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ORIGINAL ARTICLE • Braz. J. Food Technol. 27 • 2024 • https://doi.org/10.1590/1981-6723.10823 linkcopy

Open-access Chemical properties and sensory preference related to umami flavor from crude hydrolysate of Indonesian snails (Filopaludina javanica)

AuthorshipSCIMAGO INSTITUTIONS RANKINGS

Abstract

Flavor enhancers are considered important in the food industry for their ability to add umami taste, which is recognized as the fifth basic taste. Although monosodium glutamate (MSG) is the most common umami provider, it contains a sodium molecule that can negatively affect consumer health. This made flavor enhancers derived from natural resources to become an interesting alternative. Therefore, this study aimed to examine the potential of peptides produced from the digestion of the Filopaludina javanica (FJ) snails using two enzymes, particularly, bromelain and trypsin, to determine their umami characteristics. Several enzymatic digestion conditions, including enzyme-to-substrate ratios (E/S) of 1/10, 1/20, and 1/00 (w/v), as well as hydrolysis times of 3, 6, 9, 10, 12, 15, and 18 hours, were examined. The study also assessed the degree of hydrolysis (DH), total peptides, and amino acid content. To evaluate the sensory properties of the resulting peptides, hedonic testing, and principal component analysis were conducted to determine taste, aroma, and color attributes. The results showed that the highest total acceptance was obtained from bromelain and trypsin hydrolysis with an E/S ratio of 1/10 but at different hydrolysis times of 18 and 15 hours, respectively. These samples had DH values of 51.26% and 56.10%, total peptide contents of 8.26 mg/mL and 6.63 mg/mL, and amino acid contents of 88.16 ppm and 79.85 ppm, respectively. The most preferred samples were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) and database-assisted identification to determine their peptide compositions. Among the peptides identified, the top three from bromelain digestion were GPLGPLGPQGIPG, GLPGLPGLPGPKG, and PKEVNVAMIVGL VVAG, while the peptides from trypsin hydrolysate included FDNAGAMMSV, LGGSLLGL, and ATKALAMD CEMVGVGRNGEESVLARVSLVNQHGNCIYDKFVKA. In conclusion, the results showed that snail hydrolysate peptides held promise as natural flavor enhancers.

Keywords:
Indonesian snails; Flavor enhancers; Protein hydrolysate; LC-MS/MS; Sensory analysis

Highlights

The best protease hydrolysis conditions of Filopaludina javanica were found to be an E/S ratio of 1:10 at 15 – 18 hours

Both bromelain and trypsin hydrolysis could provide peptide hydrolysate with a sensory score over 3.5 out of 5

Umami peptides from the snail hydrolysate were reported through the use of LC-MS/MS

1 Introduction

Umami is officially recognized as the fifth sensory taste, alongside sweet, bitter, sour, and salty (Zhang et al., 2017). As a result, it serves as an important flavor enhancer in the food industry. The T1R receptor, consisting of T1R1 and T1R3, has been identified as the key receptor for both sweet and umami taste (Dang et al., 2014; Liu et al., 2019). Commercialized umami enhancers are available in a variety of forms such as monosodium glutamate (MSG), monoammonium glutamate (MAG), disodium inosinate (IMP), and disodium guanylate (GMP) (Jinap & Hajeb, 2010). Moreover, the combination of different flavor enhancers can dramatically increase the umami taste. For instance, Kuninaka reported that the use of glutamate with inosinate or guanylate could greatly enhance the intensity of umami taste (Kuninaka 1960). Among these compounds, MSG is the most widely used umami source worldwide. Although flavor enhancers are important in the food industry, most products are produced through fermentation or chemical processes. Several adverse effects of MSG such as asthma, obesity, reproductive abnormalities, anxiety, and Alzheimer’s disease, have been reported (Husarova & Ostatnikova, 2013; Niaz et al., 2018). Previous reviews also reported that excessive sodium consumption from MSG could negatively affect the human kidney. This shows there is a need to search for alternative natural umami sources that are safe for consumption but still provide a good level of umami taste.

