Fermentative aminopyrrolnitrin production by metabolically engineered Corynebacterium glutamicum.

Autor: Putri VRM; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea., Jung MH; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea., Lee JY; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea., Kwak MH; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea., Mariyes TC; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea., Kerbs A; Faculty of Biology and Center for Biotechnology, Bielefeld University, Bielefeld, Germany., Wendisch VF; Faculty of Biology and Center for Biotechnology, Bielefeld University, Bielefeld, Germany., Kong HJ; Biotechnology Research Division, National Institute of Fisheries Science, Busan, 46083, Republic of Korea., Kim YO; Biotechnology Research Division, National Institute of Fisheries Science, Busan, 46083, Republic of Korea., Lee JH; Department of Food Science & Biotechnology, BB21+, Kyungsung University, Busan, 48434, Republic of Korea. jhlee83@ks.ac.kr.
Jazyk: angličtina
Zdroj: Microbial cell factories [Microb Cell Fact] 2024 May 23; Vol. 23 (1), pp. 147. Date of Electronic Publication: 2024 May 23.
DOI: 10.1186/s12934-024-02424-y
Abstrakt: Aminopyrrolnitrin (APRN), a natural halogenated phenylpyrrole derivative (HPD), has strong antifungal and antiparasitic activities. Additionally, it showed 2.8-fold increased photostability compared to pyrrolnitrin, a commercially available HPD with antimicrobial activity. For microbial production of APRN, we first engineered anthranilate phosphoribosyltransferase encoded by trpD from Corynebacterium glutamicum, resulting in a TrpD A162D mutation that exhibits feedback-resistant against L-tryptophan and higher substrate affinity compared to wild-type TrpD. Plasmid-borne expression of trpD A162D in C. glutamicum TP851 strain with two copies of trpD A162D in the genome led to the production of 3.1 g/L L-tryptophan in flask culture. Subsequent step for L-tryptophan chlorination into 7-chloro-L-tryptophan was achieved by introducing diverse sources of genes encoding tryptophan 7-halogenase (PrnA or RebH) and flavin reductase (Fre, PrnF, or RebF). The combined expression of prnA from Serratia grimesii or Serratia plymuthica with flavin reductase gene from Escherichia coli, Pseudomonas fluorescens, or Lechevalieria aerocolonigenes yielded higher production of 7-chloro-L-tryptophan in comparison to other sets of two-component systems. In the next step, production of putative monodechloroaminopyrrolnitrin (MDAP) from 7-chloro-L-tryptophan was achieved through the expression of prnB encoding MDAP synthase from S. plymuthica or P. fluorescens. Finally, an artificial APRN biosynthetic pathway was constructed by simultaneously expressing genes coding for tryptophan 7-halogenase, flavin reductase, MDAP synthase, and MDAP halogenase (PrnC) from different microbial sources within the L-tryptophan-producing TP851 strain. As prnC from S. grimesii or S. plymuthica was introduced into the host strain, which carried plasmids expressing prnA from S. plymuthica, fre from E. coli, and prnB from S. plymuthica, APN3639 and APN3638 accumulated 29.5 mg/L and 28.1 mg/L of APRN in the culture broth. This study represents the first report on the fermentative APRN production by metabolically engineered C. glutamicum.
(© 2024. The Author(s).)
Databáze: MEDLINE
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