Identification of peptides with antioxidant and antihypertensive capacities by RP-HPLC-Q-TOF-MS in dry fermented camel sausages inoculated with different starter cultures and ripening times

2017 ◽  
Vol 100 ◽  
pp. 708-716 ◽  
Author(s):  
Lobna Mejri ◽  
Romy Vásquez-Villanueva ◽  
Mnasser Hassouna ◽  
María Luisa Marina ◽  
María Concepción García
Keyword(s):  
Starter Cultures ◽  
Rp Hplc ◽  
Tof Ms

scholarly journals Insights into the Potential of Sourdough-Related Lactic Acid Bacteria to Degrade Proteins in Wheat

2020 ◽  
Vol 8 (11) ◽  
pp. 1689 ◽  
Author(s):  
Vera Fraberger ◽  
Martin Ladurner ◽  
Alexandra Nemec ◽  
Clemens Grunwald-Gruber ◽  
Lisa M. Call ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Competitive Elisa ◽  
Starter Cultures ◽  
Bakery Products ◽  
Wheat Sensitivity ◽  
Immunogenic Proteins ◽  
High Degree ◽  
Selection Of ◽  
Tof Ms

Sourdough processing contributes to better digestible wheat-based bakery products, especially due to the proteolytic activity of lactic acid bacteria (LAB). Therefore, sourdough-related LAB were screened for their capacity to degrade immunogenic proteins like gluten and alpha-amylase-trypsin inhibitors (ATIs). Firstly, the growth of 87 isolates was evaluated on a gluten-based medium. Further, the breakdown capacity of selected isolates was determined for gluten with a focus on gliadins by measuring acidification parameters and MALDI-TOF MS protein profiles. ATI degradation after 72 h of incubation within an ATI-based medium was investigated by means of acidification, HPLC, and competitive ELISA. All isolates exhibited the potential to degrade ATIs to a high degree, whereas the gliadin degradation capacity varied more greatly among tested LAB, with Lacticaseibacillus paracasei Lpa4 exhibiting the strongest alterations of the gliadin pattern, followed by Lactiplantibacillus plantarum Lpl5. ATI degradation capacities ranged from 52.3% to 85.0% by HPLC and 22.2% to 70.2% by ELISA, with Lacticaseibacillus paracasei Lpa4 showing superior breakdown properties. Hence, a selection of specific starter cultures can be used in sourdough processing for wheat-based bakery products with reduced gluten and ATI content and, further, better tolerated products for patients suffering from non-celiac wheat sensitivity (NCWS).

scholarly journals Development of an Optimized MALDI-TOF-MS Method for High-Throughput Identification of High-Molecular-Weight Glutenin Subunits in Wheat

Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4347
Author(s):  
You-Ran Jang ◽  
Kyoungwon Cho ◽  
Se Won Kim ◽  
Susan B. Altenbach ◽  
Sun-Hyung Lim ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Throughput ◽  
Glutenin Subunits ◽  
Maldi Tof Ms ◽  
Molecular Weights ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Group 2 ◽  
Tof Ms

Because high-molecular-weight glutenin subunits (HMW-GS) are important contributors to wheat end-use quality, there is a need for high-throughput identification of HMW-GS in wheat genetic resources and breeding lines. We developed an optimized method using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) to distinguish individual HMW-GS by considering the effects of the alkylating reagent in protein extraction, solvent components, dissolving volume, and matrix II components. Using the optimized method, 18 of 22 HMW-GS were successfully identified in standard wheat cultivars by differences in molecular weights or by their associations with other tightly linked subunits. Interestingly, 1Bx7 subunits were divided into 1Bx7 group 1 and 1Bx7 group 2 proteins with molecular weights of about 82,400 and 83,000 Da, respectively. Cultivars containing the 1Bx7 group 2 proteins were distinguished from those containing 1Bx7OE using well-known DNA markers. HMW-GS 1Ax2* and 1Bx6 and 1By8 and 1By8*, which are difficult to distinguish due to very similar molecular weights, were easily identified using RP-HPLC. To validate the method, HMW-GS from 38 Korean wheat varieties previously evaluated by SDS-PAGE combined with RP-HPLC were analyzed by MALDI-TOF-MS. The optimized MALDI-TOF-MS method will be a rapid, high-throughput tool for selecting lines containing desirable HMW-GS for breeding efforts.