Alternative flavor enhancer sources that have been reported are rohu head (Bruno et al., 2019), straw mushroom (Xu et al., 2019), morel mushroom (Gao et al., 2021), kokimi (Rhyu et al., 2020), puffer fish (Zhang et al., 2012), sunflower seed (Bao et al., 2020), and tempeh (Amin et al., 2020). Among these compounds, peptides are a particularly promising form of flavor enhancers. Peptides, which are short-chain amino acids from 2 to 50 units, can be obtained by breaking down proteins through biological or chemical processes. Although the specific methods by which peptides improve flavor are still not entirely understood, factors such as amino acid composition, amino acid sequence, hydrophilic properties, and molecular weight are believed to play a significant role. Peptides have been found to have the ability to develop unique taste and odor characteristics, thereby improving mouthfulness and aftertaste (Beauchamp, 2009). In addition, there was a significant increase in investigations on the use of peptide derivatives as umami ingredients (Zhang et al., 2017, 2019; Qi et al., 2022). Given their natural origin and potential for modification, peptides represent an interesting challenge to explore for new flavor enhancers.

In the past decade, the use of snails has significantly increased. Snails are not only well-known as a delicacy in Europe but also valued for mucins, which contain active compounds used in skincare products, surgical glues, and wound healing agents (McDermott et al., 2021). Furthermore, the shells of snails can be used to produce chitosan and chitooligosaccharides that possess antioxidant and anticholesterol activities (Larasati et al., 2023; Rosida et al., 2023). Filopaludina javanica (FJ) is one of the 50 dominant snail species found in Indonesia (Priawandiputra et al., 2017, 2020). This edible gastropod found in freshwater areas, is a rich source of protein containing abundant numbers of essential amino acids, calcium, and a modest quantity of fat (Putra et al., 2020; Puspitasari et al., 2022). Given its high protein content, snails have the potential to produce peptides that can serve as flavor enhancers under proteolytic conditions. Previous reviews have used proteolytic enzymes such as bromelain, trypsin, and papain to release umami substances derived from Pila ampullacea (Rosida et al., 2021, 2022; Putra et al., 2021). Several factors that influence the production of peptides derived from edible protein include the type of enzyme, the ratio of enzyme-to-substrate (E/S), hydrolysis time, and temperature (Priyanto et al., 2015).

Based on the above description, this study aims to focus on analyzing peptides generated from the digestion of FJ using two different enzymes, including bromelain and trypsin. The E/S ratio and hydrolysis time were observed, along with the degree of hydrolysis (DH), amino acid concentration, and total peptide contents. Hydrolysate was also evaluated for sensory characteristics. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) associated with a computer database was used to determine the best hydrolysis condition and to identify the most potent umami peptide fragments. The results showed that proteolytic peptide hydrolysate from FJ has the potential to be used as a natural flavor enhancer.

2 Materials and methods

2.1 Materials and chemical

Fresh FJ snails were purchased from a local market in Surabaya City, East Java, Indonesia. Bromelain (EC 3.4.22.33) was manually obtained from fresh pineapple fruit using the method described by (Mohan et al., 2016), resulting in an enzymatic activity of 3.32 ± 0.06 U/mL. Trypsin (EC 3.4.21.4), Probumin® bovine serum albumin (BSA) powder (life science grade), and L-glutamic acid were obtained from Sigma-Aldrich (St. Louis, MO, USA). Other chemicals, including potassium sodium tartrate (KNaC4H4O6.4H2O), sodium hydroxide (NaOH), sodium carbonate (Na2SO3), trichloroacetic acid (TCA), formic acid (FA), acetonitrile (ACN), and Folin-Ciocalteu reagent (FCR), were purchased from Merck KGaA (Darmstadt, Germany). A Pierce BCA™ Protein Assay kit and reagents were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Furthermore, the deionized water was filtered through a Milli-Q® (Millipore) water purification system (Billerica, MA, USA).

