Production of trans-2,3-dihydro-3-hydroxyanthranilic acid by engineered Pseudomonas chlororaphis GP72

Abstract

Trans-2,3-dihydro-3-hydroxyanthranilic acid (DHHA) is a cyclic β-amino acid that can be used for the synthesis of chiral materials and nonnatural peptides. The aim of this study was to accumulate DHHA by engineering Pseudomonas chlororaphis GP72, a nonpathogenic strain that produces phenazine-1-carboxylic acid and 2-hydroxyphenazine. First, the phzF deletion mutant DA1 was constructed, which produced 1.91 g/L DHHA. Moreover, rpeA and pykF were disrupted and then ppsA and tktA were co-expressed in strain DA1. The resulting strain DA4 increased DHHA concentration to 4.98 g/L, which is 2.6-fold than that of DA1. The effects of the addition of glucose, glycerol, L-tryptophan, and Feon DHHA production were also investigated. Strain DA4 produced 7.48 g/L of DHHA in the culture medium in the presence of 12 g/L glucose and 3 mM Fe, which was 1.5-fold higher than the strain in the original fermentation conditions. These results indicate the potential of P. chlororaphis GP72 as a DHHA producer.

Adsorption/desorption characteristics, separation and purification of phenazine‐1‐carboxylic acid from fermentation extract by macroporous adsorbing resins

“…PCA and PCA‐producing strains are substantially employed as biocontrol agents because of their potential fungicidal efficiency, minimum toxicity status, and biodegradability. In recent years, an array of studies have been focused on engineering high‐yielding strains and fermentation processes optimization for induced production of phenazine and its derivatives . However, the isolation and purification of PCA in biotechnological production remains to be addressed.…”

Synthesis of cinnabarinic acid by metabolically engineered Pseudomonas chlororaphis GP72

“…Therefore, to construct a synthetic pathway to generate CA from HAA, it is necessary to develop a novel and efficient strategy to synthesize HAA. HAA is structurally similar to trans ‐2,3‐dihydro‐3‐hydroxyanthranilic acid (DHHA), which is a natural intermediate for the synthesis of phenazines in some types of Pseudomonas strains (Blankenfeldt et al, ; Hu et al, ; Yue et al, ). Dehydrogenation at the C2 and C3 position of DHHA into benzene ring structure may be a feasible means of generating HAA.…”

“…Pseudomonas chlororaphis GP72 (hereafter referred to as GP72) is a biocontrol strain that produces phenazine‐1‐carboxylic acid (PCA) and 2‐hydroxyphenazine (2‐OH‐PHZ) in high yields (Huang et al, ; Liu, Hu, Wang, & Zhang, ). In our previous studies, we constructed an engineered strain of GP72 that could produce 10.06 g/L of DHHA from glycerol (Hu et al, ; Yue et al, ), which is a major byproduct of biodiesel. Therefore, strain GP72 may be a unique platform to synthesize HAA and CA from renewable resources.…”

Enhanced trans‐2,3‐dihydro‐3‐hydroxyanthranilic acid production by pH control and glycerol feeding strategies in engineered Pseudomonas chlororaphis GP72

“…In addition, DHHA is an important intermediate of the phenazine biosynthetic pathway in phenazine‐producing Pseudomonas strains (Fig. ) …”

“…Since GP72 ensures adequate metabolite flux in the phenazine biosynthesis pathway, higher production of DHHA may be achieved by this strain. In our previous study, we constructed an engineered strain DA4 from wild type GP72 by the deletion of phzF , pykF and rpeA and overexpression of tktA and ppsA genes and 7.48 g L ‐1 of DHHA was obtained in a shake flask fermentation …”