Subunit Characteristics of Pig Pancreas Ferritin Revealed by MALDI-TOF MS and RP-HPLC

2008 ◽  
Vol 24 (5) ◽  
pp. 550-556 ◽  
Author(s):  
L HUANG ◽  
Z LIN ◽  
Q LIN ◽  
L LUO ◽  
H HUANG
Keyword(s):  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Tof Ms

Determination and identification of hydrophilic and hydrophobic arsenic species in methanol extract of fresh cod liver by RP-HPLC with simultaneous ICP-MS and ESI-Q-TOF-MS detection

2013 ◽  
Vol 141 (3) ◽  
pp. 3093-3102 ◽  
Author(s):  
Uriel Arroyo-Abad ◽  
Susanne Lischka ◽  
Christian Piechotta ◽  
Jürgen Mattusch ◽  
Thorsten Reemtsma
Keyword(s):  
Methanol Extract ◽  
Arsenic Species ◽  
Ms Detection ◽  
Rp Hplc ◽  
Icp Ms ◽  
Tof Ms

scholarly journals Comparison of MALDI-TOF-MS and RP-HPLC as Rapid Screening Methods for Wheat Lines With Altered Gliadin Compositions

2020 ◽  
Vol 11 ◽  
Author(s):  
You-Ran Jang ◽  
Kyoungwon Cho ◽  
Sewon Kim ◽  
Jae-Ryeong Sim ◽  
Su-Bin Lee ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Mutation Breeding ◽  
Rapid Screening ◽  
Food Allergies ◽  
Screening Methods ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Rp Hplc ◽  
Wheat Lines ◽  
Tof Ms

The wheat gliadins are a complex group of flour proteins that can trigger celiac disease and serious food allergies. As a result, mutation breeding and biotechnology approaches are being used to develop new wheat lines with reduced immunogenic potential. Key to these efforts is the development of rapid, high-throughput methods that can be used as a first step in selecting lines with altered gliadin contents. In this paper, we optimized matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and reversed-phase high-performance liquid chromatography (RP-HPLC) methods for the separation of gliadins from Triticum aestivum cv. Chinese Spring (CS). We evaluated the quality of the resulting profiles using the complete set of gliadin gene sequences recently obtained from this cultivar as well as a set of aneuploid lines in CS. The gliadins were resolved into 13 peaks by MALDI-TOF-MS. α- or γ-gliadins that contain abundant celiac disease epitopes and are likely targets for efforts to reduce the immunogenicity of flour were found in several peaks. However, other peaks contained multiple α- and γ-gliadins, including one peak with as many as 12 different gliadins. In comparison, separation of proteins by RP-HPLC yielded 28 gliadin peaks, including 13 peaks containing α-gliadins and eight peaks containing γ-gliadins. While the separation of α- and γ-gliadins gliadins achieved by RP-HPLC was better than that achieved by MALDI-TOF-MS, it was not possible to link peaks with individual protein sequences. Both MALDI-TOF-MS and RP-HPLC provided adequate separation of ω-gliadins. While MALDI-TOF-MS is faster and could prove useful in studies that target specific gliadins, RP-HPLC is an effective method that can be applied more broadly to detect changes in gliadin composition.

scholarly journals Use of Maldi-Tof MS biosensor in microbial assessment of Brazilian kefir grains

Revista CERES ◽  
2019 ◽  
Vol 66 (1) ◽  
pp. 72-76
Author(s):  
Roberta Oliveira Viana ◽  
Karina Teixeira Magalhães-Guedes ◽  
Disney Ribeiro Dias ◽  
Rosane Freitas Schwan
Keyword(s):  
Microbial Population ◽  
Luria Bertani ◽  
Starter Cultures ◽  
Accurate Identification ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Microbiological Control ◽  
Identification Of Species ◽  
Kefir Grains ◽  
Tof Ms

ABSTRACT The aim of this study was to evaluate the use of Maldi-Tof MS biosensor in microbial assessment of Brazilian kefir grains. Maldi-Tof MS is a new methodology for the rapid diagnosis of microorganisms. A total of 358 microorganisms were isolated, 31 were yeasts and 327 were bacteria (divided into lactic and acetic bacteria). Microbial colonies were grown in Luria-Bertani agar medium and incubated at 35 °C for 18h and used in the identification of species by Maldi-Tof MS. The microbial population identified in Brazilian kefir grains was Lactobacillus paracasei, Saccharomyces cerevisiae, Lactobacillus plantarum, Acetobacter pasteurianus, and Acetobacter syzygii. This study demonstrated a rapid and accurate identification of the Brazilian kefir grains microorganisms using the Maldi-Tof MS biosensor. In conclusion, the Maldi-Tof MS technology can facilitate the microbiological control in a fermentation process using kefir grains as starter cultures.

