Commercial riboflavin production by recombinant Bacillus subtilis : down-stream processing and comparison of the composition of riboflavin produced by fermentation or chemical synthesis

Bretzel, Werner; Schurter, W.; Ludwig, Bernd; Kupfer, Ernst; Doswald, S; Pfister, Martin; Loon, A. P. G. M. Van

doi:10.1038/sj.jim.2900604

Cited by 42 publications

(16 citation statements)

References 7 publications

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“…# 83885) (Supplementary material S2). The crystal shape of the yellow riboflavin precipitate agrees with previous Bretzel's observation (Bretzel et al, ). Indeed, although clostridia species is known to produce riboflavin (Lim et al, ), as illustrated in Figure for the simplified metabolic network used for the model, but at concentrations that are close‐to‐under the detection limit to the best of our knowledge (Arzberger, ; Hickey, ; Lim et al, ).…”

Section: Resultssupporting

confidence: 91%

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

Zhao

Kasbi

Chen

et al. 2017

Biotech & Bioengineering

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The present study reveals that supplementing sodium acetate (NaAc) strongly stimulates riboflavin production in acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum ATCC 824 with xylose as carbon source. Riboflavin production increased from undetectable concentrations to ∼0.2 g L (0.53 mM) when supplementing 60 mM NaAc. Of interest, solvents production and biomass yield were also promoted with fivefold acetone, 2.6-fold butanol, and 2.4-fold biomass adding NaAc. A kinetic metabolic model, developed to simulate ABE biosystem, with riboflavin production, revealed from a dynamic metabolic flux analysis (dMFA) simultaneous increase of riboflavin (ribA) and GTP (precursor of riboflavin) (PurM) synthesis flux rates under NaAc supplementation. The model includes 23 fluxes, 24 metabolites, and 72 kinetic parameters. It also suggested that NaAc condition has first stimulated the accumulation of intracellular metabolite intermediates during the acidogenic phase, which have then fed the solventogenic phase leading to increased ABE production. In addition, NaAc resulted in higher intracellular levels of NADH during the whole culture. Moreover, lower GTP-to-adenosine phosphates (ATP, ADP, AMP) ratio under NaAc supplemented condition suggests that GTP may have a minor role in the cell energetic metabolism compared to its contribution to riboflavin synthesis.

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Section: Resultssupporting

confidence: 91%

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

Zhao

Kasbi

Chen

et al. 2017

Biotech & Bioengineering

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“…The latter organism has been successfully employed at a commercial scale to produce riboflavin for feed and food fortification purposes. However, since this production procedure involved a genetically recombinant organism regulatory approval was required where substantial equivalence of the product to non-recombinant riboflavin had to be established, while no DNA from the production strain was allowed to be present in the final product [ 34 ]. The purpose of the present study was not to develop strains which would challenge these already available processes, but rather to look at the potential to isolate strains with an improved riboflavin production phenotype that could replace the riboflavin-consuming parent strains in traditional food fermentation processes thus improving the bioavailability of riboflavin.…”

Section: Discussionmentioning

confidence: 99%

A general method for selection of riboflavin-overproducing food grade micro-organisms

et al. 2006

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“…The main industrial producer strains of riboflavin include two yeast-like fungi Eremothecium ashbyii and Ashbya gossypii, as well as the bacteria Bacillus subtilis [8]. Compared to fungal fermentation, bacterial fermentation has several advantages for riboflavin biosynthesis, such as a short fermentation time, simple culture media, and the mature genetic engineering technologies applied to prokaryotic bacteria [9,10]. Lactic acid bacteria (LAB) are a group of probiotic bacteria that are closely related to human life, and they have been widely used in the food industry to expand the source and diversity of food, as well as to improve the nutritional value of food.…”

Section: Introductionmentioning

confidence: 99%

Screening and Spontaneous Mutation of Pickle-Derived Lactobacillus plantarum with Overproduction of Riboflavin, Related Mechanism, and Food Application

Zhang

Corke

et al. 2020

Foods

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Riboflavin, also known as vitamin B2, plays an important role in human cell metabolism and participates in various redox reactions and in energy utilization. In this study, 90 riboflavin-producing lactic acid bacteria (LAB) were screened out from pickle juices. The yields of riboflavin in these LAB were about 0.096–0.700 mg/L, and one strain, Lactobacillus plantarum RYG-YYG-9049, was found to produce the highest riboflavin content. Next, roseoflavin was used to induce the spontaneous mutation of RYG-YYG-9049, and selected roseoflavin-resistant colonies generally produced higher riboflavin contents, ranging from 1.013 to 2.332 mg/L. The No. 10 mutant, L. plantarum RYG-YYG-9049-M10, had the highest riboflavin content. Next, the molecular mechanism of enhancing riboflavin production in RYG-YYG-9049-M10 was explored, leading to the finding that roseoflavin treatment did not change the rib operons including the ribA, ribB, ribC, ribH, and ribG genes. Unexpectedly, however, this mechanism did induce an insertion of a 1059-bp DNA fragment in the upstream regulatory region of the rib operon, as compared to the wild-type RYG-YYG-9049. To the best of our knowledge, this is the first report that roseoflavin could induce an insertion of DNA fragment in LAB to increase riboflavin content, representing a new mutation type that is induced by roseoflavin. Finally, in order to fortify riboflavin content in soymilk, RYG-YYG-9049 and RYG-YYG-9049-M10 were used to ferment soymilk, and several fermentation parameters were optimized to obtain the fermented soymilk with riboflavin contents of up to 2.920 mg/L. In general, roseoflavin induction is an economical and feasible biotechnological strategy to induce riboflavin-overproducing LAB, and this strategy can be used to develop LAB-fermented functional foods that are rich in riboflavin.

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Commercial riboflavin production by recombinant Bacillus subtilis : down-stream processing and comparison of the composition of riboflavin produced by fermentation or chemical synthesis

Cited by 42 publications

References 7 publications

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

A dynamic metabolic flux analysis of ABE (acetone‐butanol‐ethanol) fermentation by Clostridium acetobutylicum ATCC 824, with riboflavin as a by‐product

A general method for selection of riboflavin-overproducing food grade micro-organisms

Screening and Spontaneous Mutation of Pickle-Derived Lactobacillus plantarum with Overproduction of Riboflavin, Related Mechanism, and Food Application

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