2.2 Enzymatic hydrolysis of FJ

Snail flesh was separated from the shells and cleaned under running tap water to remove any mucilage. The cleaned snail flesh was then blended with distilled water at a ratio of 1:2 (w/v) using a blender (Philip HR2115, Amsterdam, Netherlands). Bromelain or trypsin was added to the snail paste at different E/S ratios of 1:10, 1:20, and 1:100 (w/v). The enzymatic reactions were carried out at 54 °C in a Memmert UF55 incubator (Memmert GmbH + Co. KG, Schwabach, Germany) for varying durations of 3, 6, 9, 12, 15, and 18 hours, based on the protease actions of bromelain and trypsin. The hydrolysis reaction was stopped by using high-temperature conditions at 90 oC for 10 minutes. The resulting mixture was centrifuged at room temperature at 3,000 rpm for 30 minutes, and the supernatant was collected for further analysis. These digestion products were further referred to as FJ hydrolysate.

2.3 DH assessment

The level of DH was evaluated using a modified method described by Hoyle and Merrit (Hoyle & Merrit, 1994). A 1 mL aliquot of snail hydrolysate was mixed with 1 mL of 10% TCA solution and incubated for 30 minutes. Subsequently, the mixture was centrifuged at 3,000 rpm for 15 minutes. The supernatant was collected, and the soluble protein content was calculated using the Lowry protein assay method (Lowry et al., 1951), with partial modification by Hartree (Hartree, 1972) and a BSA standard curve. The DH was calculated using the following Equation 1.

DH (%)= [soluble protein content in 10% TCA (mg) / total protein content (mg)] x 100 (1)

2.4 Total peptide content quantification

Total peptide content was quantified using a modified Folin-phenol method (Ledoux & Lamy, 1986). Snail hydrolysate was mixed with 15% TCA at a volume ratio of 2:1 (w/v) and incubated for 1 hour at 25 oC. After the incubation, the mixture was centrifuged at 5,000 rpm for 10 minutes. The color density of the supernatant was measured at a wavelength of 680 nm using a spectrophotometer.

2.5 Determination of amino acid content

Amino acid content was quantified using the Moore and Stein method as described by (Moore & Stein, 1948). To perform the analysis, 5 mL of snail hydrolysate was thoroughly mixed with 2.5 mL of 40% ethanol and 0.5 mL of ninhydrin reagent. The mixture was then heated in boiling water for 20 minutes, and the resulting purple color, which showed the presence of amino acid, was measured at a wavelength of 570 nm using a spectrophotometer. The measured value was interpreted against an L-glutamic acid standard curve to determine the amino acid content.

2.6 Statistical analysis

All results were shown as the mean value ± standard deviation. DH, peptide contents, and amino acid concentrations were statistically analyzed using a two-way analysis of variance (ANOVA), followed by a post hoc test with a false discovery rate of 5%. Dunnet’s test for multiple comparisons (DMRT) was used to analyze the interaction between the E/S ratio and hydrolysis time. Fisher’s least significant difference (LSD) test was used to calculate the significance of only one variable when no interaction was found between the two variables. The result was statistically different when the p-value was less than 0.05. All statistical analyses were performed using Minitab software (version 19.1 2019).

2.7 Sensory evaluation

Sensory evaluation was conducted by a group of well-trained panelists who were capable of identifying five basic tastes such as sweet, bitter, sour, salty, and umami. This group of panelists included 20 students from the Department of Food Technology, Universitas Pembangunan Nasional Veteran Jawa Timur, aged between 20 and 22 years. Umami flavor of FJ hydrolysate was assessed using a 5-point hedonic scale ranging from dislike it extremely to like it extremely. The hedonic parameters were performed based on taste, aroma, color, and total acceptability. Peptide sequences in FJ hydrolysate were then identified using LC-MS/MS, followed by a database search engine to study and summarize the strong potent fragments.

