Assessment of Enzymatic Improvers in Flours Using LC–MS/MS Detection of Marker Tryptic Peptides

Prandi, Barbara; Lambertini, Francesca; Varani, M.; Faccini, Andrea; Suman, Michele; Leporati, Andrea; Tedeschi, Tullia; Sforza, Stefano

doi:10.1021/jasms.9b00109

Cited by 14 publications

(12 citation statements)

References 25 publications

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“…The approach using discovery-driven proteomics underlined that amylase preparations used in baking generally contain a small number of functional proteins responsible for the hydrolytic activity of the preparation. This was also shown before by Picariello et al and Prandi et al [28,29]. The iBAQ evaluation is suitable for the relative quantitation of several different proteins in one sample [25,30] and it showed differences in the protein composition between the preparations.…”

Section: Discussionsupporting

confidence: 74%

Impact of exogenous α-amylases on sugar formation in straight dough wheat bread

Rebholz

Sebald

Dirndorfer

et al. 2020

Eur Food Res Technol

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The use of bacterial or fungal α-amylases is common in wheat bread production to improve several quality-related parameters such as loaf volume, crust color or staling behavior. To study the impact of exogenous α-amylases on straight dough wheat bread, we quantitated mono-, di- and oligosaccharides and residual α-amylase activity in bread crumb during storage for up to 96 h. Discovery-driven proteomics of the five α-amylase preparations studied showed that only a few different amylases per preparation were responsible for the hydrolytic effect. Compared to the control, the supplementation with α-amylase from Bacillus amyloliquefaciens in wheat dough preparation led to major changes in the sugar composition of bread crumb during storage with the formation of oligosaccharides like maltopentaose, maltohexaose, maltoheptaose, and maltooctaose. A residual activity corresponding to 4.0% of the applied activity was determined in the breads prepared with α-amylase from B. amyloliquefaciens, but no residual activity was detected for any of the other fungal or bacterial α-amylases from Aspergillus oryzae or Thermoactinomyces vulgaris. Whether the detected residual activity is related to the characteristics of bread staling or bread crumb properties must be clarified in further studies.

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Section: Discussionsupporting

confidence: 74%

Impact of exogenous α-amylases on sugar formation in straight dough wheat bread

Rebholz

Sebald

Dirndorfer

et al. 2020

Eur Food Res Technol

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“…For the group of maltotetraogenic amylase and the group of maltogenic α-amylase preparations, one amylase each was identified as being responsible for the amylolytic activity. Our results are in accordance with Prandi et al who also identified P19531, a maltogenic α-amylase from G. stearothermophilus, as being responsible for the amylolytic activity in maltogenic α-amylase preparations applied in baking [28]. To the best of our knowledge, we are the first to analyze maltotetraogenic amylase preparations with proteomics tools and we identified a single amylase P22963 as cause of the amylolytic activity.…”

Section: Discussionsupporting

confidence: 92%

Impact of exogenous maltogenic α-amylase and maltotetraogenic amylase on sugar release in wheat bread

Rebholz

Sebald

Dirndorfer

et al. 2021

Eur Food Res Technol

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The use of exogenous maltogenic α-amylases or maltotetraogenic amylases of bacterial origin is common in wheat bread production, mainly as antistaling agents to retard crumb firming. To study the impact of maltogenic α-amylase and maltotetraogenic amylase on straight dough wheat bread, we performed a discovery-driven proteomics approach with commercial enzyme preparations and identified the maltotetraogenic amylase P22963 from Pelomonas saccharophila and the maltogenic α-amylase P19531 from Geobacillus stearothermophilus, respectively, as being responsible for the amylolytic activity. Quantitation of mono-, di- and oligosaccharides and residual amylase activity in bread crumb during storage for up to 96 h clarified the different effects of residual amylase activity on the sugar composition. Compared to the control, the application of maltogenic α-amylase led to an increased content of maltose and especially higher maltooligosaccharides during storage. Residual amylase activity was detectable in the breads containing maltogenic α-amylase, whereas maltotetraogenic amylase only had a very low residual activity. Despite the residual amylase activities and changes in sugar composition detected in bread crumb, our results do not allow a definite evaluation of a potential technological function in the final product. Rather, our study contributes to a fundamental understanding of the relation between the specific amylases applied, their residual activity and the resulting changes in the saccharide composition of wheat bread during storage.

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“…The dry protein extracts were dissolved in 0.1 M HCl (50 mg in 10 ml) or in 75 mM HCl in 25% acetonitrile and 75% water (25 mg in 4 ml) depending on their solubility. Electrophoresis and identification of the protein bands were carried out as described in a previous work ( 15 ). Briefly, a sample volume corresponding to 30 μg of protein was dried under nitrogen flow and reconstituted with reducing sample buffer (sample buffer XT 4× and reducing agent XT 20×, suitably diluted with distilled water; Biorad, Hercules, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Targeting the Nutritional Value of Proteins From Legumes By-Products Through Mild Extraction Technologies

et al. 2021

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Legumes have been known for centuries for their good nutritional properties. Unfortunately, during processing, from 5 to 25% of this production is wasted, generating by-products that can still be a rich source of useful compounds, such as proteins, which can still be used in food and feed formulations. The choice of the extraction technique is important to preserve the nutritional value of proteins since drastic conditions of pH and/or temperature could damage them. In this work, two mild extraction techniques (direct assisted extraction—DAE and enzymatic assisted extraction—EAE) were applied for protein extraction from legume by-products obtained from agro-industrial processes. The quality of proteins was evaluated considering protein integrity [SDS-PAGE, degree of hydrolysis (DH), free amino acid content, racemization degree] and nutritional features [amino acid score (AAS), digestibility]. Direct assisted extraction is the technique that has best preserved protein integrity (1–5% DH and free amino acid content <1%), The digestibility of proteins extracted with EAE is higher (no protein bands detected in SDS-PAGE) than with the one of DAE extracts, making this technique particularly suitable for those food and feed formulation were a high digestibility of proteins is required.

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Assessment of Enzymatic Improvers in Flours Using LC–MS/MS Detection of Marker Tryptic Peptides

Cited by 14 publications

References 25 publications

Impact of exogenous α-amylases on sugar formation in straight dough wheat bread

Impact of exogenous α-amylases on sugar formation in straight dough wheat bread

Impact of exogenous maltogenic α-amylase and maltotetraogenic amylase on sugar release in wheat bread

Targeting the Nutritional Value of Proteins From Legumes By-Products Through Mild Extraction Technologies

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