Genome mining and physiological analyses uncover adaptation strategies and biotechnological potential of Virgibacillus dokdonensis T4.6 isolated from high-salt shrimp paste.

Virgibacillus spp. stand out as a potent starter culture for accelerating the fermention of fish sauces and shrimp pastes. However, the underlying molecular mechanisms responsible for their adaptation and biotechnological potential remain elusive. Therefore, the present study focuses on phenotypic and genomic analyses of a halophilic bacterium Virgibacillus dokdonensis T4.6, derived from Vietnamese high-salt fermented shrimp paste. The draft genome contained 4,096,868 bp with 3780 predicted coding sequences. Genome mining revealed the presence of 143 genes involved in osmotic adaptation explaining its resistant phenotype to 24% (w/v) NaCl. Among them, 37 genes making up the complete ectoine metabolism pathway, confirmed its ability to produce 4.38 ± 0.29 wt% ectoine under 12.5% NaCl stress. A significant finding was the identification of 39 genes responsible for an entire degradation pathway of the toxic biogenic amine histamine, which was in agreement with its histamine degradation rate of 42.7 ± 2.1% in the HA medium containing 5 mM histamine within 10 days at 37 °C. Furthermore, 114 proteolytic and 19 lipolytic genes were detected which might contribute to its survival as well as the nutrient quality and flavor of shrimp paste. Of note, a putative gene vdo2592 was found as a possible novel lipase/esterase due to its unique Glycine-Aspartate-Serine-Leucine (GDSL) sequence motif. This is the first report to reveal the adaptative strategies and related biotechnological potential of Virgibacillus associated with femented foods. Our findings indicated that V. dokdonensis T4.6 is a promising starter culture for the production of fermented shrimp paste products.

Metabolome analysis to investigate the effect of controlled fermentation on taste-related metabolites in terasi.

INTRODUCTION: Terasi is a fermented shrimp paste unique to Indonesia and is used in dishes to add umami and saltiness. In a previous study, the controlled fermentation of terasi was optimized using starters containing three bacterial isolates: Staphylococcus saprophyticus, Bacillus subtilis, and Lactobacillus murinus. However, the influence of controlled fermentation using these starters on the metabolites in terasi has not yet been studied. OBJECTIVES: Therefore, this study aimed to investigate the effect of controlled fermentation on taste-related metabolites in terasi using a metabolomics approach. RESULTS: Non-targeted analysis indicated that amino acids contributed to variations during fermentation. Subsequently, targeted analysis of amino acids revealed that terasi subjected to controlled fermentation using a starter with a 2:1:2 ratio of S. saprophyticus, B. subtilis, and L. murinus, respectively, resulted in a product containing D-amino acids, such as D-Asp, D-Gln, and D-Leu that was unique when compared to other terasi products prepared using controlled fermentation. Genetic analysis of isolates from the terasi produced using controlled fermentation was also carried out, and this is the first study to suggest that Staphylococcus spp. has the potential to produce D-amino acids. CONCLUSION: In conclusion, the ratio of bacterial species in starter cultures used in controlled fermentation influenced the amino acid profile of the product and starters with a higher ratio of Staphylococcus spp. may result in the production of D-amino acids.

Allobacillus salarius sp. nov., and Allobacillus saliphilus sp. nov., isolated from shrimp paste (ka-pi) in Thailand.

Two strains of moderately halophilic, Gram-stain-positive and spore-forming rods, designated as SKP4-8T and SKP8-2T isolated from shrimp paste (Ka-pi), were taxonomic studied based on polyphasic approach. Strain SKP4-8T grew at pH 6.0-9.0 (optimum 7.0), at 25-45 °C (optimum 37 °C) and in 1-16% (w/v) NaCl (optimum 5-10%). Strain SKP8-2T grew at pH 6.0-9.0 (optimum 8.0), at 25-45 °C (optimum 37 °C) and in 0-20% (w/v) NaCl (optimum 3-10%). The strains contained meso-diaminopimelic acid in cell-wall peptidoglycan and the major menaquinone was MK-7. Strain SKP4-8T contained iso-C15:0, anteiso-C15:0 and iso-C17:0; and strain SKP8-2T contained anteiso-C15:0, iso-C15:0, iso-C16:0 and antesio-C17:0 as major cellular fatty acids. Phosphatidylglycerol, diphosphatidylglycerol, unknown phospholipids and an unknown glycolipid were detected as major polar lipids. On the basis of 16S rRNA gene sequence analysis, strains SKP4-8T and SKP8-2T belonged to the genus Allobacillus and were closely related to Allobacillus halotolerans LMG 24826T with 98.8% and 99.3% similarity, respectively. The comparative genome analysis based on average nucleotide identity (ANI) and digital DNA-DNA hybridization revealed that both strains showed the values below 95 and 70%, from each other and from Allobacillus halotolerans LMG 24826T, respectively. Based on the data from this polyphasic study, strains SKP4-8T and SKP8-2T represent novel species of the genus Allobacillus and the name Allobacillus salarius sp. nov. was proposed for SKP4-8T (= KCTC 33905T = LMG 30016T = TISTR 2499T); and Allobacillus saliphilus sp. nov. for SKP8-2T (= KCTC 33906T = LMG 29682T = TISTR 2558T).

