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Title: The complex physiology of Cellvibrio japonicus xylan degradation relies on a single cytoplasmic β-xylosidase for xylo-oligosaccharide utilization

Abstract

Lignocellulose degradation by microbes plays a central role in global carbon cycling, human gut metabolism, and renewable energy technologies. While considerable effort has been put into understanding the biochemical aspects of lignocellulose degradation, much less work has been done to understand how these enzymes work in an in vivo context. Here in this paper, we report a systems level study of xylan degradation in the saprophytic bacterium Cellvibrio japonicus. Transcriptome analysis indicated seven genes that encode carbohydrate active enzymes were up-regulated during growth with xylan containing media. In-frame deletion analysis of these genes found that only gly43F is critical for utilization of xylo-oligosaccharides, xylan, and arabinoxylan. Heterologous expression of gly43F was sufficient for the utilization of xylo-oligosaccharides in Escherichia coli. Additional analysis found that the xyn11A, xyn11B, abf43L, abf43K, and abf51A gene products were critical for utilization of arabinoxylan. Furthermore, a predicted transporter (CJA_1315) was required for effective utilization of xylan substrates, and we propose this unannotated gene be called xntA (xylan transporter A). Our major findings are 1) C. japonicus employs both secreted and surface associated enzymes for xylan degradation, which differs from the strategy used for cellulose degradation, and 2) a single cytoplasmic β-xylosidase is essential for themore » utilization of xylo-oligosaccharides.« less

Authors:
 [1];  [1];  [1];  [1]; ORCiD logo [1]
  1. Univ. of Maryland Baltimore County (UMBC), Baltimore, MD (United States). Dept. of Biological Sciences
Publication Date:
Research Org.:
Univ. of Maryland Baltimore County (UMBC), Baltimore, MD (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23). Biological Systems Science Division; USDOE
OSTI Identifier:
1415270
Alternate Identifier(s):
OSTI ID: 1416401
Grant/Contract Number:  
SC0014183
Resource Type:
Accepted Manuscript
Journal Name:
Molecular microbiology
Additional Journal Information:
Journal Volume: 107; Journal Issue: 5; Journal ID: ISSN 0950-382X
Publisher:
Wiley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; β-xylosidase; Cellvibrio japonicus; polysaccharide degradation; xylan; xylanase

Citation Formats

Blake, Andrew D., Beri, Nina R., Guttman, Hadassa S., Cheng, Raymond, and Gardner, Jeffrey G. The complex physiology of Cellvibrio japonicus xylan degradation relies on a single cytoplasmic β-xylosidase for xylo-oligosaccharide utilization. United States: N. p., 2017. Web. doi:10.1111/mmi.13903.
Blake, Andrew D., Beri, Nina R., Guttman, Hadassa S., Cheng, Raymond, & Gardner, Jeffrey G. The complex physiology of Cellvibrio japonicus xylan degradation relies on a single cytoplasmic β-xylosidase for xylo-oligosaccharide utilization. United States. doi:10.1111/mmi.13903.
Blake, Andrew D., Beri, Nina R., Guttman, Hadassa S., Cheng, Raymond, and Gardner, Jeffrey G. Thu . "The complex physiology of Cellvibrio japonicus xylan degradation relies on a single cytoplasmic β-xylosidase for xylo-oligosaccharide utilization". United States. doi:10.1111/mmi.13903. https://www.osti.gov/servlets/purl/1415270.
@article{osti_1415270,
title = {The complex physiology of Cellvibrio japonicus xylan degradation relies on a single cytoplasmic β-xylosidase for xylo-oligosaccharide utilization},
author = {Blake, Andrew D. and Beri, Nina R. and Guttman, Hadassa S. and Cheng, Raymond and Gardner, Jeffrey G.},
abstractNote = {Lignocellulose degradation by microbes plays a central role in global carbon cycling, human gut metabolism, and renewable energy technologies. While considerable effort has been put into understanding the biochemical aspects of lignocellulose degradation, much less work has been done to understand how these enzymes work in an in vivo context. Here in this paper, we report a systems level study of xylan degradation in the saprophytic bacterium Cellvibrio japonicus. Transcriptome analysis indicated seven genes that encode carbohydrate active enzymes were up-regulated during growth with xylan containing media. In-frame deletion analysis of these genes found that only gly43F is critical for utilization of xylo-oligosaccharides, xylan, and arabinoxylan. Heterologous expression of gly43F was sufficient for the utilization of xylo-oligosaccharides in Escherichia coli. Additional analysis found that the xyn11A, xyn11B, abf43L, abf43K, and abf51A gene products were critical for utilization of arabinoxylan. Furthermore, a predicted transporter (CJA_1315) was required for effective utilization of xylan substrates, and we propose this unannotated gene be called xntA (xylan transporter A). Our major findings are 1) C. japonicus employs both secreted and surface associated enzymes for xylan degradation, which differs from the strategy used for cellulose degradation, and 2) a single cytoplasmic β-xylosidase is essential for the utilization of xylo-oligosaccharides.},
doi = {10.1111/mmi.13903},
journal = {Molecular microbiology},
number = 5,
volume = 107,
place = {United States},
year = {2017},
month = {12}
}

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Figures / Tables:

Fig. 1. C. japonicus CAZyme genes up-regulated during exponential growth using xylan. Fig. 1. C. japonicus CAZyme genes up-regulated during exponential growth using xylan.: C. japonicus CAZyme genes up-regulated during exponential growth using xylan. The volcano plot summarizes RNAseq comparative analysis of gene expression for xylan grown cells versus glucose grown cells. Fold change (log2 scale) is on the x-axis, with positivemore » values depicting genes up-regulated on xylan-containing media. The vertical dashed lines indicate a 2-fold cut-off. The p-value of the RNAseq analysis (-log10 scale) is on the y-axis, and the horizontal dashed line indicates a p≤0.01. The seven up regulated CAZyme genes are shown in color, with associated gene names in the key. Exact fold-change and p-values for these seven genes can be found in Table S1A.« less

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