Comprehensive Profiling of Proteome Changes Provide Insights of Industrial Penicillium chrysogenum During Pilot and Industrial Penicillin G Fermentation

Abstract

The intracellular proteomes of the Penicillium chrysogenum throughout pilot and industrial processes were investigated by using 2-DE combined with MALDI-TOF-TOF MS, respectively. We detected a total of 223 spots corresponding to 154 proteins and 231 spots corresponding to 157 proteins throughout pilot and industrial processes, respectively. The levels of glyceraldehyde-3-phosphate dehydrogenase increased (5.1- and 2.5-fold) under the pilot process, while its levels were no significant changes under the industrial process at 140 and 170 h when compared with that at 2 h. The levels of isocitrate lyase and fumarate hydratase were increased significantly under the industrial process, while their levels had no obvious changes after 20 h of fermentation throughout the pilot process. These results indicate that there were remarkable differences in carbohydrate metabolism (including glycolysis, gluconeogenesis, pentose phosphate pathway, and tricarboxylic acid cycle) of P. chrysogenum during the pilot and industrial fermentations, which likely result in alterations of the primary metabolism and penicillin biosynthesis. Moreover, the differences in the levels of proteins involved in amino acid metabolisms (including valine, cysteine, and α-aminoadipic acid biosynthesis) indicated that the pilot and industrial processes influenced the supplies of penicillin precursors. Compared with that at 2 h, the maximum levels of superoxide (6.9-fold, at 32 h) and catalase (9-fold, at 80 h) during the industrial process and the maximum levels of superoxide (1.2-fold, at 20 h) and catalase (7.7-fold at 128 h) during the pilot process revealed the significant difference in cell redox homeostasis and stress responses during scale-up fermentation. Particularly, 10 spots corresponding to isopenicillin N synthetase and 4 spots corresponding to isopenicillin N (IPN) acyltransferase in pilot and industrial processes were identified, respectively. The levels of IPN acyltransferase (spots 197 and 198) and CoA ligase at 80 h during the industrial process were around 2-fold of that during the pilot process, indicating that the industrial process with a higher penicillin production per cell might provide available environments to induce over-expression of IPN acyltransferase and accelerate penicillin formation. These results provide new insights into the globally potential responses of P. chrysogenum to variations of environments in different fermentation scales so as to consequently regulate the penicillin production.

Omics Approaches Applied to Penicillium chrysogenum and Penicillin Production: Revealing the Secrets of Improved Productivity

“…These results suggest that the industrial process might provide a friendly environment for amino acid metabolism to enhance the penicillin-precursor production (valine, cysteine, and α-aminoadipic acid). Furthermore, Cheng and co-workers [ 83 ] found an overproduction of the IAT and the CoA ligase during the industrial fermentation, which supports the higher penicillin production at industrial scale.…”

“…Proteome analysis of different strains of P. chrysogenum has given Proteomics the chance to enhance the understanding of the penicillin production physiology by means of a reverse engineering process as recently reviewed by Barreiro and García-Estrada [ 3 ]. Thus, data collected from the wild-type strain ( P. chrysogenum NRRL 1951), the reference laboratory strain ( P. chrysogenum Wisconsin 54-1255) and several high penicillin production strains ( P. chrysogenum AS-P-78, P. chrysogenum DS17690; P. chrysogenum QMSHB-11-06) have boosted the proteome analyses and knowledge of penicillin production [ 64 , 65 , 82 , 83 ].…”

“…Finally, the functional proteomics that provides information about protein functions through the identification of interacting protein partners. Expression proteomics analyses have been carried out in penicillin-producer strains to compare between different them or between different culture conditions [ 64 , 65 , 83 , 87 , 88 , 89 , 90 ]. However, structural proteomics have been used to achieve reference maps of different sub-proteomes (e.g.…”

“…In parallel to an increase in penicillin titers due to the screening of new penicillin-producer strains, the production processes were also focused on deep-tank fermentation and nutrient supply [ 3 , 103 ]. The proteome analysis of the changes between pilot and industrial fermentation scales of P. chrysogenum was the fourth expression proteomics published [ 83 ]. The strain QMSHB-11-06 of P. chrysogenum grown in 50-L fermenters (pilot scale) and 150,000-L fermenters (industrial scale) was used to analyze its proteomes.…”

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

“…Some of the strains developed by companies for industrial production have recently had their genomes sequenced [ 26 , 27 , 28 ]. In addition, transcriptomic, proteomic, and metabolomic studies have been performed trying to elucidate the metabolic changes underwent by industrial strains that led to the high productivity they possess [ 20 , 21 , 26 , 29 , 30 , 31 , 32 , 33 , 34 , 35 ]. Some recent reviews summarize and make a comprehensive account of the main results obtained in these studies [ 25 , 36 , 37 , 38 , 39 , 40 ].…”