Acid Hydrolysis of Protein in a Microcapillary Tube for the Recovery of Tryptophan

Adebiyi, Abayomi P.; Jin, Dong Hao; Ogawa, Tomoya; Muramoto, Koji

doi:10.1271/bbb.69.255

Cited by 24 publications

(9 citation statements)

References 6 publications

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“…The amino acid (AA) profile of raw FMs and MMs was assessed after the hydrolysis step using liquid chromatography coupled with mass spectrometry. The hydrolysis of the raw material was carried out according to the method described by Adebiyi et al [52] with minor modifications. In brief, 2 g of dry powder of each sample was suspended in 15 mL of 0.1 M HCl and homogenized using ULTRA-TURRAX T25 (IKA ® -Werke GmbH & Co. KG, Staufen, Germany) for 90 s at 4 • C. Then, 50 µL of the suspension was mixed with 150 µL of a freshly prepared solution of 6 N HCl, 3% phenol, and 1% 2-mercaptoethanol and incubated for 30 min at 160 • C in a glass-sealed vial to obtain complete hydrolysis of protein material, as this temperature allows rapid hydrolysis of the protein component with a recovery mean values of the AAs greater than 98%; however, cysteine cannot be detected under these conditions [53,54].…”

Section: Amino Acid Profilementioning

confidence: 99%

An Alternative Approach to Evaluate the Quality of Protein-Based Raw Materials for Dry Pet Food

Montegiove

Pellegrino

Emiliani

et al. 2021

Animals

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The majority of dry pet food currently on the market is produced using fresh meats (FMs) and especially meat meals (MMs) as the main protein source. The transport and storage conditions of the raw materials, together with thermal and mechanical treatments in the case of MMs, may result in undesirable alterations of food products and their protein content. This study was conducted to analyze the protein component of three different kinds of raw materials used for dry pet food production, i.e., chicken, pork, and salmon. The quantitative analysis of the protein component was determined using the traditional Kjeldahl method and near-infrared (NIR) spectroscopy, and an alternative method, i.e., the Bradford assay, while the qualitative analysis was performed through SDS-PAGE, followed by Coomassie Blue staining. The amino acid (AA) profile was also evaluated by quadrupole time-of-flight liquid chromatography/mass spectrometry (Q-TOF LC/MS). In addition, the digestibility was tested through in vitro gastric and small intestine digestion simulation. Statistical analysis was performed by the Student’s t-test, and data are reported as mean ± SEM, n = 10 (p < 0.05). The results showed that the MMs are lower in quality compared to FMs, both in terms of protein bioavailability and digestibility, having a lower soluble protein (SP) content (chicken MM = 8.6 g SP/100 g dry sample; pork MM = 6.2 g SP/100 g dry sample; salmon MM = 7.9 g SP/100 g dry sample) compared to FMs (chicken FM = 14.6 g SP/100 g dry sample; pork FM = 15.1 g SP/100 g dry sample; salmon FM = 13.7 g SP/100 g dry sample). FMs appear, therefore, to be higher-quality ingredients for pet food production. Moreover, the Bradford assay proved to be a quick and simple method to better estimate protein bioavailability in the raw materials used for dry pet food production, thanks to its correlation with the in vitro digestibility.

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Section: Amino Acid Profilementioning

confidence: 99%

An Alternative Approach to Evaluate the Quality of Protein-Based Raw Materials for Dry Pet Food

Montegiove

Pellegrino

Emiliani

et al. 2021

Animals

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“…The advantage of using methane sulfonic acid in comparison to HCl is that tryptophan is not destroyed and methionine is determined as methionine sulfoxide. To reduce the losses incurred by the acid hydrolysis certain protective agents such as phenol, thioglycolic acid, mercapto ethanol, indole or tryptamine are added to the sample Adebiyi et al [117] used a single step protein hydrolysis process with 4 M methanesulfonic acid at 115°C for 22 h in the presence of 0.02% tryptamine, in which even tryptophan and cysteine were determined.…”

Section: Acid Hydrolysis Of Fish Proteinsmentioning

confidence: 99%

Fish Processing Wastes as a Potential Source of Proteins, Amino Acids and Oils: A Critical Review