2.8 Peptide identification by LC-MS/MS and database searching

A 5 μL sample of snail hydrolysate containing 0.1% FA was injected into an LC-MS/MS machine (Thermo Scientific Dionex Ultimate 3000 RSLCnano) associated with electrospray ionization (Compact™ quadrupole time-of-flight, Bruker Daltonik, Bremen, Germany). Liquid chromatography (LC) was conducted using a reversed-phase column (Acclaim PepMap RSLC Column C18 NanoViper, 75 µm × 150 mm, particle size 2 µm). The mobile phase consisted of 0.1% FA in deionized water (Solution A) and 80% acetonitrile (ACN) in deionized water (Solution B). The flow rate was maintained at 0.3 μL/min under a linear gradient condition of 2% to 85% solution B in 15 minutes. A mass spectrometry scan was performed in the m/z range of 300 to 1,500, and the tandem mass spectrometry (MS/MS) spectra were generated using Bruker qTOF Control Software and analyzed through the Mascot Server (Matrix Science). The database search engine parameters were arranged in the following ways, (1) peptides and proteins matched the UniProt database of UniProt architaenioglossa, (2) no enzyme cleavage specificity, (3) fixed modification of carbamidomethyl at cysteine residues and variable modification of oxidation at methionine residues, (4) peptides charge of 1+, 2+, and 3+, (5) peptides tolerance of ±1.2 Da, and (6) MS/MS fragment tolerance of ±0.6 Da. Proteins were considered significant when the mascot scores were greater than a threshold, and the scores were fixed with a p-value < 0.05.

3 Results and discussion

3.1 Effect of different proteolysis conditions on the DH in FJ hydrolysate

FJ hydrolysate was obtained through proteolytic digestion using bromelain and trypsin. The value of DH was used to identify the most effective hydrolytic enzyme because it directly showed the extent of peptide bond cleavage in the protein hydrolysate. Figure 1 presented the DH value obtained from bromelain and trypsin digestion of FJ snail flesh. It was evident that long incubation time would provide a high level of DH value for both enzymes. The highest DH value (61.58%) was found with trypsin digestion at an E/S ratio of 1:10 for 18 hours. For trypsin digestion, it was also obvious that increasing the E/S ratio of 1:10 to 1:100 would increase DH results in all incubation times. DH was found to be positively related to umami taste (Wang et al., 2016), showing that high DH value was expected. Therefore, the results suggested that using trypsin at high concentration (1:10 E/S) and long incubation (18 hours) would be more appropriate than using bromelain enzyme. It was observed that bromelain could cleave not only protein but also ester and amide groups (Manzoor et al., 2016). These additional cleavage activities might improve the flavor and taste of food products than trypsin digestion.

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Figure 1
Effect of hydrolysis time and E/S ratio on the DH values (%) of diverse FJ hydrolysate generated from proteolytic digestion of bromelain (continuous line) and trypsin (dash line). The different E/S ratios were 1:100 (Ο), 1:20 (□), and 1:10 (Δ). The results represented the means ± SD of triplication. The marked alphabet in the same line was significantly different (p< 0.05).

3.2 Effect of different proteolysis conditions on the total peptide content in FJ hydrolysate

Figure 2 presented the total peptide content in FJ hydrolysate derived from bromelain and trypsin digestion. Both enzymes were effective in hydrolyzing FJ protein, but the most dominant result was observed from trypsin digestion at an E/S ratio of 1:10 for 18 hours, resulting in peptide contents of 8.7 ± 0.25 mg/mL. An increase in incubation time would result in higher total peptide contents for both enzymes. Furthermore, increasing the E/S ratio generally led to higher total peptide content. Although the highest content of total peptide was achieved with tryptic hydrolysis, bromelain produced a higher peptide content when compared to trypsin digestion. This difference might be attributed to the specificity of FJ protein hydrolysate to the active site of bromelain. Peptide content after enzymatic digestion was an important parameter, as a higher peptide content signified a greater chance of obtaining umami peptides from the sample. Based on the total peptide content, trypsin appeared to be more appropriate for umami peptide preparation from FJ. However, sensory evaluation was necessary to confirm the presence of umami peptide after enzymatic digestion.

