Cloning, DNA sequencing, and expression of the gene encoding Clostridium thermocellum cellulase CelJ, the largest catalytic component of the cellulosome

Ahsan, Mohammad Mainui; Kimura, Takeshi; Karita, Shuichi; Sakka, Kazuo; Ohmiya, Kunio

doi:10.1128/jb.178.19.5732-5740.1996

Cited by 72 publications

(44 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For example, in C. thermocellum, the dockerin of Cel9D-Cel44A (formerly CelJ [1]) does not seem to bind to cohesins 1, 4, and 7 (136) or to cohesins 2 and 3 (362) of CipA. Its binding specificity remains to be determined.…”

Section: Assembly Of the Cellulosomementioning

confidence: 99%

“…CelF, the CelS equivalent in C. cellulolyticum, also displays significant activity in reducing the viscosity of a CMC solution (271). Some of the cloned endoglucanase genes are celA (26), celB (105), celC (298), celD (143,326), celE (113), celF (245), celG (188), celH (361), celI (101,123,373), celJ (encoding component S2 of the complex) (1,8), celM (162), celN (373), celQ (9), celT (172), and celX (113).…”

Section: Genes and Enzymesmentioning

confidence: 99%

See 1 more Smart Citation

Cellulase, Clostridia, and Ethanol

Demain

Newcomb

2005

Microbiol Mol Biol Rev

810

579

View full text Add to dashboard Cite

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.

show abstract

Section: Assembly Of the Cellulosomementioning

confidence: 99%

Section: Genes and Enzymesmentioning

confidence: 99%

Cellulase, Clostridia, and Ethanol

Demain

Newcomb

2005

Microbiol Mol Biol Rev

810

579

View full text Add to dashboard Cite

show abstract

“…In addition to cell surface proteins, the PKD domain is found in many biopolymer hydrolases, such as chitinases (4,5), celluloses (6), and proteases (7)(8)(9), suggesting that it may play an important role in biopolymer degradation. The structures of three PKD domains have been solved, which show that, though their sequences are different, they all adopt a ␤-helix fold and a conserved sequence area with two Trp residues in the hydrophobic core (2,3).…”

mentioning

confidence: 99%

Mechanistic Insight into the Function of the C-terminal PKD Domain of the Collagenolytic Serine Protease Deseasin MCP-01 from Deep Sea Pseudoalteromonas sp. SM9913

Wang

Zhao

et al. 2010

Journal of Biological Chemistry

View full text Add to dashboard Cite

Deseasin MCP-01 is a bacterial collagenolytic serine protease. Its catalytic domain alone can degrade collagen, and its C-terminal PKD domain is a collagen-binding domain (CBD) that can improve the collagenolytic efficiency of the catalytic domain by an unknown mechanism. Here, scanning electron microscopy (SEM), atomic force microscopy (AFM), zeta potential, and circular dichroism spectroscopy were used to clarify the functional mechanism of the PKD domain in MCP-01 collagenolysis. The PKD domain observably swelled insoluble collagen. Its collagenswelling ability and its improvement to the collagenolysis of the catalytic domain are both temperature-dependent. SEM observation showed the PKD domain swelled collagen fascicles with an increase of their diameter from 5.3 m to 8.8 m after 1 h of treatment, and the fibrils forming the fascicles were dispersed. AFM observation directly showed that the PKD domain bound collagen, swelled the microfibrils, and exposed the monomers. The PKD mutant W36A neither bound collagen nor disturbed its structure. Zeta potential results demonstrated that PKD treatment increased the net positive charges of the collagen surface. PKD treatment caused no change in the content or the thermostability of the collagen triple helix. Furthermore, the PKD-treated collagen could not be degraded by gelatinase. Therefore, though the triple helix monomers were exposed, the PKD domain could not unwind the collagen triple helix. Our study reveals the functional mechanism of the PKD domain of the collagenolytic serine protease MCP-01 in collagen degradation, which is distinct from that of the CBDs of mammalian matrix metalloproteases. The polycystic kidney disease (PKD)3 domain is an 80 -90-amino acid module originally found in the human PKD1 gene encoding the cell surface glycoprotein polycystin-1 (1), and later, in many surface layer proteins of archaebacteria (2, 3). In addition to cell surface proteins, the PKD domain is found in many biopolymer hydrolases, such as chitinases (4, 5), celluloses (6), and proteases (7-9), suggesting that it may play an important role in biopolymer degradation. The structures of three PKD domains have been solved, which show that, though their sequences are different, they all adopt a ␤-helix fold and a conserved sequence area with two Trp residues in the hydrophobic core (2, 3). However, the functional mechanism of the PKD domain in biopolymer hydrolases is largely unknown, except that the PKD domains in chitinase ChiA and collagenolytic serine protease deseasin MCP-01 are both reported to function as a binding domain (5, 10). Genome sequence analysis and experimental results show that the PKD domain is common in the hydrolases of marine heterotrophic bacteria. For example, Gramella forsetii has 14 exported proteins with PKD domains (11). Chitin-binding protease AprIV, deseasin MCP-01, and many other marine deseasins contain one or two PKD domains (5, 9). Therefore, elucidating the function and the mechanism of action of the PKD domains in these hydrolases would hav...

