Introducing glutathione biosynthetic capability into Lactococcus lactis subsp. cremoris NZ9000 improves the oxidative-stress resistance of the host.
This study describes how a metabolic engineering approach can be used to improve bacterial stress resistance. Some Lactococcus lactis strains are capable of taking up glutathione, and the imported glutathione protects this organism against H(2)O(2)-induced oxidative stress. L. lactis subsp. cremoris NZ9000, a model organism of this species that is widely used in the study of metabolic engineering, can neither synthesize nor take up glutathione. The study described here aimed to improve the oxidative-stress resistance of strain NZ9000 by introducing a glutathione biosynthetic capability. We show that the glutathione produced by strain NZ9000 conferred stronger resistance on the host following exposure to H(2)O(2) (150 mM) and a superoxide generator, menadione (30 microM). To explore whether glutathione can complement the existing oxidative-stress defense systems, we constructed a superoxide dismutase deficient mutant of strain NZ9000, designated as NZ4504, which is more sensitive to oxidative stress, and introduced the glutathione biosynthetic capability into this strain. Glutathione produced by strain NZ4504(pNZ3203) significantly shortens the lag phase of the host when grown aerobically, especially in the presence of menadione. In addition, cells of NZ4504(pNZ3203) capable of producing glutathione restored the resistance of the host to H(2)O(2)-induced oxidative stress, back to the wild-type level. We conclude that the resistance of L. lactis subsp. cremoris NZ9000 to oxidative stress can be increased in engineered cells with glutathione producing capability.
Fu
Rui-Yan
RY
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Southern Yangtze University, Wuxi 214036, People's Republic of China.
Bongers
Roger S
RS
van Swam
Iris I
II
Chen
Jian
J
Molenaar
Douwe
D
Kleerebezem
Michiel
M
Hugenholtz
Jeroen
J
Li
Yin
Y
D001709:Biotechnology / Q000379:methods*; D017931:DNA Primers; D005818:Genetic Engineering / Q000379:methods*; D005978:Glutathione / Q000096:biosynthesis* / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000633:toxicity; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D013482:Superoxide Dismutase / Q000172:deficiency; D024483:Vitamin K 3
D017931:DNA Primers; D024483:Vitamin K 3; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D005978:Glutathione