Halobacillus fulvus sp. nov., a moderately halophilic bacterium isolated from shrimp paste (Ka-pi) in Thailand.

An aerobic, Gram-stain-positive, endospore-forming, rod-shaped and moderately halophilic strain SKP4-6T, was isolated from shrimp paste (Ka-pi) collected from Samut Sakhon Province, Thailand. Phylogenetic analysis revealed that strain SKP4-6T belonged to the genus Halobacillus and was most closely related to Halobacillus salinus JCM 11546T (98.6 %), Halobacillus locisalis KCTC 3788T (98.6 %) and Halobacillus yeomjeoni KCTC 3957T (98.6 %) based on 16S rRNA gene sequence similarity. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strain SKP4-6T and its related species were 18.2-19.3 % and 69.84-84.51 %, respectively, which were lower than the threshold recommended for species delineation. The strain grew optimally at 30-40 °C, at pH 7.0 and with 10-15 % (w/v) NaCl. It contained l-Orn-d-Asp in the cell wall peptidoglycan. The DNA G+C content was 44.8 mol%. The major fatty acids were iso-C15 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The predominant isoprenoid quinone was MK-7. Phosphatidylglycerol and diphosphatidylglycerol were present as major polar lipids. Based on this polyphasic approach, digital DNA-DNA relatedness and ANI values, strain SKP4-6T represents a novel species of the genus Halobacillus, for which the name Halobacillus fulvus sp. nov. is proposed. The type strain is SKP4-6T (=JCM 32624T=TISTR 2595T).

Lentibacillus lipolyticus sp. nov., a moderately halophilic bacterium isolated from shrimp paste (Ka-pi).

A Gram-stain-positive, aerobic, spore-forming, moderately halophilic bacterium, SSKP1-9T, was isolated from traditional salted shrimp paste (Ka-pi) produced in Samut Sakhon Province, Thailand. This strain grew optimally at 37-40 °C, pH 7.0 and in the presence of 8-16 % (w/v) NaCl. The 16S rRNA gene sequence similarity values between strain SSKP1-9T and Lentibacillus juripiscarius TISTR 1535T and Lentibacillus halophilus TISTR 1549T were 98.7 and 97.2 %, respectively. Based on 16S rRNA gene sequence similarity, strain SSKP1-9T represents a distinct novel species, as shown by phenotypic traits, DNA-DNA hybridization and average nucleotide identity values. In addition, the whole-cell protein profile confirmed the novelty of the taxon. The genomic DNA G+C content was 44.6 mol%. The major isoprenoid quinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. Polar lipid analysis revealed the presence of phosphatidylglycerol, diphosphatidylglycerol, four unidentified lipids, an unidentified phospholipid and an unidentified glycolipid. The major cellular fatty acids were anteiso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The results of phenotypic and chemotaxonomic characteristics and whole-genome analysis support that strain SSKP1-9T represents a novel species of Lentibacillus, for which the name Lentibacilluslipolyticus sp. nov. is proposed. The type strain is SSKP1-9T (=JCM 32625T=TISTR 2597T).

Bacillus salacetis sp. nov., a slightly halophilic bacterium from Thai shrimp paste (Ka-pi).

