Intensifying niacin-based biosynthesis of NAD + to enhance 3-hydroxypropionic acid production in Klebsiella pneumoniae
- PDF / 1,364,752 Bytes
- 12 Pages / 547.087 x 737.008 pts Page_size
- 7 Downloads / 144 Views
(0123456789().,-volV) ( 01234567 89().,-volV)
ORIGINAL RESEARCH PAPER
Intensifying niacin-based biosynthesis of NAD+ to enhance 3-hydroxypropionic acid production in Klebsiella pneumoniae Shimin Wu . Peng Zhao . Qingyang Li . Pingfang Tian
Received: 7 March 2020 / Accepted: 19 September 2020 Ó Springer Nature B.V. 2020
Abstract Objective Glycerol-based biosynthesis of 3-hydroxypropionic acid (3-HP) in Klebsiella pneumoniae involves two reactions: glycerol conversion to 3-hydroxypropionaldehyde (3-HPA) by glycerol dehydratase, and 3-HPA conversion to 3-HP by aldehyde dehydrogenase (ALDH). The ALDH catalysis consumes a lot of cofactor nicotinamide adenine dinucleotide (NAD?), which constrains 3-HP production. Results Here we report that intensifying niacinbased biosynthesis of NAD? can substantially enhance 3-HP production. We constructed tac promoter-driven NAD? synthesis pathway in K. pneumoniae. The strain only overexpressing nicotinate phosphoribosyltransferase (PncB) showed 14.24% increase in the production of NAD? relative to the stain harboring an empty vector. When PncB was coexpressed with PuuC (one of native ALDHs), the
recombinant strain exhibited increased ALDH activity but slightly reduced 3-HP production due to plasmid burden. When 30 mg niacin l-1 (a substrate for biosynthesis of NAD?) was added into shake flask, the strain produced 0.55 g 3-HP l-1, which was 2.75 times that of the control. In a 5-L bioreactor, replenishment of niacin led to 36.43% increase of 3-HP production. Conclusions These results indicated that intensifying niacin-based biosynthesis of NAD? boosts 3-HP production. Keywords Aldehyde dehydrogenase Glycerol 3Hydroxypropionic acid Klebsiella pneumonia NAD?
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10529-020-03011-y) contains supplementary material, which is available to authorized users. S. Wu P. Zhao P. Tian (&) College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China e-mail: [email protected] Q. Li School of Food Science and Engineering, South China University of Technology, Guangzhou 510641, China
3-Hydroxypropionic acid (3-HP) is a commercially valuable platform compound from which an array of bulk chemicals can be produced (Tokuyama et al. 2014; Li et al. 2013). 3-HP can be naturally synthesized by a clade of microbes such as Lactobacillus reuteri (Talarico et al. 1988), Alcaligenes faecalis (Ansede et al. 1999), Phomopsis phaseoli (Schwarz et al. 2004), Melanconium betulinum (Schwarz et al. 2004), Meyerozyma guilliermondii (Zhang et al. 2017) and Klebsiella pneumoniae (Kumar and Park 2018).
123
Biotechnol Lett
Of these microbes, K. pneumoniae is highly attractive because of its striking biochemical properties, such as fast growth and extraordinary ability to utilize glycerol. In particular, K. pneumoniae can naturally generate vitamin B12 (Ashok et al. 2013), which is the cofactor of glycerol dehydratase (GDHt, EC: 4.2.1.30, encoded by d
Data Loading...
