PMID: 9537366Apr 16, 1998Paper

Roles of the catalytic domain and two cellulose binding domains of Thermomonospora fusca E4 in cellulose hydrolysis

Journal of Bacteriology
D IrwinD B Wilson

Abstract

Thermomonospora fusca E4 is an unusual 90.4-kDa endocellulase comprised of a catalytic domain (CD), an internal family IIIc cellulose binding domain (CBD), a fibronectinlike domain, and a family II CBD. Constructs containing the CD alone (E4-51), the CD plus the family IIIc CBD (E4-68), and the CD plus the fibronectinlike domain plus the family II CBD (E4-74) were made by using recombinant DNA techniques. The activities of each purified protein on bacterial microcrystalline cellulose (BMCC), filter paper, swollen cellulose, and carboxymethyl cellulose were measured. Only the whole enzyme, E4-90, could reach the target digestion of 4.5% on filter paper. Removal of the internal family IIIc CBD (E4-51 and E4-74) decreased activity markedly on every substrate. E4-74 did bind to BMCC but had almost no hydrolytic activity, while E4-68 retained 32% of the activity on BMCC even though it did not bind. A low-activity mutant of one of the catalytic bases, E4-68 (Asp55Cys), did bind to BMCC, although E4-51 (Asp55Cys) did not. The ratios of soluble to insoluble reducing sugar produced after filter paper hydrolysis by E4-90, E4-68, E4-74, and E4-51 were 6.9, 3.5, 1.3, and 0.6, respectively, indicating that the family IIIc CBD is important f...Continue Reading

References

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Jun 5, 1983·Journal of Molecular Biology·D Hanahan
Mar 14, 1995·Biochemistry·S ZhangD B Wilson
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Citations

Oct 25, 2002·The Journal of Biological Chemistry·Henri-Pierre FierobeJean-Pierre Bélaïch
Apr 10, 2004·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·David B Wilson
Feb 27, 2007·The Journal of Biological Chemistry·Breeanna R UrbanowiczJocelyn K C Rose
Oct 2, 2007·Applied and Environmental Microbiology·Florence MingardonHenri-Pierre Fierobe
Oct 15, 2008·Journal of Molecular Recognition : JMR·Osmair V OliveiraRoberto D Lins
Dec 21, 2010·Applied and Environmental Microbiology·Florence MingardonFrances H Arnold
Jun 21, 2017·Preparative Biochemistry & Biotechnology·Cristian Alarid-García, Eleazar Maximo Escamilla-Silva
Jul 9, 2003·European Journal of Biochemistry·Diana C IrwinDavid B Wilson
Mar 21, 2007·Applied and Environmental Microbiology·Yongchao LiDavid B Wilson
Jun 17, 2008·Biotechnology and Bioengineering·Felix MoserDavid B Wilson
Mar 10, 2005·Microbiology and Molecular Biology Reviews : MMBR·Arnold L DemainJ H David Wu
Jul 1, 2008·FEBS Letters·Morten Munch NielsenBirte Svensson
Mar 30, 2013·Critical Reviews in Microbiology·Eva Maria Gomez del Pulgar, Anas Saadeddin
Dec 4, 2012·Applied and Environmental Microbiology·Fei Zheng, Shaojun Ding
Jan 15, 2015·Parasites & Vectors·Sahreena LakhundiNaveed Ahmed Khan
Jul 15, 2015·Applied Microbiology and Biotechnology·Cong ZhangXuemei Lu
Aug 6, 2019·Applied Biochemistry and Biotechnology·Shanshan Wu, Shufang Wu
Feb 29, 2008·Biotechnology and Bioengineering·Yongchao Li, David B Wilson
Dec 18, 2004·Biotechnology Letters·Lars Hildén, Gunnar Johansson
Oct 19, 2004·Annual Review of Microbiology·Edward A BayerRaphael Lamed
Mar 22, 2014·Applied Microbiology and Biotechnology·Cong ZhangXuemei Lu

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