Fine-tuning of xylose metabolism in genetically engineered Saccharomyces cerevisiae by scattered integration of xylose assimilation genes.
Manipulation of multiple genes is a common experience in metabolic engineering and synthetic biology studies. Chromosome integration of multiple genes in one single position is always performed, however, there is so far no study on the integration of multiple genes separately in various positions (here in after referred to as "scattered integration") and its effect on fine-tuning of cellular metabolism. In this study, scattered integration of the xylose assimilation genes PsXR, PsXDH and ScXK was investigated in Saccharomyces cerevisiae, and transcription analysis of these genes as well as their enzyme activities were compared with those observed when the genes were integrated into one single site (defined as "tandem integration" here). Not only notable differences in transcription levels and enzyme activities were observed when the genes were integrated by the two strategies, but also change of the cofactor preference of PsXR gene was validated. Xylose fermentation was further studied with the strains developed with these strategies, and elevated xylose utilization rate was obtained in the scattered integration strain. These results proved that by positioning multiple genes on different chromosomes, fine-tuning of cellular metabolism could be achieved in recombinant S. cerevisiae.
School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China.
Qi
Q
Zuo
Xin-Qing
XQ
Zhao
Liang
L
Xiong
Hai-Jun
HJ
Liu
You-Hai
YH
Xu
Shi-Yang
SY
Hu
Zhong-Yi
ZY
Ma
Qing-Wei
QW
Zhu
Feng-Wu
FW
Bai
D000449:Aldehyde Reductase / Q000096:biosynthesis / Q000235:genetics*; D015825:Chromosomes, Fungal / Q000235:genetics; D050542:D-Xylulose Reductase / Q000096:biosynthesis / Q000235:genetics*; D018274:Electroporation; D005285:Fermentation; D060847:Metabolic Engineering / Q000379:methods*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis / Q000235:genetics*; D010843:Pichia / Q000201:enzymology / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D014994:Xylose / Q000378:metabolism*
D029701:Saccharomyces cerevisiae Proteins; D014994:Xylose; D000449:Aldehyde Reductase; D050542:D-Xylulose Reductase; D017853:Phosphotransferases (Alcohol Group Acceptor); C032700:xylulokinase
Saccharomyces cerevisiae; Scattered chromosomal integration; Tandem chromosomal integration; Xylose assimilation genes