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Figure 2
Effect of hydrolysis time and E/S ratio on the peptide content (mg/mL) of diverse FJ hydrolysate generated from proteolytic digestion of bromelain (continuous line) and trypsin (dash line). The different E/S ratios were 1:100 (Ο), 1:20 (□), and 1:10 (Δ). The results represented the means ± SD of triplication. The marked alphabet in the same line was significantly different (p< 0.05).

3.3 Effect of different proteolysis conditions on the amino acid content in FJ hydrolysate

In addition to peptides, amino acids also had the properties of flavor enhancers. Disodium inosinate (IMP), disodium guanylate (GMP), and lysine were examples of amino acids widely examined for their use as flavor enhancers in many meat products (Campagnol et al., 2012, 2017; Freitas et al., 2017). Therefore, it was important to study amino acid content in FJ hydrolysate after digestion by enzymes. The observation showed that trypsin hydrolysate generally released more amino acids than bromelain hydrolysate, consistent with the results for total peptide content. Although the highest amino acid was witnessed from bromelain digestion at 88.16 ppm (1:10 E/S and 18 hours of incubation time), the total profile of released amino acid content was more dominant with trypsin digestion (Figure 3.). This difference could be attributed to the specificity of the enzyme active site in relation to the primary and secondary structures of the FJ protein chain. Prolonged exposure of FJ protein to the enzyme resulted in increased amino acid content in hydrolysate for both enzymes. A significant increase in amino acids was observed after 9 hours of bromelain digestion and 12 hours of trypsin digestion. This phenomenon occurred because the enzyme primarily acted on the outer area of protein molecules during the initial stage (Lowenthal et al., 2014). As the tertiary structure of the protein unfolded, the enzyme gained access to the inner parts of the protein chain where bromelain could find more cutting sites than trypsin. At the end of the reaction, the highest amino acid content was found from bromelain hydrolysate.

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Figure 3
Effect of hydrolysis time and E/S ratio on the amino acid content (ppm) of diverse FJ hydrolysate generated from proteolytic digestion of bromelain (continuous line) and trypsin (dash line). The different E/S ratios were 1:100 (Ο), 1:20 (□), and 1:10 (Δ). The results represented the means ± SD of triplication. The marked alphabet in the same line was significantly different (p< 0.05).

3.4 Effect of different proteolysis conditions on the hedonic sensory characteristics of FJ hydrolysate

Sensory evaluation of FJ hydrolysate obtained from different enzymatic digestion conditions was conducted using a panel of trained participants. Panelists were asked to rate the samples based on three attributes, including taste, aroma, and color, using s 5-point hedonic scale. The hedonic test showed the level of consumer preference and satisfaction concerning the food product attributes (Lim, 2011). The evaluation results clearly showed that samples with high protease concentration and prolonged incubation time received higher scores in all attributes. This was in line with total peptides and amino acid content results, where higher levels of both components were expected to enhance the intensity of umami taste. The highest scores for color, aroma, and taste were 3.95, 4.10, and 3.95, respectively (Figure 4). The scores were achieved by hydrolysate sample obtained from trypsin digestion at an E/S ratio of 1:10 and an incubation time of 15 hours. For samples digested with bromelain, the most preferred one was produced under conditions of an E/S ratio of 1:10 and an 18-hour incubation time (Figure 5). Moreover, the most acceptable samples from both types of enzymatic digestion received scores significantly above the neutral value of 3.0. The results showed that proteolytic enzymes played a crucial role in enhancing flavor by hydrolyzing proteins to be peptides and amino acids. High enzyme concentrations and long enzymatic exposure time were particularly effective in generating umami taste in protein hydrolysate.