A Gram-stain-positive, slightly halophilic, endospore-forming, strictly aerobic, rod-shaped bacterium, designated SKP7-4T, was isolated from shrimp paste collected from Samut Sakhon province, Thailand. Strain SKP7-4T grew at pH 6.0-9.0 (optimum, 7.5), at 20-40 °C (37 °C) and in 0-15 % (w/v) NaCl (1-3 %). The diamino acid found in the cell-wall peptidoglycan was meso-diaminopimelic acid. Menaquinone with seven isoprene units was the major isoprenoid quinone. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and three unidentified phospholipids were detected as polar lipids. It contained iso-C15 : 0 and anteiso-C15 : 0 as major cellular fatty acids. On the basis of 16S rRNA gene sequence analysis, strain SKP7-4T belonged to the genus Bacillus and was closely related to Bacillus vietnamensis JCM 11124T, Bacillus marisflavi JCM 11544T, Bacillus aquimaris JCM 11545T and Bacillusoryzaecorticis JCM 19602T, with 98.7, 97.9, 97.8 and 97.8 % similarity, respectively. The draft genome of SKP7-4T was 4.68 Mb with 5208 coding sequences with an average G+C content of 43.2 mol%. The ANIb and ANIm values of strain SKP7-4T were 70.0 and 84.3 %, respectively, and the digital DNA-DNA hybridization value was 20 % in comparison with the draft genome of B. vietnamensis JCM 11124T. On the basis of the results of phenotypic, chemotaxonomic and phylogenetic analyses, the strain should represent a novel species of the genus Bacillus and the name Bacillus salacetis sp. nov. is proposed. The type strain is SKP7-4T (=JCM 33205T=KCTC 43014T=TISTR 2596T).

Characterization and mutation analysis of a halotolerant serine protease from a new isolate of Bacillus subtilis.

OBJECTIVES: A bacterial halotolerant enzyme was characterized to understand the molecular mechanism of salt adaptation and to explore its protein engineering potential. RESULTS: Halotolerant serine protease (Apr_No16) from a newly isolated Bacillus subtilis strain no. 16 was characterized. Multiple alignments with previously reported non-halotolerant proteases, including subtilisin Carlsberg, indicated that Apr_No16 has eight acidic or polar amino acid residues that are replaced by nonpolar amino acids in non-halotolerant proteases. Those residues were hypothesized to be one of the primary contributors to salt adaptation. An eightfold mutant substituted with Ala residues exhibited 1.2- and 1.8-fold greater halotolerance at 12.5% (w/v) NaCl than Apr_No16 and Carlsberg, respectively. Amino acid substitution notably shifted the theoretical pI of the eightfold mutant, from 6.33 to 9.23, compared with Apr_No16. The resulting protein better tolerated high salt conditions. CONCLUSIONS: Changing the pI of a bacterial serine protease may be an effective strategy to improve the enzyme's halotolerance.

Microbiological and chemical changes of shrimp Acetes vulgaris during Kapi production.

Microbiological and chemical changes in shrimp Acetes vulgaris during production of Kapi (salted shrimp paste of Thailand) including salting, drying and fermentation were monitored. Moisture content of samples decreased rapidly after salting and drying steps. The lower water activity was found in the final product (0.694). The pH decreased within the first 10 days of fermentation and continuously increased as fermentation progressed. Protein underwent degradation throughout Kapi production as indicated by increasing TCA-soluble peptides and degree of hydrolysis. The increases in peroxide value as well as thiobarbituric acid reactive substances value revealed that lipid oxidation occurred throughout all processes. Total viable count, halophilic, proteolytic and lipolytic bacteria counts increased continuously during Kapi production, while lactic acid bacteria count slightly decreased at the final stage of fermentation. Thus, proteolysis and lipolysis took place throughout Kapi production, and contributed to the characteristics of finished product. These changes were governed by both endogenous and microbial enzymes.

Chemical quality assessment of traditional salt-fermented shrimp paste from Northern Mindanao, Philippines.

BACKGROUND: Shrimp paste is an important fermented commodity in the Philippines, but so far its quality parameters have hardly been characterized. In this study, paste samples procured in the province of Agusan del Norte, Philippines from three different traditional manufacturers and from a commercial supermarket were analyzed for their chemical composition. RESULTS: Both traditional and commercial shrimp pastes varied in their content of protein (12.9-15.3 g per 100 g), fat (0.50-1.94 g per 100 g), saturated fatty acids (32.6-39.1 g per 100 g fatty acid methyl esters (FAME)), monounsaturated fatty acids (15.1-18.7 g per 100 g FAME) and polyunsaturated fatty acids (30.7-40.8 g per 100 g FAME). Their pH ranged between 6.8 and 7.7. The samples were microbiologically stable owing to their low water activity (0.70-0.74) and high NaCl content (4.04-5.15 g per 100 g). Although all samples were processed in the same country and under similar conditions, differences were observed in some parameters: thiobarbituric acid-reactive substances (2.32-5.03 µg malondialdehyde g(-1)), total non-protein nitrogen (3.07-5.15 g N per 100 g), free non-protein nitrogen (1.17-2.39 g N per 100 g), biogenic amines and mineral content. The biogenic amine index varied between 0 and 976 for the different samples; only one sample could be considered as class 1 quality. CONCLUSION: The results showed that there is a high variation in the quality of the product which could be linked to differences in the fermentation process and hygienic quality.

