Physiological characterisation of Penicillium chrysogenum strains expressing the expandase gene from Streptomyces clavuligerus during batch cultivations. Growth and adipoyl-7-aminodeacetoxycephalosporanic acid production.

Robin, Jarno Jacky Christian; Jakobsen, Mogens; Beyer, Michael; Noorman, Henk; Nielsen, Jens

doi:10.1007/s002530100787

Cited by 27 publications

(3 citation statements)

References 8 publications

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“…In case of pellet formation the cultures grew only exponentially in the beginning of the cultivation prior to pellet formation. The obtained growth rates are reasonable starting points for industrial fermentations, for example the penicillin producing P. chrysogenum , has a maximum growth rate under comparable conditions of around 0.2 h −1 [ 20 , 21 ]. An interesting observation is that the species with the highest growth rates, P. decumbens and P. coprophilum , have the smallest genome size [ 15 ].…”

Section: Discussionmentioning

confidence: 99%

Physiological characterization of secondary metabolite producing Penicillium cell factories

Grijseels

Nielsen

et al. 2017

Fungal Biol Biotechnol

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Background Penicillium species are important producers of bioactive secondary metabolites. However, the immense diversity of the fungal kingdom is only scarcely represented in industrial bioprocesses and the upscaling of compound production remains a costly and labor intensive challenge. In order to facilitate the development of novel secondary metabolite producing processes, two routes are typically explored: optimization of the native producer or transferring the enzymatic pathway into a heterologous host. Recent genome sequencing of ten Penicillium species showed the vast amount of secondary metabolite gene clusters present in their genomes, and makes them accessible for rational strain improvement. In this study, we aimed to characterize the potential of these ten Penicillium species as native producing cell factories by testing their growth performance and secondary metabolite production in submerged cultivations.ResultsCultivation of the fungal species in controlled submerged bioreactors showed that the ten wild type Penicillium species had promising, highly reproducible growth characteristics in two different media. Analysis of the secondary metabolite production using liquid chromatography coupled with high resolution mass spectrometry proved that the species produced a broad range of secondary metabolites, at different stages of the fermentations. Metabolite profiling for identification of the known compounds resulted in identification of 34 metabolites; which included several with bioactive properties such as antibacterial, antifungal and anti-cancer activities. Additionally, several novel species–metabolite relationships were found.ConclusionsThis study demonstrates that the fermentation characteristics and the highly reproducible performance in bioreactors of ten recently genome sequenced Penicillium species should be considered as very encouraging for the application of native hosts for production via submerged fermentation. The results are particularly promising for the potential development of the ten analysed Penicillium species for production of novel bioactive compounds via submerged fermentations.Electronic supplementary materialThe online version of this article (doi:10.1186/s40694-017-0036-z) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Physiological characterization of secondary metabolite producing Penicillium cell factories

Grijseels

Nielsen

et al. 2017

Fungal Biol Biotechnol

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“…The secondary structures of residues 81-90 and 167-177 were highly disordered and were not determined [84]. The cylinders, arrows, and black lines represent -helices, -sheets, and coil structures, respectively expression in E. coli [28,34,49], S. lividans [22,33], Penicillium chrysogenum [13,24,68,[70][71][72][73], Pseudomonas putida [47], and Pichia pastoris [3]. High level expression of recombinant scDAOCS has often been carried out in E. coli cells together with the use of traditional extraction to purify scDAOCS from E. coli cell lysate for biochemical studies [57,58,90].…”

Section: Expression Of Recombinant S Clavuligerus Daocsmentioning

confidence: 99%

Directed evolution and rational approaches to improving Streptomyces clavuligerus deacetoxycephalosporin C synthase for cephalosporin production

