Crz1p Regulates pH Homeostasis in Candida glabrata by Altering Membrane Lipid Composition.
The asexual facultative aerobic haploid yeastCandida glabratais widely used in the industrial production of various organic acids. To elucidate the physiological function of theC. glabratatranscription factor Crz1p (CgCrz1p) and its role in tolerance to acid stress, we deleted or overexpressed the corresponding gene,CgCRZ1Deletion ofCgCRZ1resulted in a 60% decrease in the dry weight of cells (DCW) and a 50% drop in cell viability compared with those of the wild type at pH 2.0. Expression of lipid metabolism-associated genes was also significantly downregulated. Consequently, the proportion of C18:1fatty acids, the ratio of unsaturated to saturated fatty acids, and the ergosterol content decreased by 30%, 46%, and 30%, respectively. Additionally, membrane integrity, fluidity, and H+-ATPase activity were reduced by 45%, 9%, and 50%, respectively. In contrast, overexpression of CgCrz1p increased C18:1and ergosterol contents by 16% and 40%, respectively. Overexpression also enhanced membrane integrity, fluidity, and H+-ATPase activity by 31%, 6%, and 20%, respectively. Moreover, in the absence of pH buffering, the DCW and pyruvate titers increased by 48% and 60%, respectively, compared to that of the wild type. Together, these results suggest that CgCrz1p regulates tolerance to acidic conditions by altering membrane lipid composition inC. glabrataIMPORTANCEThis study provides insight into the metabolism ofCandida glabrataunder acidic conditions, such as those encountered during the industrial production of organic acids. We found that overexpression of the transcription factor CgCrz1p improved viability, biomass, and pyruvate yields at a low pH. Analysis of plasma membrane lipid composition indicated that CgCrz1p might play an important role in its integrity and fluidity and that it enhanced the pumping of protons in acidic environments. We propose that altering the structure of the cell membrane may provide a successful strategy for increasingC. glabrataproductivity at a low pH.
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Dongni
http://orcid.org/0000-0002-1797-4368
D
Yan
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Xiaobao
X
Lin
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Yanli
Y
Qi
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Hui
H
Liu
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Xiulai
X
Chen
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, China mingll@jiangnan.edu.cn.|Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.|Laboratory of Food Microbial-Manufacturing Engineering, Jiangnan University, Wuxi, Jiangsu, China.
Liming
L
Liu
Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu, China.
Jian
J
Chen
Gene. 1991 May 15;101(1):149-52
2060792
Microb Cell Fact. 2012 Feb 15;11:24
22335940
Int J Food Microbiol. 2015 Jul 16;205:112-8
25897995
Mol Microbiol. 2002 Dec;46(5):1319-33
12453218
Evolution. 2012 Sep;66(9):2961-8
22946816
Biotechnol Bioeng. 2000 Jan 20;67(2):165-76
10592514
Lett Appl Microbiol. 2004;39(2):199-206
15242462
Antimicrob Agents Chemother. 2001 Dec;45(12):3366-74
11709310
Curr Genet. 1994 May;25(5):469-71
8082194
J Clin Microbiol. 1990 Jul;28(7):1509-13
2199515
Biotechnol Adv. 2000 Nov 1;18(7):581-99
14538096
J Biol Chem. 2011 Apr 22;286(16):13859-68
21343301
Mol Biol Cell. 2012 Jul;23(13):2402
22745339
J Microbiol Methods. 2009 Jan;76(1):70-4
18840479
Science. 2010 Aug 27;329(5995):1085-8
20798321
Wei Sheng Wu Xue Bao. 2011 Mar;51(3):340-5
21604548
FEBS Lett. 1998 Mar 27;425(2):323-8
9559673
Microbiology. 2012 Aug;158(Pt 8):2133-43
22628481
Antimicrob Agents Chemother. 2010 Apr;54(4):1639-43
20100876
Appl Microbiol Biotechnol. 2010 Jul;87(4):1507-16
20445974
Appl Environ Microbiol. 1995 May;61(5):1904-9
7646027
Appl Microbiol Biotechnol. 2007 Jan;73(6):1387-93
17043823
Mol Microbiol. 2006 Mar;59(5):1429-51
16468987
Metab Eng. 2013 Sep;19:10-6
23707987
J Biosci Bioeng. 2009 Jan;107(1):1-6
19147100
Appl Environ Microbiol. 2008 Feb;74(4):1136-44
18156322
J Ind Microbiol Biotechnol. 2012 Jul;39(7):1031-9
22366811
Biochem J. 2014 Mar 1;458(2):323-34
24341558
Eur J Biochem. 1991 Sep 1;200(2):487-93
1889413
Biochim Biophys Acta. 2012 Apr;1821(4):647-53
22342273
Biochemistry. 2009 Oct 13;48(40):9360-71
19725516
Appl Environ Microbiol. 2007 Aug;73(15):4824-31
17557846
PLoS One. 2015 Sep 29;10(9):e0139306
26418026
Mol Microbiol. 2011 Feb;79(4):900-13
21299646
Yeast. 2010 Aug;27(8):473-8
20641012
J Biol Chem. 2011 Jun 10;286(23):20977-90
21487010
Eukaryot Cell. 2014 Jun;13(6):694-705
24681686
Appl Microbiol Biotechnol. 2013 Apr;97(7):3239-51
23435986
Appl Microbiol Biotechnol. 2015 Jun;99(12):5227-36
25683664
Front Microbiol. 2015 Aug 18;6:843
26347728
Mol Microbiol. 2007 Nov;66(4):858-71
17927701
Biotechnol Bioeng. 2015 Jan;112(1):156-67
25060134
J Biol Chem. 2002 Aug 23;277(34):31079-88
12058033
J Biol Chem. 2006 Nov 10;281(45):34135-45
16980694
FEMS Microbiol Rev. 2000 Oct;24(4):469-86
10978547
Biotechnol Bioeng. 2006 Dec 5;95(5):992-1002
16807925
G3 (Bethesda). 2012 Jun;2(6):675-91
22690377
Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2319-24
18252819
FEMS Yeast Res. 2011 Aug;11(5):430-9
21457451
Curr Genet. 1992 Apr;21(4-5):269-73
1525853