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Figure 4
The hedonic analysis of FJ hydrolysate was conducted with diverse hydrolysis times and E/S ratio of each attribute using different proteolytic digestion of (A) bromelain and (B) trypsin. The annotations used in the analysis described different hydrolysis conditions, including 100B3H/100T3H = E/S 1:100 (w/v) for 3 hours, 100B6H/100T6H = E/S 1:100 (w/v) for 6 hours, 100B9H/100T9H = E/S 1:100 (w/v) for 9 hours, 100B12H/100T12H = E/S 1:100 (w/v) for 12 hours, 100B15H/100T15H = E/S 1:100 (w/v) for 15 hours, 100B18H/100T18H = E/S 1:100 (w/v) for 18 hours, 20B3H/20T3H = E/S 1:20 (w/v) for 3 hours, and 20B6H/20T6H = E/S 1:20 (w/v) for 6 hours. These annotations also consisted of 20B9H/20T9H = E/S 1:20 (w/v) for 9 hours, 20B12H/20T12H = E/S 1:20 (w/v) for 12 hours, 20B15H/20T15H = E/S 1:20 (w/v) for 15 hours, 20B18H/20T18H = E/S 1:20 (w/v) for 18 hours, 10B3H/10T3H = E/S 1:10 (w/v) for 3 hours, 10B6H/10T6H = E/S 1:10 (w/v) for 6 hours, 10B9H/10T9H = E/S 1:10 (w/v) for 9 hours, 10B12H/10T12H = E/S 1:10 (w/v) for 12 hours, 10B15H/10T15H = E/S 1:10 (w/v) for 15 hours, and 10B18H/10T18H =E/S 1:10 (w/v) for 18 hours. A 5-point hedonic scale was used to assess the liking of sensory attributes of FJ hydrolysate, with scores ranging from like extremely to dislike extremely. The bar marked with different alphabet showed significant differences in mean values (p-value < 0.05).
thumbnail of Figure 5
Figure 5
Sensory attribute of overall acceptance of FJ hydrolysate with diverse hydrolysis time and E/S ratio using different proteolytic digestions of bromelain and trypsin.

The total acceptance of FJ hydrolysate derived from both enzymes was in accordance with the survey results of individual sensory attributes. The most preferred sample of trypsin and bromelain hydrolysate was obtained at an E/S ratio of 1:10, with an incubation time of 15 and 18 hours, respectively. Previous reviews showed that short peptides with hydrophobic amino acids at the end of their chains tended to have a pronounced bitterness (Temussi, 2011; Huang et al., 2019). In addition, a significant number of peptides containing residues with hydrophobic side chains were known for their distinct bitter taste (Imai et al., 2019). Several investigations have also shown that certain hydrophobic amino acids such as glycine (Gly), alanine (Ala), valine (Val), and phenylalanine (Phe), contributed to umami taste (Lioe et al., 2006; Dang et al., 2015; Salles et al., 1995). In this study, the identified peptides in the bromelain hydrolysate contained amino acid residues that contributed to umami taste content of 58.75%, while the trypsin hydrolysate contained 52.19%. These results could contribute to the acceptability of sensory evaluations.

3.5 Identified peptide sequences of FJ hydrolysate from the selected proteolysis conditions

Numerous investigations have examined the effect of peptides in protein hydrolysate on umami taste. It was further reported that amino acid sequence in peptides, particularly at the N-terminal and C-terminal, played a key role in determining flavor characteristics and intensity (Zhang et al., 2019). In line with these experimental results, peptide content greatly affected umami taste. Therefore, the examination of the amino acid sequence was conducted using LC-MS/MS associated with a database search engine. The results were expressed as MASCOT scores, where higher scores showed greater reliability of amino acid sequence identification. The identified peptides in this study were based on MASCOT scores not lower than 30 and were summarized in Tables 1 and 2 for FJ hydrolysate prepared by bromelain and trypsin, respectively.

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Table 1
Identified peptides from Hydrolysate of FJ using bromelain with E/S 1:10 (w/v) for 18 hours.
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Table 2
Identified peptides from Hydrolysate of FJ using trypsin with E/S 1:10 (w/v) for 15 hours.