Production of bacteriocin by Virgibacillus salexigens isolated from "terasi": a traditionally fermented shrimp paste in Indonesia.

A natural antibacterial-substance-producing gram-positive bacterium was isolated from terasi shrimp paste, a popular fermented product in Indonesia. This strain, a spore-forming and strictly aerobic bacterium, was identified as Virgibacillus salexigens by 16S rRNA gene sequence analysis. The antibacterial substance purified from the precipitated product in the culture supernatant of the strain using ammonium sulfate showed a broad inhibition spectrum against gram-positive bacteria, including a typical foodborne bacterium, namely, Listeria monocytogenes. The antibacterial activity of the substance was inactivated by treatments with various proteolytic enzymes. It was stable after heating or pH treatment, and approximately 60% of the initial activity remained even after heating at 121 °C for 15 min. In addition, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis indicated that its monoisotopic mass weight was 5318.4 Da (M+H)(+). On the basis of the results obtained by the automated Edman degradation technique and MALDI-TOF MS analysis, the substance can be classified as a member of Class IId bacteriocins, but it could not be identified as any of the previously purified substances except for the putative bacteriocin predicted from the draft genome sequence data of gram-positive bacteria such as Virgibacillus and Bacillus strains.

Characterization and in vitro biological activities of Thai traditional fermented shrimp pastes.

In this work, chemical and biological characteristics of two types of Thai fermented shrimp paste, Kapi Ta Dam and Kapi Ta Deang, at different fermentation periods and their raw materials were investigated. Kapi had low water activity and high proteins with high glutamic acid and lysine. Both Kapis, which had different sources, showed similar characteristics. The number of lactic acid bacteria in the products increased during the early stages of fermentation. Free α-amino acid contents in the products increased with the fermentation time. The water extracts from Kapi products showed strong antioxidative activities against ABTS(+) radical, and ACE inhibitory activity but they did not exhibit antimicrobial activity against Staphylococcus aureus, Bacillus cereus, Escherichia coli and Salmonella Typhimurium. Biological activities in Kapi could be developed by fermentation process, enzymatic hydrolysis of proteins and non-enzymatic browning reactions. Kapi could, thus, serve as a potential source of natural bioactive substances.

Changes in antioxidant activities and physicochemical properties of Kapi, a fermented shrimp paste, during fermentation.

Changes in chemical composition, physical properties and antioxidant activities of Kapi were monitored during fermentation for 12 months. DPPH (2, 2-diphenyl-1-picryl hydrazyl), ABTS (2, 2 - azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging activity as well as ferric reducing antioxidant power (FRAP) gradually increased as the fermentation time increased, particularly during the first 8 months (P < 0.05). Thereafter, the decreases in DPPH and ABTS radical scavenging activities were observed (P < 0.05), whereas FRAP remained constant (P > 0.05). The continuous increases in ammonia nitrogen, formaldehyde nitrogen and amino nitrogen contents were noticeable within the first 8 months (P < 0.05), indicating the formation of peptides and free amino acids via the hydrolysis of protein by both microbial and indigenous proteases. Browning intensity most likely caused by the formation of Maillard reaction products (MRPs) were concomitantly observed throughout fermentation, as evidenced by the decreases in lightness (L*-value), but the increases in redness (a*-value) and yellowness (b*-value). Low level of thiobarbituric acid reactive substances in Kapi was found during 12 months. Antioxidant activities of Kapi were more likely governed by the low molecular weight peptides, amino acids as well as Maillard reaction products generated during fermentation.

Effects of 'bask in sunlight and dewed at night' on the formation of fermented flavor in shrimp paste after maturation.