Goo

Chua

Sim

2009

J Ind Microbiol Biotechnol

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It is approximately 60 years since the discovery of cephalosporin C in Cephalosporium acremonium. Streptomycetes have since been found to produce the structurally related cephamycin C. Studies on the biosynthetic pathways of these two compounds revealed a common pathway including a step governed by deacetoxycephalosporin C synthase which catalyses the ring-expansion of penicillin N to deacetoxycephalosporin C. Because of the therapeutic importance of cephalosporins, this enzyme has been extensively studied for its ability to produce these antibiotics. Although, on the basis of earlier studies, its substrate specificity was believed to be extremely narrow, relentless efforts in optimizing the in-vitro enzyme assay conditions showed that it is able to convert a wide range of penicillin substrates differing in their side chains. It is a member of 2-oxoglutarate-dependent dioxygenase protein family, which requires the iron(II) ion as a co-factor and 2-oxoglutarate and molecular oxygen as co-substrates. It has highly conserved HXDX( n ) H and RXS motifs to bind the co-factor and co-substrate, respectively. With advances in technology, the genes encoding this enzyme from various sources have been cloned and heterologously expressed for comparative analyses and mutagenesis studies. A high level of recombinant protein expression has also enabled crystallization of this enzyme for structure determination. This review will summarize some of the earlier biochemical characterization and describe the mechanistic action of this enzyme revealed by recent structural studies. This review will also discuss some of the approaches used to identify the amino acid residues involved in binding the penicillin substrate and to modify its substrate preference for possible industrial application.

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“…We consider this finding as escape phenomenon at the high sucrose concentration. According to other studies (Nilsson and Bjurman, 1998;Robin et al, 2001;Park and Gander, 1998;El-Kady et al, 1995) molds are well known for their ability to adapt to high osmolarity environments through the polyols intracellular accumulation. Penicillium species accumulate glycerol as major osmoregulation substance (Blomberg and Adler, 1992;Harris, 1981;Hocking, 1986).…”

Section: Metabolic Responsementioning

confidence: 99%

Effect of Sucrose on the Physiology and Terrestric Acid Production of Penicillium Aurantiogriseum

Bouhoudan¹,

Chidi²,

Khaddor³

2020

AgricultForest

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Penicillium aurantiogriseum (P. aurantiogriseum) is a post-harvest pathogen that causes significant losses in agricultural production during storage. It plays an important role in food and feed spoilage, and it contaminates agricultural products with mycotoxins that are potentially harmful to human and animal health. P. aurantiogriseum is one of the most toxic species in the genus Penicillium, and it is often isolated from foods, vegetables, fruits and permafrost sediments from the Arctic and Antarctic. It has also been isolated from the marine environment. Thus, it is resistant to several types of stress related to nutrients and growing conditions. This study aimed to determine the effect of sucrose on the physiology of P. aurantiogriseum in order to control its growth and toxigenesis. Mycotoxin production was determined by TLC technique. Our results show a close relationship between the physiological state of P. aurantiogriseum and the secretion of mycotoxins under carbon stress conditions. The physiological state of the pathogen reveals a correlation between increased sucrose concentration and the intensity of aging signs. Aging signs begin to disappear at a sucrose concentration of 400 g/l, which allows the normal characteristics of P. aurantiogriseum to reappear. It is suggested that this transformation is meant to avoid the action of sucrose. Terrestric acid production was recorded at the time of appearance of aging signs. Terrestric acid is always maintained, even after returning to a normal physiological state, but its production was diminished. The growth of P. aurantiogriseum can be controlled by modifying the sucrose concentration in growth medium. This allowed us to determine the critical concentration at which the pathogen suffered and thus reached the phase of decline earlier while mycotoxin production was minimal.

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Physiological characterisation of Penicillium chrysogenum strains expressing the expandase gene from Streptomyces clavuligerus during batch cultivations. Growth and adipoyl-7-aminodeacetoxycephalosporanic acid production.

Cited by 27 publications

References 8 publications

Physiological characterization of secondary metabolite producing Penicillium cell factories

Physiological characterization of secondary metabolite producing Penicillium cell factories

Directed evolution and rational approaches to improving Streptomyces clavuligerus deacetoxycephalosporin C synthase for cephalosporin production

Effect of Sucrose on the Physiology and Terrestric Acid Production of Penicillium Aurantiogriseum

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