The results showed that peptides obtained at the 15-hour hydrolysis time were more abundant compared to those obtained after 18 hours of hydrolysis. Furthermore, peptide fragments during the 18-hour hydrolysis were shorter than those from the 15-hour time. Several amino acids have been recognized for their contribution to umami taste, including tyrosine (Tyr) and Phe (Salles et al., 1995), histidine (His), Val, alanine (Ala) (Dang et al., 2015), serine (Ser), Gly, and threonine (Thr) (Lioe et al., 2006). It was found that bromelain and trypsin hydrolysate contained the amino acids responsible for umami taste at levels of 55.37% and 51.85%, respectively. Interestingly, Phe and Val were also reported for their bitter taste effect (Zhang et al., 2012), which could increase sensory acceptance. In FJ hydrolysate, the proportions of the bitter-related amino acids were 16.95% for bromelain and 9.49% for trypsin digestion.

Recent investigation showed that a Gly residue at the N-terminal of peptides played a crucial role in umami taste due to its strong binding with receptors (Amin et al., 2020). In the FJ bromelain hydrolysate, two out of 10 identified peptides (20.00%) contained Gly at the N-terminal, specifically GPLGPLGPQGIPG and GLPGLPGLPGPKG. On the other hand, only 2 out of 21 peptides (9.52%) from trypsin hydrolysis contained Gly at the N-terminal, including GTSDIIIP and GLLGPKALYVF. While Gly at the N-terminal could strongly bind with umami receptors, arginine (Arg) residue at the C-terminal could perform a similar binding activity (Zhang et al., 2012). Both bromelain and trypsin hydrolysate was found to have only one identified peptide ended with Arg at C-terminal which were PLLPQTAADLQAMLRVVCYYHSFVPSQILKGAVELR, and AESLTYLSLGANNLTLMPELR, respectively.

Although FJ hydrolysate produced by trypsin had fewer types of umami peptides, it achieved a higher acceptance level than the bromelain hydrolysate under the same enzymatic digestion condition, as indicated by sensory evaluation results. This could be attributed to the higher concentration of amino acids in trypsin hydrolysate compared to those in bromelain digestion (Figure 3).

4 Conclusion

In conclusion, FJ snails could be used as a source of umami peptides through protease digestion with bromelain or trypsin. The results showed that hydrolysis conditions greatly affected the total acceptance score, specifically a high concentration of E/S ratio and extended protease exposure time increased the score. The optimal hydrolysis condition was found to be at an E/S ratio of 1:10 for 15-18 hours. Chemical analysis showed that peptides and amino acid contents were important factors contributing to umami taste in the hydrolysate. Furthermore, an investigation into peptide sequences identified key amino acids, such as Gly at the amino-terminal and Arg at the carboxylic end, which played significant roles in enhancing umami taste. Sensory analysis showed the most preferred samples were those with an E/S ratio of 1:10 and hydrolyzed for 15 and 18 hours. Therefore, FJ snails became a great source for extracting flavor enhancer peptides to be used in the food industry.

Acknowledgements

The authors acknowledge the financial support for this study from the Institute of Research and Community Service (LPPM) UPN Veteran Jawa Timur under the international study collaboration grants program for Dedin Finatsiyatull Rosida (SPP/42/UN.63.8/LT/V/2020). They are also grateful to Ms. Supitcha Pannengpetch from the Center for Research and Innovation, Faculty of Medical Technology, Mahidol University (Salaya Campus), Nakonpathum, Thailand, for her valuable assistance in providing LC-MS/MS proteomics services and peptide identification.

  • Cite as:
    Kongpichitchoke, T., Rosida, D. F., Havanapan, P., Putra, A. Y. T., & Priyanto, A. D. (2024). Chemical properties and sensory preference related to umami flavor from crude hydrolysate of Indonesian snails (Filopaludina javanica). Brazilian Journal of Food Technology, 27, e2023108. https://doi.org/10.1590/1981-6723.10823
  • Funding:
    None.

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Edited by

  • Associate Editor:
    Maria Eugenia de Oliveira Mamede.

Publication Dates

  • Publication in this collection
    16 Dec 2024
  • Date of issue
    2024

History

  • Received
    13 Sept 2023
  • Accepted
    03 Oct 2024
Creative Common - by 4.0
This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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