caa a22 4500 606211977 CHVBK 20210128101040.0 cr unu---uuuuu 210128e20150101xx s 000 0 eng 10.1007/s10295-014-1530-4 doi (NATIONALLICENCE)springer-10.1007/s10295-014-1530-4 Integrative expression vectors for overexpression of xylitol dehydrogenase ( XYL2 ) in Osmotolerant yeast, Candida glycerinogenes WL2002-5 [Elektronische Daten] [Cheng Zhang, Hong Zong, Bin Zhuge, Xinyao Lu, Huiying Fang, Jian Zhuge] Yeasts are excellent hosts for the production of recombinant proteins. Candida glycerinogenes WL2002-5, an osmotolerant yeast with extremely high glycerol productivity, provides an attractive eukaryotic expression platform. The integrative vectors PURGAP-gfp and PURGPD-gfp harbouring phleomycin-resistance coding sequence and GFP coding sequence with PCgGAP, PCgGPD promoter, respectively, were constructed. The recombinant plasmid PURPpGAP-gfp with the promoter PPpGAP based on the sequence of Pichia pastoris GAPDH gene and the plasmid PURScGAP-gfp with the promoter PScGAP from Saccharomyces cerevisiae were constructed. After transformation, the copy number of gfp gene, which determined using fluorescent quantitative real-time polymerase chain reaction (FQ-RTPCR) in genome of C. glycerinogenes is 1. Expressions of gfp at different levels were conducted using different promoters by osmotic stress containing NaCl or glucose for the recombinant strains. In this study, C. glycerinogenes WL2002-5, expressing xylitol dehydrogenase (XYL2) gene from Pichia stipitis, has the ability to produce glycerol from xylose entered into pentose phosphate pathway. Two recombinant strains of PURGAPX, PURGPDX with XYL2 overexpression were constructed to ferment a mixture of glucose and xylose simultaneously in batch fermentation. Compared to C. glycerinogenes WL2002-5 strain, glycerol production from xylose in strains PURGAPX, PURGPDX were increased by 95.9 and 121.1%, respectively. Society for Industrial Microbiology and Biotechnology, 2014 Candida glycerinogenes nationallicence Integrative vector nationallicence Phleomycin nationallicence Green fluorescent protein nationallicence Genetic transformation system nationallicence Xylitol dehydrogenase nationallicence Zhang Cheng The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Zong Hong The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Zhuge Bin The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Lu Xinyao The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Fang Huiying The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Zhuge Jian The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/1(2015-01-01), 113-124 1367-5435 42:1<113 2015 42 10295 https://doi.org/10.1007/s10295-014-1530-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-014-1530-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Cheng The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Zong Hong The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Zhuge Bin The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Lu Xinyao The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Fang Huiying The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 700 1- Zhuge Jian The Key Lab of Industrial Biotechnology of Ministry of Education, Research Centre of Industrial Microorganisms, School of Biotechnology, Jiangnan University, 214122, Wuxi, Jiangsu, People's Republic of China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/1(2015-01-01), 113-124 1367-5435 42:1<113 2015 42 10295