show abstract

“…These observations are consistent with the phenomenon that rXyn11ADoc containing the "ST" motif can interact with the cohesin domains of C. josui CipA in addition to C. thermocellum CipA, although it is unclear why only the dockerin domain of Xyn11A can recognize the noncognate cohesin domains as counterparts. Before starting this study, we expected that the dockerin domain of Cel9D-Cel44A would interact with the cohesin domains of C. josui CipA, because Cel9D-Cel44A was shown not to react with the second and third cohesin polypeptides from C. thermocellum CipA (26) and the amino acid sequence of its dockerin domain was unusual (14), i.e. it contained A and V in the conserved positions of the first segment.…”

Section: Discussionmentioning

confidence: 99%

“…These observations suggested that the dockerin domains of C thermocellum Cel9A, Cel5C, and Cel48A could bind to any cohesin domains in the cognate CipA with a similar affinity. On the other hand, the second and third cohesin domains of C. thermocellum CipA interacted with all the catalytic subunits, except for the largest catalytic subunit now known as Cel9D-Cel44A (formerly CelJ) (14), of the cellulosome in an affinity blotting analysis (14), suggesting that generally there is no selectivity in the interactions among cohesin and dockerin domains within the same species, but that there may be at least one exception. Because Cel9D-Cel44A is known as a component of the cellulosome, the cohesin domains, except for the second and third ones, should associate with the dockerin domain of Cel9D-Cel44A, i.e.…”

mentioning

confidence: 98%

Cohesin-Dockerin Interactions within and between Clostridium josui and Clostridium thermocellum

Jindou

Soda

Karita

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

The cellulosome components are assembled into the cellulosome complex by the interaction between one of the repeated cohesin domains of a scaffolding protein and the dockerin domain of an enzyme component. We prepared five recombinant cohesin polypeptides of the Clostridium thermocellum scaffolding protein CipA, two dockerin polypeptides of C. thermocellum Xyn11A and Xyn10C, four cohesin polypeptides of Clostridium josui CipA, and two dockerin polypeptides of C. josui Aga27A and Cel8A, and qualitatively and quantitatively examined the cohesin-dockerin interactions within C. thermocellum and C. josui, respectively, and the species specificity of the cohesin-dockerin interactions between these two bacteria. Surface plasmon resonance (SPR) analysis indicated that there was a certain selectivity, with a maximal 34-fold difference in the K D values, in the cohesin-dockerin interactions within a combination of C. josui, although this was not detected by qualitative analysis. Affinity blotting analysis suggested that there was at least one exception to the species specificity in the cohesin-dockerin interactions, although species specificity was generally conserved among the cohesin and dockerin polypeptides from C. thermocellum and C. josui, i.e. the dockerin polypeptides of C. thermocellum Xyn11A exceptionally bound to the cohesin polypeptides from C. josui CipA. SPR analysis confirmed this exceptional binding. We discuss the relationship between the species specificity of the cohesin-dockerin binding and the conserved amino acid residues in the dockerin domains.

show abstract

Cloning, DNA sequencing, and expression of the gene encoding Clostridium thermocellum cellulase CelJ, the largest catalytic component of the cellulosome

Cited by 72 publications

References 53 publications

Cellulase, Clostridia, and Ethanol

Cellulase, Clostridia, and Ethanol

Mechanistic Insight into the Function of the C-terminal PKD Domain of the Collagenolytic Serine Protease Deseasin MCP-01 from Deep Sea Pseudoalteromonas sp. SM9913

Cohesin-Dockerin Interactions within and between Clostridium josui and Clostridium thermocellum

Contact Info

Product

Resources

About