixon, J E, Ferreira, L M, Durrant, A J, Laurie, J I, Hazlewood, G P, Gilbert, H J

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA constructed in pUC18 and expressed in Escherichia coli was screened for recombinants expressing 4-methylumbelliferyl beta-D-glucoside...

A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain.

Ferreira, L M, Wood, T M, Williamson, G, Faulds, C, Hazlewood, G P, Black, G W, ...

The 5' regions of genes xynB and xynC, coding for a xylanase and arabinofuranosidase respectively, are identical and are reiterated four times within the Pseudomonas fluorescens subsp. cellulosa...

The non-catalytic cellulose-binding domain of a novel cellulase from Pseudomonas fluorescens subsp. cellulosa is important for the efficient hydrolysis of Avicel.

Hall, J, Black, G W, Ferreira, L M, Millward-Sadler, S J, Ali, B R, Hazlewood, G P, ...

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA, constructed in lambda ZAPII, was screened for carboxymethyl-cellulase activity. The pseudomonad insert from a recombinant phage...

Novel cellulose-binding domains, NodB homologues and conserved modular architecture in xylanases from the aerobic soil bacteria Pseudomonas fluorescens subsp. cellulosa and Cellvibrio mixtus.

Millward-Sadler, S J, Davidson, K, Hazlewood, G P, Black, G W, Gilbert, H J, Clarke, J H

To test the hypothesis that selective pressure has led to the retention of cellulose-binding domains (CBDs) by hemicellulase enzymes from aerobic bacteria, four new xylanase (xyn) genes from two...

A non-modular endo-beta-1,4-mannanase from Pseudomonas fluorescens subspecies cellulosa.

Braithwaite, K L, Black, G W, Hazlewood, G P, Ali, B R, Gilbert, H J

Pseudomonas fluorescens subsp. cellulosa when cultured in the presence of carob galactomannan degraded the polysaccharide. To isolate gene(s) from P. fluorescens subsp. cellulosa encoding...

Evidence for a general role for non-catalytic thermostabilizing domains in xylanases from thermophilic bacteria.

Fontes, C M, Hazlewood, G P, Morag, E, Hall, J, Hirst, B H, Gilbert, H J

A genomic library of Clostridium thermocellum DNA constructed in lambda ZAPII was screened for xylanase-expressing clones. Cross-hybridization experiments revealed a new xylanase gene isolated from...

A modular xylanase containing a novel non-catalytic xylan-specific binding domain.

Black, G W, Hazlewood, G P, Millward-Sadler, S J, Laurie, J I, Gilbert, H J

Xylanase D (XYLD) from Cellulomonas fimi contains a C-terminal cellulose-binding domain (CBD) and an internal domain that exhibits 65% sequence identity with the C-terminal CBD. Full-length XYLD...

Intronless celB from the anaerobic fungus Neocallimastix patriciarum encodes a modular family A endoglucanase.

Zhou, L, Xue, G P, Orpin, C G, Black, G W, Gilbert, H J, Hazlewood, G P

The cDNA designated celB from the anaerobic rumen fungus Neocallimastix patriciarum contained a single open reading frame of 1422 bp coding for a protein (CelB) of M(r) 53,070. CelB expressed by...

Xylanase B from Neocallimastix patriciarum contains a non-catalytic 455-residue linker sequence comprised of 57 repeats of an octapeptide.

Black, G W, Hazlewood, G P, Xue, G P, Orpin, C G, Gilbert, H J

A Neocallimastix patriciarum cDNA library was screened for xylanase-expressing clones, which were distinct from the previously characterized N. patriciarum xynA cDNA encoding xylanase A. A single...

Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes.

Kellett, L E, Poole, D M, Ferreira, L M, Durrant, A J, Hazlewood, G P, Gilbert, H J

The complete nucleotide sequence of the Pseudomonas fluorescens subsp. cellulosa xynB gene, encoding an endo-beta-1,4-xylanase (xylanase B; XYLB) has been determined. The structural gene consists of...

The non-catalytic C-terminal region of endoglucanase E from Clostridium thermocellum contains a cellulose-binding domain.

Durrant, A J, Hall, J, Hazlewood, G P, Gilbert, H J

Mature endoglucanase E (EGE) from Clostridium thermocellum consists of 780 amino acid residues and has an Mr of 84,016. The N-terminal 334 amino acids comprise a functional catalytic domain....

