Inoculation‐density‐dependent responses and pathway shifts in <i>Saccharomyces cerevisiae</i>

Cheng, Jun; Ding, Ming-Zhu; Tian, Hong-Chi; Yang, Yuan

doi:10.1002/pmic.200900249

Cited by 20 publications

(16 citation statements)

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“…Previous studies have shown that the initial cell concentration not only influences cellular and molecular behaviors [10,11] but also changes the lag phase of cell growth, growth and fermentation rates, final product concentration, and yield [12][13][14][15]. This study examined the effects of the initial cell concentration of MA-R4 on fermentation parameters using complex medium containing xylose or mixtures of glucose and xylose.…”

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confidence: 98%

Effect of Initial Cell Concentration on Ethanol Production by Flocculent Saccharomyces cerevisiae with Xylose-Fermenting Ability

Matsushika

Sawayama

2010

Appl Biochem Biotechnol

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Different initial cell concentrations of a recombinant flocculent Saccharomyces cerevisiae MA-R4 were evaluated for their effects on xylose fermentation and glucose-xylose cofermentation. A high initial cell concentration greatly increased both the substrate utilization and ethanol production rates. During xylose fermentation, the highest rates of xylose consumption (2.58 g/L h) and ethanol production (0.83 g/L h) were obtained at an initial cell concentration of 13.1 g/L. During cofermentation, the highest rates of glucose consumption (14.4 g/L h), xylose consumption (2.79 g/L h), and ethanol production (6.68 g/L h) were obtained at an initial cell concentration of 12.7 g/L. However, a high initial cell density had no positive effect on the maximum ethanol concentration and ethanol yield mainly due to the increased amount of by-products including xylitol. The ethanol yield remained almost constant (0.34 g/g) throughout xylose fermentation (initial cell concentration range, 1.81-13.1 g/L), while it was slightly lower at high initial cell concentrations (9.87 and 12.7 g/L) during cofermentation. The determination of the appropriate initial cell concentration is necessary for the improvement of substrate utilization and ethanol yield.

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confidence: 98%

Effect of Initial Cell Concentration on Ethanol Production by Flocculent Saccharomyces cerevisiae with Xylose-Fermenting Ability

Matsushika

Sawayama

2010

Appl Biochem Biotechnol

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“…However, in Actinomycetes YJ1 strain, the maximum antibiotic activity against Sclerotinia sclerotiorum was obtained at the inoculation volume of 5 %, while the antibiotic activity was not enhanced with the increases of pitching ratio (Song et al 2012). It was also demonstrated that the high pitching ratio (30 %, v/v) significantly enhanced the streptolydigin production of S. lydicus E9 (Cheng et al 2009). Herein, the most detected metabolites including amino acids were significant variations under different pitching ratios (Fig.…”

Section: Discussionmentioning

confidence: 78%

“…In S. cerevisiae, high inoculation size (40 gL −1 ) significantly shortened the fermentation and enhanced final ethanol production (Cheng et al 2009). It was reported that the high cell density fermentations could be achieved by improving the oxygen conditions without affecting drastically the cellular physiological conditions (Verbelen et al 2009).…”

Section: Discussionmentioning

confidence: 99%

“…In Streptomyces coelicolor, concordance analysis was applied to detect reliable changes for 97 putatively identified metabolics that are significantly concordant and reproducible across the time series (Jankevics et al 2011). Previous researches demonstrated that the pathway shifts and variations of metabolite levels in Saccharomyces cerevisiae responded the stresses triggered by high cell density (Cheng et al 2009;Ding et al 2009a). Comparative LC-MS analysis in S. lydicus under 10 % pitching ration was rapid, sensitive, and suitable for characterizing intermediates, metabolites, and pathways for elucidation of the metabolic alterations during strain improvement (Li et al 2006).…”

Section: Introductionmentioning

confidence: 99%

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Metabolic analysis reveals the amino acid responses of Streptomyces lydicus to pitching ratios during improving streptolydigin production

Cheng

Liang

Ding

et al. 2013

Appl Microbiol Biotechnol

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Pitching ratio has been reported to impact not only on the primary metabolism, but also the secondary metabolism. Comparative metabolomics was used to explore the metabolic responses of Streptomyces lydicus E9 to pitching ratios (1, 10, and 30%, v/v). We identified more than 120 metabolites involved in glycolysis, tricarboxylic acid cycle, and amino acid and secondary metabolism, of which there are significant differences in the quantified 32 metabolites under different pitching ratios by gas chromatography coupled to time-of-flight mass spectrometry. The intracellular levels of most amino acids (e.g., valine, alanine, and isoleucine) declined with the increases of pitching ratios. Especially, the relative abundances of glutamate and proline were not only decreased with the increases of pitching rations, but also had much low level at stages II and III, which might be related to the significant enhancement in streptolydigin of S. lydicus E9 under 30% high pitching ratio. Moreover, principal component analysis revealed that eight metabolites, including glucopyranoside, maltose, cAMP, glycine, proline, lysine, isoleucine, and valine, were considered as potential biomarkers to distinguish the influences of pitching ratios on streptolydigin production. Further investigations demonstrated that the additions of exogenous glutamate and proline (100 mgL⁻¹) enhanced significantly the accumulation of streptolydigin, indicating that glutamate was the synthetic precursor of streptolydigin, while proline in S. lydicus E9 was converted into glutamate and consequently improved streptolydigin biosynthesis. Therefore, these findings provide new insights into the amino acid responses of S. lydicus E9 to pitching ratios and provide potential strategies to improve streptolydigin production.

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“…Intracellular protein extraction was carried out as described previously [20]. At the determined time (2,8,20,32,50,80,98,128,140, and 170 h), the fermentation broths were rapidly harvested and then centrifuged for 5 min at 3000 × g. The cells were collected and washed three times with ice-cooled Milli-Q water containing 1 mM phenylmethanesulfonyl fluoride (PMSF) to remove the extracellular components.…”

Section: Protein Extraction and Two-dimensional Gel Electrophoresismentioning

confidence: 99%

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

Cheng

Zhao

Qiao

et al. 2016

Appl Biochem Biotechnol

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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.

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Inoculation‐density‐dependent responses and pathway shifts in Saccharomyces cerevisiae

Cited by 20 publications

References 35 publications

Effect of Initial Cell Concentration on Ethanol Production by Flocculent Saccharomyces cerevisiae with Xylose-Fermenting Ability

Effect of Initial Cell Concentration on Ethanol Production by Flocculent Saccharomyces cerevisiae with Xylose-Fermenting Ability

Metabolic analysis reveals the amino acid responses of Streptomyces lydicus to pitching ratios during improving streptolydigin production

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

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