Research Article
Genetic Engineering and Improvement of a Zymomonas mobilis for Arabinose Utilization and Its Performance on Pretreated Corn Stover Hydrolyzate
Yat-Chen Chou*, Jeffrey Linger, Shihui Yang and Min ZhangNational Bioenergy Center, National Renewable Energy Laboratory, Golden, CO 80401, USA
- Corresponding Author:
- Yat-Chen Chou
National Bioenergy Center
National Renewable Energy Laboratory
15013 Denver West Parkway Golden, CO 80401,USA
Tel: 3033847766
E-mail: yat.chen.chou@nrel.gov
Received date:: March 18, 2015; Accepted date:: April 21, 2015; Published date:: April 28, 2015
Citation: Chou YC, Linger J, Yang S, Zhang M (2015) Genetic Engineering and Improvement of a Zymomonas mobilis for Arabinose Utilization and Its Performance on Pretreated Corn Stover Hydrolyzate. J Biotechnol Biomater 5:179. doi:https://dx.doi.org/10.4172/biotechnology-biomaterials.1000179
Copyright: © 2015 Chou YC, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
A glucose, xylose and arabinose utilizing Zymomonas mobilis strain was constructed by incorporating arabinose catabolic pathway genes, araBAD encoding L-ribulokinase, L-arabinose isomerase and L-ribulose-5-phosphate- 4-epimerase in a glucose, xylose co-fermenting host, 8b, using a transposition integration approach. Further improvement on this arabinose-capable integrant, 33C was achieved by applying a second transposition to create a genomic knockout (KO) mutant library. Using arabinose as a sole carbon source and a selection pressure, the KO library was subjected to a growth-enrichment process involving continuous sub-culturing for over 120 generations. Strain 13-1-17, isolated from such process demonstrated significant improvement in metabolizing arabinose in a dilute acid pretreated, saccharified corn stover slurry. Through Next Generation Sequencing (NGS) analysis, integration sites of the transposons were identified. Furthermore, multiple additional point mutations (SNPs: Single Nucleotide Polymorphisms) were discovered in 13-1-17, affecting genes araB and RpiB in the genome. We speculate that these mutations may have impacted the expression of the enzymes coded by these genes, ribulokinase and Ribose 5-P-isomerase, thus attributing to the improvement of the arabinose utilization.