A new series of long-chain dicarboxylic acids with vicinal dimethyl branching found as major components of the lipids of Butyrivibrio spp.

Klein, R A, Hazlewood, G P, Kemp, P, Dawson, R M

1. Some members of the genus Butyrivibrio, including a general fatty acid auxotroph (strain S2), contain as a major part of their complex lipids a high-molecular-weight component that is probably...

Complex lipids of a lipolytic and general-fatty-acid-requiring Butyrivibrio sp. isolated from the ovine rumen.

Hazlewood, G P, Clarke, N G, Dawson, R M

The complex lipids of the naturally-occurring general-fatty-acid-auxotroph Butyrivibrio S2 [Hazlewood & Dawson (1979) J. Gen. Microbiol. 112, 15-27] grown with palmitic acid as sole fatty-acid...

Structure of diabolic acid-containing phospholipids isolated from Butyrivibrio sp.

Clarke, N G, Hazlewood, G P, Dawson, R M

The structures of the diabolic acid-containing phospholipids of Buryrivibrio S2 grown in the presence of palmitic acid have been investigated. Generally they consist of two conventional bacterial...

Intermolecular transacylation of phosphatidylethanolamine by a Butyrivibrio sp.

Hazlewood, G P, Dawson, R M

1. Washed cells and supernatant from a culture of a Butyrivibrio sp. carry out the intermolecular transacylation reaction 2 phosphatidylethanolamine leads to...

A phospholipid-deacylating system of bacteria active in a frozen medium.

Hazlewood, G P, Dawson, R M

A phosphatidylcholine-deacylating system present in a Butyrivibrio species (probably fibrisolvens) shows appreciable activity at low temperatures with a maximum hydrolysis rate at--10 degrees C. 2....

A protein targeting signal that functions in polarized epithelial cells in vivo.

Ali, S, Hall, J, Hazlewood, G P, Hirst, B H, Gilbert, H J

Eukaryotic membrane-associated polypeptides often contain a glycosylphosphatidylinositol (GPI) anchor that signals the attachment of GPI lipids to these proteins. The GPI anchor can function as a...

Evidence that linker sequences and cellulose-binding domains enhance the activity of hemicellulases against complex substrates.

Black, G W, Rixon, J E, Clarke, J H, Hazlewood, G P, Theodorou, M K, Morris, P, ...

Xylanase A (XYLA) and arabinofuranosidase C (XYLC) from Pseudomonas fluorescens subsp. cellulosa are modular enzymes consisting of discrete cellulose-binding domains (CBDs) and catalytic domains...

Arabinanase A from Pseudomonas fluorescens subsp. cellulosa exhibits both an endo- and an exo- mode of action.

McKie, V A, Black, G W, Millward-Sadler, S J, Hazlewood, G P, Laurie, J I, Gilbert, H J

Pseudomonas fluorescens subsp. cellulosa expressed arabinanase activity when grown on media supplemented with arabinan or arabinose. Arabinanase activity was not induced by the inclusion of other...

Pseudomonas cellulose-binding domains mediate their effects by increasing enzyme substrate proximity.

Bolam, D N, Ciruela, A, McQueen-Mason, S, Simpson, P, Williamson, M P, Rixon, J E, ...

To investigate the mode of action of cellulose-binding domains (CBDs), the Type II CBD from Pseudomonas fluorescens subsp. cellulosa xylanase A (XYLACBD) and cellulase E (CELECBD) were expressed as...

Homologous xylanases from Clostridium thermocellum: evidence for bi-functional activity, synergism between xylanase catalytic modules and the presence of xylan-binding domains in enzyme complexes.

Fernandes, A C, Fontes, C M, Gilbert, H J, Hazlewood, G P, Fernandes, T H, Ferreira, L M

Clostridium thermocellum produces a consortium of plant-cell-wall hydrolases that form a cell-bound multi-enzyme complex called the cellulosome. In the present study two similar xylanase genes, xynU...

The type II and X cellulose-binding domains of Pseudomonas xylanase A potentiate catalytic activity against complex substrates by a common mechanism.

Gill, J, Rixon, J E, Bolam, D N, McQueen-Mason, S, Simpson, P J, Williamson, M P, ...

Xylanase A (Pf Xyn10A), in common with several other Pseudomonas fluorescens subsp. cellulosa polysaccharidases, consists of a Type II cellulose-binding domain (CBD), a catalytic domain (Pf...