The purpose of the study was to illustrate the roles of three primary indexes, namely sunlight, ventilation and stirring, in the 'bask in sunlight and dewed at night' technique on the quality of shrimp paste, through a laboratory-scale design. The results showed that changes in the post-ripening fermentation conditions, especially sunlight, was instrumental in the physicochemical properties of the shrimp paste. E-nose and SPME-GC-MS were employed to assess the volatile flavor of post-ripening fermentation. A total of 29 key volatile aroma components played a crucial role in the development of post-ripening flavor in shrimp paste with or without sunlight. Lipidomic analysis revealed that sunlight promoted the oxidative degradation of FA, resulting in the production of a diverse range of flavor compounds that imparted the unique aroma of shrimp paste. The findings of this study will establish a theoretical basic for better control of the post-ripening fermentation of traditional shrimp paste.

Evaluation of the biogenic amines in low-salt shrimp paste cooked under various conditions.

Shrimp paste, a type of traditional Chinese food prepared from shrimp fermented with salt, contains biogenic amines (BAs). In this study, the BA content, salinity, and pH of eight commercial low-salt shrimp pastes were analyzed. In addition, the influences of various cooking conditions on the BA content of it were evaluated by HPLC. The total BA amount per product ranged between 32.39 and 1051.16 mg/kg. The salinity and pH were found significantly inversely correlated with the total BA amount. Of the cooking methods tested, after microwave heating and stir-frying, the total BA amount of shrimp paste, which showed the highest BA amount among 8 samples, declined from 1051.16 to 598.48 and 650.49 mg/kg, respectively; however, boiling or steaming showed no significant effects on the total BA amount. These results indicated possible health risk of low-salt shrimp paste whereas the risk could be reduced by choosing appropriate cooking method.

A multi-omics-based investigation into the flavor formation mechanisms during the fermentation of traditional Chinese shrimp paste.

The fermentation process of traditional shrimp paste is closely associated with the production of flavor substances, but the formation mechanism of key aroma components is still unclear. In this study, a comprehensively flavor profile analysis of traditional fermented shrimp paste was carried out by E-nose and SPME-GC-MS. A total of 17 key volatile aroma components with OAV > 1 contributed greatly to the overall flavor formation of shrimp paste. In addition, high-throughput sequencing (HTS) analysis revealed that Tetragenococcus was the dominant genera in the whole fermentation process. Moreover, metabolomics analysis showed that the oxidation and degradation of lipids, protein, organic acids and amino acids produced a large number of flavor substances and intermediates, which laid the foundation for the Maillard reaction in term of generating the distinct aroma of the traditional shrimp paste. This work will provide theoretical support for the realization of flavor regulation and quality control in traditional fermented foods.

Effects of a Novel Starter Culture on Quality Improvement and Putrescine, Cadaverine, and Histamine Inhibition of Fermented Shrimp Paste.

Fermented shrimp paste is a popular food in Asian countries. However, biogenic amines (BAs) are a typically associated hazard commonly found during the fermentation of shrimp paste and pose a food-safety danger. In this work, an autochthonic salt-tolerant Tetragenococcus muriaticus TS (T. muriaticus TS) strain was used as a starter culture for grasshopper sub shrimp paste fermentation. It was found that with the starter culture, putrescine, cadaverine, and histamine concentrations were significantly lower (p < 0.05) with a maximal reduction of 19.20%, 14.01%, and 28.62%, respectively. According to high-throughput sequencing data, T. muriaticus TS could change the interactions between species and reduce the abundance of bacterial genera positively associated with BAs, therefore inhibiting the BA accumulation during shrimp paste fermentation. Moreover, the volatile compounds during the fermentation process were also assessed by HS-SPME-GC-MS. With the starter added, the content of pyrazines increased, while the off-odor amines decreased. The odor of the shrimp paste was successfully improved. These results indicate that T. muriaticus TS can be used as an appropriate starter culture for improving the safety and quality of grasshopper sub shrimp paste.

Draft genome sequencing data of the moderately halophilic bacterium, Allobacillus halotolerans SKP2-8 from shrimp paste (ka-pi).

A moderately halophilic, Gram-stain-positive, spore-forming rod-shaped bacterium, designated SKP2-8 was isolated from a traditional fermented shrimp paste (Ka-pi) collected from the market in Samut Sakhon province, Thailand. This isolate SKP2-8 was closely related to Allobacillus halotolerans LMG 24826T with 99.56% similarity based on 16S rRNA gene sequence. The draft genome of SKP2-8 was 2.53 Mb with 2,515 coding sequences with an average G+C content of 39.5 mol%. The ANIb, ANIm, AAI and the digital DNA-DNA hybridization values of isolate SKP2-8 were 97.22%, 97.64%, 97.75% and 78.0%, respectively, compared with A. halotolerans LMG 24826T. Based on the phenotypic characteristics, DNA-DNA relatedness and phylogenomic analysis, it was identified as Allobacillus halotolerans. The genome sequence data of this isolate provide information for further analysis of the potential biotechnological use of this microorganism and guide the characterization. The draft genome was deposited at DDBJ/ EMBL/GenBank (DNA Databank of Japan/European Molecular Biology Laboratory/Genbank) (VMHF00000000).