scholarly journals Sintesis Tetrapeptida PADY menggunakan Metode Fasa Padat dan Aktivitas Antioksidannya

2019 ◽  
Vol 5 (1) ◽  
Author(s):  
Rani Maharani Ph.D ◽  
Dadan Sumiarsa ◽  
Christina Marpaung ◽  
Achmad Zainuddin ◽  
Ace Tatang Hidayat ◽  
...  
Keyword(s):  
Antioxidant Activity ◽  
Peptide Synthesis ◽  
1H Nmr ◽  
Solid Phase ◽  
Solid Phase Peptide Synthesis ◽  
Synthesis Method ◽  
Antioxidant Assay ◽  
Rp Hplc ◽  
Amur Sturgeon ◽  
Tof Ms

Peptida antioksidan merupakan kelompok peptida yang berperan penting karena dapat menetralkan radikal bebas, sehingga dapat mencegah dan mengobati penyakit kronis. Salah satu senyawa peptida antioksidan alami yang telah ditemukan peneliti sebelumnya adalah senyawa tetrapeptida PAGY (Pro-Ala-Gly-Tyr) yang diisolasi dari gelatin kulit ikan amur sturgeon (Acipenser schrenckii) dan dilaporkan memiliki aktivitas antioksidannya dengan IC50 5.38 mg/mL dalam uji DPPH dan 0,008 mg/mL dalam uji ABTS berturut-turut. Kelompok kami telah berhasil mensintesis PAGY bersama dengan analognya yakni PSGY, PFFY, PAFY, dan PAIY dengan menggunakan metode sintesis peptida fase padat (SPPS). Pengujian aktivitas antioksidan pada senyawa hasil sintesis menunjukkan bahwa PSGY memiliki aktivitas antioksidan lebih baik dari PAGY. Pencarian analog tetrapeptida antioksidan yang lebih baik hingga saat ini masih terus dilakukan. Pada penelitian ini telah berhasil disintesis analog tetrapeptida lainnya PADY (Pro-Ala-Asp-Tyr) dengan metode sintesis peptida fase padat menggunakan strategi Fmoc/t-Bu pada resin 2-klorotritilklorida dilanjutkan dengan pengujian aktivitas antioksidannya. HATU/HOAT digunakan sebagai reagen pengkopling dalam sintesis PADY. Pemurnian krud PADY dilakukan menggunakan RP-HPLC preparatif sehingga diperoleh PADY murni seberat 14.7 mg (12.6%). Penentuan struktur peptida hasil sintesis dianalisis dengan menggunakan spektroskopi 1H-NMR dan TOF-MS. Pada pengujian aktivitas antioksidan dengan metode DPPH, PADY hasil sintesis memberikan nilai IC50 sebesar 1.850 mg/mL, yang mengindikasikan bahwa PADY menunjukkan aktivitas antioksidan yang lebih rendah daripada PAGY hasil sintesis peneliti sebelumnya. Kata kunci: Antioksidan, tetrapeptida, sintesis peptida fase padat. Antioxidant peptide is a class of peptides that play an important in neutralizing free radicals, therefore this compound can be used to prevent and treat chronic diseases. One of the natural antioxidant peptides reported by previous researcher is PAGY (Pro-Ala-Gly-Tyr), which is isolated from amur sturgeon fish (Acipenser schrenckii) gelatin that showed antioxidant activity with IC50 5.38 and 0.008 mg/mL using DPPH and ABTS assay, respectively. Our group has successfully synthesized PAGY, along with its analogues of PSGY, PFFY, PAFY, and PAIY using solid phase peptide synthesis method (SPPS). Antioxidant assay on synthesised compounds showed that PSGY has better antioxidant activity than PAGY. The search on the analogues of the antioxidant tetrapeptide was continued. From this study, a tetrapeptide analogue PADY (Pro-Ala-Asp-Tyr) has been successfully synthesised by solid phase peptide synthesis method with Fmoc/t-Bu strategy on 2-chlorotrityl chloride resin and tested for its antioxidant activity. HATU and HOAt reagents were used as the coupling reagent for the synthesis of PADY. The resulting PADY peptide solid was then purified using preparative RP-HPLC yielding PADY of 14.7 mg (12.6%). Characterisation of the synthesized compound was analysed by  1H-NMR and TOF-MS. On the antioxidant assay using DPPH method, PADY showed IC50 value of 1.850 mg/mLindicating a lower activity than the synthetic PAGY. Keywords: Antioxidant, tetrapeptide, solid phase peptide synthesis (SPPS).

Sign in / Sign up

Export Citation Format

Share Document