A modular cinnamoyl ester hydrolase from the anaerobic fungus Piromyces equi acts synergistically with xylanase and is part of a multiprotein cellulose-binding cellulase-hemicellulase complex.

Fillingham, I J, Kroon, P A, Williamson, G, Gilbert, H J, Hazlewood, G P

A collection of clones, isolated from a Piromyces equi cDNA expression library by immunoscreening with antibodies raised against affinity purified multienzyme fungal cellulase-hemicellulase complex,...

Characterization of hybrid proteins consisting of the catalytic domains of Clostridium and Ruminococcus endoglucanases, fused to Pseudomonas non-catalytic cellulose-binding domains.

Poole, D M, Durrant, A J, Hazlewood, G P, Gilbert, H J

The N-terminal 160 or 267 residues of xylanase A from Pseudomonas fluorescens subsp. cellulosa, containing a non-catalytic cellulose-binding domain (CBD), were fused to the N-terminus of the...

The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains.

Ferreira, L M, Hazlewood, G P, Barker, P J, Gilbert, H J

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was constructed in pUC18 and Escherichia coli recombinants expressing 4-methylumbelliferyl beta-D-cellobioside-hydrolysing activity...

Growth and Survival of Genetically Manipulated Lactobacillus plantarum in Silage

Sharp, R., O'Donnell, A. G., Gilbert, H. G., Hazlewood, G. P.

The growth and persistence of two genetically manipulated forms of Lactobacillus plantarum NCDO (National Collection of Dairy Organisms) 1193 have been monitored in grass silage. Both recombinants...

Expression of a Clostridium thermocellum endoglucanase gene in Lactobacillus plantarum.

Bates, E E, Gilbert, H J, Hazlewood, G P, Huckle, J, Laurie, J I, Mann, S P

Recombinant plasmid pM25 containing the celE gene of Clostridium thermocellum, which codes for an enzymatically active endoglucanase, was transformed into Lactobacillus plantarum by electroporation....

Isolation of proteolytic rumen bacteria by use of selective medium containing leaf fraction 1 protein (ribulosebisphosphate carboxylase).

Hazlewood, G P, Orpin, C G, Greenwood, Y, Black, M E

The principle proteolytic bacteria isolated from bovine rumen contents by virtue of the ability to obtain nitrogen from the proteolysis of leaf fraction 1 protein (ribulosebisphosphate carboxylase,...

Spatial separation of protein domains is not necessary for catalytic activity or substrate binding in a xylanase.

Ferreira, L M, Durrant, A J, Hall, J, Hazlewood, G P, Gilbert, H J

Xylanase A (XYLA) from Pseudomonas fluorescens subspecies cellulosa shows sequence conservation with two endoglucanases from the same organism. The conserved sequence in XYLA, consisting of the...

Characterization of the gene celD and its encoded product 1,4-beta-D-glucan glucohydrolase D from Pseudomonas fluorescens subsp. cellulosa.

Rixon, J E, Ferreira, L M, Durrant, A J, Laurie, J I, Hazlewood, G P, Gilbert, H J

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA constructed in pUC18 and expressed in Escherichia coli was screened for recombinants expressing 4-methylumbelliferyl beta-D-glucoside...

A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain.

Ferreira, L M, Wood, T M, Williamson, G, Faulds, C, Hazlewood, G P, Black, G W, ...

The 5' regions of genes xynB and xynC, coding for a xylanase and arabinofuranosidase respectively, are identical and are reiterated four times within the Pseudomonas fluorescens subsp. cellulosa...

The non-catalytic cellulose-binding domain of a novel cellulase from Pseudomonas fluorescens subsp. cellulosa is important for the efficient hydrolysis of Avicel.

Hall, J, Black, G W, Ferreira, L M, Millward-Sadler, S J, Ali, B R, Hazlewood, G P, ...

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA, constructed in lambda ZAPII, was screened for carboxymethyl-cellulase activity. The pseudomonad insert from a recombinant phage...

Novel cellulose-binding domains, NodB homologues and conserved modular architecture in xylanases from the aerobic soil bacteria Pseudomonas fluorescens subsp. cellulosa and Cellvibrio mixtus.

