Nonribosomal peptide synthetases and their biotechnological potential in Penicillium rubens

Abstract

Nonribosomal peptide synthetases (NRPSs) are large multimodular enzymes that synthesize a diverse variety of peptides. Many of these are currently used as pharmaceuticals, thanks to their activity as antimicrobials (penicillin, vancomycin, daptomycin, echinocandin), immunosuppressant (cyclosporin) and anticancer compounds (bleomycin). Because of their biotechnological potential, NRPSs have been extensively studied in the past decades. In this review, we provide an overview of the main structural and functional features of these enzymes, and we consider the challenges and prospects of engineering NRPSs for the synthesis of novel compounds. Furthermore, we discuss secondary metabolism and NRP synthesis in the filamentous fungus Penicillium rubens and examine its potential for the production of novel and modified β-lactam antibiotics.

Emulating nonribosomal peptides with ribosomal biosynthetic strategies

“…NRPS modules have been re-engineered to produce novel peptide analogues and general strategies for engineering NRPSs (or NRPS/PKS hybrids) have been reviewed elsewhere. 203–206 In brief, the specificity-conferring code of A domains can be altered to change substrate selection by high throughput methods such as yeast surface display, 207,208 or complete modules, multiple domains or exchange units (XUs) can be swapped between different NRPSs to generate new peptides. 209,210 Despite recent successes and advances in NRPS redesign, it remains challenging to engineer and clone a functional hybrid NRPS, and thereby produce novel NRP analogues, due to the mere size of these megasynthetases and inter-domain and inter-module protein interactions that have to remain intact for a functional NRPS.…”

Penicillium chrysogenum, a Vintage Model with a Cutting-Edge Profile in Biotechnology

“…In summary, the CRISPR/Cas9 technique, in combination with other techniques, will allow engineering P. chrysogenum in ways that have not yet been fully explored, which include, but are not limited to, the replacement of natural gene promoters in BGCs by engineered strong and adjustable promoters to improve production yields of endogenous or exogenous compounds [ 323 ], the engineering of modular enzymes such as PKS and NRPS by module swapping to obtain modified secondary metabolites (discussed in [ 235 ] and [ 359 ]), and the combination of biosynthetic genes from different organisms to produce structurally diverse “unnatural products” [ 360 ].…”

Transcriptional Activation of Biosynthetic Gene Clusters in Filamentous Fungi

“…To increase the titers of penicillin produced, early strains of P. rubens have been subjected to decades of random mutagenesis and selection processes, collectively known as the classical strain improvement program. This has also led to increased capabilities to grow in submerged cultivation conditions and in defined media, features that are very desirable in the industry (Harris et al, 2009;Guzmán-Chávez et al, 2018;Iacovelli et al, 2021a). The CSI program also led to a major reduction of the expression and/or mutational inactivation of nonpenicillin BGCs.…”