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Increased ethanol tolerance associated with the pntAB locus of Oenococcus oeni and Lactobacillus buchneri.

Abstract

Lactobacillus buchneri and Oenococcus oeni are two unique ethanol-tolerant Gram-positive bacteria species. Genome comparison analyses revealed that L. buchneri and O. oeni possess a pntAB locus that was absent in almost all other lactic acid bacteria (LAB) genomes. Our hypothesis is that the pntAB locus contributes to the ethanol tolerance trait of these two distinct ethanol-tolerant organisms. The pntAB locus, consisting of the pntA and pntB genes, codes for NADP(H) transhydrogenase subunits. This membrane-bound transhydrogenase catalyzes the reduction of NADP+ and is known as an important enzyme in maintaining cellular redox balance. In this study, the transhydrogenase operon from L. buchneri NRRL B-30929 and O. oeni PSU-1 were cloned and analyzed. The LbpntB shared 71.0% identity with the O. oeni (OopntB). The entire pntAB locus was expressed in Lactococcus lactis ssp. lactis IL1403 resulting in an increased tolerance to ethanol (6%), butanol (1.8%) and isopropanol (1.8%) when compared to the control strain. However, the recombinant E. coli cells carrying the entire pntAB locus did not show any improved ethanol tolerance. Independent expression of OopntB and LbpntB in recombinant E. coli BL21(DE3)pLysS host demonstrated higher tolerance to ethanol when compared with a control E. coli BL21(DE3)pLysS strain carrying pET28b vector. Ethanol tolerance comparison of E. coli strains carrying LbpntB and OopntB showed that LbpntB conferred higher ethanol tolerance (4.5%) and resulted in greater biomass, while the OopntB conferred lower ethanol tolerance (4.0%) resulted lower biomass. Therefore, the pntB gene from L. buchneri is a better choice in generating higher ethanol tolerance. This is the first study to uncover the role of pntAB locus on ethanol tolerance.

Authors+Show Affiliations

USDA-ARS-NCAUR RPT, Peoria, USA. Siqing.liu@ars.usda.gov.USDA-ARS-NCAUR RPT, Peoria, USA.University of California, Davis, Davis, USA.University of California, Davis, Davis, USA.USDA-ARS-NCAUR Bioenergy Research, Peoria, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31289974

Citation

Liu, Siqing, et al. "Increased Ethanol Tolerance Associated With the pntAB Locus of Oenococcus Oeni and Lactobacillus Buchneri." Journal of Industrial Microbiology & Biotechnology, 2019.
Liu S, Skory C, Liang X, et al. Increased ethanol tolerance associated with the pntAB locus of Oenococcus oeni and Lactobacillus buchneri. J Ind Microbiol Biotechnol. 2019.
Liu, S., Skory, C., Liang, X., Mills, D., & Qureshi, N. (2019). Increased ethanol tolerance associated with the pntAB locus of Oenococcus oeni and Lactobacillus buchneri. Journal of Industrial Microbiology & Biotechnology, doi:10.1007/s10295-019-02209-y.
Liu S, et al. Increased Ethanol Tolerance Associated With the pntAB Locus of Oenococcus Oeni and Lactobacillus Buchneri. J Ind Microbiol Biotechnol. 2019 Jul 9; PubMed PMID: 31289974.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Increased ethanol tolerance associated with the pntAB locus of Oenococcus oeni and Lactobacillus buchneri. AU - Liu,Siqing, AU - Skory,Chris, AU - Liang,Xiaojin, AU - Mills,David, AU - Qureshi,Nasib, Y1 - 2019/07/09/ PY - 2019/03/14/received PY - 2019/06/21/accepted PY - 2019/7/11/entrez KW - Butanol tolerance KW - Ethanol tolerance KW - Lactobacillus buchneri KW - Oenococcus oeni KW - pntAB JF - Journal of industrial microbiology & biotechnology JO - J. Ind. Microbiol. Biotechnol. N2 - Lactobacillus buchneri and Oenococcus oeni are two unique ethanol-tolerant Gram-positive bacteria species. Genome comparison analyses revealed that L. buchneri and O. oeni possess a pntAB locus that was absent in almost all other lactic acid bacteria (LAB) genomes. Our hypothesis is that the pntAB locus contributes to the ethanol tolerance trait of these two distinct ethanol-tolerant organisms. The pntAB locus, consisting of the pntA and pntB genes, codes for NADP(H) transhydrogenase subunits. This membrane-bound transhydrogenase catalyzes the reduction of NADP+ and is known as an important enzyme in maintaining cellular redox balance. In this study, the transhydrogenase operon from L. buchneri NRRL B-30929 and O. oeni PSU-1 were cloned and analyzed. The LbpntB shared 71.0% identity with the O. oeni (OopntB). The entire pntAB locus was expressed in Lactococcus lactis ssp. lactis IL1403 resulting in an increased tolerance to ethanol (6%), butanol (1.8%) and isopropanol (1.8%) when compared to the control strain. However, the recombinant E. coli cells carrying the entire pntAB locus did not show any improved ethanol tolerance. Independent expression of OopntB and LbpntB in recombinant E. coli BL21(DE3)pLysS host demonstrated higher tolerance to ethanol when compared with a control E. coli BL21(DE3)pLysS strain carrying pET28b vector. Ethanol tolerance comparison of E. coli strains carrying LbpntB and OopntB showed that LbpntB conferred higher ethanol tolerance (4.5%) and resulted in greater biomass, while the OopntB conferred lower ethanol tolerance (4.0%) resulted lower biomass. Therefore, the pntB gene from L. buchneri is a better choice in generating higher ethanol tolerance. This is the first study to uncover the role of pntAB locus on ethanol tolerance. SN - 1476-5535 UR - https://www.unboundmedicine.com/medline/citation/31289974/Increased_ethanol_tolerance_associated_with_the_pntAB_locus_of_Oenococcus_oeni_and_Lactobacillus_buchneri L2 - http://dx.doi.org/10.1007/s10295-019-02209-y DB - PRIME DP - Unbound Medicine ER -