Millward-Sadler, S J, Davidson, K, Hazlewood, G P, Black, G W, Gilbert, H J, Clarke, J H

To test the hypothesis that selective pressure has led to the retention of cellulose-binding domains (CBDs) by hemicellulase enzymes from aerobic bacteria, four new xylanase (xyn) genes from two...

A non-modular endo-beta-1,4-mannanase from Pseudomonas fluorescens subspecies cellulosa.

Braithwaite, K L, Black, G W, Hazlewood, G P, Ali, B R, Gilbert, H J

Pseudomonas fluorescens subsp. cellulosa when cultured in the presence of carob galactomannan degraded the polysaccharide. To isolate gene(s) from P. fluorescens subsp. cellulosa encoding...

Evidence for a general role for non-catalytic thermostabilizing domains in xylanases from thermophilic bacteria.

Fontes, C M, Hazlewood, G P, Morag, E, Hall, J, Hirst, B H, Gilbert, H J

A genomic library of Clostridium thermocellum DNA constructed in lambda ZAPII was screened for xylanase-expressing clones. Cross-hybridization experiments revealed a new xylanase gene isolated from...

A modular xylanase containing a novel non-catalytic xylan-specific binding domain.

Black, G W, Hazlewood, G P, Millward-Sadler, S J, Laurie, J I, Gilbert, H J

Xylanase D (XYLD) from Cellulomonas fimi contains a C-terminal cellulose-binding domain (CBD) and an internal domain that exhibits 65% sequence identity with the C-terminal CBD. Full-length XYLD...

Intronless celB from the anaerobic fungus Neocallimastix patriciarum encodes a modular family A endoglucanase.

Zhou, L, Xue, G P, Orpin, C G, Black, G W, Gilbert, H J, Hazlewood, G P

The cDNA designated celB from the anaerobic rumen fungus Neocallimastix patriciarum contained a single open reading frame of 1422 bp coding for a protein (CelB) of M(r) 53,070. CelB expressed by...

Xylanase B from Neocallimastix patriciarum contains a non-catalytic 455-residue linker sequence comprised of 57 repeats of an octapeptide.

Black, G W, Hazlewood, G P, Xue, G P, Orpin, C G, Gilbert, H J

A Neocallimastix patriciarum cDNA library was screened for xylanase-expressing clones, which were distinct from the previously characterized N. patriciarum xynA cDNA encoding xylanase A. A single...

Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes.

Kellett, L E, Poole, D M, Ferreira, L M, Durrant, A J, Hazlewood, G P, Gilbert, H J

The complete nucleotide sequence of the Pseudomonas fluorescens subsp. cellulosa xynB gene, encoding an endo-beta-1,4-xylanase (xylanase B; XYLB) has been determined. The structural gene consists of...

The non-catalytic C-terminal region of endoglucanase E from Clostridium thermocellum contains a cellulose-binding domain.

Durrant, A J, Hall, J, Hazlewood, G P, Gilbert, H J

Mature endoglucanase E (EGE) from Clostridium thermocellum consists of 780 amino acid residues and has an Mr of 84,016. The N-terminal 334 amino acids comprise a functional catalytic domain....

Characterization of hybrid proteins consisting of the catalytic domains of Clostridium and Ruminococcus endoglucanases, fused to Pseudomonas non-catalytic cellulose-binding domains.

Poole, D M, Durrant, A J, Hazlewood, G P, Gilbert, H J

The N-terminal 160 or 267 residues of xylanase A from Pseudomonas fluorescens subsp. cellulosa, containing a non-catalytic cellulose-binding domain (CBD), were fused to the N-terminus of the...

The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains.

Ferreira, L M, Hazlewood, G P, Barker, P J, Gilbert, H J

A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was constructed in pUC18 and Escherichia coli recombinants expressing 4-methylumbelliferyl beta-D-cellobioside-hydrolysing activity...

A new series of long-chain dicarboxylic acids with vicinal dimethyl branching found as major components of the lipids of Butyrivibrio spp.

Klein, R A, Hazlewood, G P, Kemp, P, Dawson, R M

1. Some members of the genus Butyrivibrio, including a general fatty acid auxotroph (strain S2), contain as a major part of their complex lipids a high-molecular-weight component that is probably...

Complex lipids of a lipolytic and general-fatty-acid-requiring Butyrivibrio sp. isolated from the ovine rumen.