Halophilic bacteria as starter cultures: A new strategy to accelerate fermentation and enhance flavor of shrimp paste.

Retaining the traditional flavor while shortening the fermentation cycle is the current research focus for shrimp paste fermentation technology. The present study investigated the effect of combined use of halophilic bacteria as starters on the sensory and flavor characteristics of rapidly fermented shrimp paste. Sensory evaluation indicated that the starter-inoculated samples had high texture, appearance, and overall quality scores. Headspace gas chromatography-ion mobility spectrometry/mass spectrometry (HS-GC-IMS/MS) identified 95 volatile compounds, the fingerprint profiles of the starter-inoculated samples were similar to those of the traditional sample. Notably, the content of benzaldehyde, phenylethylaldehyde, and 3-methylbutyraldehyde increased significantly in the starter-inoculated samples (p < 0.05), which may provide an intense malt, caramel, and pleasant odor. Although the content of certain flavor substances in the starter-inoculated samples was lower than those of traditional sample, the use significantly reduced the fermentation time and mimicked the flavor profile of traditional shrimp paste to some extent.

Variations in the physicochemical properties and bacterial community composition during fermentation of low-salt shrimp paste.

The main purpose of the present study was to investigate the influence of two fermentation temperatures (10 °C and 20 °C) on the physicochemical properties and bacterial community composition of the low-salt shrimp paste. The physicochemical properties pH values, total volatile basic nitrogen (TVBN), amino acid nitrogen (AAN) and biogenic amines (BAs) of low-salt shrimp paste were analyzed and compared. The bacterial community composition was analyzed by high-throughput sequencing technology. Results indicate that the pH value fluctuated between 8.50 and 7.95, and the AAN, TVBN and total BAs levels of low-salt shrimp paste increased gradually after fermentation started, but the AAN content fluctuated steadily after 12 w. The contents of TVBN and total BAs in shrimp paste fermented at 10 °C were lower than those at 20 °C. Thus, the appropriate fermentation temperature and period were 10 °C and 12 w, respectively. Carnobacteriaceae and Staphylococaceae were the predominant family for the fermented shrimp paste. At the genus level, Alkalibacterium was predominantly present at 8 wk (96.36%), while Staphylococcus became the predominant one at 20 wk (58.71%). Correlation analysis shows that Alkalibacterium was positively associated with tyramine (R2 = 0.59), and Staphylococcus and Tetragenococcus were positively associated with AAN and TVBN. These findings are used to facilitate the quality control and improvement of low-salt shrimp paste production process in the future.

Dynamic Changes in the Bacterial Community and Metabolic Profile during Fermentation of Low-Salt Shrimp Paste (Terasi).

Low-salt shrimp paste, or terasi, is an Indonesian fermented food made from planktonic shrimp mixed with a low concentration of salt. Since high daily intake of sodium is deemed unhealthy, reduction of salt content in shrimp paste production is desired. Until now, there is no reported investigation on the bacterial population and metabolite composition of terasi during fermentation. In this study, the bacterial community of terasi was assessed using high-throughput sequencing of the 16S rRNA V3-V4 region. From this analysis, Tetragenococcus, Aloicoccus, Alkalibacillus, Atopostipes, and Alkalibacterium were found to be the dominant bacterial genus in low-salt shrimp paste. GC/MS-based metabolite profiling was also conducted to monitor the metabolite changes during shrimp paste fermentation. Results showed that acetylated amino acids increased, while glutamine levels decreased, during the fermentation of low-salt shrimp paste. At the start of shrimp paste fermentation, Tetragenococcus predominated with histamine and cadaverine accumulation. At the end of fermentation, there was an increase in 4-hydroxyphenyl acetic acid and indole-3-acetic acid levels, as well as the predominance of Atopostipes. Moreover, we found that aspartic acid increased during fermentation. Based on our findings, we recommend that fermentation of low-salt shrimp paste be done for 7 to 21 days, in order to produce shrimp paste that has high nutritional content and reduced health risk.