Brooks¹

2013

Microb Biochem Technol

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The fish processing industry is a major exporter of seafood and marine products in many countries. About 70% of the fish is processed before final sale. Processing of fish involves stunning, grading, slime removal, deheading, washing, scaling, gutting, cutting of fins, meat bone separation and steaks and fillets. During these steps significant amount of waste (20-80% depending upon the level of processing and type of fish) is generated which can be utilized as fish silage, fishmeal and fish sauce. Fish waste can also be used for production of various value added products such as proteins, oil, amino acids, minerals, enzymes, bioactive peptides, collagen and gelatin. The fish proteins are found in all parts of the fish. There are three types of proteins in fish: structural proteins, sacroplasmic proteins and connective tissue proteins. The fish proteins can be extracted by chemical and enzymatic process. In the chemical method, salts (NaCl and LiCl) and solvents (isopropanol and aezotropic isopropanol) are used, whereas during the enzymatic extraction, enzymes (alcalase, neutrase, protex, protemax and flavorzyme) are used to extract proteins from fish. These fish proteins can be used as a functional ingredient in many food items because of their properties (water holding capacity, oil absorption, gelling activity, foaming capacity and emulsifying properties). They can also be used as milk replacers, bakery substitutes, soups and infant formulas. The amino acids are the building blocks of protein. There are 16-18 amino acids present in fish proteins. The amino acids can be produced from fish protein by enzymatic or chemical processes. The enzymatic hydrolysis involves the use of direct protein substrates and enzymes such as alcalase, neutrase, carboxypeptidase, chymotrypsin, pepsin and trypsin. In the chemical hydrolysis process, acid or alkali is used for the breakdown of protein to extract amino acids. The main disadvantage of this method is the complete destruction of tryptophan and cysteine and partial destruction of tyrosine, serine and threonine. The amino acids present in the fish can be utilized in animal feed in the form of fishmeal and sauce or can be used in the production of various pharmaceuticals. The fish oil contains two important polyunsaturated fatty acids called EPA and DHA or otherwise called as omega-3 fatty acids. These omega-3 fatty acids have beneficial bioactivities including prevention of atherosclerosis, protection against maniac-depressive illness and various other medicinal properties. Fish oil can also be converted to non-toxic, biodegradable, environment friendly biodiesel using chemical or enzymatic transesterification.

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“…(Refer to Supplementary Figure S2 and Supplementary Table S1 for complete results of amino acid analysis of the purified CFTR standard in amphipol.) Alanine (1015.112 pmoles were present in the 100 μL standard) was chosen to determine the concentration of CFTR standard as amino acid hydrolysis with various solvents resulted in 100% recovery of alanine [66] and there were no alanine residues in the FLAG ® peptide that was also present in the standard. The calculation of the concentration of the CFTR standard is as follows:weight Alaweight CFTR=number of Ala∗MW AlaMW CFTR 72065 pgweight CFTR=83∗71 pg/pmol170507.57 pg/pmol…”

Section: Methodsmentioning

confidence: 99%

Cholesterol Interaction Directly Enhances Intrinsic Activity of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)

Chin

Ramjeesingh

Hung

et al. 2019

Cells

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The recent cryo-electron microscopy structures of zebrafish and the human cystic fibrosis transmembrane conductance regulator (CFTR) provided unprecedented insights into putative mechanisms underlying gating of its anion channel activity. Interestingly, despite predictions based on channel activity measurements in biological membranes, the structure of the detergent purified, phosphorylated, and ATP-bound human CFTR protein did not reveal a stably open conduction pathway. This study tested the hypothesis that the functional properties of the detergent solubilized CFTR protein used for structural determinations are different from those exhibited by CFTR purified under conditions that retain associated lipids native to the membrane. It was found that CFTR purified together with phospholipids and cholesterol using amphipol: A8-35, exhibited higher rates of catalytic activity, phosphorylation dependent channel activation and potentiation by the therapeutic compound, ivacaftor, than did CFTR purified in detergent. The catalytic activity of phosphorylated CFTR detergent micelles was rescued by the addition of phospholipids plus cholesterol, but not by phospholipids alone, arguing for a specific role for cholesterol in modulating this function. In summary, these studies highlight the importance of lipid interactions in the intrinsic activities and pharmacological potentiation of CFTR.

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Acid Hydrolysis of Protein in a Microcapillary Tube for the Recovery of Tryptophan

Cited by 24 publications

References 6 publications

An Alternative Approach to Evaluate the Quality of Protein-Based Raw Materials for Dry Pet Food

An Alternative Approach to Evaluate the Quality of Protein-Based Raw Materials for Dry Pet Food

Fish Processing Wastes as a Potential Source of Proteins, Amino Acids and Oils: A Critical Review

Cholesterol Interaction Directly Enhances Intrinsic Activity of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR)

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