• Title/Summary/Keyword: {TEX}$F_{1}${/TEX}{TEX}$F_{0}${/TEX}ATPase

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Deletion of cg1360 Affects ATP Synthase Function and Enhances Production of L-Valine in Corynebacterium glutamicum

  • Wang, Xiaochen;Yang, Hongyu;Zhou, Wei;Liu, Jun;Xu, Ning
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1288-1298
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    • 2019
  • Bacterial ATP synthases drive ATP synthesis by a rotary mechanism, and play a vital role in physiology and cell metabolism. Corynebacterium glutamicum is well known as an industrial workhorse for amino acid production, and its ATP synthase operon contains eight structural genes and two adjacent genes, cg1360 and cg1361. So far, the physiological functions of Cg1360 (GenBank CAF19908) and Cg1361 (GenBank CAF19909) remain unclear. Here, we showed that Cg1360 was a hydrophobic protein with four transmembrane helices (TMHs), while no TMH was found in Cg1361. Deletion of cg1360, but not cg1361, led to significantly reduced cell growth using glucose and acetic acid as carbon sources, reduced F1 portions in the membrane, reduced ATP-driven proton-pumping activity and ATPase activity, suggesting that Cg1360 plays an important role in ATP synthase function. The intracellular ATP concentration in the ${\Delta}cg1360$ mutant was decreased to 72% of the wild type, while the NADH and NADPH levels in the ${\Delta}cg1360$ mutant were increased by 29% and 26%, respectively. However, the ${\Delta}cg1361$ mutant exhibited comparable intracellular ATP, NADH and NADPH levels with the wild-type strain. Moreover, the effect of cg1360 deletion on L-valine production was examined in the L-valine-producing V-10 strain. The final production of L-valine in the $V-10-{\Delta}cg1360$ mutant reached $9.2{\pm}0.3g/l$ in shake flasks, which was 14% higher than that of the V-10 strain. Thus, Cg1360 can be used as an effective engineering target by altering energy metabolism for the enhancement of amino acid production in C. glutamicum.

Dietary phosphorus deficiency impaired growth, intestinal digestion and absorption function of meat ducks

  • Xu, Huimin;Dai, Shujun;Zhang, Keying;Ding, Xuemei;Bai, Shiping;Wang, Jianping;Peng, Huanwei;Zeng, Qiufeng
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1897-1906
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    • 2019
  • Objective: An experiment was conducted to investigate the effects of dietary non-phytate phosphorus (nPP) deficiency on intestinal pH value, digestive enzyme activity, morphology, nutrient utilization, and gene expression of NaPi-IIb in meat ducks from 1 to 21 d of age. Methods: A total of 525 one-d-old Cherry Valley ducklings were fed diets (with 7 pens of 15 ducklings, or 105 total ducklings, on each diet) with five levels of nPP (0.22%, 0.34%, 0.40%, 0.46%, or 0.58%) for 21 d in a completely randomized design. Five experimental diets contained a constant calcium (Ca) content of approximately 0.9%. Body weight (BW), body weight gain (BWG), feed intake (FI), and feed to gain ratio (F:G) were measured at 14 and 21 d of age. Ducks were sampled for duodenum and jejunum digestion and absorption function on 14 and 21 d. Nutrient utilization was assessed using 25- to 27-d-old ducks. Results: The results showed ducks fed 0.22% nPP had lower (p<0.05) growth performance and nutrient utilization and higher (p<0.05) serum Ca content and alkaline phosphatase (ALP) activity. When dietary nPP levels were increased, BW (d 14 and 21), BWG and FI (all intervals), and the serum phosphorus (P) content linearly and quadratically increased (p<0.05); and the jejunal pH value (d 14), duodenal muscle layer thickness (d 14), excreta dry matter, crude protein, energy, Ca and total P utilization linearly increased (p<0.05); however, the serum ALP activity, jejunal $Na^+-K^+$-ATPase activity, and duodenal NaPi-IIb mRNA level (d 21) linearly decreased (p<0.05). Conclusion: The results indicated that ducks aged from 1 to 21 d fed diets with 0.22% nPP had poor growth performance related to poor intestinal digestion and absorption ability; but when fed diets with 0.40%, 0.46%, and 0.58% nPP, ducks presented a better growth performance, intestinal digestion and absorption function.