Interactions of Endoglucanases with Amorphous Cellulose Films Resolved by Neutron Reflectometry and Quartz Crystal Microbalance with Dissipation Monitoring

Journal Article · Thu May 03 04:00:00 EDT 2012 · Langmuir

DOI:https://doi.org/10.1021/la300955q· OSTI ID:1042855

Cheng, Gang ^[1]; Datta, Supratim ^[2]; Liu, Zelin ^[3]; Wang, Chao ^[3]; Murton, Jaclyn K. ^[4]; Brown, Page A. ^[4]; Jablin, Michael S. ^[5]; Dubey, Manish ^[5]; Majewski, Jaroslaw ^[5]; Halbert, Candice E. ^[6]; Browning, James F. ^[6]; Esker, Alan R. ^[3]; Watson, Brian J. ^[7]; Zhang, Haito ^[7]; Hutcheson, Steven W. ^[7]; Huber, Dale L. ^[8]; Sale, Kenneth L. ^[1]; Simmons, Blake A. ^[1]; Kent, Michael S. ^[1]

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1. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States); Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
2. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
3. Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States). Dept. of Chemistry
4. Sandia National Lab. (SNL-CA), Livermore, CA (United States); Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
5. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
6. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS)
7. Univ. of Maryland, College Park, MD (United States). Dept. of Cell Biology and Molecular Genetics
8. Sandia National Lab. (SNL-CA), Livermore, CA (United States); Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

A study of the interaction of four endoglucanases with amorphous cellulose films by neutron reflectometry (NR) and quartz crystal microbalance with dissipation monitoring (QCM-D) is reported. The endoglucanases include a mesophilic fungal endoglucanase (Cel45A from H. insolens), a processive endoglucanase from a marine bacterium (Cel5H from S. degradans), and two from thermophilic bacteria (Cel9A from A. acidocaldarius and Cel5A from T. maritima). The use of amorphous cellulose is motivated by the promise of ionic liquid pretreatment as a second generation technology that disrupts the native crystalline structure of cellulose. The endoglucanases displayed highly diverse behavior. Cel45A and Cel5H, which possess carbohydrate-binding modules (CBMs), penetrated and digested within the bulk of the films to a far greater extent than Cel9A and Cel5A, which lack CBMs. While both Cel45A and Cel5H were active within the bulk of the films, striking differences were observed. With Cel45A, substantial film expansion and interfacial broadening were observed, whereas for Cel5H the film thickness decreased with little interfacial broadening. Lastly, these results are consistent with Cel45A digesting within the interior of cellulose chains as a classic endoglucanase, and Cel5H digesting predominantly at chain ends consistent with its designation as a processive endoglucanase.

• Research Organization: Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source
• Sponsoring Organization: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
• DOE Contract Number: AC05-00OR22725; AC02-05CH11231; W-7405-ENG-36; SC0001090; AC52-06NA25396; AC04-94AL85000
• OSTI ID: 1042855
• Journal Information: Langmuir, Journal Name: Langmuir Journal Issue: 22 Vol. 28; ISSN 0743-7463
• Publisher: American Chemical Society
• Country of Publication: United States
• Language: English

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59 BASIC BIOLOGICAL SCIENCES