Hazlewood, G P, Clarke, N G, Dawson, R M

The complex lipids of the naturally-occurring general-fatty-acid-auxotroph Butyrivibrio S2 [Hazlewood & Dawson (1979) J. Gen. Microbiol. 112, 15-27] grown with palmitic acid as sole fatty-acid...

Structure of diabolic acid-containing phospholipids isolated from Butyrivibrio sp.

Clarke, N G, Hazlewood, G P, Dawson, R M

The structures of the diabolic acid-containing phospholipids of Buryrivibrio S2 grown in the presence of palmitic acid have been investigated. Generally they consist of two conventional bacterial...

Intermolecular transacylation of phosphatidylethanolamine by a Butyrivibrio sp.

Hazlewood, G P, Dawson, R M

1. Washed cells and supernatant from a culture of a Butyrivibrio sp. carry out the intermolecular transacylation reaction 2 phosphatidylethanolamine leads to...

A phospholipid-deacylating system of bacteria active in a frozen medium.

Hazlewood, G P, Dawson, R M

A phosphatidylcholine-deacylating system present in a Butyrivibrio species (probably fibrisolvens) shows appreciable activity at low temperatures with a maximum hydrolysis rate at--10 degrees C. 2....

A protein targeting signal that functions in polarized epithelial cells in vivo.

Ali, S, Hall, J, Hazlewood, G P, Hirst, B H, Gilbert, H J

Eukaryotic membrane-associated polypeptides often contain a glycosylphosphatidylinositol (GPI) anchor that signals the attachment of GPI lipids to these proteins. The GPI anchor can function as a...

Evidence that linker sequences and cellulose-binding domains enhance the activity of hemicellulases against complex substrates.

Black, G W, Rixon, J E, Clarke, J H, Hazlewood, G P, Theodorou, M K, Morris, P, ...

Xylanase A (XYLA) and arabinofuranosidase C (XYLC) from Pseudomonas fluorescens subsp. cellulosa are modular enzymes consisting of discrete cellulose-binding domains (CBDs) and catalytic domains...

Arabinanase A from Pseudomonas fluorescens subsp. cellulosa exhibits both an endo- and an exo- mode of action.

McKie, V A, Black, G W, Millward-Sadler, S J, Hazlewood, G P, Laurie, J I, Gilbert, H J

Pseudomonas fluorescens subsp. cellulosa expressed arabinanase activity when grown on media supplemented with arabinan or arabinose. Arabinanase activity was not induced by the inclusion of other...

Pseudomonas cellulose-binding domains mediate their effects by increasing enzyme substrate proximity.

Bolam, D N, Ciruela, A, McQueen-Mason, S, Simpson, P, Williamson, M P, Rixon, J E, ...

To investigate the mode of action of cellulose-binding domains (CBDs), the Type II CBD from Pseudomonas fluorescens subsp. cellulosa xylanase A (XYLACBD) and cellulase E (CELECBD) were expressed as...

Homologous xylanases from Clostridium thermocellum: evidence for bi-functional activity, synergism between xylanase catalytic modules and the presence of xylan-binding domains in enzyme complexes.

Fernandes, A C, Fontes, C M, Gilbert, H J, Hazlewood, G P, Fernandes, T H, Ferreira, L M

Clostridium thermocellum produces a consortium of plant-cell-wall hydrolases that form a cell-bound multi-enzyme complex called the cellulosome. In the present study two similar xylanase genes, xynU...

The type II and X cellulose-binding domains of Pseudomonas xylanase A potentiate catalytic activity against complex substrates by a common mechanism.

Gill, J, Rixon, J E, Bolam, D N, McQueen-Mason, S, Simpson, P J, Williamson, M P, ...

Xylanase A (Pf Xyn10A), in common with several other Pseudomonas fluorescens subsp. cellulosa polysaccharidases, consists of a Type II cellulose-binding domain (CBD), a catalytic domain (Pf...

A modular cinnamoyl ester hydrolase from the anaerobic fungus Piromyces equi acts synergistically with xylanase and is part of a multiprotein cellulose-binding cellulase-hemicellulase complex.

Fillingham, I J, Kroon, P A, Williamson, G, Gilbert, H J, Hazlewood, G P

A collection of clones, isolated from a Piromyces equi cDNA expression library by immunoscreening with antibodies raised against affinity purified multienzyme fungal cellulase-hemicellulase complex,...