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Title: Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries

Abstract

Complete hydrolysis of cellulose has been noted as a key characteristic of biomass technology due to the limitation of industrial production hosts to use cellodextrin, the partial hydrolysis product of cellulose. Cellobiose, a β-1,4-linked glucose dimer, is a major cellodextrin of the enzymatic hydrolysis (via endoglucanase and exoglucanase) of cellulose. Conversion of cellobiose to glucose is executed by β-glucosidase. The complete extracellular hydrolysis of celluloses has several critical barriers in biomass technology. An alternative bioengineering strategy to make the bioprocessing less challenging is to engineer microbes with the abilities to hydrolyze and assimilate the cellulosic-hydrolysate cellodextrin. Microorganisms engineered to metabolize cellobiose rather than the monomeric glucose can provide several advantages for lignocellulose-based biorefineries. This review describes the recent advances and challenges in engineering efficient intracellular cellobiose metabolism in industrial hosts. This review also describes the limitations of and future prospectives in engineering intracellular cellobiose metabolism.

Authors:
 [1];  [1];  [2];  [1];  [3]
  1. Ulsan National Institute of Science and Technology (UNIST), Ulsan (Korea, Republic of)
  2. Joint BioEnergy Inst., Emeryville, CA (United States)
  3. Joint BioEnergy Inst., Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Technical Univ. of Denmark, Horsholm (Denmark). Novo Nordisk Foundation Center for Biosustainability; Synthetic Biology Engineering Research Center (Synberc), Berkeley, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1436614
DOE Contract Number:
AC02-05CH11231
Resource Type:
Journal Article
Resource Relation:
Journal Name: Bioresource Technology; Journal Volume: 239; Journal Issue: C
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Parisutham, Vinuselvi, Chandran, Sathesh-Prabu, Mukhopadhyay, Aindrila, Lee, Sung Kuk, and Keasling, Jay D. Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries. United States: N. p., 2017. Web. doi:10.1016/j.biortech.2017.05.001.
Parisutham, Vinuselvi, Chandran, Sathesh-Prabu, Mukhopadhyay, Aindrila, Lee, Sung Kuk, & Keasling, Jay D. Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries. United States. doi:10.1016/j.biortech.2017.05.001.
Parisutham, Vinuselvi, Chandran, Sathesh-Prabu, Mukhopadhyay, Aindrila, Lee, Sung Kuk, and Keasling, Jay D. Thu . "Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries". United States. doi:10.1016/j.biortech.2017.05.001.
@article{osti_1436614,
title = {Intracellular cellobiose metabolism and its applications in lignocellulose-based biorefineries},
author = {Parisutham, Vinuselvi and Chandran, Sathesh-Prabu and Mukhopadhyay, Aindrila and Lee, Sung Kuk and Keasling, Jay D.},
abstractNote = {Complete hydrolysis of cellulose has been noted as a key characteristic of biomass technology due to the limitation of industrial production hosts to use cellodextrin, the partial hydrolysis product of cellulose. Cellobiose, a β-1,4-linked glucose dimer, is a major cellodextrin of the enzymatic hydrolysis (via endoglucanase and exoglucanase) of cellulose. Conversion of cellobiose to glucose is executed by β-glucosidase. The complete extracellular hydrolysis of celluloses has several critical barriers in biomass technology. An alternative bioengineering strategy to make the bioprocessing less challenging is to engineer microbes with the abilities to hydrolyze and assimilate the cellulosic-hydrolysate cellodextrin. Microorganisms engineered to metabolize cellobiose rather than the monomeric glucose can provide several advantages for lignocellulose-based biorefineries. This review describes the recent advances and challenges in engineering efficient intracellular cellobiose metabolism in industrial hosts. This review also describes the limitations of and future prospectives in engineering intracellular cellobiose metabolism.},
doi = {10.1016/j.biortech.2017.05.001},
journal = {Bioresource Technology},
number = C,
volume = 239,
place = {United States},
year = {Thu May 04 00:00:00 EDT 2017},
month = {Thu May 04 00:00:00 EDT 2017}
}
  • The cellulolytic ruminal bacterium Ruminococcus flavefaciens FD-1 utilizes cellobiose but not glucose as a substrate for growth. Cellobiose uptake by R. flavefaciens FD-1 was measured under anaerobic conditions (N{sub 2}), using (G-{sup 3}H)cellobiose. The rate of cellobiose uptake for early- or late-log-phase cellobiose-grown cells was 9 nmol/min per mag of whole-cell protein. Cellobiose uptake was inhibited by electron transport inhibitors, iron-reactive compounds, proton ionophores, sulfhydryl inhibitors, N,N-dicyclohexylcarbodiimide, and NaF, as well as lasalocid and monensin. The results support the existence of an active transport system for cellobiose. Transport of (U-{sup 14}C)glucose was not detected with this system. Phosphorylation of cellobiosemore » was not by a phosphoenolpyruvate-dependent system. Cellobiose phosphorylase activity was detected by both a coupled spectrophotometric assay and a discontinuous assay. The enzyme was produced constitutively in cellobiose-grown cells at a specific activity of 329 nmol/min per mg of cell-free extract protein.« less
  • Liberation of fermentable sugars from recalcitrant biomass is among the most costly steps for emerging cellulosic ethanol production. Here we compared two pretreatment methods (dilute acid, DA, and cellulose solvent and organic solvent lignocellulose fractionation, COSLIF) for corn stover. At a high cellulase loading [15 filter paper units (FPUs) or 12.3 mg cellulase per gram of glucan], glucan digestibilities of the corn stover pretreated by DA and COSLIF were 84% at hour 72 and 97% at hour 24, respectively. At a low cellulase loading (5 FPUs per gram of glucan), digestibility remained as high as 93% at hour 24 formore » the COSLIF-pretreated corn stover but reached only {approx}60% for the DA-pretreated biomass. Quantitative determinations of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a non-hydrolytic recombinant protein TGC were measured for the first time. The COSLIF-pretreated corn stover had a CAC of 11.57 m{sup 2}/g, nearly twice that of the DA-pretreated biomass (5.89 m{sup 2}/g). These results, along with scanning electron microscopy images showing dramatic structural differences between the DA- and COSLIF-pretreated samples, suggest that COSLIF treatment disrupts microfibrillar structures within biomass while DA treatment mainly removes hemicellulose. Under the tested conditions COSLIF treatment breaks down lignocellulose structure more extensively than DA treatment, producing a more enzymatically reactive material with a higher CAC accompanied by faster hydrolysis rates and higher enzymatic digestibility.« less